Benzene Toxicological Profile tp3 3v PDF

TOXICOLOGICAL PROFILE FOR
BENZENE
U.S. DEPARTMENT OF HEALTH AND HUMAN SERVICES
Public Health Service
Agency for Toxic Substances and Disease Registry
August 2007
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DISCLAIMER
The use of company or product name(s) is for identification only and does not imply endorsement by the
Agency for Toxic Substances and Disease Registry.
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UPDATE STATEMENT
A Toxicological Profile for Benzene, Draft for Public Comment was released in August 2005. This
edition supersedes any previously released draft or final profile.
Toxicological profiles are revised and republished as necessary. For information regarding the update
status of previously released profiles, contact ATSDR at:
Division of Toxicology and Environmental Medicine/Applied Toxicology Branch
1600 Clifton Road NE
Mailstop F-32
Atlanta, Georgia 30333
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FOREWORD
This toxicological profile is prepared in accordance with guidelines developed by the Agency for
Toxic Substances and Disease Registry (ATSDR) and the Environmental Protection Agency (EPA). The
original guidelines were published in the Federal Register on April 17, 1987. Each profile will be revised
and republished as necessary.
The ATSDR toxicological profile succinctly characterizes the toxicologic and adverse health
effects information for the hazardous substance described therein. Each peer-reviewed profile identifies
and reviews the key literature that describes a hazardous substance's toxicologic properties. Other
pertinent literature is also presented, but is described in less detail than the key studies. The profile is not
intended to be an exhaustive document; however, more comprehensive sources of specialty information
are referenced.
The focus of the profiles is on health and toxicologic information; therefore, each toxicological
profile begins with a public health statement that describes, in nontechnical language, a substance's
relevant toxicological properties. Following the public health statement is information concerning levels
of significant human exposure and, where known, significant health effects. The adequacy of information
to determine a substance's health effects is described in a health effects summary. Data needs that are of
significance to protection of public health are identified by ATSDR and EPA.
Each profile includes the following:
(A) The examination, summary, and interpretation of available toxicologic information and
epidemiologic evaluations on a hazardous substance to ascertain the levels of significant
human exposure for the substance and the associated acute, subacute, and chronic health
effects;
(B) A determination of whether adequate information on the health effects of each substance is
available or in the process of development to determine levels of exposure that present a
significant risk to human health of acute, subacute, and chronic health effects; and
(C) Where appropriate, identification of toxicologic testing needed to identify the types or levels
of exposure that may present significant risk of adverse health effects in humans.
The principal audiences for the toxicological profiles are health professionals at the Federal, State,
and local levels; interested private sector organizations and groups; and members of the public.
This profile reflects ATSDRs assessment of all relevant toxicologic testing and information that
has been peer-reviewed. Staff of the Centers for Disease Control and Prevention and other Federal
scientists have also reviewed the profile. In addition, this profile has been peer-reviewed by a
nongovernmental panel and is being made available for public review. Final responsibility for the
contents and views expressed in this toxicological profile resides with ATSDR.
vi
The toxicological profiles are developed in response to the Superfund Amendments and
Reauthorization Act (SARA) of 1986 (Public Law 99-499) which amended the Comprehensive
Environmental Response, Compensation, and Liability Act of 1980 (CERCLA or Superfund). This
public law directed ATSDR to prepare toxicological profiles for hazardous substances most commonly
found at facilities on the CERCLA National Priorities List and that pose the most significant potential
threat to human health, as determined by ATSDR and the EPA. The availability of the revised priority
list of 275 hazardous substances was announced in the Federal Register on December 7, 2005 (70 FR
72840). For prior versions of the list of substances, see Federal Register notices dated April 17, 1987
(52 FR 12866); October 20, 1988 (53 FR 41280); October 26, 1989 (54 FR 43619); October 17, 1990 (55
FR 42067); October 17, 1991 (56 FR 52166); October 28, 1992 (57 FR 48801); February 28, 1994 (59
FR 9486); April 29, 1996 (61 FR 18744); November 17, 1997 (62 FR 61332); October 21, 1999 (64 FR
56792); October 25, 2001 (66 FR 54014); and November 7, 2003 (68 FR 63098). Section 104(i)(3) of
CERCLA, as amended, directs the Administrator of ATSDR to prepare a toxicological profile for each
substance on the list.
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QUICK REFERENCE FOR HEALTH CARE PROVIDERS

Toxicological Profiles are a unique compilation of toxicological information on a given hazardous
substance. Each profile reflects a comprehensive and extensive evaluation, summary, and interpretation
of available toxicologic and epidemiologic information on a substance. Health care providers treating
patients potentially exposed to hazardous substances will find the following information helpful for fast
answers to often-asked questions.
Primary Chapters/Sections of Interest
Chapter 1: Public Health Statement: The Public Health Statement can be a useful tool for educating
patients about possible exposure to a hazardous substance. It explains a substances relevant
toxicologic properties in a nontechnical, question-and-answer format, and it includes a review of
the general health effects observed following exposure.
Chapter 2: Relevance to Public Health: The Relevance to Public Health Section evaluates, interprets,
and assesses the significance of toxicity data to human health.
Chapter 3: Health Effects: Specific health effects of a given hazardous compound are reported by type
of health effect (death, systemic, immunologic, reproductive), by route of exposure, and by length
of exposure (acute, intermediate, and chronic). In addition, both human and animal studies are
reported in this section.
NOTE: Not all health effects reported in this section are necessarily observed in the clinical
setting. Please refer to the Public Health Statement to identify general health effects observed
following exposure.
Pediatrics: Four new sections have been added to each Toxicological Profile to address child health
issues:
Section 1.6
How Can (Chemical X) Affect Children?
Section 1.7
How Can Families Reduce the Risk of Exposure to (Chemical X)?
Section 3.7
Childrens Susceptibility
Section 6.6
Exposures of Children
Other Sections of Interest:

Section 3.8
Biomarkers of Exposure and Effect
Section 3.11 Methods for Reducing Toxic Effects
ATSDR Information Center

Phone: 1-800-CDC-INFO (800-232-4636) or 1-888-232-6348 (TTY) Fax: (770) 488-4178
E-mail: cdcinfo@cdc.gov
Internet: http://www.atsdr.cdc.gov
The following additional material can be ordered through the ATSDR Information Center:
Case Studies in Environmental Medicine: Taking an Exposure HistoryThe importance of taking an
exposure history and how to conduct one are described, and an example of a thorough exposure
history is provided. Other case studies of interest include Reproductive and Developmental
Hazards; Skin Lesions and Environmental Exposures; Cholinesterase-Inhibiting Pesticide
Toxicity; and numerous chemical-specific case studies.
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Managing Hazardous Materials Incidents is a three-volume set of recommendations for on-scene

(prehospital) and hospital medical management of patients exposed during a hazardous materials
incident. Volumes I and II are planning guides to assist first responders and hospital emergency
department personnel in planning for incidents that involve hazardous materials. Volume III
Medical Management Guidelines for Acute Chemical Exposuresis a guide for health care
professionals treating patients exposed to hazardous materials.
Fact Sheets (ToxFAQs) provide answers to frequently asked questions about toxic substances.
Other Agencies and Organizations
The National Center for Environmental Health (NCEH) focuses on preventing or controlling disease,
injury, and disability related to the interactions between people and their environment outside the
workplace. Contact: NCEH, Mailstop F-29, 4770 Buford Highway, NE, Atlanta,
GA 30341-3724 Phone: 770-488-7000 FAX: 770-488-7015.
The National Institute for Occupational Safety and Health (NIOSH) conducts research on occupational
diseases and injuries, responds to requests for assistance by investigating problems of health and
safety in the workplace, recommends standards to the Occupational Safety and Health
Administration (OSHA) and the Mine Safety and Health Administration (MSHA), and trains
professionals in occupational safety and health. Contact: NIOSH, 200 Independence Avenue,
SW, Washington, DC 20201 Phone: 800-356-4674 or NIOSH Technical Information Branch,
Robert A. Taft Laboratory, Mailstop C-19, 4676 Columbia Parkway, Cincinnati, OH 45226-1998
Phone: 800-35-NIOSH.
The National Institute of Environmental Health Sciences (NIEHS) is the principal federal agency for
biomedical research on the effects of chemical, physical, and biologic environmental agents on
human health and well-being. Contact: NIEHS, PO Box 12233, 104 T.W. Alexander Drive,
Research Triangle Park, NC 27709 Phone: 919-541-3212.
Referrals
The Association of Occupational and Environmental Clinics (AOEC) has developed a network of clinics
in the United States to provide expertise in occupational and environmental issues. Contact:
AOEC, 1010 Vermont Avenue, NW, #513, Washington, DC 20005 Phone: 202-347-4976
FAX: 202-347-4950 e-mail: AOEC@AOEC.ORG Web Page: http://www.aoec.org/.
The American College of Occupational and Environmental Medicine (ACOEM) is an association of
physicians and other health care providers specializing in the field of occupational and
environmental medicine. Contact: ACOEM, 25 Northwest Point Boulevard, Suite 700, Elk
Grove Village, IL 60007-1030 Phone: 847-818-1800 FAX: 847-818-9266.
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CONTRIBUTORS
CHEMICAL MANAGER(S)/AUTHOR(S):
Sharon Wilbur, M.A.
Sam Keith, M.S., C.H.P.
Obaid Faroon, Ph.D.
ATSDR, Division of Toxicology and Environmental Medicine, Atlanta, GA
David Wohlers, Ph.D.
Julie Stickney, Ph.D.
Sari Paikoff, Ph.D.
Gary Diamond, Ph.D
Antonio Quiones-Rivera, Ph.D.
Syracuse Research Corporation, North Syracuse, NY
THE PROFILE HAS UNDERGONE THE FOLLOWING ATSDR INTERNAL REVIEWS:

1.
Health Effects Review. The Health Effects Review Committee examines the health effects
chapter of each profile for consistency and accuracy in interpreting health effects and classifying
end points.
2.
Minimal Risk Level Review. The Minimal Risk Level Workgroup considers issues relevant to
substance-specific Minimal Risk Levels (MRLs), reviews the health effects database of each
profile, and makes recommendations for derivation of MRLs.
3.
Data Needs Review. The Applied Toxicology Branch reviews data needs sections to assure
consistency across profiles and adherence to instructions in the Guidance.
4.
Green Border Review. Green Border review assures the consistency with ATSDR policy.
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PEER REVIEW
A peer review panel was assembled for benzene. The panel consisted of the following members:
1.
Dr. Jeffrey Fisher, Professor and Department Head, Department of Environmental Health
Science, University of Georgia, Athens, Georgia;
2.
Dr. Tee Guidotti, Chair and Professor, Department of Occupational and Environmental Health,
School of Public Health and Sciences, The George Washington University Medical Center,
Washington, DC; and
3.
Dr. Rogene Henderson, Senior Scientist, Lovelace Respiratory Research Institute, Albuquerque,
New Mexico.
These experts collectively have knowledge of benzene's physical and chemical properties, toxicokinetics,
key health end points, mechanisms of action, human and animal exposure, and quantification of risk to
humans. All reviewers were selected in conformity with the conditions for peer review specified in
Section 104(I)(13) of the Comprehensive Environmental Response, Compensation, and Liability Act, as
amended.
Scientists from the Agency for Toxic Substances and Disease Registry (ATSDR) have reviewed the peer
reviewers' comments and determined which comments will be included in the profile. A listing of the
peer reviewers' comments not incorporated in the profile, with a brief explanation of the rationale for their
exclusion, exists as part of the administrative record for this compound.
The citation of the peer review panel should not be understood to imply its approval of the profile's final
content. The responsibility for the content of this profile lies with the ATSDR.
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CONTENTS
DISCLAIMER ..............................................................................................................................................ii
UPDATE STATEMENT .............................................................................................................................iii
FOREWORD ................................................................................................................................................ v
QUICK REFERENCE FOR HEALTH CARE PROVIDERS....................................................................vii
CONTRIBUTORS.......................................................................................................................................ix
PEER REVIEW ...........................................................................................................................................xi
CONTENTS...............................................................................................................................................xiii
LIST OF FIGURES ..................................................................................................................................xvii
LIST OF TABLES.....................................................................................................................................xix
1. PUBLIC HEALTH STATEMENT.......................................................................................................... 1
1.1
WHAT IS BENZENE?................................................................................................................ 1
1.2
WHAT HAPPENS TO BENZENE WHEN IT ENTERS THE ENVIRONMENT? .................. 2
1.3
HOW MIGHT I BE EXPOSED TO BENZENE? ....................................................................... 3
1.4
HOW CAN BENZENE ENTER AND LEAVE MY BODY? .................................................... 4
1.5
HOW CAN BENZENE AFFECT MY HEALTH? ..................................................................... 4
1.6
HOW CAN BENZENE AFFECT CHILDREN? ........................................................................ 6
1.7
HOW CAN FAMILIES REDUCE THE RISK OF EXPOSURE TO BENZENE? .................... 7
1.8
IS THERE A MEDICAL TEST TO DETERMINE WHETHER I HAVE BEEN
EXPOSED TO BENZENE?........................................................................................................ 7
1.9
WHAT RECOMMENDATIONS HAS THE FEDERAL GOVERNMENT MADE TO
PROTECT HUMAN HEALTH?................................................................................................. 8
1.10 WHERE CAN I GET MORE INFORMATION? ....................................................................... 9
2. RELEVANCE TO PUBLIC HEALTH ................................................................................................. 11
2.1
BACKGROUND AND ENVIRONMENTAL EXPOSURES TO BENZENE IN THE
UNITED STATES..................................................................................................................... 11
2.2
SUMMARY OF HEALTH EFFECTS...................................................................................... 12
2.3
MINIMAL RISK LEVELS (MRLs) ......................................................................................... 21
3. HEALTH EFFECTS.............................................................................................................................. 29
3.1
INTRODUCTION ..................................................................................................................... 29
3.2
DISCUSSION OF HEALTH EFFECTS BY ROUTE OF EXPOSURE .................................. 29
3.2.1
Inhalation Exposure .............................................................................................................. 30
3.2.2
Oral Exposure...................................................................................................................... 104
3.2.3
Dermal Exposure................................................................................................................. 137
3.3
GENOTOXICITY ................................................................................................................... 141
3.4
TOXICOKINETICS................................................................................................................ 154
3.4.1
Absorption........................................................................................................................... 155
3.4.2
Distribution ......................................................................................................................... 163
3.4.3
Metabolism.......................................................................................................................... 166
3.4.4
Elimination and Excretion................................................................................................... 175
3.4.5
Physiologically Based Pharmacokinetic (PBPK)/Pharmacodynamic (PD) Models ........... 181
3.5
MECHANISMS OF ACTION ................................................................................................ 200
3.5.1
Pharmacokinetic Mechanisms............................................................................................. 200
3.5.2
Mechanisms of Toxicity...................................................................................................... 201
3.5.3
Animal-to-Human Extrapolations ....................................................................................... 204
3.6
TOXICITIES MEDIATED THROUGH THE NEUROENDOCRINE AXIS ........................ 205
3.7
CHILDRENS SUSCEPTIBILITY ......................................................................................... 206
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3.8
BIOMARKERS OF EXPOSURE AND EFFECT .................................................................. 208
3.8.1
Biomarkers Used to Identify or Quantify Exposure to Benzene......................................... 209
3.8.2
Biomarkers Used to Characterize Effects Caused by Benzene ........................................... 211
3.9
INTERACTIONS WITH OTHER CHEMICALS .................................................................. 212
3.10
POPULATIONS THAT ARE UNUSUALLY SUSCEPTIBLE ............................................. 214
3.11
METHODS FOR REDUCING TOXIC EFFECTS................................................................. 216
3.11.1
Reducing Peak Absorption Following Exposure ............................................................ 216
3.11.2
Reducing Body Burden................................................................................................... 217
3.11.3
Interfering with the Mechanism of Action for Toxic Effects ......................................... 217
3.12
ADEQUACY OF THE DATABASE...................................................................................... 218
3.12.1
Existing Information on Health Effects of Benzene ....................................................... 219
3.12.2
Identification of Data Needs ........................................................................................... 221
3.12.3
Ongoing Studies.............................................................................................................. 236
4. CHEMICAL AND PHYSICAL INFORMATION.............................................................................. 239
4.1
CHEMICAL IDENTITY......................................................................................................... 239
4.2
PHYSICAL AND CHEMICAL PROPERTIES...................................................................... 239
5. PRODUCTION, IMPORT/EXPORT, USE, AND DISPOSAL .......................................................... 243
5.1
PRODUCTION ....................................................................................................................... 243
5.2
IMPORT/EXPORT ................................................................................................................. 244
5.3
USE.......................................................................................................................................... 248
5.4
DISPOSAL .............................................................................................................................. 249
6. POTENTIAL FOR HUMAN EXPOSURE ......................................................................................... 251
6.1
OVERVIEW............................................................................................................................ 251
6.2
RELEASES TO THE ENVIRONMENT ................................................................................ 253
6.2.1
Air ....................................................................................................................................... 253
6.2.2
Water ................................................................................................................................... 257
6.2.3
Soil ...................................................................................................................................... 257
6.3
ENVIRONMENTAL FATE.................................................................................................... 258
6.3.1
Transport and Partitioning................................................................................................... 258
6.3.2
Transformation and Degradation ........................................................................................ 259
6.4
LEVELS MONITORED OR ESTIMATED IN THE ENVIRONMENT ............................... 265
6.4.1
Air ....................................................................................................................................... 265
6.4.2
Water ................................................................................................................................... 269
6.4.3
Sediment and Soil ............................................................................................................... 271
6.4.4
Other Environmental Media................................................................................................ 271
6.5
GENERAL POPULATION AND OCCUPATIONAL EXPOSURE ..................................... 273
6.6
EXPOSURES OF CHILDREN ............................................................................................... 281
6.7
POPULATIONS WITH POTENTIALLY HIGH EXPOSURES ........................................... 283
6.8
6.8.1
Identification of Data Needs ............................................................................................... 284
6.8.2
Ongoing Studies .................................................................................................................. 288
7. ANALYTICAL METHODS ............................................................................................................... 291
7.1
BIOLOGICAL MATERIALS................................................................................................. 291
7.2
ENVIRONMENTAL SAMPLES............................................................................................ 295
7.3
7.3.1
Identification of Data Needs ............................................................................................... 302
7.3.2
Ongoing Studies .................................................................................................................. 304
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8. REGULATIONS AND ADVISORIES ............................................................................................... 307

9. REFERENCES .................................................................................................................................... 313
10. GLOSSARY ...................................................................................................................................... 377
APPENDICES
A. ATSDR MINIMAL RISK LEVELS AND WORKSHEETS ............................................................. A-1
B. USERS GUIDE.................................................................................................................................. B-1
C. ACRONYMS, ABBREVIATIONS, AND SYMBOLS...................................................................... C-1
D. INDEX ................................................................................................................................................ D-1
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LIST OF FIGURES
3-1. Levels of Significant Exposure to Benzene Inhalation.................................................................... 66
3-2. Levels of Significant Exposure to Benzene Oral........................................................................... 122
3-3. Metabolic Pathways for Benzene ..................................................................................................... 167
3-4. Conceptual Representation of a Physiologically Based Pharmacokinetic (PBPK) Model for a
Hypothetical Chemical Substance.................................................................................................... 183
3-5. General Structure of Physiologically Based Pharmacokinetic Models of Benzene ......................... 184
3-6. Existing Information on Health Effects of Benzene......................................................................... 220
6-1. Frequency of NPL Sites with Benzene Contamination .................................................................... 252
6-2. Environmental Transformation Products of Benzene in Various Media.......................................... 260
6-3. Benzene Emissions and Exposures................................................................................................... 275
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LIST OF TABLES
3-1. Levels of Significant Exposure to Benzene Inhalation.................................................................... 33
3-2. Levels of Significant Exposure to Benzene Oral........................................................................... 106
3-3. Levels of Significant Exposure to Benzene Dermal ...................................................................... 139
3-4. Genotoxicity of Benzene In Vivo...................................................................................................... 142
3-5. Genotoxicity of Benzene In Vitro..................................................................................................... 146
3-6. Summary Comparison of Physiologically Based Pharmacokinetic Models for Benzene ................ 185
3-7. Ongoing Studies on the Health Effects of Benzene.......................................................................... 237
4-1. Chemical Identity of Benzene .......................................................................................................... 240
4-2. Physical and Chemical Properties of Benzene ................................................................................. 241
5-1. Facilities that Produce, Process, or Use Benzene ............................................................................. 245
5-2. Current U.S. Manufacturers of Benzene........................................................................................... 247
6-1. Releases to the Environment from Facilities that Produce, Process, or Use Benzene...................... 254
6-2. Benzene Levels in Air Samples........................................................................................................ 266
6-3. Benzene in Food ............................................................................................................................... 272
6-4. Percentage of Employees Exposed to Benzene by Exposure Level and Industry Division ............. 280
6-5. Ongoing Studies on the Potential for Human Exposure to Benzene ................................................ 289
7-1. Analytical Methods for Determining Benzene in Biological Samples............................................. 292
7-2. Analytical Methods for Determining Metabolites of Benzene in Urine........................................... 294
7-3. Analytical Methods for Determining Benzene in Environmental Samples...................................... 296
7-4. Ongoing Studies on Benzene, Analytical Methods .......................................................................... 305
8-1. Regulations and Guidelines Applicable to Benzene......................................................................... 308
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BENZENE
1. PUBLIC HEALTH STATEMENT

This public health statement tells you about benzene and the effects of exposure to it.
The Environmental Protection Agency (EPA) identifies the most serious hazardous waste sites in
the nation. These sites are then placed on the National Priorities List (NPL) and are targeted for
long-term federal clean-up activities. Benzene has been found in at least 1,000 of the
1,684 current or former NPL sites. Although the total number of NPL sites evaluated for this
substance is not known, the possibility exists that the number of sites at which benzene is found
may increase in the future as more sites are evaluated. This information is important because
these sites may be sources of exposure and exposure to this substance may harm you.
When a substance is released either from a large area, such as an industrial plant, or from a
container, such as a drum or bottle, it enters the environment. Such a release does not always
lead to exposure. You can be exposed to a substance only when you come in contact with it.
You may be exposed by breathing, eating, or drinking the substance, or by skin contact.
If you are exposed to benzene, many factors will determine whether you will be harmed. These
factors include the dose (how much), the duration (how long), and how you come in contact with
it. You must also consider any other chemicals you are exposed to and your age, sex, diet,
family traits, lifestyle, and state of health.
1.1
WHAT IS BENZENE?
Benzene, also known as benzol, is a colorless liquid with a sweet odor. Benzene evaporates into
air very quickly and dissolves slightly in water. Benzene is highly flammable. Most people can
begin to smell benzene in air at approximately 60 parts of benzene per million parts of air (ppm)
and recognize it as benzene at 100 ppm. Most people can begin to taste benzene in water at 0.5
4.5 ppm. One part per million is approximately equal to one drop in 40 gallons. Benzene is
found in air, water, and soil. Benzene comes from both industrial and natural sources.
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Industrial Sources and Uses. Benzene was first discovered and isolated from coal tar in the
1800s. Today, benzene is made mostly from petroleum. Because of its wide use, benzene ranks
in the top 20 in production volume for chemicals produced in the United States. Various
industries use benzene to make other chemicals, such as styrene (for Styrofoam and other
plastics), cumene (for various resins), and cyclohexane (for nylon and synthetic fibers). Benzene
is also used in the manufacturing of some types of rubbers, lubricants, dyes, detergents, drugs,
and pesticides.
Natural Sources. Natural sources of benzene, which include gas emissions from volcanoes and
forest fires, also contribute to the presence of benzene in the environment. Benzene is also
present in crude oil and gasoline and cigarette smoke. For more information on characteristics
and uses of benzene, see Chapters 4 and 5.
1.2
WHAT HAPPENS TO BENZENE WHEN IT ENTERS THE ENVIRONMENT?
Benzene is commonly found in the environment. Industrial processes are the main sources of
benzene in the environment. Benzene levels in the air can be elevated by emissions from
burning coal and oil, benzene waste and storage operations, motor vehicle exhaust, and
evaporation from gasoline service stations. Tobacco smoke is another source of benzene in air,
particularly indoors. Industrial discharge, disposal of products containing benzene, and gasoline
leaks from underground storage tanks release benzene into water and soil.
Benzene can pass into air from water and soil surfaces. Once in the air, benzene reacts with
other chemicals and breaks down within a few days. Benzene in the air can also be deposited on
the ground by rain or snow.
Benzene in water and soil breaks down more slowly. Benzene is slightly soluble in water and
can pass through the soil into underground water. Benzene in the environment does not build up
in plants or animals. For more information on what happens to benzene after it gets into the
environment, see Chapters 5 and 6.
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1.3
HOW MIGHT I BE EXPOSED TO BENZENE?
Everyone is exposed to a small amount of benzene every day. You are exposed to benzene in the
outdoor environment, in the workplace, and in the home. Exposure of the general population to
benzene mainly occurs through breathing air that contains benzene. The major sources of
benzene exposure are tobacco smoke, automobile service stations, exhaust from motor vehicles,
and industrial emissions. Vapors (or gases) from products that contain benzene, such as glues,
paints, furniture wax, and detergents, can also be a source of exposure. Auto exhaust and
industrial emissions account for about 20% of the total national exposure to benzene. About half
of the exposure to benzene in the United States results from smoking tobacco or from exposure
to tobacco smoke. The average smoker (32 cigarettes per day) takes in about 1.8 milligrams
(mg) of benzene per day. This amount is about 10 times the average daily intake of benzene by
nonsmokers.
Measured levels of benzene in outdoor air have ranged from 0.02 to 34 parts of benzene per
billion parts of air (ppb) (1 ppb is 1,000 times less than 1 ppm). People living in cities or
industrial areas are generally exposed to higher levels of benzene in air than those living in rural
areas. Benzene levels in the home are usually higher than outdoor levels. People may be
exposed to higher levels of benzene in air by living near hazardous waste sites, petroleum
refining operations, petrochemical manufacturing sites, or gas stations.
For most people, the level of exposure to benzene through food, beverages, or drinking water is
not as high as through air. Drinking water typically contains less than 0.1 ppb benzene. Benzene
has been detected in some bottled water, liquor, and food. Leakage from underground gasoline
storage tanks or from landfills and hazardous waste sites that contain benzene can result in
benzene contamination of well water. People with benzene-contaminated tap water can be
exposed from drinking the water or eating foods prepared with the water. In addition, exposure
can result from breathing in benzene while showering, bathing, or cooking with contaminated
water.
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Individuals employed in industries that make or use benzene may be exposed to the highest
levels of benzene. As many as 238,000 people may be occupationally exposed to benzene in the
United States. These industries include benzene production (petrochemicals, petroleum refining,
and coke and coal chemical manufacturing), rubber tire manufacturing, and storage or transport
of benzene and petroleum products containing benzene. Other workers who may be exposed to
benzene include coke oven workers in the steel industry, printers, rubber workers, shoe makers,
laboratory technicians, firefighters, and gas station employees. For more information on how
you might be exposed to benzene, see Chapter 6.
1.4
HOW CAN BENZENE ENTER AND LEAVE MY BODY?
Benzene can enter your body through your lungs, gastrointestinal tract, and across your skin.
When you are exposed to high levels of benzene in air, about half of the benzene you breathe in
passes through the lining of your lungs and enters your bloodstream. When you are exposed to
benzene in food or drink, most of the benzene you take in by mouth passes through the lining of
your gastrointestinal tract and enters your bloodstream. A small amount will enter your body by
passing through your skin and into your bloodstream during skin contact with benzene or
benzene-containing products. Once in the bloodstream, benzene travels throughout your body
and can be temporarily stored in the bone marrow and fat. Benzene is converted to products,
called metabolites, in the liver and bone marrow. Some of the harmful effects of benzene
exposure are caused by these metabolites. Most of the metabolites of benzene leave the body in
the urine within 48 hours after exposure. For more information on how benzene can enter and
leave your body, see Chapter 3.
1.5
HOW CAN BENZENE AFFECT MY HEALTH?
Scientists use many tests to protect the public from harmful effects of toxic chemicals and to find
ways for treating persons who have been harmed.
One way to learn whether a chemical will harm people is to determine how the body absorbs,
uses, and releases the chemical. For some chemicals, animal testing may be necessary. Animal
testing may also help identify health effects such as cancer or birth defects. Without laboratory
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animals, scientists would lose a basic method for getting information needed to make wise
decisions that protect public health. Scientists have the responsibility to treat research animals
with care and compassion. Scientists must comply with strict animal care guidelines because
laws today protect the welfare of research animals.
After exposure to benzene, several factors determine whether harmful health effects will occur,
as well as the type and severity of such health effects. These factors include the amount of
benzene to which you are exposed and the length of time of the exposure. Most information on
effects of long-term exposure to benzene are from studies of workers employed in industries that
make or use benzene. These workers were exposed to levels of benzene in air far greater than
the levels normally encountered by the general population. Current levels of benzene in
workplace air are much lower than in the past. Because of this reduction and the availability of
protective equipment such as respirators, fewer workers have symptoms of benzene poisoning.
Brief exposure (510 minutes) to very high levels of benzene in air (10,00020,000 ppm) can
result in death. Lower levels (7003,000 ppm) can cause drowsiness, dizziness, rapid heart rate,
headaches, tremors, confusion, and unconsciousness. In most cases, people will stop feeling
these effects when they are no longer exposed and begin to breathe fresh air.
Eating foods or drinking liquids containing high levels of benzene can cause vomiting, irritation
of the stomach, dizziness, sleepiness, convulsions, rapid heart rate, coma, and death. The health
effects that may result from eating foods or drinking liquids containing lower levels of benzene
are not known. If you spill benzene on your skin, it may cause redness and sores. Benzene in
your eyes may cause general irritation and damage to your cornea.
Benzene causes problems in the blood. People who breathe benzene for long periods may
experience harmful effects in the tissues that form blood cells, especially the bone marrow.
These effects can disrupt normal blood production and cause a decrease in important blood
components. A decrease in red blood cells can lead to anemia. Reduction in other components
in the blood can cause excessive bleeding. Blood production may return to normal after
exposure to benzene stops. Excessive exposure to benzene can be harmful to the immune
BENZENE
6
system, increasing the chance for infection and perhaps lowering the body's defense against
cancer.
Long-term exposure to benzene can cause cancer of the blood-forming organs. This condition is
called leukemia. Exposure to benzene has been associated with development of a particular type
of leukemia called acute myeloid leukemia (AML). The Department of Health and Human
Services has determined that benzene is a known carcinogen (can cause cancer). Both the
International Agency for Cancer Research and the EPA have determined that benzene is
carcinogenic to humans.
Exposure to benzene may be harmful to the reproductive organs. Some women workers who
breathed high levels of benzene for many months had irregular menstrual periods. When
examined, these women showed a decrease in the size of their ovaries. However, exact exposure
levels were unknown, and the studies of these women did not prove that benzene caused these
effects. It is not known what effects exposure to benzene might have on the developing fetus in
pregnant women or on fertility in men. Studies with pregnant animals show that breathing
benzene has harmful effects on the developing fetus. These effects include low birth weight,
delayed bone formation, and bone marrow damage.
We do not know what human health effects might occur after long-term exposure to food and
water contaminated with benzene. In animals, exposure to food or water contaminated with
benzene can damage the blood and the immune system and can cause cancer. See Chapters 2
and 3 for more information on the health effects resulting from benzene exposure.
1.6
HOW CAN BENZENE AFFECT CHILDREN?
This section discusses potential health effects in humans from exposures during the period from
conception to maturity at 18 years of age.
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7
Children can be affected by benzene exposure in the same ways as adults. Benzene can pass
from the mothers blood to a fetus. It is not known if children are more susceptible to benzene
poisoning than adults.
1.7 HOW CAN FAMILIES REDUCE THE RISK OF EXPOSURE TO BENZENE?
If your doctor finds that you have been exposed to substantial amounts of benzene, ask whether
your children might also have been exposed. Your doctor might need to ask your state health
department to investigate.
Gasoline and cigarette smoke are two main sources of human exposure to benzene. Benzene
exposure can be reduced by limiting contact with these sources. People are exposed to benzene
from both active and passive second-hand smoke. Average smokers take in about 10 times more
benzene than nonsmokers each day. Families are encouraged not to smoke in their house, in
enclosed environments, or near their children.
Benzene is a major component of gasoline and used in many manufacturing processes.
Increased levels of benzene can be found at fueling stations, and in air emissions from
manufacturing plants and hazardous waste sites. Living near gasoline fueling stations or
hazardous waste sites may increase exposure to benzene. People are advised not to have their
families play near fueling stations, manufacturing plants, or hazardous waste sites.
1.8 IS THERE A MEDICAL TEST TO DETERMINE WHETHER I HAVE BEEN
EXPOSED TO BENZENE?
Several tests can show whether you have been exposed to benzene. Some of these tests may be
available at your doctor's office. All of these tests are limited in what they can tell you. The test
for measuring benzene in your breath must be done shortly after exposure. This test is not very
helpful for detecting very low levels of benzene in your body. Benzene can be measured in your
blood. However, because benzene rapidly disappears in the blood, measurements may be useful
only for recent exposures.
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8
In the body, benzene is converted to products called metabolites. Certain metabolites of

benzene, such as phenol, muconic acid, and S-phenylmercapturic acid can be measured in the
urine. The amount of phenol in urine has been used to check for benzene exposure in workers.
The test is useful only when you are exposed to benzene in air at levels of 10 ppm or greater.
However, this test must also be done shortly after exposure, and it is not a reliable indicator of
how much benzene you have been exposed to, because phenol is present in the urine from other
sources (diet, environment). Measurements of muconic acid or S-phenylmercapturic acid in the
urine are more sensitive and reliable indicators of benzene exposure. The measurement of
benzene in blood or of metabolites in urine cannot be used for making predictions about whether
you will experience any harmful health effects. Blood counts of all components of the blood and
examination of bone marrow are used to determine benzene exposure and its health effects.
For people exposed to relatively high levels of benzene, complete blood analyses can be used to
monitor possible changes related to exposure. However, blood analyses are not useful when
exposure levels are low. For more information on tests for benzene exposure, see Chapters 3
and 7.
1.9 WHAT RECOMMENDATIONS HAS THE FEDERAL GOVERNMENT MADE TO
PROTECT HUMAN HEALTH?
The federal government develops regulations and recommendations to protect public health.
Regulations can be enforced by law. The EPA, the Occupational Safety and Health
Administration (OSHA), and the Food and Drug Administration (FDA) are some federal
agencies that develop regulations for toxic substances. Recommendations provide valuable
guidelines to protect public health, but cannot be enforced by law. The Agency for Toxic
Substances and Disease Registry (ATSDR) and the National Institute for Occupational Safety
and Health (NIOSH) are two federal organizations that develop recommendations for toxic
substances.
Regulations and recommendations can be expressed as not-to-exceed levels, that is, levels of a
toxic substance in air, water, soil, or food that do not exceed a critical value that is usually based
on levels that affect animals; they are then adjusted to levels that will help protect humans.
BENZENE
9
Sometimes these not-to-exceed levels differ among federal organizations because they used
different exposure times (an 8-hour workday or a 24-hour day), different animal studies, or other
factors.
Recommendations and regulations are also updated periodically as more information becomes
available. For the most current information, check with the federal agency or organization that
provides it. Some regulations and recommendations for benzene include the following:
EPA has set 5 ppb as the maximum permissible level of benzene in drinking water. EPA has set
a goal of 0 ppb for benzene in drinking water and in water such as rivers and lakes because
benzene can cause leukemia. EPA estimates that 10 ppb benzene in drinking water that is
consumed regularly or exposure to 0.4 ppb in air over a lifetime could cause a risk of one
additional cancer case for every 100,000 exposed persons. EPA recommends 200 ppb as the
maximum permissible level of benzene in water for short-term exposures (10 days) for children.
EPA requires that the National Response Center be notified following a discharge or spill into
the environment of 10 pounds or more of benzene.
OSHA regulates levels of benzene in the workplace. The maximum allowable amount of
benzene in workroom air during an 8-hour workday, 40-hour workweek is 1 ppm. Because
benzene can cause cancer, NIOSH recommends that all workers wear special breathing
equipment when they are likely to be exposed to benzene at levels exceeding the recommended
(8-hour) exposure limit of 0.1 ppm. For more information on federal regulations, see Chapter 8.
1.10
WHERE CAN I GET MORE INFORMATION?
If you have any more questions or concerns, please contact your community or state health or
environmental quality department, or contact ATSDR at the address and phone number below.
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10
ATSDR can also tell you the location of occupational and environmental health clinics. These
clinics specialize in recognizing, evaluating, and treating illnesses that result from exposure to
hazardous substances.
Toxicological profiles are also available on-line at www.atsdr.cdc.gov and on CD-ROM. You
may request a copy of the ATSDR ToxProfilesTM CD-ROM by calling the toll-free information
and technical assistance number at 1-800-CDCINFO (1-800-232-4636), by e-mail at
cdcinfo@cdc.gov, or by writing to:
Division of Toxicology and Environmental Medicine
1600 Clifton Road NE
Mailstop F-32
Atlanta, GA 30333
Fax: 1-770-488-4178
Organizations for-profit may request copies of final Toxicological Profiles from the following:
National Technical Information Service (NTIS)
5285 Port Royal Road
Springfield, VA 22161
Phone: 1-800-553-6847 or 1-703-605-6000
Web site: http://www.ntis.gov/
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2. RELEVANCE TO PUBLIC HEALTH
2.1 BACKGROUND AND ENVIRONMENTAL EXPOSURES TO BENZENE IN THE UNITED

STATES
Benzene is widely distributed in the environment. The exposure scenario of most concern to the general
public is low-level inhalation over long periods. This is because the general population is exposed to
benzene mainly through inhalation of contaminated air, particularly in areas of heavy traffic and around
gas stations, and through inhalation of tobacco smoke from both active and passive smoking. Smoking
has been identified as the single most important source of benzene exposure for the estimated 40 million
U.S. smokers. Smoking accounts for approximately half of the total benzene exposure of the general
population. Individuals employed in industries that make or use benzene, or products containing benzene,
are probably exposed to the highest concentrations of atmospheric benzene. In addition, benzene is a
common combustion product, providing high inhalation exposure potential for firefighters. Of the
general population, those residing around certain chemical manufacturing sites or living near waste sites
containing benzene or near leaking gasoline tanks may be exposed to concentrations of benzene that are
higher than background air concentrations. In private homes, benzene levels in the air have been shown
to be higher in homes with attached garages, or where the inhabitants smoke inside the house.
Although low levels of benzene have been detected in certain foods, beverages, and tap water, these do
not constitute major sources of exposure for most people. However, leakage from underground gasoline
storage tanks and seepage from landfills and hazardous waste sites have resulted in significant benzene
contamination of well water. People with contaminated tap water can be exposed from drinking the water
or eating foods prepared with it. In addition, exposure can also occur via inhalation during showering,
bathing, or cooking with contaminated tap water. Showering and bathing with benzene-contaminated
water can also contribute significantly to dermal exposure.
The Benzene Subregistry Baseline Technical Report of the National Exposure Registry contains
information on 1,143 persons who had documented exposure to benzene in their drinking water and were
exposed for at least 30 days. No causal relationship has been proposed for health conditions identified in
the base subregistry or continued follow-up of the population.
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12
2.2
SUMMARY OF HEALTH EFFECTS
The carcinogenicity of benzene is well documented in exposed workers. Epidemiological studies and
case reports provide clear evidence of a causal relationship between occupational exposure to benzene
and benzene-containing solvents and the occurrence of acute myelogenous leukemia (AML). The
epidemiological studies are generally limited by confounding chemical exposures and methodological
problems, including inadequate or lack of exposure monitoring and low statistical power, but a consistent
excess risk of leukemia across studies indicates that benzene is the causal factor.
In vivo and in vitro data from both humans and animals indicate that benzene and/or its metabolites are
genotoxic. Chromosomal aberrations (hypo- and hyperdiploidy, deletions, breaks, and gaps) in peripheral
lymphocytes and bone marrow cells are the predominant effects seen in humans.
Damage to both the humoral and cellular components of the immune system has been known to occur in
humans following inhalation exposure. This is manifested by decreased levels of antibodies and
decreased levels of leukocytes in workers. Animal data support these findings.
The most characteristic systemic effect resulting from intermediate and chronic benzene exposure is
arrested development of blood cells. Early biomarkers of exposure to relatively low levels of benzene
include depressed numbers of one or more of the circulating blood cell types. A common clinical finding
in benzene hematotoxicity is cytopenia, which is a decrease in various cellular elements of the circulating
blood manifested as anemia, leukopenia, or thrombocytopenia in humans and in animals. Benzeneassociated cytopenias vary and may involve a reduction in one (unicellular cytopenias) to all three
(pancytopenia) cellular elements of the blood.
Benzene also causes a life-threatening disorder called aplastic anemia in humans and animals. This
disorder is characterized by reduction of all cellular elements in the peripheral blood and in bone marrow,
leading to fibrosis, an irreversible replacement of bone marrow. Benzene has also been associated with
acute non-lymphocytic leukemia in humans, and aplastic anemia may be an early indicator of developing
acute non-lymphocytic leukemia in some cases.
Limited information is available on other systemic effects reported in humans and is associated with
highlevel benzene exposure. Respiratory effects have been noted after acute exposure of humans to
benzene vapors. Cardiovascular effects, particularly ventricular fibrillation, have been suggested as the
BENZENE
13
cause of death in fatal exposures to benzene vapor. Gastrointestinal effects have been noted in humans
after fatal inhalation exposure (congestive gastritis), and ingestion (toxic gastritis and pyloric stenosis), of
benzene. Myelofibrosis (a form of aplastic anemia) was reported by a gasoline station attendant who had
been exposed to benzene by inhalation, and probably also through dermal contact. Myalgia was also
reported in steel plant workers exposed to benzene vapors. Reports of renal effects in humans after
benzene exposure consist of kidney congestion after fatal inhalation exposure. Dermal and ocular effects
including skin irritation and burns, and eye irritation have been reported after exposure to benzene vapors.
Swelling and edema have been reported to occur in a human who swallowed benzene. Studies in animals
show systemic effects after inhalation exposure, including cardiovascular effects. Oral administration of
benzene to animals has yielded information concerning hepatic effects. A study conducted in rabbits
lends support to the finding that benzene is irritating and damaging to the skin and also shows that it is
irritating and damaging to the eyes following dermal or ocular application.
Neurological effects have been commonly reported in humans following high-level exposure to benzene.
Fatal inhalation exposure has been associated with vascular congestion in the brain. Chronic inhalation
exposure has been associated with distal neuropathy, difficulty in sleeping, and memory loss. Oral
exposure results in symptoms similar to inhalation exposure. Studies in animals suggest that inhalation
exposure to benzene results in depressed electrical activity in the brain, loss of involuntary reflexes and
narcosis, decrease in hind-limb grip strength and tremors, and narcosis, among other symptoms. Oral
exposure to benzene has not been shown to cause significant changes in behavior. No neurological
effects have been reported after dermal exposure to liquid benzene in either humans or animals.
Acute inhalation and oral exposures of humans to high concentrations of benzene have caused death.
These exposures are also associated with central nervous system depression. Chronic low-level exposures
have been associated with peripheral nervous system effects. Abnormalities in motor conduction velocity
were noted in four of six pancytopenic individuals occupationally exposed to adhesives containing
benzene.
Evidence of an effect of benzene exposure on human reproduction is not sufficient to demonstrate a
causal association. Some animal studies provide limited evidence that benzene affects reproductive
organs following inhalation exposure. Results from studies of benzene administered orally to rats and
mice indicate no adverse effect on male or female reproductive organs at 17 weeks, but at 2 years,
endometrial polyps were observed in female rats, preputial gland lesions were observed in male mice, and
ovarian lesions were observed in female mice. Results are conflicting or inconclusive as to whether
BENZENE
14
inhalation of benzene vapors reduces the number of live fetuses and/or the incidences of pregnancy.
Other studies are negative for effects on reproductive competence.
Epidemiological studies implicating benzene as a developmental toxicant have many limitations, and
thus, it is not possible to assess the effect of benzene on the human fetus. Results of inhalation studies
conducted in animals are fairly consistent across species and demonstrate that, at levels >47 ppm, benzene
is fetotoxic as evidenced by decreased fetal weight and/or minor skeletal variants. Benzene has also been
shown to reduce pup body weight in mice. A persistent decrease in the number of erythroid precursors
was found in mice exposed in utero. Benzene has not been shown to be teratogenic, but has been shown
to be fetotoxic in animals at high concentrations that are maternally toxic.
Cancer.
The strongest evidence for the leukemogenic potential of benzene comes from series of cohort
mortality studies on workers exposed to benzene in Ohio (the Pliofilm study) and China (the NCI/CAPM
study). The Pliofilm study investigated workers exposed to benzene in three rubber hydrochloride
(Pliofilm) manufacturing plants. Mortality from all leukemias was increased but declined after
additional years of follow-up, suggesting that the excess risk diminished with time since exposure.
Exposures in the most recent 10 years were most strongly associated with leukemia risk, and there was no
significant relation between leukemia death and benzene exposures received more than 20 years
previously. AML accounted for most of the increased leukemia, and the risk of AML increased with
increasing cumulative exposure above 200 ppm-years.
The NCI/CAPM study, a collaboration between the National Cancer Institute and the Chinese Academy
of Preventive Medicine, evaluated lymphohematopoietic malignancies and other hematologic disorders in
74,828 benzene-exposed workers employed in 672 factories in 12 cities in China. Findings included
increased risks for all leukemias, acute nonlymphocytic leukemia (ANLL), and combined ANLL and
precursor myelodysplastic syndromes. These risks were increased at average exposure levels of 10
24 ppm and cumulative exposure levels of 4099 ppm-years, and tended to increase with increasing
average and cumulative levels of exposure.
The results of the Pliofilm and NCI/CAPM studies are consistent with epidemiologic studies and case
reports showing increased incidences of leukemia in shoe factory and rotogravure plant workers exposed
to high benzene levels during its use as a solvent. No significant increases in leukemia or other
lymphohematopoietic malignancies were found in chemical industry workers or petroleum industry
workers exposed to lower levels of benzene.
BENZENE
15
Possible associations between occupational exposure to benzene and non-Hodgkins lymphoma (NHL)
and multiple myeloma have been suggested. The risk for mortality from NHL increased with increasing
level and duration of benzene exposure the NCI/CAPM study. The significance of this finding is unclear
because NHL mortality was not significantly elevated in the cohort overall, concerns regarding the
adequacy of the data have been raised, and increases in NHL were not found in other cohort mortality
studies or in case-control studies of benzene-exposed workers.
The risk of mortality from multiple myeloma was increased in one of the early assessments of the
Pliofilm cohort. The implication of this finding is unclear because the risk declined to non-significant
levels in subsequent follow-up studies, and was not supported by the findings of other cohort mortality
studies. Additionally, population-based and hospital-based case-control studies indicate that benzene
exposure is not likely to be causally related to the risk of multiple myeloma. A meta-analysis of casecontrol studies found no significant association between occupational exposure to benzene and benzenecontaining products and risk of multiple myeloma from sources categorized as benzene and/or organic
solvents, petroleum, or petroleum products.
Animal studies provide supporting evidence for the carcinogenicity of benzene. Benzene has been shown
to be a multiple site carcinogen in rats and mice following inhalation and oral exposure. Tumors that
were increased in rats that were exposed to 200 or 300 ppm benzene by inhalation for 47 hours/day,
5 days/week for up to 104 weeks included carcinomas of the Zymbal gland and oral cavity. Mice that
were exposed to 100 or 300 ppm benzene for 6 hours/day, 5 days/week for 16 weeks and observed for
18 months or life developed a variety of tumors, including thymic lymphomas, myelogenous leukemias,
and Zymbal gland, ovarian, and lung tumors.
In oral bioassays conducted by the National Toxicology Program, benzene was administered to rats and
mice by gavage at dose levels of 25200 mg/kg/day on 5 days/week for 103 weeks. Tumors that were
induced in the rats included Zymbal gland carcinomas and squamous cell papillomas and carcinomas of
the oral cavity and skin. In the mice, benzene caused tumors that included malignant lymphomas,
Zymbal gland carcinomas, lung alveolar/bronchiolar adenomas and carcinomas, Harderian gland
adenomas, preputial gland squamous cell carcinomas, and mammary gland carcinomas. Similar effects
occurred in rats exposed to 50500 mg/kg/day benzene by gavage on 45 days/week for up to 104 weeks
and observed for life; induced tumors included carcinomas of the Zymbal gland, oral cavity, forestomach,
BENZENE
16
nasal cavity, and skin. Mice that were similarly exposed to 500 mg/kg/day for 52 or 78 weeks developed
Zymbal gland carcinomas, mammary carcinomas, and lung adenomas.
Application of benzene to the skin of animals has not produced evidence of carcinogenicity, although
most of the dermal studies were inadequate for cancer evaluation. Many dermal carcinogenicity studies
of other chemicals used benzene as a vehicle and treated large numbers of control animals (mice) with
benzene alone. None of these studies indicated that benzene induced skin tumors; however, all possible
tumor sites usually were not examined.
EPA, IARC, and the Department of Health and Human Services have concluded that benzene is a human
carcinogen. The Department of Health and Human Services determined that benzene is a known
carcinogen based on human evidence showing a causal relationship between exposure to benzene and
cancer. Two studies classified benzene in Group 1 (carcinogenic to humans) based on sufficient evidence
in both humans and animals. EPA classified benzene in Category A (known human carcinogen) based on
convincing evidence in humans supported by evidence from animal studies. Under EPAs most recent
guidelines for carcinogen risk assessment, benzene is characterized as a known human carcinogen for all
routes of exposure based on convincing human evidence as well as supporting evidence from animal
studies. Based on human leukemia data, EPA derived a range of inhalation unit risk values of 2.2x10-6
7.8x10-6 (g/m3)-1 for benzene. For risks ranging from 1x10-4 to 1x10-7, the corresponding air
concentrations range from 13.045.0 g/m3 (414 ppb) to 0.0130.045 g/m3 (0.0040.014 ppb),
respectively.
The consensus conclusion that benzene is a human carcinogen is based on sufficient inhalation data in
humans supported by animal evidence, including the oral studies in animals. The human cancer induced
by inhalation exposure to benzene is predominantly acute nonlymphocytic (myelocytic) leukemia,
whereas benzene is a multiple site carcinogen in animals by both the inhalation and oral routes. Due to
the lack of oral carcinogenicity data in humans, as well as the lack of a well-demonstrated and
reproducible animal model for leukemia from benzene exposure, EPA extrapolated an oral slope factor
from the inhalation unit risk range. The oral slope factor ranges from 1.5x10-2 to 5.5x10-2 (mg/kg/day)-1,
and for cancer risks from 1x10-4 to 1x10-7, the corresponding dose levels are 6.7x10-31.8x10-3 to
6.7x10-61.8x10-6 mg/kg/day, respectively.
Hematological Effects.
Both human and animal studies have shown that benzene exerts toxic effects
on various parts of the hematological system. All of the major types of blood cells are susceptible
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17
(erythrocytes, leukocytes, and platelets). In the less severe cases of toxicity, specific deficiencies occur in
individual types of blood elements. A more severe effect occurs when there is hypoplasia of the bone
marrow, or hypercellular marrow exhibiting ineffective hematopoiesis so that all types of blood cells are
found in reduced numbers. This is known as pancytopenia. A biphasic response (i.e., a hyperplastic
effect in addition to destruction of the bone marrow cells) has been observed. Severe damage to the bone
marrow involving cellular aplasia is known as aplastic anemia and can occur with prolonged exposure to
benzene. This condition can lead to leukemia.
Numerous earlier studies of benzene-exposed workers demonstrated that chronic exposure to benzene air
concentrations of 10 ppm or more resulted in adverse hematological effects, which increased in severity
with increasing benzene exposure levels. Animal studies support the findings in humans. Significantly
reduced counts for all three blood factors (white blood cells [WBCs], red blood cells [RBCs], and
platelets); and other evidence of adverse effects on blood-forming units (reduced bone marrow cellularity,
bone marrow hyperplasia and hypoplasia, granulocytic hyperplasia, decreased numbers of colony-forming
granulopoietic stem cells and erythroid progenitor cells, damaged erythrocytes and erythroblast-forming
cells) have been observed in animals at benzene concentrations in the range of 10300 ppm and above.
Several more recent epidemiological studies have demonstrated hematological effects (including
significant reductions in WBC, RBC, and platelet counts) in workers chronically exposed to benzene
levels below 10 ppm, and even as low as 1 ppm or less. Results of one of these studies served as the basis
for a chronic-duration inhalation MRL for benzene. Other reports demonstrated the lack of clinical signs
of hematotoxicity following long-term, low-level occupational exposure to benzene levels below
approximately 0.5 ppm (8-hour time-weighted average [TWA]). These investigators utilized a defined
range of clinically normal hematological values and compared the prevalence of abnormal results
between benzene-exposed workers and unexposed controls. The normal range for certain hematological
parameters is necessarily broad due to large interindividual differences in clinical status. Restricting the
comparison of benzene-exposed and nonexposed populations to only those values considered clinically
abnormal or adverse may reduce the sensitivity of a particular study to detect meaningful changes at the
population level.
Only one study was found that described hematological effects in humans after oral exposure to benzene.
No reports describing hematological effects in humans following direct dermal exposure to benzene were
found. However, intermediate- and chronic-duration animal studies show that loss of blood elements
occurs in animals exposed to benzene in drinking water or by gavage at doses as low as 825 mg/kg/day.
BENZENE
18
Based on information found in the literature, it is reasonable to expect that adverse hematological effects
might occur in humans after inhalation, oral, or dermal exposure, since absorption of benzene through any
route of exposure would increase the risk of damage to blood elements. Studies show that the
hematological system is susceptible to chronic exposure at low levels, so people living in and around
hazardous waste sites that may be exposed to contaminated air, drinking water, soil, or food may be at an
increased risk for adverse hematological effects. Deficiencies in various types of blood cells lead to other
disorders, such as hemorrhagic conditions from a lack of platelets, susceptibility to infection from the lack
of leukocytes, and increased cardiac output from the lack of erythrocytes.
Immunological and Lymphoreticular Effects.
Benzene has been shown to have adverse
immunological effects in humans following inhalation exposure for intermediate and chronic durations.
Adverse immunological effects in animals occur following both inhalation and oral exposure for acute,
intermediate, and chronic durations. The effects include damage to both humoral (antibody) and cellular
(leukocyte) responses. Human studies of intermediate and chronic duration have shown that benzene
causes decreases in the levels of circulating leukocytes in workers at low levels (30 ppm) of exposure and
decreases in levels of circulating antibodies in workers exposed to benzene at 37 ppm. Other studies
have shown decreases in human lymphocytes and other blood elements after exposure; these effects have
been seen at occupational exposure levels as low as 1 ppm or less. Animal data support these findings.
Both humans and rats have shown increases in leukocyte alkaline phosphatase activity. No studies
regarding effects from oral or dermal exposure in humans were located. However, exposure to benzene
through ingestion or dermal contact could cause immunological effects similar to those seen after
inhalation exposure in humans and inhalation and oral exposure in animals.
Animal studies have also shown that benzene decreases circulating leukocytes and decreases the ability of
lymphoid tissue to produce the mature lymphocytes necessary to form antibodies. This has been
demonstrated in animals exposed for acute, intermediate, or chronic periods via the inhalation route. This
decrease in lymphocyte numbers is reflected in impaired cell-mediated immune functions in mice
following intermediate inhalation exposure to 100 ppm of benzene. The impaired cellular immunity after
benzene treatment was observed both in vivo and in vitro. Mice exposed to 100 ppm for a total of
100 days were challenged with 104 polyoma virus-induced tumor cells (PYB6). Nine of 10 mice had
reduced tumor resistance resulting in the development of lethal tumors. In the same study, lymphocytes
were obtained from spleens of benzene-treated mice and tested for their immune capacity in vitro. The
results showed that two other immune functions, alloantigen response (capacity to respond to foreign
BENZENE
19
antigens) and cytotoxicity, were also impaired. Similar effects were noted in mice exposed to benzene via
the oral route for intermediate time periods, and in rats and mice exposed for chronic time periods. A
decrease in spleen weight was observed in mice after acute-duration exposure to benzene at 25 ppm, the
same dose levels at which a decrease in circulating leukocytes was observed. Similar effects on spleen
weight and circulating leukocytes were observed in mice exposed to 12 ppm benzene 2 hours/day for
30 days. The acute-duration inhalation MRL was based on a study showing decreased mitogen-induced
blastogenesis of B-lymphocytes following exposure of mice to benzene vapors at a concentration of
10 ppm, 6 hours/day for 6 days. The intermediate-duration inhalation MRL was based on a study
showing delayed splenic lymphocyte reaction to foreign antigens evaluated by in vitro mixed lymphocyte
culture following exposure of mice to benzene vapors at a concentration of 10 ppm, 6 hours/day,
5 days/week for a total of 20 exposures.
Based on information found in the literature, it is reasonable to expect that adverse immunological effects
might occur in humans after inhalation, oral, or dermal exposure, since absorption of benzene through any
route of exposure would increase the risk of damage to the immunological system. Studies show that the
immunological system is susceptible to chronic exposure at low levels, so people living in and around
hazardous waste sites who may be exposed to contaminated air, drinking water, soil, or food may be at an
increased risk for adverse immunological effects.
Neurological Effects.
In humans, results of occupational studies indicate that there is a cause-and-
effect relationship between acute inhalation of very high concentrations of benzene and symptoms
indicative of central nervous system toxicity. These symptoms, observed following both acute nonlethal
and lethal exposures, include drowsiness, dizziness, headache, vertigo, tremor, delirium, and loss of
consciousness. These symptoms are reversible when symptomatic workers are transferred from the
problem area. Comparable toxicity in humans has been reported following ingestion of benzene at doses
of 125 mg/kg and above. Occupational exposure to benzene has also been reported to produce
neurological abnormalities in humans. Electromyographical and motor conduction velocity examinations
were conducted on six patients with aplastic anemia, all of whom worked in environments where
adhesives containing benzene were used (in one case, air concentrations bracketed around 210 ppm).
Abnormalities in motor conduction velocity were noted in four of the six pancytopenic individuals and
were thought to result from a direct effect of benzene on the peripheral nerves and/or spinal cord.
In its acute stages, benzene toxicity appears to be due primarily to the direct effects of benzene on the
central nervous system, whereas the peripheral nervous system appears to be the target following chronic
BENZENE
20
low-level exposures. In addition, because benzene may induce an increase in brain catecholamines, it
may also have a secondary effect on the immune system via the hypothalamus-pituitary-adrenal axis.
Increased metabolism of catecholamines can result in increased adrenal corticosteroid levels, which are
immunosuppressive.
Animal studies provide additional support that benzene affects the nervous system following acute
inhalation and oral exposures, albeit at extremely high acute exposure levels. Effects reported include
narcosis, nervous system depression, tremors, and convulsions. Acute and intermediate inhalation
exposures have also been reported to produce adverse neurological effects in animals including a
reduction in hind-limb grip strength and evoked electrical activity in the brain, and behavioral
disturbances. Effects of benzene on learning were investigated in male hooded rats of the SpragueDawley strain given 550 mg/kg of benzene in corn oil or corn oil without benzene, intraperitoneally,
on days 9, 11, and 13 postpartum. The rats exposed to benzene exhibited a significantly impaired
learning ability when tested on problems of the closed-field, maze-learning task. This sign of
neurotoxicity was not observed in control animals. In another study, 47-day-old juvenile cotton rats were
maintained on one of two isocaloric diets containing either 4 or 16% crude protein for a 26-day
experimental period. Animals were treated intraperitoneally with either 0 (corn oil), 100, 500, or
1,000 mg/kg benzene in corn oil for 3 consecutive days. The first dose was administered on days 15
17 of the experimental period. Animals were terminated on day 27. During the experimental period,
severe loss of coordination was observed in some rats on the low protein diet immediately after exposure
to benzene, but this subsided.
Intermediate oral exposures resulted in changes in the levels of monoamine transmitters in the brain
without treatment-related behavioral changes. Mice exposed to 3 ppm for 2 hours/day for 30 days
exhibited increased levels of acetylcholinesterase in the brain. In vitro studies suggest that benzene may
have a direct effect on brain cells. Primary astrocyte cultures prepared from neonatal rat cerebella were
treated with 3, 6, or 9 mmol/L benzene for 1 hour. ATPase and Mg2+-ATPase activity were inhibited in a
dose-related manner, and were detected at 7892% of control values for ATPase, and 6074% of control
values for Mg2+-ATPase.
These data suggest that humans exposed to benzene in the occupational setting for acute, intermediate, or
chronic durations via the inhalation and oral routes are at risk of developing neurological effects.
However, benzene levels in ambient air, drinking water, and at hazardous waste sites are lower and not
likely to be of concern.
BENZENE
21
2.3
MINIMAL RISK LEVELS (MRLs)
Estimates of exposure levels posing minimal risk to humans (MRLs) have been made for benzene. An
MRL is defined as an estimate of daily human exposure to a substance that is likely to be without an
appreciable risk of adverse effects (noncarcinogenic) over a specified duration of exposure. MRLs are
derived when reliable and sufficient data exist to identify the target organ(s) of effect or the most sensitive
health effect(s) for a specific duration within a given route of exposure. MRLs are based on
noncancerous health effects only and do not consider carcinogenic effects. MRLs can be derived for
acute, intermediate, and chronic duration exposures for inhalation and oral routes. Appropriate
methodology does not exist to develop MRLs for dermal exposure.
Although methods have been established to derive these levels (Barnes and Dourson 1988; EPA 1990),
uncertainties are associated with these techniques. Furthermore, ATSDR acknowledges additional
uncertainties inherent in the application of the procedures to derive less than lifetime MRLs. As an
example, acute inhalation MRLs may not be protective for health effects that are delayed in development
or are acquired following repeated acute insults, such as hypersensitivity reactions, asthma, or chronic
bronchitis. As these kinds of health effects data become available and methods to assess levels of
significant human exposure improve, these MRLs will be revised.
Inhalation MRLs
An MRL of 0.009 ppm has been derived for acute-duration inhalation exposure (14 days or less)
to benzene.
The acute-duration inhalation MRL of 0.009 ppm was derived from a lowest-observed-adverse-effect
level (LOAEL) value of 10.2 ppm for reduced lymphocyte proliferation following mitogen stimulation in
mice (Rozen et al. 1984). The concentration was adjusted for intermittent exposure by multiplying the
LOAEL (10.2 ppm) by 6 hours/24 hours to correct for less than a full day of exposure. The resulting
adjusted LOAEL, 2.55 ppm, was then converted to a human equivalent concentration (HEC) according to
EPA (1994b) methodology for calculating a HEC for extrarespiratory effects of a category 3 gas (such as
benzene) as follows:
LOAELHEC
= LOAELADJ x ([Hb/g]A/[Hb/g]H)
BENZENE
22
where:
LOAELHEC
= the LOAEL dosimetrically adjusted to a human equivalent concentration
LOAELADJ
= the LOAEL adjusted from intermittent to continuous exposure
[Hb/g]A/[Hb/g]H = the ratio of the blood:gas partition coefficient of the chemical for the laboratory animal
species to the human value
If the animal blood:gas partition coefficient is greater than the human blood:gas partition coefficient, a
default value of 1 is used for the ratio. According to Wiester et al. (2002), blood:gas partition coefficients
for benzene in mice and humans are 17.44 and 8.12, respectively. Therefore, the default value of 1 is
applied, in which case, the LOAEL[HEC] is equivalent to the LOAEL[ADJ].
Therefore:
LOAELHEC = LOAELADJ = 2.55 ppm
The resulting LOAEL(HEC) of 2.55 ppm was then divided by an uncertainty factor of 300 (10 for the use of
LOAEL, 3 for extrapolation from animals to humans using dosimetric conversion, and 10 for human
variability) to yield the MRL value of 0.009 ppm (see Appendix A). An increased number of
micronucleated polychromatic erythrocytes (MN-PCEs), decreased numbers of granulopoietic stem cells
(Toft et al. 1982), lymphopenia (Cronkite et al. 1985), lymphocyte depression, and increased
susceptibility to bacterial infection (Rosenthal and Snyder 1985) are among the adverse hematological
and immunological effects observed in several other acute-duration inhalation studies. The study by
Rozen et al. (1984) shows benzene immunotoxicity (reduced mitogen-induced lymphocyte proliferation)
at a slightly lower exposure level than these other studies. C57BL/6J mice were exposed to 0, 10.2, 31,
100, and 301 ppm benzene for 6 hours/day for 6 days. Control mice were exposed to filtered, conditioned
air only. Lymphocyte counts were depressed at all exposure levels; erythrocyte counts were elevated at
10.2 ppm, equal to controls at 31 ppm, and depressed at 100 and 301 ppm. Femoral B-lymphocyte and
splenic B-lymphocyte numbers were reduced at 100 ppm. Levels of circulating lymphocytes and
mitogen-induced blastogenesis of femoral B-lymphocytes were depressed after exposure to 10.2 ppm
benzene for 6 days. Mitogen-induced blastogeneses of splenic T-lymphocytes were depressed after
exposure to 31 ppm of benzene for 6 days. In another study, mice exhibited a 50% decrease in the
BENZENE
23
population of colony-forming granulopoietic stem cells (CFU-E) after exposure to 10 ppm benzene for
6 hours/day for 5 days (Dempster and Snyder 1991). In a study by Wells and Nerland (1991), groups of
45 male Swiss-Webster mice were exposed to 3, 25, 55, 105, 199, 303, 527, 1,150, or 2,290 ppm
benzene for 6 hours/day for 5 days. The number of leukocytes in peripheral blood and spleen weights
were significantly decreased compared with untreated controls at all concentrations >25 ppm. Therefore,
3 ppm was the no-observed-adverse-effect level (NOAEL) and 25 ppm was the LOAEL for these effects.
These data support the choice of Rozen et al. (1984) as the study from which to derive the MRL.
An MRL of 0.006 ppm has been derived for intermediate-duration inhalation exposure (15
364 days) to benzene.
The intermediate-duration inhalation MRL of 0.006 ppm was derived from a LOAEL value of 10 ppm for
significantly delayed splenic lymphocyte reaction to foreign antigens evaluated in in vitro mixed
lymphocyte reaction following the exposure of male C57Bl/6 mice to benzene vapors 6 hours/day,
5 days/week for 20 exposure days (Rosenthal and Snyder 1987). The concentration was adjusted for
intermittent exposure by multiplying the LOAEL (10 ppm) by 6 hours/24 hours to correct for less than a
full day of exposure and by 5 days/7days to correct for less than a full week of exposure. The resulting
adjusted LOAEL, 1.8 ppm, was then converted to a HEC according to EPA (1994b) methodology for
calculating a HEC for extrarespiratory effects of a category 3 gas (such as benzene) as follows:
LOAELHEC
= LOAELADJ x ([Hb/g]A/[Hb/g]H)
where:
LOAELHEC
= the LOAEL dosimetrically adjusted to a human equivalent concentration
LOAELADJ
= the LOAEL adjusted from intermittent to continuous exposure
[Hb/g]A/[Hb/g]H = the ratio of the blood:gas partition coefficient of the chemical for the laboratory animal
species to the human value
default value of 1 is used for the ratio. According to Wiester et al. (2002), blood:gas partition coefficients
for benzene in mice and humans are 17.44 and 8.12, respectively. Therefore, the default value of 1 is
applied, in which case, the LOAEL[HEC] is equivalent to the LOAEL[ADJ].
BENZENE
24
Therefore:
LOAELHEC = LOAELADJ = 1.8 ppm
The resulting LOAEL(HEC) of 1.8 ppm was then divided by an uncertainty factor of 300 (10 for the use of
LOAEL, 3 for extrapolation from animals to humans using dosimetric conversion, and 10 for human
variability) to yield the MRL value of 0.006 ppm (see Appendix A).
Results of several studies support the choice of Rosenthal and Snyder (1987) as the basis for the
intermediate-duration inhalation MRL for benzene (Baarson et al. 1984; Green et al. 1981a, 1981b),
although the supporting studies employed a single exposure level, which precluded consideration as
critical studies for MRL derivation. Exposure of C57BL mice to 10 ppm benzene for 6 hours/day,
5 days/week caused significant depressions in numbers of lymphocytes (ca. 30% lower than controls) as
early as exposure day 32; this effect was also noted at the other scheduled periods of testing (exposure
days 66 and 178) (Baarson et al. 1984). Splenic RBCs were significantly reduced (ca. 15% lower than
controls) at exposure days 66 and 178. The failure of the erythrons of benzene-exposed mice to support
normal red cell mass was illustrated by the significant reduction in peripheral red cell numbers in these
animals at 66 and 178 days of benzene exposure. Green et al. (1981a, 1981b) exposed male CD-1 mice to
benzene vapors at concentrations of 0 or 9.6 ppm for 6 hours/day, 5 days/week for 50 days and assessed
the effects of exposure on cellularity in the spleen, bone marrow, and peripheral blood. Exposure-related
effects included a 90% increase in numbers of multipotential hematopoietic stem cells (CFU-S) (Green et
al. 1981a), approximately 25% increase in spleen weight and total splenic nucleated cellularity (Green et
al. 1981b), and 80% increase in nucleated RBCs (Green et al. 1981b).
One intermediate-duration inhalation study reported increased rapid response to an electrical shock in
mice at 0.78 ppm (Li et al. 1992). However, this study was not selected as the critical study for deriving
an intermediate-duration inhalation MRL for benzene due to apparent discrepancies between reported and
actual benzene exposure levels. Other animal studies identified neurological effects only at much higher
exposure levels (Carpenter et al. 1944; Dempster et al. 1984; Evans et al. 1981; Frantik et al. 1994; Green
et al. 1978).
An MRL of 0.003 ppm has been derived for chronic-duration inhalation exposure (365 days or
more) to benzene.
BENZENE
25
This MRL is based on statistically significantly decreased counts of B-lymphocytes in workers of shoe
manufacturing industries in Tianjin, China (Lan et al. 2004a, 2004b), using a benchmark dose (BMD)
analysis. The 250 benzene-exposed workers had been employed for an average of 6.12.9 years.
Controls consisted of 140 age- and gender-matched workers in clothing manufacturing facilities in which
measurable benzene concentrations were not found (detection limit 0.04 ppm). Benzene exposure was
monitored by individual organic vapor monitors (full shift) 5 or more times during 16 months prior to
phlebotomy. Benzene-exposed workers were categorized into four groups (140 controls, 109 at <1 ppm,
110 at 1<10 ppm, and 31 at 10 ppm) according to mean benzene exposure levels measured during
1 month prior to phlebotomy. Complete blood count (CBC) and differential were analyzed mechanically.
Coefficients of variation for all cell counts were <10%.
Mean 1-month benzene exposure levels in the four groups (controls, <1 ppm, 1<10 ppm, and 10 ppm)
were <0.04, 0.570.24, 2.852.11, and 28.7320.74 ppm, respectively. Hematological values were
adjusted to account for potential confounding factors (i.e., age, gender, cigarette smoking, alcohol
consumption, recent infection, and body mass index). All types of WBCs and platelets were significantly
decreased in the lowest exposure group (<1 ppm), ranging in magnitude from approximately 8 to 15%
lower than controls. Although similar statistical analyses for the mid- and high-exposure groups were not
included in the study report, decreases in all types of WBCs and platelets were noted at these exposure
levels as well; the decreases in the highest exposure group ranged in magnitude from 15 to 36%.
Lymphocyte subset analysis revealed significantly decreased CD4+-T cells, CD4+/CD8+ ratio, and
B cells. Hemoglobin concentrations were significantly decreased only within the highest (10 ppm)
exposure group. Tests for a linear trend using benzene air level as a continuous variable were significant
for platelets and all WBC measures except monocytes and CD8+-T cells. Upon restricting the linear
trend analyses to workers exposed to <10 ppm benzene, excluding controls, inverse associations remained
for total WBCs, granulocytes, lymphocytes, B cells, and platelets. In order to evaluate the effect of past
benzene exposures on the hematological effects observed in this study, the authors compared findings for
a group of workers who had been exposed to <1 ppm benzene over the previous year (n=60) and a subset
who also had <40 ppm-years lifetime cumulative benzene exposure (n=50). The authors stated that the
same cell types were significantly reduced in these groups, but did not provide further information of the
magnitude (i.e., percent change) of the hematological effects observed. These data suggest that the
1-month benzene exposure results could be used as an indicator of longer-term, low-level benzene
hematotoxicity. To demonstrate that the observed effects were attributable to benzene, significantly
decreased levels of WBCs, granulocytes, lymphocytes, and B cells were noted in a subgroup of the
<1-ppm group for which exposure to other solvents was negligible.
BENZENE
26
As shown in Table A-1 of Appendix A, exposure-response relationships were noted for several blood
factors. Benzene-induced decreased B cell count was selected as the critical effect for BMD modeling
because it represented the highest magnitude of effect (i.e., B cell count in the highest exposure group was
approximately 36% lower than that of controls). A BMD modeling approach was selected to identify the
point of departure because the critical study (Lan et al. 2004a, 2004b) identified a LOAEL in the absence
of a NOAEL.
As discussed in detail in Appendix A, the BMD analysis selected a point of departure (BMCL0.25sd) of
0.1 ppm, which was adjusted from the 8-hour TWA to a continuous exposure concentration
(BMCL0.25sdADJ) using the default occupational minute volume (EPA 1994b). The resulting BMCL0.25sdADJ
of 0.03 ppm was divided by an uncertainty factor of 10 (for human variability) to yield the chronicduration inhalation MRL value of 0.003 ppm.
Several recent epidemiological studies provide supporting information to the Lan et al. (2004a, 2004b)
findings of hematotoxicity in workers chronically exposed to relatively low levels of benzene (Qu et al.
2002, 2003a, 2003b; Rothman et al. 1996a, 1996b; Ward et al. 1996). Qu et al. (2002, 2003a, 2003b)
compared hematologic values among 105 healthy workers (51 men, 54 women) in industries with a
history of benzene usage (Tianjin, China) and 26 age-and gender-matched workers in industries that did
not use benzene. A LOAEL of 2.26 ppm was identified for significantly reduced total WBCs,
neutrophils, and RBCs; there was also an indication of benzene-induced changes in some hematological
values at exposure levels lower than the current industry 8-hour TWA of 1 ppm. Rothman et al. (1996a,
1996b) identified an 8-hour TWA LOAEL of 7.6 ppm for a group of 11 benzene-exposed workers in a
cross-sectional study of 44 healthy workers in Chinese (Shanghai) industries with a history of benzene
usage and age- and gender-matched workers in industries that did not use benzene. In a nested casecontrol study of a cohort of workers in the Pliofilm production departments of a rubber products
manufacturer in Ohio (Ward et al. 1996), a strong exposure-response relationship was correlated with low
WBC count. A weak positive exposure-response relationship was observed for RBCs, which was
significant for cumulative exposure up until the blood test date. The study authors noted that there was no
evidence for a threshold for hematologic effects and suggested that exposure to benzene levels <5 ppm
may result in hematologic suppression.
BENZENE
27
Oral MRLs
No acute-duration oral MRL was derived due to a lack of appropriate data on the effects of acute oral
exposure to benzene.
In a study by Thienes and Haley (1972), central nervous system symptoms of giddiness, vertigo, muscular
incoordination, unconsciousness, and death of humans have been reported when doses of 125 mg
kg/body weight benzene were ingested. The only lower doses in the acute oral database are
50 mg/kg/day, at which pregnant rats experienced alopecia of hindlimbs and trunk (Exxon 1986), an end
point not suitable for MRL derivation for benzene because the toxicological significance is not clear; and
88 mg/kg/day, a concentration resulting in slight non-dose-related central nervous system depression in
rats (Cornish and Ryan 1965).
No intermediate-duration oral MRL was derived. Examination of the literature indicated that studies by
Hsieh et al. (1988b, 1991) and Wolf et al. (1956) should be considered for derivation of an intermediateduration oral MRL. Adult male CD-1 mice were exposed to 0, 8, 40, or 180 mg/kg/day benzene in the
drinking water for 4 weeks (Hsieh et al. 1988b, 1991). Serious hematological and immunological effects
(leukopenia, erythrocytopenia, lymphopenia, enhanced splenic lymphocyte proliferation) occurred at the
lowest dose, precluding its use for MRL derivation. In the study by Wolf et al. (1956), a NOAEL of
1 mg/kg/day and a LOAEL of 50 mg/kg/day for leukopenia were identified in rats given benzene in oil by
gavage for 6 months. However, closer examination of the study by Wolf et al. (1956) revealed that the
results were not adequately supported by the data and analysis presented in the paper. Therefore, Wolf et
al. (1956) was not selected for derivation of the MRL.
An MRL of 0.0005 mg/kg/day has been derived for chronic-duration oral exposure (365 days or
more) to benzene.
No human data are available to evaluate hematological effects following oral exposure to benzene. No
adequate oral data were located from which a chronic-duration oral MRL for benzene could be derived,
although a 2-year carcinogenesis bioassay of orally-exposed rats and mice is available (NTP 1986). Male
rats were given 50, 100, or 200 mg/kg/day and female rats and mice of both sexes were given 0, 25, 50,
and 100 mg/kg/day benzene in corn oil for 2 years. The dose of 25 mg/kg/day was a LOAEL for
hematotoxicity and immunotoxicity in rats and mice, and is higher than the serious LOAEL of
8 mg/kg/day in the intermediate-duration database. Therefore, the threshold for hematological and
immunological effects of benzene was not identified.
BENZENE
28
However, results of toxicokinetic studies of inhaled benzene in humans (Nomiyama and Nomiyama
1974a; Pekari et al. 1992; Srbova et al. 1950) and inhaled and orally-administered benzene in rats and
mice (Sabourin et al. 1987) indicate that absorption of benzene at relatively low levels of exposure is
approximately 50% of an inhaled dose and essentially 100% of an oral dose. Based on these assumptions,
inhalation data can be used to estimate equivalent oral doses that would be expected to similarly affect the
critical targets of benzene toxicity. Therefore, the point of departure for the chronic-duration inhalation
MRL for benzene, namely the BMCL0.25sdADJ of 0.03 ppm for decreased B cell counts in benzene-exposed
workers (Lan et al. 2004a, 2004b), serves as the point of departure for deriving the chronic-duration oral
MRL as well.
The point of departure (in ppm) was converted to mg/m3 using the molecular weight of 78.11 for benzene
and assuming 25 C and 760 mm Hg:
BMCL0.25sdADJ of 0.03 ppm x 78.11/24.45 = 0.096 mg/m3
The BMCL0.25sdADJ of 0.096 mg/m3 for inhaled benzene was converted to an equivalent BMDL0.25sdADJ for
ingested benzene using EPA (1988b) human reference values for inhalation rate (20 m3/day) and body
weight (70 kg) and a factor of 0.5 to adjust for differences in absorption of benzene following inhalation
versus oral exposure (50 versus 100%, respectively) as follows:
BMDL0.25sdADJ = BMCL0.25sdADJ of 0.096 mg/m3 x 20 m3/day x 0.5 70 kg = 0.014 mg/kg/day
A total uncertainty factor of 30 (10 for human variability and 3 for uncertainty in route-to-route
extrapolation) was applied to the BMDL0.25sdADJ of 0.014 mg/kg/day; the resulting chronic-duration oral
MRL is 0.0005 mg/kg/day.
BENZENE
29
3. HEALTH EFFECTS
3.1
INTRODUCTION
The primary purpose of this chapter is to provide public health officials, physicians, toxicologists, and
other interested individuals and groups with an overall perspective on the toxicology of benzene. It
contains descriptions and evaluations of toxicological studies and epidemiological investigations and
provides conclusions, where possible, on the relevance of toxicity and toxicokinetic data to public health.
A glossary and list of acronyms, abbreviations, and symbols can be found at the end of this profile.
3.2
DISCUSSION OF HEALTH EFFECTS BY ROUTE OF EXPOSURE
To help public health professionals and others address the needs of persons living or working near
hazardous waste sites, the information in this section is organized first by route of exposure (inhalation,
oral, and dermal) and then by health effect (death, systemic, immunological, neurological, reproductive,
developmental, genotoxic, and carcinogenic effects). These data are discussed in terms of three exposure
periods: acute (14 days or less), intermediate (15364 days), and chronic (365 days or more).
Levels of significant exposure for each route and duration are presented in tables and illustrated in
figures. The points in the figures showing no-observed-adverse-effect levels (NOAELs) or lowestobserved-adverse-effect levels (LOAELs) reflect the actual doses (levels of exposure) used in the studies.
LOAELs have been classified into "less serious" or "serious" effects. "Serious" effects are those that
evoke failure in a biological system and can lead to morbidity or mortality (e.g., acute respiratory distress
or death). "Less serious" effects are those that are not expected to cause significant dysfunction or death,
or those whose significance to the organism is not entirely clear. ATSDR acknowledges that a
considerable amount of judgment may be required in establishing whether an end point should be
classified as a NOAEL, "less serious" LOAEL, or "serious" LOAEL, and that in some cases, there will be
insufficient data to decide whether the effect is indicative of significant dysfunction. However, the
Agency has established guidelines and policies that are used to classify these end points. ATSDR
believes that there is sufficient merit in this approach to warrant an attempt at distinguishing between
"less serious" and "serious" effects. The distinction between "less serious" effects and "serious" effects is
considered to be important because it helps the users of the profiles to identify levels of exposure at which
major health effects start to appear. LOAELs or NOAELs should also help in determining whether or not
BENZENE
30
3. HEALTH EFFECTS
the effects vary with dose and/or duration, and place into perspective the possible significance of these
effects to human health.
The significance of the exposure levels shown in the Levels of Significant Exposure (LSE) tables and
figures may differ depending on the user's perspective. Public health officials and others concerned with
appropriate actions to take at hazardous waste sites may want information on levels of exposure
associated with more subtle effects in humans or animals (LOAELs) or exposure levels below which no
adverse effects (NOAELs) have been observed. Estimates of levels posing minimal risk to humans
(Minimal Risk Levels or MRLs) may be of interest to health professionals and citizens alike.
Levels of exposure associated with carcinogenic effects (Cancer Effect Levels, CELs) of benzene are
indicated in Tables 3-1 and 3-2 and Figures 3-1 and 3-2. Because cancer effects could occur at lower
exposure levels, Figures 3-1 and 3-2 also shows a range for the upper bound of estimated excess risks,
ranging from a risk of 1 in 10,000 to 1 in 10,000,000 (10-4 to 10-7), as developed by EPA.
A User's Guide has been provided at the end of this profile (see Appendix B). This guide should aid in
the interpretation of the tables and figures for Levels of Significant Exposure and the MRLs.
3.2.1
Inhalation Exposure
Although occupational or environmental exposure to benzene or benzene-containing materials may

include inhalation, oral, and dermal exposure routes, the inhalation and dermal routes are usually of
primary concern in such scenarios. Data regarding occupational or environmental exposure in which
inhalation is considered to have been the primary exposure route are summarized in this section
(Section 3.2.1). Information regarding adverse health effects following oral or dermal exposure to
benzene or benzene-containing materials is summarized in Sections 3.2.2 and 3.2.3, respectively.
3.2.1.1 Death
Case reports of fatalities due to acute benzene exposures have appeared in the literature since the early
1900s (Cronin 1924; Greenburg 1926; Hamilton 1922). Deaths occurred suddenly or within several hours
after exposure (Avis and Hutton 1993; Cronin 1924; Greenburg 1926; Hamilton 1922; Winek et al. 1967).
The benzene concentrations encountered by the victims were not often known. However, it has been
estimated that 510 minutes of exposure to 20,000 ppm benzene in air is usually fatal (Flury 1928).
Lethality in humans has been attributed to asphyxiation, respiratory arrest, central nervous system
BENZENE
31
3. HEALTH EFFECTS
depression, or suspected cardiac collapse (Avis and Hutton 1993; Hamilton 1922; Winek and Collom
1971; Winek et al. 1967). Cyanosis, hemolysis, and congestion or hemorrhage of organs were reported in
the cases for which there were autopsy reports (Avis and Hutton 1993; Greenburg 1926; Hamilton 1922;
Winek et al. 1967). No studies were located regarding noncancer-related mortality in humans following
long-term inhalation exposure to benzene. Cancer-related mortality data for chronic-duration human
occupational exposure to benzene are presented in Section 3.2.1.7.
In animals, acute inhalation exposure to high concentrations of benzene has caused death. An inhalation
LC50 value for rats was calculated as 13,700 ppm for a 4-hour exposure (Drew and Fouts 1974).
Additionally, 4 of 6 rats died following a 4-hour exposure to 16,000 ppm benzene (Smyth et al. 1962).
However, in a study by Green et al. (1981b), male CD-1 mice exposed by inhalation to doses of benzene
up to 4,862 ppm, 6 hours/day for 5 days showed no lethality. Lower doses (up to 400 ppm) for longer
periods of time (2 weeks) did not cause death in mice (Cronkite et al. 1985). Lethality in monkeys and
cats exposed to unspecified concentrations has been ascribed to ventricular fibrillation due to increased
release of adrenaline (Nahum and Hoff 1934). Exposure of rabbits to 45,000 ppm of benzene for
approximately 30 minutes caused narcosis that was followed by the death of all exposed animals
(Carpenter et al. 1944). Furthermore, early deaths of rats and mice have occurred from intermediate and
chronic exposure to air concentrations of 200 or 300 ppm of benzene in cancer studies (Cronkite et al.
1989; Farris et al. 1993; Maltoni et al. 1982a, 1983). Intermediate exposures (6 hours/day, 5 days/week
for 50 days) of male CD-1 mice to benzene at doses of 9.6 ppm caused no increase in mortality, although
mice exposed to 302 ppm benzene under the same regimen for a total of 26 weeks showed mortality
approaching 50% (Green et al. 1981b). Mortality was observed in 97% of the CBA/Ca mice exposed to
300 ppm benzene for 16 weeks, as compared to 20% mortality in sham-exposed mice (Cronkite 1986). In
Sprague-Dawley rats that received 300 ppm benzene vapor for 6 hours/day, 5 days/week for 691 days, the
calculated median survival time was shown to be 51 weeks as compared to 65 weeks for controls (Snyder
et al. 1978a). However, Snyder et al. (1984) reported a median survival time of 546 days for male
Sprague-Dawley rats exposed to 100 ppm benzene for 5 days/week, 6 hours/day for life, compared to
560 days for air-exposed controls. It is not clear whether the difference in survival was significant or due
to benzene exposure since both controls and exposed rats experienced early mortality from respiratory
infections. Companion studies were also conducted with AKR and C57BL mice exposed to 300 ppm
benzene (Snyder et al. 1978a, 1980). The calculated median survival time for AKR and C57BL mice
exposed to 300 ppm benzene was shown to be 11 and 41 weeks compared to 39 and 75 weeks,
respectively, for controls. For AKR mice exposed to 100 ppm, the calculated median survival time was
39 weeks (number of deaths not shown) as compared to 41 weeks for controls (Snyder et al. 1980).
BENZENE
32
3. HEALTH EFFECTS
The LC50 value and all reliable LOAEL values for each species and duration category are recorded in
Table 3-1 and plotted in Figure 3-1.
3.2.1.2 Systemic Effects
No studies were located regarding endocrine, metabolic, or body weight effects in humans or
gastrointestinal, musculoskeletal, endocrine, metabolic, or dermal effects in animals following inhalation
exposure to benzene. Available data pertaining to systemic effects are presented below.
The highest NOAEL values and all reliable LOAEL values for systemic effects in each species and
duration category are recorded in Table 3-1 and plotted in Figure 3-1.
Respiratory Effects.
Respiratory effects have been reported in humans after acute exposure to
benzene vapors (Avis and Hutton 1993; Midzenski et al. 1992; Winek and Collom 1971; Winek et al.
1967; Yin et al. 1987b). Fifteen male workers employed in removing residual fuel from shipyard tanks
were evaluated for benzene exposure (Midzenski et al. 1992). Mucous membrane irritation was noted in
80% and dyspnea was noted in 67% of the workers at occupational exposures of >60 ppm for up to
3 weeks. Nasal irritation and sore throat were reported by male and female workers exposed to 33 and
59 ppm benzene, respectively, for more than 1 year (Yin et al. 1987b). After a fatal occupational
exposure to benzene vapors on a chemical cargo ship for only minutes, autopsy reports on three victims
revealed hemorrhagic, edematous lungs (Avis and Hutton 1993). Acute granular tracheitis, laryngitis,
bronchitis, and massive hemorrhages of the lungs were observed at autopsy of an 18-year-old male who
died of benzene poisoning after intentional inhalation of benzene (Winek and Collom 1971). Similarly,
acute pulmonary edema was found during the autopsy of a 16-year-old who died after sniffing glue
containing benzene (Winek et al. 1967).
Snyder et al. (1978a, 1984) reported no treatment-related effects on lung tissue in male Sprague-Dawley
rats exposed to 0, 100, or 300 ppm benzene 5 days/week, 6 hours/day for life. In addition, no adverse
histopathological effects on lung tissue were observed in AKR/J or C57BL/65 mice exposed to 300 ppm
benzene for life (Snyder et al. 1978a, 1980).
Table 3-1 Levels of Significant Exposure to Benzene - Inhalation
BENZENE
a
Key to Species
Figure (Strain)
Exposure/
Duration/
Frequency
(Route)
LOAEL
NOAEL
System
Less Serious
(ppm)
Serious
(ppm)
(ppm)
Reference
Chemical Form
Comments
ACUTE EXPOSURE
Death
1
Human
1d
5-10 min
20000
(death)
Flury 1928
(LC50)
Drew and Fouts 1974
(4/6 died)
Smyth et al. 1962
(death in 36.2 min)
Carpenter et al. 1944
20000
12
Rat
(SpragueDawley)
4 hr
13700
13700
14
Rat
(NS)
4 hr
16000
16000
13
Rabbit
(NS)
3.7-36.2 min
45000
45000
226
Systemic
Human
5
1-21 d
2.5-8 hr/d
Resp
Midzenski et al. 1992
60 M (mucous membrane
irritation, dyspnea)
3. HEALTH EFFECTS
60
419
Hemato
60 M (leukopenia, anemia,
thrombocytopenia, MCV
elevation)
60
Dermal
60 M (skin irritation)
60
Rat
(SpragueDawley)
Gd 6-15
6 hr/d
Bd Wt
2200 F (decreased maternal

body weight)
300 F
300
Green et al. 1978
2200
815
Rat
(SpragueDawley)
Gd 6-15
7 hr/d
Bd Wt
body weight and weight
gain)
10 F
10
Kuna and Kapp 1981
50
33
818
Rat
(Wistar)
7d
8 hr/d
BENZENE
a
Key to Species
Figure (Strain)
Exposure/
Duration/
Frequency
(Route)
(continued)
LOAEL
NOAEL
System
Less Serious
(ppm)
Hemato
(ppm)
Reference
Chemical Form
Comments
Li et al. 1986
100 F (leukopenia)
50 F
50
Serious
(ppm)
100
910
Rat
(Wistar)
15 min
Cardio
3526 M (ventricular arrhythmia)
Magos et al. 1990
3526
423
10
Rat
(CFY)
Gd 7-14
24 hr/d
Hepatic
Tatrai et al. 1980a
125 F
125
696

weight gain of <22.08%
of controls)
125
11
Rat
(CFY)
Gd 7-14
24 hr/d
Hepatic
141 F (increased relative liver

weight)
47 F
47
Tatrai et al. 1980b
141
824
Bd Wt
3. HEALTH EFFECTS
Bd Wt
weight gain)
47
12
Rat
(SpragueDawley)
2 wk
5 d/wk
6 hr/d
Hemato
300
30
30
(decrease in leukocytes,
males; decrease in
lymphocytes)
Ward et al. 1985
300
951
13
Mouse
(BALB/c)
7d
6 hr/d
Hemato
211 M (depressed WBC count)
47 M
47
Aoyama 1986
211
670
Bd Wt
47 M
211 M (16% decrease in body

weight)
47
211
34
14
Mouse
(BALB/c)
14 d
6 hr/d
BENZENE
a
Key to Species
Figure (Strain)
Exposure/
Duration/
Frequency
(Route)
(continued)
LOAEL
NOAEL
System
Less Serious
(ppm)
Serious
(ppm)
Hemato
(ppm)
Reference
Chemical Form
Comments
Aoyama 1986
48 M (depressed WBC count)

48
671
Bd Wt
48 M
208 M (18% decrease in body

weight)
48
208
15
Mouse
(DBA/2)
2 wk
6 hr/d
5 d/wk
300
877
Bd Wt
300 M (15% decrease)

300
16
Mouse
11 d
(Hale- Stoner) 5 d/wk
6 hr/d
Hemato
400 M (decreased erythrocytes

and leukocytes)
Cronkite et al. 1982
100
3. HEALTH EFFECTS
300 M (hematocrit decreased by Chertkov et al. 1992

26%, leukocytes
decreased by 80%, bone
marrow cellularity
decreased by 93%)
Hemato
400
381
17
Mouse
(C57B1/6
BNL)
2 wk
5 d/wk
6 hr/d
Hemato
25
25
(decreased hematocrit,
hemolytic anemia)
100
330
18
Mouse
2d
6 hr/d
Hemato
400 M (decreased CFU-E cells)
400
821
19
Mouse
8d
(C57BL/6BNL)6 hr/d
Hemato
3000 F (decreased marrow

cellularity)
3000
828
35
20
Mouse
(DBA/2J)
5d
6 hr/d
BENZENE
a
Key to Species
Figure (Strain)
Exposure/
Duration/
Frequency
(Route)
(continued)
LOAEL
NOAEL
System
Less Serious
(ppm)
Serious
(ppm)
Hemato
(ppm)
Reference
Chemical Form
Comments
Dempster and Snyder 1991
10 M (50% decrease in CFU-E

numbers)
10
885
21
Mouse
(C57B1/6)
2-8 d
24 hr/d
Hemato
100 M (leukopenia; decrease in

marrow cellularity)
Gill et al. 1980
103 M (decreased marrow

cellularity;
granulocytopenia,
lymphocytopenia;
decreased
polymorphonucleucytes)
Green et al. 1981b
300 M (reduced bone marrow

cellularity and CFU-E
development)
Neun et al. 1992
100
911
22
5d
6 hr/d
Hemato
9.9 M
9.9
103
396
Bd Wt
4862 M
3. HEALTH EFFECTS
Mouse
(CD-1)
4862
23
Mouse
(Swiss
Webster,
C57B1/6J)
2 wk
4 d/wk
6 h/d
Hemato
300
807
24
Mouse
(Hybrid)
5d
5 d/wk
6 hr/d
Hemato
900 F (CFU-E depression)
300 F
300
Plappert et al. 1994a
900
945
36
25
Mouse
(C57BI/6J)
6d
6 hr/d
BENZENE
a
Key to Species
Figure (Strain)
Exposure/
Duration/
Frequency
(Route)
(continued)
LOAEL
NOAEL
System
Less Serious
(ppm)
Serious
(ppm)
Hemato
(ppm)
Reference
Chemical Form
Comments
Rozen et al. 1984
10.2 M (depressed lymphocyte

counts; elevated RBCs)
10.2
845
26
Mouse
(NMRI)
1-10 d
24 hr/d
Toft et al. 1982

cellularity; increased
polychromatic
erythrocytes; decreased
granulopoietic stem cells)
Hemato
21
27
Mouse
(NMRI)
1 wk
Hemato
Toft et al. 1982
14 M
14
302
28
Mouse
(NMRI)
2 wk
5 d/wk
8 hr/d
Hemato
Toft et al. 1982
21 M (increased
micronucleated
polychromatic
erythrocytes; decreased
10.5 M
10.5
3. HEALTH EFFECTS
25
21
327
29
Mouse
(CD-1)
2 wk
5 d/wk
6 hr/d
Hemato
300
30
30
(anemia, decreased
hemoglobin,erythrocytes
and hematocrit;
hypoplasia of bone
marrow; leukopenia)
Ward et al. 1985
300
346
Bd Wt
300
300
37
30
Mouse
(SwissWebster)
BENZENE
a
Key to Species
Figure (Strain)
Exposure/
Duration/
Frequency
(Route)
(continued)
LOAEL
NOAEL
System
5d
6 hr/d
Hemato
Gd 7-20
24 hr/d
Bd Wt
Less Serious
(ppm)
(ppm)
3M
3
Serious
(ppm)
Reference
Chemical Form
25 M (decrease in WBC count)
Wells and Nerland 1991
313 F (reduced maternal weight

gain)
Ungvary and Tatrai 1985
100 F (leukopenia; increased

leukocyte alkaline
phosphatase)
Li et al. 1986
400 M (29% reduction in total

splenic cells, 28% lower
thymus weight)
Robinson et al. 1997
Comments
25
894
31
Rabbit
156.5 F
156.5
313
826
50 F
50
100
15
33
Rat
(SpragueDawley)
2 wk
5 d/wk
6 hr/d
200 M
200
3. HEALTH EFFECTS
Immuno/ Lymphoret
Rat
7d
32
8 hr/d
(Wistar)
400
1012
34
Mouse
(BALB/c)
7d
6 hr/d
47 M (depressed T- and
B-lymphocytes;
decreased spleen weight
and WBC count)
Aoyama 1986
48 M (depressed T- and
B-lymphocytes;
decreased spleen and
thymus weights and
WBC count)
Aoyama 1986
47
255
35
Mouse
(BALB/c)
14 d
6 hr/d
48
672
38
36
Mouse
(DBA/2)
BENZENE
a
Key to Species
Figure (Strain)
Exposure/
Duration/
Frequency
(Route)
(continued)
LOAEL
NOAEL
System
Less Serious
(ppm)
Serious
(ppm)
(ppm)
2 wk
6 hr/d
5 d/wk
Reference
Chemical Form
Comments
300 M (leukocytes decreased by Chertkov et al. 1992

80%, bone marrow
cellularity decreased by
93%)
300
906
37
Mouse
(CBA/Ca)
2 wk
5 d/wk
6 hr/d
25
10
10
Cronkite 1986
(lymphopenia)
25
311
Mouse
11 d
6 hr/d
400 M (decreased bone marrow Cronkite et al. 1982

cellularity)
400
382
39
Mouse
(C57B1/6
BNL)
2 wk
5 d/wk
6 hr/d
25
10
10
(lymphopenia)
25
3. HEALTH EFFECTS
38
400
40
Mouse
(CBA/Ca
BNL)
2d
5 d/wk
6 hr/d
3000 M (decreased lymphocytes,

CFU-S content in
marrow)
3000
825
41
Mouse
(C57B1/6)
2-8 d
24 hr/d
100 M (leukopenia; decrease in

marrow cellularity)
Gill et al. 1980
100
998
39
42
Mouse
(CD-1)
5d
6 hr/d
BENZENE
a
Key to Species
Figure (Strain)
Exposure/
Duration/
Frequency
(Route)
(continued)
LOAEL
NOAEL
System
Less Serious
(ppm)
Serious
(ppm)
(ppm)
9.9 M
9.9
Reference
Chemical Form
103 M (decreased femoral

marrow and splenic
cellularities; reduced
splenic granulocytes)
Green et al. 1981a

cellularity)
Neun et al. 1992
Comments
103
392
43
Mouse
(Swiss
Webster,
C57B1/6J)
2 wk
4 d/wk
6 h/d
300
907
Mouse
(Hybrid)
5d
5 d/wk
6 hr/d
300 F (increased helper

lymphocytes)
100 F
100
300
946
45
Mouse
(C57BL/6)
1-12 d
6 hr/d
30 M (Listeria infection, T and

B lymphocyte
depression)
10 M
10
Rosenthal and Snyder 1985
3. HEALTH EFFECTS
44
30
37
46
Mouse
(C57BI/6J)
6d
6 hr/d
10.2 M (decreased circulating

lymphocytes and
mitogen-induced
blastogenesis of femoral
T and B-lymphocytes)
Rozen et al. 1984
10.2
847
47
Mouse
(NMRI)
2 wk
5 d/wk
8 hr/d
21 M (decreased
10.5 M
10.5
Toft et al. 1982
21
908
40
48
Mouse
(NMRI)
BENZENE
a
Key to Species
Figure (Strain)
Exposure/
Duration/
Frequency
(Route)
(continued)
LOAEL
NOAEL
System
Less Serious
(ppm)
Serious
(ppm)
1-10 d
24 hr/d
(ppm)
Reference
Chemical Form
Comments
Toft et al. 1982
21 M (decreased
21
909
49
Mouse
(CD-1)
2 wk
5 d/wk
6 hr/d
300
30
30
(leukopenia;
lymphopenia; bone
marrow hypoplasia;
histopathological lesions
in spleen, thymus,
selected lymph nodes)
Ward et al. 1985
50
Mouse
(SwissWebster)
5d
6 hr/d
25 M (decrease in spleen
weight and WBC count)
3M
3
Wells and Nerland 1991
25
439
Neurological
Human
51
30 min
300
(drowsiness, dizziness,
headaches)
3. HEALTH EFFECTS
300
347
Flury 1928
300
359
52
Human
1-21 d
2.5-8 hr/d
60 M (dizzyness, nausea,
headache, peculiar or
strong odor, chemical
taste, fatigue)
60
420
53
Rat
(SpragueDawley)
Gd 6-15
6 hr/d
2200 F (lethargy)
300 F
300
Green et al. 1978
2200
837
41
54
Mouse
(C57BL)
BENZENE
a
Key to Species
Figure (Strain)
Exposure/
Duration/
Frequency
(Route)
(continued)
LOAEL
NOAEL
System
Less Serious
(ppm)
Serious
(ppm)
1-14 d
5 d/wk
6 hr/d
(ppm)
3000 M (tremors; decreased grip

strength)
100 M (increased milk licking:

behavioral index)
100
Reference
Chemical Form
Comments
Dempster et al. 1984
3000
360
55
Mouse
(CD1,
C57BL/6J)
5d
6 hr/d
300
300
900
(hyperactivity)
(narcosis)
Evans et al. 1981
900
201
56
3.7-36.2 min
45000
Carpenter et al. 1944

(narcosis, tremors,
excitement, chewing,
loss of pupillary and blink
reflex; pupillary
contraction and
involuntary blinking)
45000
43
Reproductive
Rat
57
(SpragueDawley)
Gd 6-15
6 hr/d
100 F
Coate et al. 1984
2200 F
Green et al. 1978
125 F
Tatrai et al. 1980a
500 F
Murray et al. 1979
3. HEALTH EFFECTS
Rabbit
(NS)
100
904
58
Rat
(SpragueDawley)
Gd 6-15
6 hr/d
2200
288
59
Rat
(CFY)
Gd 7-14
24 hr/d
125
209
60
Mouse
(CF-1)
Gd 6-15
7 hr/d
500
289
42
61
Rabbit
(New
Zealand)
Gd 6-18
7 hr/d
BENZENE
a
Key to Species
Figure (Strain)
Exposure/
Duration/
Frequency
(Route)
(continued)
LOAEL
NOAEL
System
Less Serious
(ppm)
Serious
(ppm)
(ppm)
Reference
Chemical Form
Comments
Murray et al. 1979
500 F
500
291
62
Rabbit
Gd 7-20
24 hr/d
313 F (increased abortions and Ungvary and Tatrai 1985

resorptions)
156.5 F
156.5
313
372
Gd 6-15
6 hr/d
40 F
40
100 F (decreased fetal weight)
Coate et al. 1984
100 F (increased incidence of

missing sternebrae)
Green et al. 1978
100
319
64
Rat
(SpragueDawley)
Gd 6-15
6 hr/d
100
132
65
Rat
(SpragueDawley)
Gd 6-15
7 hr/d
50 F (decreased fetal weight)
10 F
10
3. HEALTH EFFECTS
Developmental
Rat
63
(SpragueDawley)
Kuna and Kapp 1981
50
136
66
Rat
(CFY)
Gd 7-14
24 hr/d
125 F (decreased mean fetal

weight; increased fetal
weight retardation;
skeletal retardation; 17%
decrease in mean
placental weight)
Tatrai et al. 1980a
125
210
43
67
Rat
(CFY)
BENZENE
a
Key to Species
Figure (Strain)
Exposure/
Duration/
Frequency
(Route)
(continued)
LOAEL
NOAEL
System
Less Serious
(ppm)
Serious
(ppm)
Gd 7-14
24 hr/d
(ppm)
141 F (significant increase in

fetal mortality)
47 F (decreased fetal weight;

signs of skeletal
retardation)
Reference
Chemical Form
Comments
Tatrai et al. 1980b
141
47
320
68
Mouse
(SwissWebster)
Gd 6-15
6 hr/d
Keller and Snyder 1988
20
103
69
Mouse
(CF-1)
Gd 6-15
7 hr/d
500 F (decreased fetal body

weight; delayed skeletal
ossifications)
Murray et al. 1979
500
57
70
Mouse
(CFLP)
Gd 6-15
12 hr/d
156.5 F (decreased fetal body

weight and skeletal
retardation)
3. HEALTH EFFECTS
20 F (decreased circulating
erythroid precursors,
elevation of granulocytic
precourser cells in
neonates and 6-week old
offspring)
10 F
10
156.5
61
71
Rabbit
(New
Zealand)
Gd 6-18
7 hr/d
500 F (increased minor skeletal

variants)
Murray et al. 1979
313 F (decreased fetal weight;

increased minor
anomalies)
500
55
72
Rabbit
(New
Zealand)
Gd 7-20
24 hr/d
156.5 F
156.5
313
335
44
BENZENE
a
Key to Species
Figure (Strain)
Exposure/
Duration/
Frequency
(Route)
(continued)
LOAEL
NOAEL
System
(ppm)
Less Serious
Serious
(ppm)
(ppm)
Reference
Chemical Form
Comments
INTERMEDIATE EXPOSURE
Death
73
Rat
(SpragueDawley)
15 wk
4-5 d/wk
4-7 hr/d
200
(death)
Maltoni et al. 1983, 1985
200
211
74
Mouse
(CBA/Ca)
16 wk
5 d/wk
6 hr/d
300 M (97% mortality)
Cronkite 1986
300
300
664
75
16 wk
5 d/wk
6 hr/d
(deaths in males during

exposure; deaths in
females shortly after
exposure)
300
832
76
Mouse
(CBA/Ca)
16 wk
5 d/wk
6 hr/d
300 M (11/125 died during first 9 Farris et al. 1993

months after initiation of
exposure)
3. HEALTH EFFECTS
Mouse
(CBA/Ca
BNL)
300
447
77
Mouse
(CD-1)
26 wk
5 d/wk
6 hr/d
302 M (50% mortality)
Green et al. 1981b
150
(pancytopenia)
Aksoy and Erdem 1978
210
(pancytopenia,
hypocellular to
hypercellular bone
marrow)
Aksoy et al. 1972
302
328
Systemic
Human
78
4 mo- 1 yr
(occup)
Hemato
4 mo -1 yr
(occup)
Hemato
150
344
79
Human
210
324
45
80
Human
1 yr
(occup)
BENZENE
a
Key to Species
Figure (Strain)
Exposure/
Duration/
Frequency
(Route)
(continued)
LOAEL
NOAEL
System
Less Serious
(ppm)
Serious
(ppm)
Hemato
40
(ppm)
Reference
Chemical Form
Comments
Cody et al. 1993
(decrease in WBC counts

in first 4 months)
40
880
81
Human
3.5 mo- 19 yr
(occup)
Hemato
3 wk
5 d/wk
6 hr/d
Hemato
29
(aplastic anemia)
Yin et al. 1987c
(decreased WBC and

lymphocytes; increased
RBC and hemoglobin)
Dow 1992
29
82
500
500
213
83
Rat
(SpragueDawley)
10 wk
Gd 0-20
Ld 5-20
5 d/wk
6 hr/d
Bd Wt
Kuna et al. 1992
300 F
300
3. HEALTH EFFECTS
Rat
(SpragueDawley)
401
84
Rat
(SpragueDawley)
13 wk
5 d/wk
6 hr/d
Hemato
300
30
30
(decrease in leukocytes,
slight decrease in
marrow cellularity)
Ward et al. 1985
300
16
Bd Wt
300
300
85
Rat
(Wistar)
204 d
5 d/wk
7 hr/d
Hemato
88 M (leukopenia)
Wolf et al. 1956
88
377
46
86
Mouse
(C57BL)
24 wk
5 d/wk
6 hr/d
BENZENE
a
Key to Species
Figure (Strain)
Exposure/
Duration/
Frequency
(Route)
(continued)
LOAEL
NOAEL
System
Less Serious
(ppm)
Serious
(ppm)
Hemato
(ppm)
Reference
Chemical Form
Comments
10 M (depressed peripheral
red blood cells, CFU-E)
Baarson et al. 1984
The use of one dose

precludes a
dose-response
assessment.
10 M (depressed splenic red

cells)
Baarson et al. 1984
The use of one dose

precludes a
dose-response
assessment.
10
287
87
Mouse
(C57BL)
24 wk
5 d/wk
6 hr/d
Hemato
10
88
Mouse
9.5 wk
6 hr/d
Hemato
400 M (decreased erythrocytes

and leukocytes,
decreased bone marrow
cellularity)
400
131
89
Mouse
(C57B1/
6BNL)
4-16 wk
5 d/wk
6 hr/d
Hemato
(stem cell depression in

bone marrow, reversible
after 2-4 weeks)
316 M (decreased stem cells in

bone marrow)
300 M (granulocytic hyperplasia

in bone marrow)
Farris et al. 1993
300
3. HEALTH EFFECTS
314
300
386
90
Mouse
(CBA/Ca)
16 wk
5 d/wk
6 hr/d
Hemato
25 M
25
316
331
91
Mouse
(CBA/Ca)
16 wk
5 d/wk
6 hr/d
Hemato
300
806
47
92
Mouse
(B6C3F1)
up to 8 wk
5 d/wk
6 hr/d
BENZENE
a
Key to Species
Figure (Strain)
Exposure/
Duration/
Frequency
(Route)
(continued)
LOAEL
NOAEL
System
Less Serious
(ppm)
Hemato
Serious
(ppm)
(ppm)
Chemical Form
Comments
Farris et al. 1997a
100 M (decreased numbers of

differentiating and
maturing hematopoietic
bone marrow cells)
10 M
10
Reference
100
1015
93
Mouse
(CD-1)
26 wk
5 d/wk
6 hr/d
302 M (decreased WBC, RBC; Green et al. 1981b

altered RBC morphology)
Hemato
302
332
302 M
3. HEALTH EFFECTS
Bd Wt
302
94
Mouse
(CD-1)
50 d
5 d/wk
6 hr/d
Hemato
Green et al. 1981b
9.6 M
9.6
399
Bd Wt
9.6 M
9.6
95
Mouse
(Kunming)
30 d
6 d/wk
2 hr/d
Hepatic
Li et al. 1992
12.52 M
12.52
Exposures conducted
under static conditions;
measured only on first
3 of 30 days.
412
Renal
12.52 M
12.52
Bd Wt
12.52 M
12.52
96
Mouse
(DBA/2,
B6C3F1,
C57B1/6)
13 wk
3 or 5 d/wk
6 hr/d
Luke et al. 1988b

300 M (depressed rate of
erythropoiesis, increased
frequency of MN-PCE
and MN-NCE)
Hemato
300
98
48
97
Mouse
(Hybrid)
8 wks
5 d/wk
6 hr/d
BENZENE
a
Key to Species
Figure (Strain)
Exposure/
Duration/
Frequency
(Route)
(continued)
LOAEL
NOAEL
System
Less Serious
(ppm)
Hemato
Serious
(ppm)
(ppm)
Chemical Form
Comments
300 F (slight anemia; BFU-E

and CFU-E depression in
bone marrow)
100 F
100
Reference
300
715
98
Mouse
(Hybrid)
8 wk
5 d/wk
6 hr/d
Hemato
300 F (decreased Hgb, Hct,

erythrocyte counts)
Plappert et al. 1994b
300
723
Mouse
(BDF1)
8 wk
5 d/wk
6 hr/d
Hemato
Seidel et al. 1989b
100 F (BFU-E and CFU-E

depression)
100
96
100
Mouse
(NMRI)
8 wk
Hemato
Toft et al. 1982
14 M
14
605
101
Mouse
(Hybrid)
6 or 7 wk
5 d/wk
6 h/d
Hemato
300 F (decreased CFU-C,

BFU-E and CFU-E)
Vacha et al. 1990
300
Ward et al. 1985
3. HEALTH EFFECTS
99
300
808
102
Mouse
(CD-1)
13 wk
5 d/wk
6 hr/d
Hemato
30
30
(pancytopenia, bone
marrow hypoplasia)
300
17
Bd Wt
300
300
103
Gn Pig
(NS)
32 or 269 d
5 d/wk
7 hr/d
Hemato
88
(leukopenia)
Wolf et al. 1956
88
379
49
104
Rabbit
(NS)
BENZENE
a
Key to Species
Figure (Strain)
Exposure/
Duration/
Frequency
(Route)
(continued)
LOAEL
NOAEL
System
243 d
5 d/wk
7 hr/d
Hemato
3 wk
5 d/wk
6 hr/d
Hemato
Less Serious
(ppm)
Serious
(ppm)
80
(ppm)
Reference
Chemical Form
(leukopenia)
Wolf et al. 1956
(decreased peripheral
WBC, and increased
erythroid cells)
Dow 1992
Comments
80
378
105
Pig
(DurocJersey)
100
20
20
100
215
210
Aksoy et al. 1972

(pancytopenia,
hypoplastic to
hyperplastic bone
marrow, enlarged spleen)
210
997
107
Human
1 yr
(occup)
40
Cody et al. 1993
(decreased lymphocytes)
40
3. HEALTH EFFECTS
Immuno/ Lymphoret
Human
4 mo- 1 yr
106
(occup)
294
108
Rat
(SpragueDawley)
3 wk
5 d/wk
6 hr/d
500
(decreased myeloid and

lymphoid cells)
Dow 1992
500
216
109
Rat
(SpragueDawley)
4 wk
5 d/wk
6 hr/d
200 M
200
400 M (29% reduction in total

splenic cells, 28% lower
thymus weight)
Robinson et al. 1997
300
Ward et al. 1985
400
1013
110
Rat
(SpragueDawley)
13 wk
5 d/wk
6 hr/d
30
30
(leukopenia and
lymphopenia)
300
353
50
111
Rat
(Wistar)
BENZENE
a
Key to Species
Figure (Strain)
Exposure/
Duration/
Frequency
(Route)
(continued)
LOAEL
NOAEL
System
Less Serious
(ppm)
Serious
(ppm)
204 d
5 d/wk
7 hr/d
(ppm)
Reference
Chemical Form
88
(leukopenia, increased
spleen weight)
Wolf et al. 1956
4570
(increased leukocyte
alkaline phosphatase,
decreased white blood
cell count)
Yin et al. 1982
Comments
88
18
112
Rat
(NS)
20 wk
6 d/wk
4 hr/d
4570
384
Mouse
(C57B1)
24 wk
5 d/wk
6 hr/d
Baarson et al. 1984
10 M (decreased number of
splenic lymphocytes)
10
603
114
Mouse
(C57B1/
6BNL)
4-16 wk
5 d/wk
6 hr/d
300
(reduced bone marrow

cellularity; stem cell
depression, reversible
after 2-4 weeks)
The use of one dose

precludes a
dose-response
assessment.
3. HEALTH EFFECTS
113
300
803
115
Mouse
(CBA/Ca
BNL)
20 d
5 d/wk
6 hr/d
316 M (decreased lymphocytes, Cronkite et al. 1989

CFU-S content in
marrow)
316
827
51
116
Mouse
(CBA/Ca)
BENZENE
a
Key to Species
Figure (Strain)
Exposure/
Duration/
Frequency
(Route)
(continued)
LOAEL
NOAEL
System
Less Serious
(ppm)
Serious
(ppm)
16 wk
5 d/wk
6 hr/d
(ppm)
Reference
Chemical Form
300 M (granulocytic
hyperplasia)
Farris et al. 1993
100 M (reduced numbers of

total bone marrow cells,
progenitor cells,
differentiating
hematopoietic cells,
peripheral blood
leukocytes and RBCs)
Farris et al. 1997a
100 M (reduced lymphocyte and

total nucleated cell
counts)
Farris et al. 1997b
Comments
300
805
117
Mouse
(B6C3F1)
up to 8 wk
5 d/wk
6 hr/d
10 M
10
3. HEALTH EFFECTS
100
1014
118
Mouse
(B6C3F1)
8 wk
5 d/wk
6 hr/d
10 M
10
100
1010
119
Mouse
(C57B1/6)
6 wk
5 d/wk
6 hr/d
1000
(leukopenia,
granulocytopenia,
lymphocytopenia)
Gill et al. 1980
1000
24
120
Mouse
(CD-1)
50 d
6 hr/d
5 d/wk
9.6 M (increased splenic

CFU-S)
9.6
Green et al. 1981a
The use of one

exposure level
precludes usefulness
for dose-response
assessment.
393
52
121
Mouse
(CD-1)
BENZENE
a
Key to Species
Figure (Strain)
Exposure/
Duration/
Frequency
(Route)
(continued)
LOAEL
NOAEL
System
Less Serious
(ppm)
Serious
(ppm)
(ppm)
26 wk
6 hr/d
5 d/wk
302 M (reduced marrow and

spleen cellularity;
decreased spleen
weight)
Reference
Chemical Form
Comments
Green et al. 1981a
302
394
122
Mouse
(CD-1)
50 d
5 d/wk
6 hr/d
Green et al. 1981b
9.6 M (increased spleen weight,

total splenic nucleated
cellularity and NRBC)
9.6
316
123
Mouse
(CD-1)
26 wk
5 d/wk
6 hr/d
Green et al. 1981b

302 M (lymphocytopenia,
anemia, decreased
spleen weight, decreased
spleen and marrow
cellularities)
3. HEALTH EFFECTS
The use of one

exposure level
precludes usefulness
for dose-response
assessment.
302
336
124
Mouse
(Kunming)
30 d
6 d/wk
2 hr/d
3.13 M
12.52 M (26% decrease in relative

spleen weight; decrease
in myelocytes,
premyelocytes,
myeloblasts, and
metamyelocytes in the
bone marrow)
3.13
Li et al. 1992
Exposures conducted
3 of 30 days.
12.52
414
53
125
Mouse
(Hybrid)
8 wks
5 d/wk
6 hr/d
BENZENE
a
Key to Species
Figure (Strain)
Exposure/
Duration/
Frequency
(Route)
(continued)
LOAEL
NOAEL
System
Less Serious
(ppm)
(ppm)
Reference
Chemical Form
Comments
300 F (increased T4/T8 ratio)
100 F
100
Serious
(ppm)
300
727
126
Mouse
(C57B1/6)
20 d
5 d/wk
6 hr/d
10 M (delayed splenic
lymphocyte reaction to
foreign antigens
evaluated in in vitro
mixed lymphocyte
reaction)
10
127
Mouse
(C57B1/6)
100 d
5 d/wk
6 hr/d
100 M (death in 9/10 mice due

to depressed
cell-mediated immunity)
100
601
128
Mouse
4-5 wk
6 hr/d
Stoner et al. 1981
200 F (suppressed antibody

response to fluid tetanus
toxoid)
50 F
50
3. HEALTH EFFECTS
600
200
34
129
Mouse
(CD-1)
13 wk
5 d/wk
6 hr/d
300
30
30
(leukocyte & lymphocyte Ward et al. 1985

depression; bone marrow
hypoplasia;
histopathologic lesions in
spleen and selected
lymph nodes)
300
352
130
Gn Pig
(NS)
32 or 269 d
5 d/wk
7 hr/d
88
Wolf et al. 1956
54
88
20
(leukopenia, increased
spleen weight)
131
Rabbit
(NS)
BENZENE
a
Key to Species
Figure (Strain)
Exposure/
Duration/
Frequency
(Route)
(continued)
LOAEL
NOAEL
System
Less Serious
(ppm)
243 d
5 d/wk
7 hr/d
Serious
(ppm)
80
(ppm)
Reference
Chemical Form
Comments
Wolf et al. 1956
(leukopenia)
80
19
132
Pig
(DurocJersey)
3 wk
5 d/wk
6 hr/d
Dow 1992
100 F (T-cell depression;

decreased peripheral
WBC; decreased total
lymphocytes)
20 F
20
100
217
3 wk
3-4 x
4 hr
929 M (calculated 30%

depression of evoked
electrical activity)
Frantik et al. 1994
856 F (calculated 30%

depression of evoked
electrical activity)
Frantik et al. 1994
3. HEALTH EFFECTS
Neurological
Rat
133
(Wistar)
929
730
134
Mouse
(H)
3 wk
3-4 x
2 hr
856
731
135
Mouse
(Kunming)
30 d
6 d/wk
2 hr/d
0.78 M (increased rapid

response)
Li et al. 1992
0.78
Exposures conducted
3 of 30 days.
413
Reproductive
Rat
136
(SpragueDawley)
10 wk
Gd 0-20
Ld 5-20
5 d/wk
6 hr/d
300 F
Kuna et al. 1992
300
899
55
137
Rat
(Wistar)
BENZENE
a
Key to Species
Figure (Strain)
Exposure/
Duration/
Frequency
(Route)
(continued)
LOAEL
NOAEL
System
Less Serious
(ppm)
Serious
(ppm)
93 d
5 d/wk
7-8 hr/d
(ppm)
Reference
Chemical Form
Comments
Wolf et al. 1956
6600 M (testicular weight

increase)
6600
73
138
Mouse
(CD-1)
13 wk
5 d/wk
6 hr/d
300
30
30
(bilateral cyst in ovaries; Ward et al. 1985

atrophy/degeneration of
testes; decrease in
spermatozoa; increase in
abnormal sperm)
300
139
Gn Pig
(NS)
32 or 269 d
5 d/wk
7-8 hr/d
88 M (testicular weight
increase)
Wolf et al. 1956
80 M (degeneration of
germinal epithelium in
testes)
Wolf et al. 1956
88
75
140
Rabbit
(NS)
243 d
5 d/wk
7-8 hr/d
3. HEALTH EFFECTS
71
80
119
Cancer
141
Human
3.5 mo- 19 yr
(occup)
29
(CEL: humanlymphocytic Yin et al. 1987c

leukemia)
29
293
142
Rat
(SpragueDawley)
15 wk
4-5 d/wk
4-7 hr/d
200
200
(CEL: hepatomas)
Maltoni et al. 1982a, 1983,

1985
219
56
143
Rat
(SpragueDawley)
BENZENE
a
Key to Species
Figure (Strain)
Exposure/
Duration/
Frequency
(Route)
(continued)
LOAEL
NOAEL
System
(ppm)
Less Serious
Serious
(ppm)
(ppm)
15 wk
5 d/wk
4-7 hr/d
200
(CEL: hepatomas; onset

of Zymbal gland
carcinoma)
Reference
Chemical Form
Comments
Maltoni et al. 1982b, 1983,

1985
200
221
144
Mouse
(CBA/Ca)
16 wk
5 d/wk
6 hr/d
100 M (CEL: leukemia)
Cronkite 1986
300
(CEL: thymic and

non-thymic lymphoma)
Cronkite et al. 1984, 1985
300

(Harderian and Zymbal
gland, squamous cell and
mammary carcinoma,
papillary
adenocarcinoma of the
lung)
100
309
Mouse
(C57BL/
6BNL)
4-16 wk
5 d/wk
6 hr/d
300
373
146
Mouse
(CBA/Ca
BNL)
16 wk
5 d/wk
6 hr/d
3. HEALTH EFFECTS
145
300
833
147
Mouse
(CBA/Ca
BNL)
16 wk
5 d/wk
6 hr/d
100 M (CEL: hepatomata,

lymphomatous and
myelogenous
neoplasms)
300 M (CEL: lymphoma in

12%)
Farris et al. 1993
100
835
148
Mouse
(CBA/Ca)
16 wk
5 d/wk
6 hr/d
300
896
57
149
Mouse
(C57BL,
CD-1)
BENZENE
a
Key to Species
Figure (Strain)
Exposure/
Duration/
Frequency
(Route)
(continued)
LOAEL
NOAEL
System
(ppm)
Less Serious
Serious
(ppm)
(ppm)
10 wk
5 d/wk
6 hr/d
1200 M (CEL: 46% lung

adenoma on CD-1 mice)
Reference
Chemical Form
Comments
Snyder et al. 1988
1200
170
CHRONIC EXPOSURE
Death
150
Rat
(SpragueDawley)
104 wk
5 d/wk
4-7 hr/d
200
(61% died, versus 46% in Maltoni et al. 1982a

controls)
200
383
Rat
(SpragueDawley)
691 d
5 d/wk
6 hr/d
300 M (median lifespan 51

weeks versus 65 weeks
in controls)
Snyder et al. 1978a
300 M (median lifespan 11-41

weeks versus 39-75
weeks in controls)
Snyder et al. 1978a, 1980
300 M (decreased survival)
Snyder et al. 1982
150
300
222
152
Mouse
(AKR/J,
C57Bl)
lifetime
5 d/wk
6 hr/d
3. HEALTH EFFECTS
151
300
1030
153
Mouse
(CD-1)
222 d
5 d/wk
6 hr/d
300
1007
Systemic
Human
154
4 mo- 15 yr
(occup)
Hemato
(pancytopenia)
150
28
58
155
Human
BENZENE
a
Key to Species
Figure (Strain)
Exposure/
Duration/
Frequency
(Route)
(continued)
LOAEL
NOAEL
System
14 yr
(Occup)
Hemato
1-3 yr
(occup)
Hemato
Less Serious
(ppm)
Serious
(ppm)
(ppm)
Reference
Chemical Form
Collins et al. 1997
0.55
0.55
Comments
The NOAEL is for

abnormal
hematological values,
not significantly altered
values that would still
fall within the range of
normal values.
999
156
Human
307
157
Human
NS
(occup)
Hemato
24
(pancytopenia,
hypoplastic bone
marrow)
Erf and Rhoads 1939
24
3. HEALTH EFFECTS
(anemia, lymphocytosis, Doskin 1971

thrombocytopenia,
leukopenia, leukocytosis)
158
Human
3-29 yr
(occup)
Hemato
Fishbeck et al. 1978
25 M (increased mean
corpuscular volume)
25
354
159
Human
0.5-5 yr
(occup)
Hemato
Goldwater 1941, Greenburg et

al. 1939
11
(anemia, macrocytosis,
thrombocytopenia)
75
(anemia and leukopenia) Kipen et al. 1989
11
303
160
Human
1-25 yr
(occup)
Hemato
20
20
75
91
59
161
Human
6.1 yr (avg)
(Occup)
BENZENE
a
Key to Species
Figure (Strain)
Exposure/
Duration/
Frequency
(Route)
(continued)
LOAEL
NOAEL
System
Less Serious
(ppm)
(ppm)
Hemato
Serious
(ppm)
Reference
Chemical Form
0.57
(reduced WBC and

platelet counts,
approximately 7-18%
lower than control
values)
Lan et al. 2004a
2.26
(reduced neutrophils and

RBC counts,
approximately 12% lower
than controls)
Qu et al. 2002, 2003
7.6
(reduced absolute
lymphocyte count,
approximately 16% lower
than controls)
Rothman et al. 1996a, 1996b
Comments
0.57
1003
162
Human
4.5-9.7 yr
(mean duration) Hemato
163
Human
6.3 yr (avg)
(Occup)
Hemato
3. HEALTH EFFECTS
2.26
1002
7.6
1001
164
Human
1-21 yr
(occup)
Hemato
>1 yr
(occup)
Hemato
Tsai et al. 1983
0.53 M
0.53
357
165
Human
0.69
(leukopenia)
Xia et al. 1995
0.69
912
60
166
Human
>1 yr
(occup)
BENZENE
a
Key to Species
Figure (Strain)
Exposure/
Duration/
Frequency
(Route)
(continued)
LOAEL
NOAEL
System
Less Serious
(ppm)
Serious
(ppm)
(ppm)
Chemical Form
Comments
Yin et al. 1987b
33 M (sore throat; nasal

irritation)
Resp
Reference
33
59 F (sore throat; nasal

irritation)
59
849
33 M
Hemato
33
59 F
59
33 M
33
59 F
59
Ocular
33 M (eye irritation)
33
59 F (eye irritation)
59
167
Rat
(SpragueDawley)
lifetime
5 d/wk
6 hr/d
Resp
300 M
3. HEALTH EFFECTS
Renal
300
1033
Hemato
100 M (anemia, leukopenia)

100
Hepatic
300 M
300
Renal
300 M
300
Bd Wt
100 M
300 M (unspecified decreased

body weight gain relative
to controls)
100
300
61
168
Mouse
(AKR/J,
C57BL/6J)
lifetime
5 d/wk
6 hr/d
BENZENE
a
Key to Species
Figure (Strain)
Exposure/
Duration/
Frequency
(Route)
(continued)
LOAEL
NOAEL
System
Less Serious
(ppm)
Resp
Serious
(ppm)
(ppm)
Reference
Chemical Form
Comments
300 M
300
145
Hemato
100 M (anemia, increased

neutrophil levels;
pancytopenia; bone
marrow hypoplasia)
100
Hepatic
300 M
300
300 M
300
Bd Wt
300 M (59% decrease in weight

gain)
300
169
Mouse
(CD-1)
222 d
5 d/wk
6 hr/d
Hemato
Snyder et al. 1982
300 M (decreased RBCs and

lymphocytes)
3. HEALTH EFFECTS
Renal
300
1008
Bd Wt
300 M (reduced body weight

gain)
300
Immuno/ Lymphoret
Human
3-5 yr
170
(occup)
11
(macrocytosis,
thrombocytopenia)
Goldwater 1941, Greenburg et

al. 1939
11
500
171
Human
1-25 yr
(occup)
75
20
20
(leukopenia)
Kipen et al. 1989
75
296
62
172
Human
>1 yr
(occup)
BENZENE
a
Key to Species
Figure (Strain)
Exposure/
Duration/
Frequency
(Route)
(continued)
LOAEL
NOAEL
System
Less Serious
(ppm)
Serious
(ppm)
0.69
(ppm)
Reference
Chemical Form
Comments
Xia et al. 1995
(leukopenia)
0.69
744
173
Rat
(SpragueDawley)
lifetime
5 d/wk
6 hr/d
100 M (decreased lymphocyte

counts, splenic
hyperplasia)
100 M (lymphocytopenia, bone

marrow hypoplasia)
300 M (lymphopenia)
Snyder et al. 1988
100
1034
174
lifetime
5 d/wk
6 hr/d
100
1031
175
Mouse
(C57BL,
CD-1)
lifetime
6 hr/d
5 d/wk
300
169
Cancer
176
Human
4-15 yr
(occup)
150
(CEL: leukemia)
(CEL: leukemia,
lymphoma)
Aksoy et al. 1987
3. HEALTH EFFECTS
Mouse
(AKR/J,
C57Bl)
150
78
177
Human
28 mo- 40 yr
362
178
Human
1-10 yr
(occup)
(occup)
Infante 1978; Infante 1977
0.3 M (CEL: leukemia)
Ott et al. 1978
10
257
179
Human
18 mo
(occup)
0.3
87
63
180
Human
1-14 yr
(occup)
BENZENE
a
Key to Species
Figure (Strain)
Exposure/
Duration/
Frequency
(Route)
(continued)
LOAEL
NOAEL
System
(ppm)
Less Serious
Serious
(ppm)
(ppm)
16
Reference
Chemical Form
Comments
Rinsky et al. 1981; Infante et

al. 1977a
253
181
Human
1-30 yr
(occup)
(CEL: leukemia)
Vigliani and Forni 1976
(CEL: chronic erythroid

leukemia)
Yin et al. 1989
200
(CEL: hepatomas)
Maltoni et al. 1982a
200
(CEL: hepatomas)
200
200
349
182
Human
>1 yr
(occup)
183
Rat
(SpragueDawley)
104 wk
5 d/wk
4-7 hr/d
200
326
184
Rat
(SpragueDawley)
104 wk
5 d/wk
4-7 hr/d
3. HEALTH EFFECTS
341
200
220
185
Rat
(SpragueDawley)
lifetime
5 d/wk
6 hr/d
Snyder et al. 1984

100 M (CEL: Zymbal gland
carcinoma, myelogenous
leukemia, liver tumors)
100
704
186
Mouse
(AKR/J,
C57BL6J)
lifetime
5 d/wk
6 hr/d
300 M (CEL: hematopoietic

neoplasms [8/40],
including 6 thymic
lymphomas)
300
144
64
187
Mouse
(C57BL,
CD-1)
BENZENE
a
Key to Species
Figure (Strain)
Exposure/
Duration/
Frequency
(Route)
(continued)
LOAEL
NOAEL
System
(ppm)
Less Serious
Serious
(ppm)
(ppm)
lifetime
every
3rd wk
7 d/wk
300 M (CEL: 35% increase of

Zymbal gland
carcinomas in C57BL
mice)
Reference
Chemical Form
Comments
Snyder et al. 1988
300
262
a The number corresponds to entries in Figure 3-1.
c Used to derive an intermediate-duration inhalation minimal risk level (MRL) of 0.006 ppm for benzene. Concentration was adjusted for intermittent exposure by multiplying by 6
hours/24 hours and 5 days/7 days and converted to a human equivalent concentration, which was divided by an uncertainty factor of 300 (10 for use of a LOAEL, 3 for extrapolation
from animals to humans using dosimetric adjustment, and 10 for human variability) (see Appendix A).
d Study results used to derive a chronic-duration inhalation minimal risk level (MRL) of 0.003 ppm for benzene, as described in detail in Appendix A. Benchmark dose (BMD)
analysis was performed on B-lymphocyte counts to select a point of departure, which was adjusted for intermittent exposure and divided by an uncertainty factor of 10 for human
variability. Study results also used to derive a chronic-duration oral minimal risk level (MRL) of 0.0005 mg/kg/day based on route-to-route extrapolation, as described in detail in
Chapter 2 and Appendix A. Benchmark dose (BMD) analysis was performed on B-lymphocyte counts in benzene-exposed workers to select a point of departure, which was adjusted
for intermittent exposure. An equivalent oral dose was estimated based on route-to-route extrapolation to determine a point of departure for deriving a chronic-duration oral MRL for
benzene, which was divided by an uncertainty factor of 30 (10 for human variability and 3 for uncertainty in route-to-route extrapolation).
3. HEALTH EFFECTS
b Used to derive an acute-duration inhalation minimal risk level (MRL) of 0.009 ppm for benzene. Concentration was adjusted for intermittent exposure by multiplying by 6 hours/24
hours and converted to a human equivalent concentration, which was divided by an uncertainty factor of 300 (10 for use of a LOAEL, 3 for extrapolation from animals to humans
using dosimetric adjustment, and 10 for human variability) (see Appendix A).
e Differences in levels of health effects and cancer effects between male and females are not indicated in Figure 3-1. Where such differences exist, only the levels of effect for the
most sensitive gender are presented.
AChE = acetylcholinesterase; Bd Wt = body weight; BFU-E = burst-forming units - erythroid; Cardio = cardiovascular; CEL = cancer effect level; CFU-E = colony-forming units erythroid progenitor cells; CFU-G = colony-forming units - granulopoietic stem cells; CFU-GM = colony-forming units - macrophages; CFU-S = colony-forming units - spleen; CNS =
central nervous system; d = day(s); F = female; Gd = gestational day; Hct = hematocrit; Hemato = hematological; Hgb = hemoglobin; hr = hour(s); LC50 = lethal concentration, 50%
kill; Ld = lactational day(s); LOAEL = lowest-observed-adverse-effect level; M = male; min = minute(s); MN-NCE = micronucleated normochromatic erythrocytes; MN-PCE =
micronucleated polychromatic erythrocytes; mo = month(s); Musc/skel = musculoskeletal; NOAEL = no-observed-adverse-effect level; NRBC = nucleated red blood cells; NS = not
specified; occup = occupational exposure; RBC = red blood cell; WBC = white blood cell; wk = week(s); yr = year(s)
65
Figure 3-1 Levels of Significant Exposure to Benzene - Inhalation

BENZENE
Acute (14 days)

Systemic
ppm
ath
pir
De
Re
lar
dio
Ca
s
va
ica
cu
r
ato
og
tol
ma
He
c
ati
He
ht
eig
l
ma
De
yW
Bo
100000
4h
1
2r
10000
3r
22m
9r
19m
6r
1000
24m
100
17m
5
13m
18m
21m
23m
29m
22m
12r
10r
8r
8r
14m
17m
10
24m
20m
22m
26m
27m
25m
29m
28m
30m
12r
13m
11r
11r
15m
14m
29m
6r
10r
13m
28m
14m
7r
7r
3. HEALTH EFFECTS
16m
15m
13m
30m
1
0.1
0.01
0.001
-Humans
k-Monkey
m-Mouse
h-Rabbit
a-Sheep
f-Ferret
n-Mink
j-Pigeon
o-Other
e-Gerbil
s-Hamster
g-Guinea Pig
Cancer Effect Level-Animals

LOAEL, More Serious-Animals
LOAEL, Less Serious-Animals
NOAEL - Animals
Cancer Effect Level-Humans

LOAEL, More Serious-Humans
LOAEL, Less Serious-Humans
NOAEL - Humans
LD50/LC50
Minimal Risk Level
for effects
other than
Cancer
66
c-Cat
d-Dog
r-Rat
p-Pig
q-Cow
Figure 3-1 Levels of Significant Exposure to Benzene - Inhalation (Continued)

BENZENE
Acute (14 days)

Systemic
ppm
dy
Bo
ho
p
ym
t
igh
al
o/L
mu
ur
Ne
Im
pr
Re
c
du
tal
en
tiv
ic
log
m
lop
ve
De
100000
56h
10000
40m
54m
1000
53r
58r
55m
31h
100
11r
41m
34m
44m
42m
44m
37m
49m
33r
33r
32r
32r
35m
37m
10
43m
45m
39m
39m
42m
45m
47m
49m
48m
46m
47m
60m
51
55m
54m
52
50m
61h
53r
57r
59r
62h
62h
69m
70m
63r
63r
71h
72h
64r
65r
66r
67r
72h
67r
68m
68m
65r
3. HEALTH EFFECTS
38m
36m
31h
50m
1
0.1
0.01
0.001
-Humans
k-Monkey
m-Mouse
h-Rabbit
a-Sheep
f-Ferret
n-Mink
j-Pigeon
o-Other
e-Gerbil
s-Hamster
g-Guinea Pig

NOAEL - Animals

NOAEL - Humans
LD50/LC50
Minimal Risk Level
for effects
other than
Cancer
67
c-Cat
d-Dog
r-Rat
p-Pig
q-Cow

BENZENE
Intermediate (15-364 days)

Systemic
ppm
ica
ath
og
tol
ma
De
He
c
ati
He
ht
al
Re
eig
yW
Bo
10000
1000
74m
75m
10
77m
73r
79
78
103g
88m
89m
90m
91m
93m
92m
80
81
90m
86m
87m
96m
97m
97m
98m
101m
99m
94m
100m
93m
84r
85r
102m
92m
82r
102m
84r
104h
105p
105p
95m
95m
95m
94m
3. HEALTH EFFECTS
100
76m
0.1
0.01
0.001
-Humans
k-Monkey
m-Mouse
h-Rabbit
a-Sheep
f-Ferret
n-Mink
j-Pigeon
o-Other
e-Gerbil
s-Hamster
g-Guinea Pig

NOAEL - Animals

NOAEL - Humans
LD50/LC50
Minimal Risk Level
for effects
other than
Cancer
68
c-Cat
d-Dog
r-Rat
p-Pig
q-Cow
Systemic
ppm
ho
t
igh
p
ym
d
Bo
o
un
g
olo
ve
cti
u
Ne
mm
ica
/L
yW
BENZENE
u
od
r*
ce
p
Re
n
Ca
10000
137r
112r
1000
119m
102m
83r
114m
106
130g
115m
121m
116m
117m
118m
123m
125m
128m
125m
127m
113m
109r
110r
111r
131h
132p
138m
136r
139g
140h
145m
146m
144m
147m
128m
107
10
133r
129m
117m
118m
120m
124m
122m
110r
138m
132p
141
126m
124m
1
3. HEALTH EFFECTS
100
84r
134m
108r
129m
109r
135m
0.1
0.01
*Doses represent the lowest dose tested per study that produced a tumorigenic
response and do not imply the existence of a threshold for the cancer endpoint.
0.001
-Humans
k-Monkey
m-Mouse
h-Rabbit
a-Sheep
f-Ferret
n-Mink
j-Pigeon
o-Other
e-Gerbil
s-Hamster
g-Guinea Pig

NOAEL - Animals

NOAEL - Humans
LD50/LC50
Minimal Risk Level
for effects
other than
Cancer
69
c-Cat
d-Dog
r-Rat
p-Pig
q-Cow

BENZENE

r*
ce
ppm
n
Ca
10000
149m
1000
148m
142r
143r
3. HEALTH EFFECTS
100
10
0.1
0.01
0.001
-Humans
k-Monkey
m-Mouse
h-Rabbit
a-Sheep
f-Ferret
n-Mink
j-Pigeon
o-Other
e-Gerbil
s-Hamster
g-Guinea Pig

NOAEL - Animals

NOAEL - Humans
LD50/LC50
Minimal Risk Level
for effects
other than
Cancer
70
c-Cat
d-Dog
r-Rat
p-Pig
q-Cow

BENZENE
Chronic (365 days)

Systemic
ppm
ry
ato
ir
sp
ath
De
a
gic
lo
ato
ati
m
He
Re
p
He
al
n
Re
lar
yW
u
Oc
ho
t
igh
/L
no
u
mm
d
Bo
p
ym
1000
152m
153m
150r
151r
168m
167r
169m
154
100
168m
160
166
157
158
159
10
167r
168m
169m
167r
166
166
163
167r
171
171
170
162
1
155
168m
167r
166
160
167r
161
164
165
172
0.1
3. HEALTH EFFECTS
156
168m
0.01
0.001
0.0001
1E-5
1E-6
-Humans
k-Monkey
m-Mouse
h-Rabbit
a-Sheep
f-Ferret
n-Mink
j-Pigeon
o-Other
e-Gerbil
s-Hamster
g-Guinea Pig

NOAEL - Animals

NOAEL - Humans
LD50/LC50
Minimal Risk Level
for effects
other than
Cancer
71
c-Cat
d-Dog
r-Rat
p-Pig
q-Cow

BENZENE
Chronic (365 days)
ho
ppm
/L
no
u
mm
p
ym
r*
ce
n
Ca
1000
175m
100
174m
173r
176
10
180
178
177
187m
181
188m
183r
184r
185r
186r
3. HEALTH EFFECTS
182
179
0.1
0.01
10-4
Estimated
0.001
Upper-Bound
10-5
Human Cancer
Risk Levels
0.0001
10-6
1E-5
1E-6
-Humans
k-Monkey
m-Mouse
h-Rabbit
a-Sheep
f-Ferret
n-Mink
j-Pigeon
o-Other
e-Gerbil
s-Hamster
g-Guinea Pig

NOAEL - Animals

NOAEL - Humans
LD50/LC50
Minimal Risk Level
for effects
other than
Cancer
72
c-Cat
d-Dog
r-Rat
p-Pig
q-Cow
10-7
BENZENE
73
3. HEALTH EFFECTS
Cardiovascular Effects.
No studies were located regarding cardiovascular effects in humans after
inhalation exposure to benzene, although ventricular fibrillation has been proposed as the cause of death
in some human poisonings (Avis and Hutton 1993; Winek and Collom 1971).
One animal study was found that investigated the effects of acute inhalation exposure to high concentrations of benzene vapor on the heart muscle of cats and monkeys (Nahum and Hoff 1934). Information
from the electrocardiograms indicated that exposure to benzene vapor caused extra systoles and
ventricular tachycardia of the prefibrillation type. Animals that had their adrenals and stellate ganglias
removed did not exhibit extra systoles or ventricular tachycardia. These findings suggest that the
arrhythmias were caused by catecholamine release and sympathetic discharge. This study is limited in
that exact levels of exposure are not available. An additional study investigated the influence of benzene
inhalation on ventricular arrhythmia in the rat (Magos et al. 1990). Rats exposed to 3,5268,224 ppm of
benzene in a closed chamber for 15 minutes exhibited an increased number of ectopic ventricular beats.
Gastrointestinal Effects.
Very few data are available describing gastrointestinal effects in humans
after inhalation exposure to benzene. In a case study involving the death of an 18-year-old boy who
intentionally inhaled benzene, the autopsy revealed congestive gastritis (Winek and Collom 1971). No
other details or data were given.
Data regarding effects on the human hematological system following acute
inhalation exposure to benzene are scant, but indicate leukopenia, anemia, and thrombocytopenia after
more than 2 days of occupational exposure to more than 60 ppm benzene (Midzenski et al. 1992).
Epidemiological studies on persons exposed to various levels of benzene in the workplace for
intermediate and chronic periods of time also indicate hematological effects. Deficiencies in most of
these studies include uncertainty in estimates of historical exposure levels, concomitant exposure to other
chemicals, and lack of appropriate control groups. However, sufficient data are available to show that the
hematopoietic system is a critical target for benzene toxicity. Studies that were conducted well and that
show effects linked to specific exposure levels are presented in Table 3-1 and Figure 3-1. Effects on
leukocytes, lymphocytes, and bone marrow are also discussed in Section 3.2.1.3.
Inhalation exposure to benzene levels in excess of regulated workplace limits (8-hour TWA of 1 ppm) for
several months to several years can result in deficits in the relative numbers of circulating blood cells,
which may be severe enough to be considered clinical pancytopenia. Continued exposure to benzene can
BENZENE
74
3. HEALTH EFFECTS
also result in aplastic anemia or leukemia (Aksoy et al. 1974; EPA 1995; Hayes et al. 1997; IARC 1982,
1987; IRIS 2007; Rinsky et al. 1987, 2002; Yin et al. 1987c, 1996a, 1996b).
Pancytopenia is the reduction in the number of all three major types of blood cells: erythrocytes (red
blood cells), thrombocytes (platelets), and leukocytes (white blood cells). In adults, all three major types
of blood cells are produced in the red bone marrow of the vertebrae, sternum, ribs, and pelvis. The red
bone marrow contains immature cells, known as multipotent myeloid stem cells, that later differentiate
into the various mature blood cells. Pancytopenia results from a reduction in the ability of the red bone
marrow to produce adequate numbers of these mature blood cells.
Aplastic anemia is a more severe effect of benzene and occurs when the bone marrow ceases to function
and the stem cells never reach maturity. Depression in bone marrow function occurs in two stages
hyperplasia (increased synthesis of blood cell elements), followed by hypoplasia (decreased synthesis).
As the disease progresses, bone marrow function decreases and the bone marrow becomes necrotic and
filled with fatty tissue. This myeloblastic dysplasia without acute leukemia has been seen in persons
exposed to benzene (Erf and Rhoads 1939). Aplastic anemia can progress to a type of leukemia known as
acute myelogenous leukemia (Aksoy 1980), which is discussed in Section 3.2.1.7.
Early biomarkers of exposure to relatively low levels of benzene include depressed numbers of one or
more of the circulating blood cell types. For example, statistically significantly decreased total red blood
cells (RBCs), white blood cells (WBCs), absolute lymphocyte count, platelets, and hematocrit were
reported for a group of 44 healthy subjects exposed to benzene in the workplace (median 8-hour timeweighted average [TWA] of 31 ppm; minimal exposure to other solvents) for an average of 6.3 years in
China (Rothman et al. 1996a, 1996b). Age- and gender-matched workers with no history of occupational
exposure to benzene served as controls. Among the 22 workers whose mean 5-day benzene exposure
levels did not exceed 31 ppm (median 8-hour TWA of 13.6 ppm), significantly depressed absolute
lymphocyte count, RBCs, and platelets were noted. Only absolute lymphocyte count was significantly
decreased in a subgroup of 11 workers with no 8-hour TWA exceeding 31 ppm (median 8-hour TWA of
7.6 ppm).
Qu et al. (2002, 2003a, 2003b) compared hematology values in a group of 130 chronically exposed
workers in China with those obtained from 51 age- and gender-matched subjects without occupational
exposure to benzene. Statistically significant trends for depressed RBCs, WBCs, and neutrophils were
observed in the benzene-exposed workers (average measured 4-week benzene exposure levels ranged
BENZENE
75
3. HEALTH EFFECTS
from 0.08 to 54.5 ppm just prior to blood testing). A subgroup of 73 of these workers, whose average
4-week benzene exposure level was 2.26 ppm, exhibited significantly depressed RBCs and neutrophils.
This study provided exposure-response data, but apparent discrepancies in reported low-concentration
results render the study of limited value for MRL derivation.
One recent cross-sectional study (Lan et al. 2004a, 2004b), performed on 250 workers exposed to benzene
in shoe manufacturing industries in Tianjin, China, and 140 age- and gender-matched workers in clothing
manufacturing facilities that did not use benzene, was of sufficient quality to serve as the basis for
deriving a chronic-duration inhalation MRL for benzene (see footnote to Table 3-1 and Appendix A).
The benzene-exposed workers had been employed for an average of 6.12.9 years. Controls consisted of
140 age-and gender-matched workers in clothing manufacturing facilities in which measurable benzene
concentrations were not found (detection limit 0.04 ppm). Benzene exposure was monitored by
individual organic vapor monitors (full shift) 5 or more times during 16 months prior to phlebotomy.
Benzene-exposed workers were categorized into four groups (controls, <1, 1<10, and 10 ppm)
according to mean benzene exposure levels measured during 1 month prior to phlebotomy. Complete
blood count (CBC) and differential were analyzed mechanically. Coefficients of variation for all cell
counts were <10%.
Mean 1-month benzene exposure levels in the four groups (controls, <1, 1<10, and 10 ppm) were
<0.04, 0.570.24, 2.852.11, and 28.7320.74 ppm, respectively. Hematological values were adjusted to
account for potential confounding factors (i.e., age, gender, cigarette smoking, alcohol consumption,
recent infection, and body mass index). All types of WBCs and platelets were significantly decreased in
the lowest exposure group (<1 ppm), ranging in magnitude from approximately 8 to 15% lower than
controls. Although similar statistical analyses for the mid- and high-exposure groups were not included
in the study report, decreases in all types of WBCs and platelets were noted at these exposure levels as
well; the decreases in the highest exposure group ranged in magnitude from 15 to 36%. Lymphocyte
subset analysis revealed significantly decreased CD4+-T cells, CD4+/CD8+ ratio, and B cells.
Hemoglobin concentrations were significantly decreased only within the highest (10 ppm) exposure
group. Tests for a linear trend using benzene air level as a continuous variable were significant for
platelets and all WBC measures except monocytes and CD8+-T cells. Upon restricting the linear trend
analyses to workers exposed to <10 ppm benzene, excluding controls, inverse associations remained for
total WBCs, granulocytes, lymphocytes, B cells, and platelets. In order to evaluate the effect of past
benzene exposures on the hematological effects observed in this study, the authors compared findings for
a group of workers who had been exposed to <1 ppm benzene over the previous year (n=60) and a subset
BENZENE
76
3. HEALTH EFFECTS
who also had <40 ppm-years lifetime cumulative benzene exposure (n=50). The authors stated that the
same cell types were significantly reduced in these groups, but did not provide further information of the
magnitude (i.e., percent change) of the hematological effects observed. These data suggest that the
1-month benzene exposure results could be used as an indicator of longer-term low-level benzene
hematotoxicity. To demonstrate that the observed effects were attributable to benzene, significantly
decreased levels of WBCs, granulocytes, lymphocytes, and B cells were noted in a subgroup (n=30; mean
1-month exposure level of 0.290.15 ppm) of the <1 ppm group for which exposure to other solvents was
negligible. Lan et al. (2004a, 2004b) also presented information on the effect of benzene on colony
forming progenitor cells (data were only presented for the mid- and high-exposure groups). Benzene
exposure was associated with a concentration-dependent decrease in colony formation and progenitor
cells were suggested to be more sensitive than circulating cells.
As described above, several epidemiology studies compared hematological variables for benzene-exposed
workers to gender- and age-matched controls and noted hematological effects at relatively low
concentrations (well below 1 ppm in some exposure groups) (Lan et al. 2004a, 2004b; Qu et al. 2002,
2003a, 2003b; Rothman et al. 1996a, 1996b). An alternative approach was used by Collins et al. (1991,
1997) and Tsai et al. (1983, 2004). This approach utilized a defined range of clinically normal
hematological values and compared the prevalence of abnormal results between benzene-exposed
workers and unexposed controls. Collins et al. (1991) found no significant correlations between benzene
exposure and the prevalence of abnormal hematological values among 200 workers exposed to benzene at
estimated concentrations ranging from 0.01 to 1.4 ppm, relative to the prevalence of abnormal
hematological values obtained from 268 unexposed workers in the same plant. In a more recent
evaluation, Collins et al. (1997) found no significant correlation between exposure to benzene at an
8-hour TWA of 0.55 ppm and prevalence of clinically-defined lymphopenia (or other measures of
hematotoxicity including mean corpuscular volume [MCV], and counts of WBCs, RBCs, hemoglobin,
and platelets) among a group of 387 workers exposed for 5 years. Tsai et al. (1983) measured benzene
levels in a Texas refinery at one time point (mean benzene concentration of 0.53 ppm) and found that
hemoglobin, hematocrit, RBCs, WBCs, and thrombocytes of workers exposed for up to 21 years were
within the range of normal values. Tsai et al. (2004) found no clinically adverse hematotoxic effects
among a large group of 1,200 petrochemical employees with mean 8-hour TWA benzene exposure levels
of 0.6 ppm from 1977 to 1988 and 0.14 ppm from 1988 to 2002. The normal range for certain
hematological parameters is necessarily broad due to large interindividual differences in clinical status.
Restricting the comparison of benzene-exposed and nonexposed populations to only those values
BENZENE
77
3. HEALTH EFFECTS
considered clinically abnormal or adverse may reduce the sensitivity of the study to detect meaningful
changes at the population level.
Many reports of hematological effects in benzene-exposed workers involve estimated exposure levels
well in excess of 1 ppm. A series of studies conducted on Turkish workers exposed to benzenecontaining adhesives in various occupations showed increased severity of effects with increased levels or
duration of exposure. The initial hematological study examined 217 male workers who were exposed for
between 4 months and 17 years to benzene-containing solvents (Aksoy et al. 1971). The concentration of
benzene in the work area ranged from 15 to 30 ppm outside work hours and reached a maximum of
210 ppm when benzene-containing adhesives were being used. Fifty-one of the workers showed clinical
hematological abnormalities such as leukopenia, thrombocytopenia, eosinophilia, and pancytopenia. An
additional cohort was identified that included 32 shoe manufacturers who had worked with benzene for
4 months to 15 years at concentrations of 1530 ppm outside work hours and 210640 ppm during the
use of benzene, and who showed pancytopenia (Aksoy et al. 1972). Examination of these people revealed
disruptions in bone marrow function, including cases of hypoplastic, acellular, hyperplastic, or
normoblastic bone marrow. The continuing assessment further identified that ineffective erythropoiesis
or increased hemolysis may have been responsible for the reticulocytosis, hyperbilirubinemia,
erythroblastemia, increase in quantitative osmotic fragility, and elevated serum lactate dehydrogenase
levels observed in some patients.
Leukopenia, lymphocytosis, a biphasic leukocyte response, and bone marrow hypercellularity were
reported for workers in a Russian industry who were exposed to a complex of hydrocarbons that included
benzene, cyclohexane, and 1,3-butadiene (Doskin 1971). Benzene levels were estimated at 3.212.8 ppm,
which are 28 times the Russian standard. Leukopenia was observed by Xia et al. (1995) in Chinese
workers exposed to 0.69140 ppm (mean=6 ppm) benzene for more than 1 year. Enlarged RBCs,
transient anemia, reduced hemoglobin concentrations, and clinically increased MCV were reported in
workers in a chemical factory who were exposed to over 25 ppm of benzene in the workplace for an
average of 9 years (Fishbeck et al. 1978).
Dosemeci et al. (1996) assessed the relative risk of abnormal hematological values with increasing
benzene levels in a study of workers employed in 672 rubber and rubber glue application facilities in
China between 1949 and 1987. Compared to workers with estimated exposures <5 ppm, relative risks of
abnormal hematological values (indicating benzene poisoning) by estimated exposure intensity at
1.5 years prior to clinically-diagnosed benzene poisoning were 2.2 (95% confidence interval [CI] 1.7
BENZENE
78
3. HEALTH EFFECTS
2.9), 4.7 (95% CI 3.46.5), and 7.2 (95% CI 5.39.8) for exposures in the ranges of 519, 2039, and
>40 ppm, respectively.
Examination of individual hematological records, demographic data, and chronological work histories of
459 workers employed at a rubber products manufacturing plant in Ohio between 1940 and 1975 revealed
significant decreases in WBC and RBC counts and hemoglobin for the period of 19401948 when the
exposure was 75 ppm (Kipen et al. 1989). The trend was not apparent during later years (19491975)
when the exposure was decreased to 1520 ppm. A more focused study of many of the same workers
during their first year of employment revealed significantly lower WBC and RBC counts in employees
exposed to benzene levels greater than the median (estimated at 4054 ppm), compared to those with
lower estimated exposure levels (Cody et al. 1993). Using a nested case-control approach at the same
rubber products manufacturing plant to assess a possible exposure-response relationship between benzene
and the risk of developing a low WBC or RBC count in workers for whom hematologic screening data
were available, Ward et al. (1996) reported a strong correlation between low WBC counts and benzene
exposure and a weak correlation between RBC count and benzene exposure.
More severe effects, including preleukemia or acute leukemia, were observed in 26 out of 28,500 benzene
workers exposed to 210650 ppm for 115 years (Aksoy et al. 1974). Clinical features of the
preleukemia included one or more of the following: anemia, leukopenia, pancytopenia, bone marrow
hyperplasia, pseudo-Pelger-Huet anomaly, and splenomegaly. A study was conducted 217 years
following the last exposure of 44 pancytopenic patients exposed to benzene (150650 ppm) in adhesives
for 4 months to 15 years (Aksoy and Erdem 1978). Of these patients, complete remission was seen in 23,
death due to complications of pancytopenia in 14, death due to myeloid metaplasia in 1, and leukemia in
6. When benzene concentrations in factories decreased in later years, less severe effects were seen. At
40 tire manufacturing plants, 231 workers were exposed for 28 months to 40 years (mean 8.8 years) to
benzene-containing solvents and thinners (Aksoy et al. 1987). The decrease in benzene content of
materials used in these workshops and the corresponding reduction in air concentration (most samples
<1 ppm) paralleled the decrease in the number of hematological abnormalities reported for benzeneexposed workers.
Another study revealed effects, ranging from mild to severe, of benzene exposure in factory workers in
China (Yin et al. 1987c). Of the 528,729 workers, 95% were exposed to mixtures of benzene, toluene,
and xylene, while 5% (26,319 workers) were exposed to benzene alone at 0.02264 ppm in air in 95% of
the work stations. Over half of the work stations had levels of benzene in the air of less than 13 ppm;
BENZENE
79
3. HEALTH EFFECTS
about 1% had levels of 13264 ppm. Benzene toxicity, as indicated by leukopenia, aplastic anemia, and
leukemia, was seen in 0.94% of the workers exposed to benzene and 0.44% of the workers exposed to the
mixtures. Similar toxicity was found in employees of 28 of the 141 shoe factories studied (124 cases in
2,740 employees) (Yin et al. 1987c). A positive correlation was observed for prevalence of adverse
benzene effects and benzene concentration in data from these 28 shoe factories. The authors determined
that the affected people were exposed to benzene concentrations >29 ppm. In one workshop, there were
4 cases of aplastic anemia in 211 workers. These workers were exposed to benzene at a mean
concentration of 324 ppm during an 8-month period of employment. The prevalence of aplastic anemia
in the shoe-making industry was about 5.8 times that in the general population. The main limitation of
this study is the lack of information on the duration of exposure.
People who were exposed to high levels of benzene vapors in the printing industry also showed severe
hematological effects. One study evaluated 332 workers who were exposed to 111,060 ppm of benzene
for 6 months to 5 years. Detailed blood studies performed on 102 of these workers revealed benzene
poisoning in 22 workers characterized by pancytopenia or other clinical signs (Goldwater 1941;
Greenburg et al. 1939). Another study reported 6 cases of pancytopenia and bone marrow dysplasia in
printers exposed to 241,060 ppm of benzene (Erf and Rhoads 1939).
Animal studies have been designed to characterize exposure-response relationships of benzene
hematotoxicity; the results provide support to the human data. Animal responses to benzene exposure are
variable and may depend on factors such as species, strain, duration of exposure, and whether exposure is
intermittent or continuous. Wide variations have also been observed in normal hematological parameters,
complicating statistical evaluation. However, the studies show that benzene exerts toxic effects at all
phases of the hematological system, from stem cell depression in the bone marrow, to pancytopenia, to
histopathological changes in the bone marrow. Effects on leukocytes, lymphocytes, and bone marrow are
also discussed in Section 3.2.1.3.
Repeated acute-, intermediate-, and chronic-duration inhalation exposure of laboratory animals (mainly
mice, but also rats, rabbits, and guinea pigs) to benzene vapor concentrations ranging from 10 to
>300 ppm demonstrate significant decreases in blood values that include RBCs, total WBCs,
lymphocytes, granulocytes, hematocrit, and hemoglobin (Aoyama 1986; Baarson et al. 1984; Chertkov et
al. 1992; Cronkite et al. 1982; Farris et al. 1997a, 1997b; Gill et al. 1980; Green et al. 1981a, 1981b; Li et
al. 1986; Rozen and Snyder 1985; Rozen et al. 1984; Snyder et al. 1982; Ward et al. 1985; Wells and
Nerland 1991; Wolf et al. 1956). Pancytopenia was noted in the study of Ward et al. (1985).
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Short-duration exposure of mice to benzene has been shown to cause both concentration- and durationrelated reductions in bone marrow cellularity (number of nucleated cells) and the number of colonyforming granulopoietic stem cells (CFU-C), and an increased frequency of micronucleated polychromatic
erythrocytes (MN-PCE) (Toft et al. 1982). Mice that were exposed continuously at benzene
concentrations 21 ppm in air for 410 days showed significant changes in all three parameters.
Intermittent exposure (8 hours/day, 5 days/week for 2 weeks) to 21 ppm significantly reduced the number
of CFU-C and elevated the frequency of MN-PCE, but did not affect bone marrow cellularity.
Intermittent exposure at concentrations 50 ppm caused significant changes in all three parameters and
decreased the ability of the spleen to form mature cells (as measured by numbers of colony-forming units
of stem cells). When mice were intermittently exposed for 2 weeks, decreased cellularity and CFU-C per
tibia were observed at 95 ppm after 68 hours/day exposure, whereas at 201 ppm benzene, decreased
cellularity (but no effect on CFU-C) was noted after 2 hours/day exposure. A decrease in cellularity and
CFU-C was observed after 48 hours/day exposure to 201 ppm benzene.
Results of numerous additional studies of laboratory animals (mainly mice, which are particularly
sensitive to benzene hematotoxicity, but also rats and pigs) support findings of benzene-induced effects
on bone marrow cellularity (hyper- and/or hypocellularity) and colony-forming stem cells, as well as
granulocytic hyperplasia, following repeated acute-, intermediate-, or chronic-duration inhalation
exposure to benzene vapors at concentrations ranging from 10 to 500 ppm (Baarson and Snyder 1991;
Baarson et al. 1984; Chertkov et al. 1992; Corti and Snyder 1996; Cronkite et al. 1982, 1985, 1989;
Dempster and Snyder 1991; Dow 1992; Farris et al. 1993, 1997b; Neun et al. 1992, 1994; Plappert et al.
1994a, 1994b; Snyder et al. 1978a, 1980, 1982; Vacha et al. 1990). For example, Dempster and Snyder
(1991) observed a 50% reduction in CFU-E (erythroid progenitor cells) in bone marrow of DBA/2 mice
exposed to 10 ppm benzene for 6 hours/day for 5 days. Farris et al. (1997b) reported benzene-induced
decreased numbers of total bone marrow forming cells, progenitor cells, and differentiating hematopoietic
cells in mice exposed to benzene vapor concentrations 200 ppm, 6 hours/day, 5 days/week for up to
8 weeks. Replication of primitive progenitor cells in the bone marrow was increased during the exposure
period, presumably as compensation for cytotoxicity. Granulocytic hyperplasia was detected in the bone
marrow of mice exposed to 300 ppm benzene in air for 6 hours/day, 5 days/week for 16 weeks, and held
18 months after the last exposure (Farris et al. 1993). Prolonged exposure to lower levels had a greater
hematotoxic effect than exposure to higher levels for a shorter period of time. Recovery from
hematotoxicity has been demonstrated following the cessation of exposure (Cronkite et al. 1982, 1985,
1989) and may be most closely associated with rate of exposure, since longer-term exposure of mice to
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316 ppm of benzene caused irreparable hematologic injury, while the hematotoxic effects caused by
shorter-term exposure to 10 times higher benzene concentrations were reversible (Cronkite et al. 1989).
One study revealed damaged erythrocytes in the peripheral blood of mice (Luke et al. 1988b). Cytotoxic
damage in the bone marrow was dependent on strain and exposure duration. Peripheral blood smears
were analyzed weekly from three strains of mice (DBA/2, B6C3F1, and C57BL/6) exposed to 300 ppm
benzene for 13 weeks (6 hours/day) for either 5 days/week (Regimen 1) or 3 days/week (Regimen 2). In
all three strains, an initial severe depression in rate of erythropoiesis was observed. The return to normal
was dependent on strain (Luke et al. 1988b) and regimen (Cronkite et al. 1989; Luke et al. 1988b). An
increase in frequency of micronucleated normochromatic erythrocytes (MN-NCE) was observed to be
dependent on strain (C57BL/6=B6C3F1>DBA/2) and regimen (Regimen 1 > Regimen 2), whereas the
increase in frequency of MN-PCE was dependent on strain (DBA/2>C57BL/6=B6C3F1) but, for the most
part, was not dependent on exposure regimen.
Damaged erythroblast-forming cells were also noted in bone marrow (Seidel et al. 1989). A substantial
decrease in erythroid colony-forming units and smaller decreases in erythroid burst-forming units
occurred in BDF1 mice intermittently exposed to 100 ppm of benzene for 8 weeks (6 hours/day,
5 days/week). Although the effects in the 100 ppm group were not apparent at 8 weeks (the end of
experiment), they did occur, and it took over 3 weeks for the erythroid burst-forming units and erythroid
colony-forming units to return to their initial values. This reduction in the number of erythroid precursors
was reflected by a slight reduction in the number of erythrocytes.
Benzene-induced hematotoxicity was also demonstrated in the spleen of rats and mice following
intermediate- or chronic-duration repeated inhalation exposure (Snyder et al. 1978a, 1984; Ward et al.
1985). Snyder et al. (1978a, 1984) reported benzene-induced increased extramedullary hematopoiesis in
the spleen. Ward et al. (1985) noted that the finding of hemosiderin in the spleen of benzene-exposed rats
could be due to erythrocyte hemolysis.
Musculoskeletal Effects.
A case of myelofibrosis was diagnosed in a 46-year-old man in October
1992 (Tondel et al. 1995). The patient worked from 1962 to 1979 as a gasoline station attendant. The
patient was referred to the Department of Hematology, University Hospital in Linkoping, Sweden, where
a bone marrow biopsy was performed. The patient described symptoms of increasing muscle pain for
1 year, fatigue for 3 weeks, night sweats, and weight loss. A bone marrow biopsy showed myelofibrosis.
The TWA concentration for gasoline station attendants was estimated to be <0.2 ppm. The occupational
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standard for benzene in Sweden was 0.5 ppm (TWA) and the Swedish short-term exposure limit was
3 ppm. Ruiz et al. (1994) reported musculoskeletal effects in employees from a steel plant of Cubatao,
S. Paulo, Brazil, who presented with neutropenia due to benzene exposure. Patients either were employed
at the steel plant (mean time of 7 years and 4 months), or were employees of a building construction
company working at repairs in the steel plant (mean time of 5 years and 5 months). Sixty percent of the
workers had nonspecific clinical complaints such as myalgia.
Hepatic Effects.
No specific reports of adverse hepatic effects of inhalation exposure to benzene in
humans were found, although Aksoy et al. (1972) reported enlarged livers in workers chronically exposed
to benzene at airborne concentrations ranging from 150 to 650 ppm.
CFY rats (20/group) were exposed to pure air, benzene (125 ppm) or benzene (400 ppm) and toluene
(265 ppm) for 24 hours/day from gestational day (Gd) 7 through 14 (Tatrai et al. 1980a). The rats were
then sacrificed on day 21 of pregnancy. Exposure to 125 ppm benzene caused a slight increase in relative
liver weight of 4.67% compared to 4.25% in controls, which was not considered adverse. In a companion
study, CFY rats were exposed to continuous benzene inhalation 24 hours/day from day 7 to day 14 of
gestation at 0, 47, 141, 470, or 939 ppm atmospheric concentrations (Tatrai et al. 1980b). At 141 ppm
benzene, there was a significant increase in relative maternal liver weight.
No treatment-related non-neoplastic histopathological effects on hepatic tissue were found in male
Sprague-Dawley rats exposed to 0, 100, or 300 ppm benzene 5 days/week, 6 hours/day for life (Snyder et
al. 1978a, 1984) or in AKR/J or C57B1/6J mice similarly exposed to 300 ppm for life (Snyder et al.
1978a, 1980).
Renal Effects.
Very little data are available describing renal effects in humans after inhalation
exposure to benzene. In a case study involving the death of an 18-year-old boy who intentionally inhaled
benzene, the autopsy revealed acute kidney congestion (Winek and Collom 1971). No other details or
data were given.
No treatment-related histopathological effects on kidney tissue were found in male Sprague-Dawley rats
were exposed to 0, 100, or 300 ppm benzene 5 days/week, 6 hours/day for life (Snyder et al. 1978a, 1984)
or in AKR/J or C57B1/6J mice similarly exposed to 300 ppm (Snyder et al. 1978a, 1980).
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Dermal Effects.
Dermal effects in humans have been reported after acute exposure to benzene vapors
(Avis and Hutton 1993). After a fatal occupational exposure to benzene vapors on a chemical cargo ship
for only minutes, autopsy reports on three victims revealed hemorrhagic respiratory tissues, and second
degree burns on the face, trunk, and limbs (Avis and Hutton 1993). Skin irritation has been noted at
occupational exposures of >60 ppm for up to 3 weeks (Midzenski et al. 1992). These effects are due to
direct contact of the skin with the vapor, and other dermal effects resulting from direct contact of the skin
are discussed in Section 3.2.3.2.
Ocular Effects.
Three hundred solvent workers who had inhalation exposures for >1 year to benzene
at 33 and 59 ppm for men and women, respectively, complained of eye irritation (Yin et al. 1987b).
Male Charles River CD rats exposed to 0, 1, 10, 30, or 300 ppm benzene 6 hours/day, 5 days/week for
10 weeks exhibited lacrimation at concentrations >10 ppm during the first 3 weeks of treatment (Shell
1980).
These effects are due to direct contact of the eyes with the vapor, and other ocular effects resulting from
direct contact of the eyes are discussed in Section 3.2.3.2.
Body Weight Effects.
Relatively few studies in animals report changes in body weight after
inhalation exposure to benzene. No change in body weight was noted in Sprague-Dawley rats or
CD-1 mice exposed to 300 ppm benzene for 13 weeks (Ward et al. 1985). No significant decrease in
body weight was observed in CD-1 mice exposed to doses up to 4,862 ppm for 6 hours/day, for 5 days, or
at lower doses of 9.6 ppm for 50 days (Green et al. 1981b). Decreases in body weight (15%) were seen in
DBA/2 mice after exposure to 300 ppm benzene in air for 6 hours/day, 5 days/week for 2 weeks
(Chertkov et al. 1992). Decreased body weight (1618%) has also been noted in mice exposed to doses
of approximately 200 ppm of benzene for 6 hours/day for 7 or 14 days (Aoyama 1986). C57BL mice
exhibited a 59% decrease in body weight gain after exposure to 300 ppm benzene, 5 hours/day,
6 days/week over their lifetime (Snyder et al. 1980). Decreased maternal body weight and weight gain
have been observed in Sprague-Dawley rats exposed to 50 ppm benzene during gestation days (Gds)
6-15 (Kuna and Kapp 1981), but not in rats exposed to doses up to 300 ppm during premating, mating,
gestation, and lactation (Kuna et al. 1992). CFY rats (20/group) were exposed to pure air, benzene
(125 ppm), or benzene (400 ppm) plus toluene (265 ppm) for 24 hours/day from Gd 7 through 14 (Tatrai
et al. 1980a). The rats were then sacrificed on day 21 of pregnancy. Exposure to 125 ppm benzene
caused decreased maternal weight gain (46.74% of starting weight as opposed to 68.82% of starting
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weight in controls). Decreased maternal weight gain was also observed in a companion experiment in
which the rats were exposed to doses as low as 47 ppm using the same study design (Tatrai et al. 1980b).
Decreased maternal body weight was also observed in rats exposed to 2,200 ppm benzene during
gestation (Green et al. 1978). Rabbits exposed to 313 ppm benzene on Gd 720 exhibited decreased
maternal weight gain (Ungvary and Tatrai 1985). Kunming mice exposed to 12.52 ppm benzene for
2 hours/day, 6 days/week for 30 days exhibited no adverse effect on body weight (Li et al. 1992).
Sprague-Dawley rats received 100 or 300 ppm benzene vapor for 6 hours/day, 5 days/week for life
(Snyder et al. 1978a, 1984). Decreased weight gain, which continued throughout the study, was observed
at 30 weeks at 300 ppm, but not at 100 ppm. AKR mice exposed to 100 or 300 ppm and C57BL mice
exposed to 300 ppm benzene vapor for 6 hours/day, 5 days/week for life had decreased weight gain at
300 ppm (Snyder et al. 1978a, 1980).
3.2.1.3 Immunological and Lymphoreticular Effects
Immunological effects have been reported in humans with occupational exposure to benzene. There are
two types of acquired immunity, humoral and cellular, and benzene damages both. First, benzene has
been shown to alter humoral immunity (i.e., to produce changes in levels of antibodies in the blood).
Painters who were exposed to benzene (37 ppm), toluene, and xylene in the workplace for 121 years
showed increased serum immunoglobulin values for IgM and decreased values for IgG and IgA (Lange et
al. 1973b). The decreased levels of immunoglobulins may represent suppression of immunoglobulinproducing cells by benzene. Other adverse reactions, characterized by a reaction between leukocytes and
agglutinins, occurred in 10 of 35 of these workers (Lange et al. 1973a). These results suggest the
occurrence of allergic blood dyscrasia in some persons exposed to benzene. However, since the workers
were exposed to multiple solvents, the specific role of benzene is uncertain.
The second type of immunity, cellular immunity, is affected by changes in circulating leukocytes and a
subcategory of leukocytes, called lymphocytes. Leukopenia was found in a series of studies of workers
exposed to benzene at levels ranging from 15 to 210 ppm in various manufacturing processes in Turkey
(Aksoy et al. 1971, 1987). Another study also noted signs of preleukemia that included loss of leukocytes
and other blood elements, bone marrow histopathology, and enlarged spleens (Aksoy et al. 1972, 1974).
Other studies in chronically exposed workers also showed losses of lymphocytes and other blood
elements (Cody et al. 1993; Goldwater 1941; Greenburg et al. 1939; Kipen et al. 1989; Ruiz et al. 1994;
Yin et al. 1987c). In these studies, benzene levels in workplace air ranged from 1 to 1,060 ppm.
Hematological effects reported in these studies are described in Section 3.2.1.2. In one study, routine
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leukocyte counts conducted every 3 months on employees of a small-scale industry in China revealed
leukopenia in workers exposed to as little as 0.69140 ppm (mean=6 ppm) for an average period of 5
6 years (Xia et al. 1995). Leukocyte alkaline phosphatase (LAP) activity was increased in benzene
workers exposed to about 31 ppm for a chronic time period (Songnian et al. 1982). Increased LAP
activity is an indicator of myelofibrosis and is associated with both decreased white blood cell counts and
with changes in bone marrow activity. The change in LAP activity could be used in the diagnosis of
benzene poisoning since it was more sensitive than the change in the leukocyte count, although it is not a
biomarker that is specific for benzene exposure. A study conducted by Li et al. (1994) during 19721987
examined 74,828 benzene-exposed workers employed in 672 factories and 35,805 unexposed workers
from 109 factories located in 12 cities in China. Estimates of gender-specific rate ratios and a comparison
of the rate ratios for females to the rate ratios for males were calculated for the incidence of hematopoietic
and lymphoproliferative (HLP) disorders, comparing all exposed workers in each of the occupational
groups to unexposed workers. Small increases in relative risks for all HLP disorders for both genders
were observed among chemical and rubber manufacturing workers, painters, and paint manufacturers. In
another study, an increase in leukocyte count and alkaline phosphatase score was observed in a pipe-fitter
who was chronically exposed to 0.9 ppm benzene in addition to other solvents (Froom et al. 1994).
Animal studies support the observations made in humans and show that benzene affects humoral and
cellular immunity. A decrease in spleen weight was observed in mice after exposure to benzene at a
concentration of 25 ppm, 6 hours/day for 5 days, the same exposure concentration at which a decrease in
circulating leukocytes was observed (Wells and Nerland 1991). Benzene decreases the formation of the
B-lymphocytes that produce the serum immunoglobulins or antibodies. Exposure to benzene at 10 ppm
and above for 6 days decreased the ability of bone marrow cells to produce mature B-lymphocytes in
C57BL/6 mice (Rozen et al. 1984). The spleen was also inhibited from forming mature T-lymphocytes at
exposure levels of 31 ppm and above. Mitogen-induced blastogenesis of B- and T-lymphocytes was
depressed at 10 ppm and above. Peripheral lymphocyte counts were depressed at all levels, whereas
erythrocyte counts were depressed only at 100 and 300 ppm. This study is the basis for the acute-duration
inhalation MRL of 0.009 ppm (see footnote to Table 3-1 and Appendix A).
A continuation of this line of studies for 6 days to 23 weeks at 300 ppm showed continued decreases in
numbers of mature B- and T-lymphocytes produced in the bone marrow, spleen, and thymus (Rozen and
Snyder 1985). Abnormalities of humoral and cell-mediated immune responses following benzene
exposure are presumably caused by a defect in the lymphoid stem cell precursors of both T- and
B-lymphocytes. Bone marrow cellularity increased 3-fold, and the number of thymic T-cells increased
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15-fold in benzene-exposed mice between the 6th and the 30th exposure. No corresponding increase in
splenic cells was noted. The marked increase in the numbers of cells in bone marrow and thymus was
interpreted by the authors to indicate a compensatory proliferation in these cell lines in response to
benzene exposure, which may play a role in the carcinogenic response of C57BL mice to inhaled
benzene. The lack of response in the spleen suggests a lack of lymphoid restorative capacity in that
organ.
Other studies have also shown similar effects on immune functions following inhalation exposure to
benzene. These include decreased numbers of circulating leukocytes and decreases in bone marrow
cellularity in mice exposed to 100 ppm and higher, 24 hours/day for up to 8 days (Gill et al. 1980);
decreased leukocytes and increased leukocyte alkaline phosphatase in rats exposed to 100 ppm for 7 days
(Li et al. 1986); decreased leukocytes and bone marrow cellularity in DBA/2 mice exposed to 300 ppm
benzene for 2 weeks (Chertkov et al. 1992); decreased leukocytes in mice exposed to 300 ppm for
10 days (Ward et al. 1985); bone marrow hyperplasia, lymphocytopenia, and anemia in mice repeatedly
exposed to 100 ppm or higher for a lifetime (Snyder et al. 1978a, 1980); and leukopenia and lymphopenia
in rats repeatedly exposed to 100 ppm for life (Snyder et al. 1978a, 1984). Aoyama (1986) noted
decreased spleen and thymus weights and decreased levels of B- and T-lymphocytes in blood and spleen
in mice exposed 6 hours/day to approximately 48 ppm of benzene for 14 days. Exposures of mice to
benzene vapor concentrations 100 ppm, 6 hours/day, 5 days/week resulted in significantly reduced
numbers of lymphocytes (ranging from 25 to 43% lower than controls) in the spleen, thymus, and femur
as early as 1 week following the initial exposure, and persisting throughout an 8-week exposure period
(Farris et al. 1997a). In rats, exposure to benzene vapors at a concentration of 400 ppm (but not 200 ppm)
for 6 hours/day, 5 days/week for 4 weeks resulted in reduced thymus weight and decreased splenic
lymphocytes (Robinson et al. 1997).
Reduced bone marrow cellularity was observed in Swiss Webster and C57BL/6J mice after exposure to
300 ppm benzene for 2 weeks (Neun et al. 1992). Decreased granulopoietic stem cells were observed at
21 ppm in NMRI mice after exposures of 10 days to 2 weeks (Toft et al. 1982). Green and colleagues
examined the effect of benzene inhalation on peripheral blood and bone marrow and spleen cells (Green
et al. 1981a, 1981b) of CD-1 mice following exposures to a number of regimens. In the acute studies,
individual test groups were exposed by inhalation to mean benzene concentrations of 0, 1.1, 9.9, 103, 306,
603, 1,276, 2,416, or 4,862 ppm, 6 hours/day for 5 days. Intermediate studies consisted of two different
exposure regimens and concentrations: 6 hours/day, 5 days/week for 10 weeks (50 days) to a mean
concentration of 0 or 9.6 ppm benzene; or 6 hours/day, 5 days/week for 26 weeks to a mean concentration
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of 0 or 302 ppm benzene. In the acute studies, marrow and splenic cellularities were reduced at 103 ppm.
Splenic granulocytes were reduced at all exposure levels except at 9.9 ppm (Green et al. 1981b). The
decrease in spleen cellularity correlated with a reduction in spleen weight at all concentrations 103 ppm.
Mean spleen weights were significantly depressed at 1.1 ppm and at doses above 9.9 ppm, but not at
9.9 ppm. In Green et al. (1981a), marrow concentration of GM-CFU-C was equivalent to or greater than
control values at all levels; however, splenic GM-CFU-C concentration was decreased at 103 ppm.
Femoral and splenic CFU-S and GM-CFU-C per organ were depressed at 103 ppm. Absolute numbers of
GM-CFU-C/femur or spleen were significantly reduced at all higher concentrations. There was no
change in the colony/cluster ratio observed in the 5-day experiment. In the 50-day experiments, increased
spleen weight, splenic nucleated cellularity, splenic nucleated erythrocytes, and CFU-S were seen at
9.6 ppm (Green et al. 1981a, 1981b). Exposure to 302 ppm for 26 weeks resulted in reduced marrow and
spleen cellularity and decreased spleen weight (Green et al. 1981a, 1981b).
Sprague-Dawley SD/Tex rats were exposed to benzene vapor at 0 or 500 ppm for 5 days/week,
6 hours/day for 3 weeks (Dow 1992). In the bone marrow differential counts, rats showed a relative
decrease in lymphoid cells at the 500 ppm dose level. There were also decreases in myeloid cells of
animals exposed to 500 ppm benzene. In a companion study, purebred Duroc-Jersey pigs were exposed
to 0, 20, 100, and 500 ppm benzene vapors 6 hours/day, 5 days/week for 3 weeks (Dow 1992). Exposure
to 500 ppm resulted in significant decreases in total white blood cells, T-cells, peripheral blood
lymphocytes, and proportion of myeloid cells in bone marrow counts.
Another series of experiments revealed that exposures as low as 25 ppm for 2 weeks caused decreases in
the numbers of circulating lymphocytes but did not affect the bone marrow cellularity in C57BL/6 or
CBA/Ca mice (Cronkite 1986; Cronkite et al. 1985). Increasing the duration (up to 16 weeks) or
concentration (up to 300400 ppm) produced the same effect of reduced peripheral blood lymphocytes as
well as decreased bone marrow cellularity that persisted for up to 8 weeks after exposure (Cronkite et al.
1982, 1985). Similar observations were made in two groups of male CBA/Ca mice exposed to a total of
6,000 ppm of benzene by inhalation using two different regimens (Cronkite et al. 1989). One group was
exposed to 316 ppm for a total of 19 times, while the second group was exposed to 3,000 ppm twice.
Although both groups had significantly decreased lymphocyte counts, the lymphocyte numbers were
much more depressed in the group of mice exposed to 316 ppm of benzene over 19 days. This suggests
that, at relatively high exposure levels, repeated or prolonged exposure is more potent than the magnitude
of exposure in causing lymphopenia. In both groups, the lymphocyte numbers had not returned to normal
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values by 214 days post exposure. The femoral bone marrow cellularity returned to normal levels 32 days
after exposure in both groups of mice.
In female BDF1 mice (C57BL/6xDBA/2F1 hybrids) exposed to 0, 300, or 900 ppm benzene 6 hours/day
for 5 days, there was a relative increase in helper lymphocytes (CD4+) at days 35, leading to an increase
of the T4/T8 (CD4+CD8+) ratio from 2 in controls to higher values at 8 weeks (Plappert et al. 1994a).
No concentration dependency was observed. Granulocytic hyperplasia of the spleen was observed in
CBA/Ca mice exposed to 300 ppm benzene for 16 weeks (Farris et al. 1993).
Leukopenia, granulocytopenia, and lymphocytopenia were observed in C57BL/6 mice exposed to
1,000 ppm benzene for 6 hours/day, 5 days/week for up to 6 weeks (Gill et al. 1980). Decreased
leukocyte counts were also noted in Sprague-Dawley rats and CD-1 mice exposed to 300 ppm benzene
6 hours/day, 5 days/week for up to 13 weeks (Ward et al. 1985). In mice, the most common compoundrelated histopathological findings were splenic periarteriolar lymphoid sheath depletion, lymphoid
depletion in the mesenteric lymph node, and plasma cell infiltration of the mandibular lymph node.
Exposure to the highest concentration caused a decrease in leukocyte and lymphocyte counts in SpragueDawley rats. Hematological changes at 300 ppm were accounted for by decreased leukocyte counts in
males on day 14 and in females on day 91. Decreases in percentage of lymphocytes in males and females
started on day 14 and lasted through day 91. Rats exhibited decreased femoral marrow cellularity as the
only histological change. Treatment-related changes were not observed at lower concentrations. Rabbits,
rats, and guinea pigs exposed to 8088 ppm for 32269 days also had decreased leukocyte counts (Wolf
et al. 1956). Leukocyte alkaline phosphatase values were increased and leukocyte counts were decreased
in rats exposed to 4,570 ppm for 20 weeks (Songnian et al. 1982). Exposure of C57BL mice to 10 ppm
benzene for 6 hours/day, 5 days/week, for 24 weeks caused depressions in the numbers of splenic
nucleated red cells and lymphocytes (Baarson et al. 1984).
Li et al. (1992) observed a 26% decrease in spleen weight in male Kunming mice exposed to 12.52 ppm
benzene 2 hours/day, 6 days/week for 30 days. Examination of the bone marrow showed decreases in
myelocytes, premyelocytes, myeloblasts, and metamyeloblasts at the same dose level.
Benzene also affects functional immune responses, as indicated by decreased resistance to infectious
agents. Pre-exposure to benzene at 30 ppm for 512 days increased the bacterial counts in mice on
day 4 of infection with Listeria monocytogenes (Rosenthal and Snyder 1985). Recovery of the immune
system was noted on day 7. The effects did not occur at 10 ppm. In addition, a concentration-dependent
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statistically significant depression was noted in T- and B-lymphocyte populations from day 1 through
day 7 at 30 ppm and above. B-cells were more sensitive to benzene than were T-cells on a percentage-ofcontrol basis. These results indicate a benzene-induced delay in immune response to L. monocytogenes.
Concentrations of 200 or 400 ppm for 45 weeks (5 days/week) suppressed the primary antibody
response to tetanus toxin in mice, but there was no effect at 50 ppm (Stoner et al. 1981). In another
intermediate-duration exposure study, no changes were noted in the numbers of splenic B-cells, T-cells,
or T-cell subsets in C57BL/6 mice exposed to 100 ppm of benzene 5 days/week for 20 days (Rosenthal
and Snyder 1987). However, when splenic T-cells from mice treated with 10 ppm and 100 ppm were
tested in vitro for their capacity to respond to foreign antigens (alloantigens) in the mixed lymphocyte
reaction (MLR), the MLR response was delayed. Further analysis showing that this delayed MLR
response was not due to the presence of benzene-induced suppressor cells and indicated that benzene
impaired the functional abilities of alloreactive T-cells. This study is the basis for the intermediateduration inhalation MRL of 0.006 ppm (see footnote to Table 3-1 and Appendix A). A similar in vitro
observation was made using T-cells from mice exposed to 100 ppm of benzene 5 days/week for 3 weeks.
These cells had a reduced tumor cytolytic activity, suggesting a benzene-induced impairment of cellmediated immunity. The impaired cell-mediated immune function was also apparent in vivo. Mice
exposed to 100 ppm for a total of 100 days were challenged with 10,000 polyoma virus-induced tumor
cells (PYB6), and 9 of 10 mice had reduced tumor resistance and developed tumors that were lethal
(Rosenthal and Snyder 1987).
The highest NOAEL values and all reliable LOAEL values for immunological effects in each species and
3.2.1.4 Neurological Effects
Following acute inhalation of benzene, humans exhibit symptoms indicative of central nervous system
effects (Cronin 1924; Flury 1928; Greenburg 1926; Midzenski et al. 1992). These symptoms, reported to
occur at levels ranging from 300 to 3,000 ppm, include drowsiness, dizziness, headache, vertigo, tremor,
delirium, and loss of consciousness. Acute exposure (510 minutes) to higher concentrations of benzene
(approximately 20,000 ppm) can result in death, which has been associated with vascular congestion in
the brain (Avis and Hutton 1993; Flury et al. 1928). Lethal exposures are also associated with
nonspecific neurological symptoms similar to those reported for nonlethal exposures. These symptoms
are similar to the consequences of exposure to multiple organic solvents and are reversible when
symptomatic workers are removed from the problem area (Kraut et al. 1988; Yin et al. 1987b). In reports
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of cases of benzene poisoning, subjects exhibited headaches, nausea, tremor, convulsions, and
unconsciousness, among other neurological effects (Cronin 1924; Greenburg 1926; Midzenski et al. 1992;
Tauber 1970).
Chronic exposure to benzene has been reported to produce neurological abnormalities in humans. Of
eight patients (six with aplastic anemia and two with preleukemia) with previous occupational exposure to
adhesives and solutions containing 988% benzene, four of the six patients with aplastic anemia showed
neurological abnormalities (global atrophy of lower extremities and distal neuropathy of upper
extremities) (Baslo and Aksoy 1982). Air concentrations in the workplace were reported to have reached
levels of 210 ppm. These findings suggest that benzene may induce toxic effects on the nervous system
involving peripheral nerves and/or spinal cord. The limitations of this study are that benzene exposure
levels were not monitored and that there was a possibility of an additional exposure to toluene (6.37
9.25%).
Chronic exposure to benzene and toluene was studied in 121 workers exposed to benzene for 29 years
(Kahn and Muzyka 1973). The air concentration of benzene between 1962 and 1965 was 615.6 ppm
(2050 mg/m3), while the toluene vapors did not exceed the 5 mg/m3 level. Subsequently (the authors do
not specify when), the air levels of both benzene and toluene have not exceeded the 5 mg/m3 level.
Seventy-four of the examined workers complained of frequent headaches (usually at the end of the work
day), became tired easily, had difficulties sleeping, and complained of memory loss. The limitations of
this study are that workers were exposed to both benzene and toluene and that the precise dose and
duration of exposure are not known.
Tondel et al. (1995) reported the case of a gasoline station attendant who had worked from 1962 to 1979.
The patient described symptoms of fatigue for 3 weeks and night sweats, among other symptoms.
The neurotoxicity of benzene has not been studied extensively in animals. Female Sprague-Dawley rats
exhibited lethargy after exposure to 2,200 ppm benzene, but not 300 ppm, on Gd 615 (Green et al.
1978). Male albino SPF rats from a Wistar-derived strain exposed to benzene for 4 hours in glass
chambers (dose not specified) exhibited depression of evoked electrical activity in the brain; the authors
calculated the 30% effect level (depressed activity) as 929 ppm (Frantik et al. 1994). When female
H strain mice were exposed to benzene for 2 hours, the 30% effect level for depression of evoked
electrical activity in the brain was 856 ppm (Frantik et al. 1994). In rabbits, symptoms that occurred
3.7 minutes following acute exposure to benzene at 45,000 ppm were relaxation and light narcosis
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(Carpenter et al. 1944). As the time after exposure progressed, so did the symptoms to include excitation,
chewing, and tremors (after 5 minutes), loss of pupillary reflex to strong light (after 6.5 minutes), loss of
blinking reflex (after 11.4 minutes), pupillary contraction (after 12 minutes), and involuntary blinking
(after 15.6 minutes). Behavioral tests of C57BL/6 mice showed significant increase in licking of
sweetened milk after 1 week of exposure to 300 ppm; a 90% decrease in hind limb grip strength after one
exposure to 1,000 or 3,000 ppm (data for 100 ppm were not reported); and tremors after one exposure to
3,000 ppm that subsided 30 minutes after the exposure (Dempster et al. 1984). In another study, designed
to reflect occupational exposure, male CD-1 and C57BL/6 mice were exposed to 300 or 900 ppm of
benzene 6 hours/day for 5 days followed by 2 weeks of no exposure after which the exposure regimen
was repeated for an unspecified amount of time (Evans et al. 1981). The following seven categories of
behavioral activities were monitored in exposed and control animals: stereotypic behavior, sleeping,
resting, grooming, eating, locomotion, and fighting. Only minimal and insignificant differences were
observed between the two strains of mice. Increased behavioral activity was observed after exposure to
benzene in both strains of mice. Mice exposed to 300 ppm of benzene had a greater increase than those
exposed to 900 ppm, probably because of narcosis-like effects induced at the higher exposure level
(Evans et al. 1981). It is not known if benzene induces behavioral changes by directly acting upon the
central nervous system. It is also not known whether these changes occur before or after hematological
changes.
Li et al. (1992) exposed male Kunming mice to 0, 0.78, 3.13, or 12.52 ppm benzene for 2 hours/day,
6 days/week for 30 days, and then monitored brain and blood acetylcholinesterase, forelimb grip strength,
locomotor activity, and rapid response. Significantly increased grip strength was observed at 0.78 ppm,
whereas at the higher doses, grip strength decreased significantly. Rapid response showed a significant
increase at the low dose; the two higher doses showed a significant depressed rapid response. Locomotor
activity increased at the low dose, was similar to control values at the middle dose, and decreased at the
high dose. However, these changes were not significantly different from the control values. A significant
decrease in acetylcholinesterase activity was noted in the brain, but it was not large enough to be
considered adverse; no change in acetylcholinesterase levels in the blood was observed. However, the
exposure levels used by Li et al. (1992) were more than 10-fold lower than those eliciting signs of
neurotoxicity in other animal studies, an indication that the actual exposure levels in the study of Li et al.
(1992) may have been higher than indicated. Uncertainty regarding actual exposure levels renders this
study of little value for purposes of risk assessment.
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All reliable LOAEL values for neurological effects in each species and duration category are recorded in
Table 3-1 and plotted in Figure 3-1.
3.2.1.5 Reproductive Effects
Data on the reproductive effects of occupational exposure to benzene suggest that benzene may impair
fertility in women (Mukhametova and Vozovaya 1972; Vara and Kinnunen 1946). However, the findings
are inconclusive due to uncertainties in exposure assessment and the limited data collected. In one study,
30 women with symptoms of benzene toxicity were examined (Vara and Kinnunen 1946). The levels of
benzene in air were not specified, but are assumed to have been much greater than the 1 ppm permitted in
today's working environment. Twelve of these women had menstrual disorders (profuse or scanty blood
flow and dysmenorrhea). Ten of the 12 women provided information on fertility. Of these 10 women,
2 had spontaneous abortions, and no births occurred during their employment, even though no
contraceptive measures had been taken. This led the investigators to suggest that benzene has a
detrimental effect on fertility at high levels of exposure. However, the study failed to provide verification
that the absence of birth was due to infertility. Gynecological examinations revealed that the scanty
menstruations of five of the patients were due to ovarian atrophy. This study is limited in that an
appropriate comparison population was not identified. Additionally, little follow-up was conducted on
the 30 women with regard to their continued work history and possible symptoms of benzene toxicity.
Disturbances of the menstrual cycle were found in women workers exposed to aromatic hydrocarbons
(benzene, toluene, xylene) (Michon 1965). The exposure levels of benzene and toluene were below
0.25 ppm. The observed group consisted of 500 women, 2040 years old. One hundred controls were
included in the study. The results showed that 21% of exposed women whose work involved sitting or
standing had irregular menstrual cycles compared to 12% in the control group. Brief (up to 2 days), long
(69 days), and prolonged (over 9 days) menstrual cycles were present in 26% of women who performed
lifting during their work as compared to 13% in the control group. Irregular amounts of menstrual flow
and pain were also observed in female workers exposed to aromatic hydrocarbons. The major limitations
of this study are that the exposure occurred from a mixture of chemicals, levels of exposure were not well
defined, duration of exposure was not stated, and activities of the controls were not provided.
Another study examined the reproductive function and incidence of gynecological effects in 360 women
exposed to petroleum (a major source of benzene) and chlorinated hydrocarbons both dermally and by
inhalation (Mukhametova and Vozovaya 1972). However, dermal exposure was considered to be
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negligible. The concentrations of benzene in the air were not well documented. When compared to
female workers with no chemical exposure, female gluers had developed functional disturbances of the
menstrual cycle. Additionally, as chemical exposure time increased, there were increases in the number
of premature interruptions of pregnancy, the percentage of cases in which the membranes ruptured late,
and the number of cases of intrauterine asphyxia of the fetus. The study limitations (including lack of
exposure history, simultaneous exposure to other substances, and lack of follow-up) make it difficult to
assess the effects of benzene on reproduction.
Reproductive competence of male workers in two organic chemical factories in France was evaluated by
Stucker et al. (1994). Analysis of 1,739 pregnancies that ended in spontaneous abortion or birth was
presented. Paternal exposure to benzene for each pregnancy was described as exposure in the 3 months
immediately before conception, and as previous job exposures. Benzene exposure was graded at two
levels: <5 ppm (low) and 5 ppm (moderate). Of the 1,739 pregnancies described, 171 ended in a
spontaneous abortion (rate=9.8%). According to exposure categories, 1,277 pregnancies were defined as
non-exposed (mate not exposed) and 270 whose mates were exposed at some time before conception. For
the 270 pregnant women, 145 of their mates were exposed during the 3 months immediately preceding
conception. The frequency of spontaneous abortion was not significantly higher for the paternal group
exposed at any time before conception than in the non-exposed group; nor was it higher for the group
exposed during the 3 months immediately before conception.
In CFY rats exposed to either pure air or benzene (125 ppm) for 24 hours/day from Gd 7 through 14, no
effect on implantation number was observed (Tatrai et al. 1980a). No changes in maternal body weight
were observed in Sprague-Dawley rats exposed to 100 ppm benzene for 6 hours/day on Gd 615 (Coate
et al. 1984). Pregnant rabbits exposed 12 hours/day to 156.5 or 313 ppm benzene on Gd 720 showed an
increase in the number of abortions and resorptions at 312 ppm (Ungvary and Tatrai 1985). However, in
other developmental toxicity studies, no effect on the number of resorptions was seen in rats at doses as
high as 2,200 ppm (Green et al. 1978), in mice at 500 ppm (Murray et al. 1979), or in rabbits at doses of
500 ppm (Murray et al. 1979).
Reproductive effects have been noted in experimental animals exposed for intermediate durations, but the
levels of exposure were higher (806,600 ppm) than those to which humans are exposed in the modern
industrial environment (Ward et al. 1985; Wolf et al. 1956). In an intermediate-duration inhalation study,
groups of male and female CD-1 mice were exposed to benzene vapor concentrations of 0, 1, 10, 30, or
300 ppm, 5 days/week, 6 hours/day to benzene vapor for 13 weeks (Ward et al. 1985). Histopathological
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changes were observed in ovaries (bilateral cysts) and testes (atrophy/degeneration, decrease in
spermatozoa, moderate increase in abnormal sperm forms) of mice exposed to 300 ppm benzene; the
severity of gonadal lesions was greater in the males. An inhalation study was conducted exposing rats
(6,600 ppm), rabbits (80 ppm), and guinea pigs (88 ppm) to benzene for 78 hours/day, 5 days/week for
93, 243, and 32 or 269 days, respectively (Wolf et al. 1956). Male rats showed an increase in testicular
weight after 93 days at the 6,600 ppm level. The guinea pigs showed a slight increase in average
testicular weight at the 88 ppm level. Rabbits showed slight histopathological testicular changes
(degeneration of the germinal epithelium) when exposed to 80 ppm. Since only one or two rabbits were
used in this study, it was not possible to draw any conclusions regarding benzene's ability to induce
testicular damage in the rabbits. Continuous exposure of female rats to 210 ppm benzene for 1015 days
before cohabitation with males and 3 weeks after cohabitation resulted in a complete absence of litters
(Gofmekler 1968). It is not known whether this was due to failure to mate, infertility, or early
preimplantation losses of fertilized ova. In a fertility study, female rats exposed up to 300 ppm benzene
for 10 weeks during premating, mating, gestation, and lactation showed no effect on indices of fertility,
reproduction, and lactation (Kuna et al. 1992).
The highest NOAEL values and all reliable LOAEL values for reproductive effects in each species and
3.2.1.6 Developmental Effects
The available human data on the developmental effects of benzene after inhalation exposure are
inconclusive. The studies designed specifically to investigate developmental effects are limited, primarily
because of concomitant exposure to other chemicals, inadequate sample size, and lack of quantification of
exposure levels (Budnick et al. 1984; Goldman et al. 1985; Heath 1983; Olsen 1983). Benzene crosses
the human placenta and is present in the cord blood in amounts equal to or greater than those in maternal
blood (Dowty et al. 1976). In a study of subjects with known benzene poisoning in Italy, Forni et al.
(1971a) reported the case of one pregnant worker exposed to benzene in the air during her entire
pregnancy. Although she had severe pancytopenia and increased chromosomal aberrations, she delivered
a healthy son with no evidence of chromosomal alterations. The following year, she delivered a healthy
daughter. However, increased frequency of chromatid and isochromatid breaks and sister chromatid
exchange was found in lymphocytes from 14 children of female workers exposed by inhalation to
benzene (dose not specified) and other organic solvents during pregnancy (Funes-Cravioto et al. 1977).
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No mention was made of whether the mothers showed signs of toxicity or whether physical abnormalities
occurred among their offspring.
There are numerous inhalation studies in which animals have been exposed to benzene during pregnancy
(Coate et al. 1984; Green et al. 1978; Kuna and Kapp 1981; Murray et al. 1979; Tatrai et al. 1980a,
1980b; Ungvary and Tatrai 1985). None of these studies demonstrated that benzene was teratogenic even
at levels that induced maternal and fetal toxicity. Fetotoxicity was evidenced by decreased body weight
and by increased skeletal variants such as missing sternebrae and extra ribs, which were not considered to
be malformations. Alterations in hematopoiesis have also been observed in the fetuses and offspring of
pregnant mice exposed to low levels of benzene (Keller and Snyder 1986, 1988). These studies are
discussed below.
Mice exposed to 500 ppm benzene for 7 hours/day on days 615 of pregnancy exhibited fetal growth
retardation (i.e., decreased fetal body weight) and increased minor skeletal variants (i.e., delayed
ossification) (Murray et al. 1979). There were no fetal malformations and no significant effect on the
incidence of pregnancy, average number of live fetuses, resorptions per litter, or maternal weight gain.
No malformations in fetuses and no significant effects on incidence of pregnancy, average number of live
fetuses, or resorptions per litter were observed when rabbits were exposed to benzene 7 hours/day at
500 ppm during gestation for 9 days, although an increase in minor skeletal variations was observed
(Murray et al. 1979).
Pregnant mice exposed 12 hours/day to 156.5 or 313 ppm benzene on Gd 615 had pups with significant
weight retardation and retardation of skeletal development, but no malformations (Ungvary and Tatrai
1985). A parallel study in rabbits showed that inhalation of benzene at 313 ppm caused fetal weight
reduction, and an increase in minor fetal anomalies (Ungvary and Tatrai 1985).
As was the case with mice and rabbits, the fetotoxicity of benzene in rats is also demonstrated by retarded
fetal weight and/or minor skeletal variants (Coate et al. 1984; Green et al. 1978; Kuna and Kapp 1981;
Tatrai et al. 1980b). In an experiment conducted by Green et al. (1978), pregnant Sprague-Dawley rats
were exposed to 100, 300, or 2,200 ppm benzene for 6 hours/day on Gd 615. Exposure to high levels of
benzene (2,200 ppm) during gestation resulted in a significant decrease in fetal weight, whereas dams
breathing air containing lower levels of benzene (100 or 300 ppm) during gestation bore young that were
similar in weight and crownrump length to control pups. Statistically significant numbers of fetuses
with delayed ossification were found in groups exposed to concentrations of 300 and 2,200 ppm. The
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litter incidence of missing sternebrae was significantly increased in the 100 and 2,200 ppm exposure
groups. Maternal toxicity, as indicated by a decrease in maternal weight gain, was evident only at the
2,200 ppm level. The female offspring appeared to be affected to a greater extent than males with respect
to delayed ossification and missing sternebrae.
Kuna and Kapp (1981) found decreased fetal weight after exposure to 50 ppm. From a group of 151 pups
examined after in utero exposure to 500 ppm benzene, a single pup exhibited exencephaly. In the same
study, of 98 pups examined for skeletal effects after in utero exposures of 500 ppm, a single pup had
angulated ribs and 2 others had nonsequential ossification of the forefeet. These anomalies were not
statistically significant and may have resulted from maternal nutritional stress.
No significant skeletal malformations occurred in pups of rats exposed during gestation to 47 ppm for
8 days, 24 hours/day (Tatrai et al. 1980b) or 100 ppm for 10 days, 6 hours/day (Coate et al. 1984).
Decreased fetal weights were seen at 47 ppm (Tatrai et al. 1980b) and 100 ppm (Coate et al. 1984), and
increased fetal mortality was observed at 141 ppm (Tatrai et al. 1980b). In CFY rats exposed to pure air
or 125 ppm benzene on Gd 714, there was a 17% decrease in placental weight, a decrease in mean fetal
weight, and evidence of skeletal retardation (Tatrai et al. 1980a). Continuous exposure of female rats to
6 concentrations of benzene ranging from 0.3 to 210 ppm for 1015 days before cohabitation with males
and 3 weeks after did not affect newborn weight or induce malformations, but there were differences in
the weights of individual organs of the dams at all exposure levels (Gofmekler 1968). There was a slight
tendency toward decreased litter sizes at 20 ppm of benzene. A complete absence of litters resulted from
exposure to 210 ppm. It is not known whether this was due to failure to mate, infertility, or early
preimplantation losses of fertilized ova.
Alterations in hematopoiesis have also been observed in the fetuses and offspring of pregnant mice
exposed to benzene (Keller and Snyder 1986). Administration of 20 ppm benzene to pregnant Swiss
Webster mice for 6 hours/day on Gd 615 caused reductions in the levels of the CFU-E of the fetuses,
whereas 5 and 10 ppm benzene caused enhancement of these colony-forming cells. In 2-day-old
neonates, CFU-E numbers in the 5 ppm group returned to control values, but the 10 ppm neonates showed
a bimodal response by litter. Granulocytic colony-forming cells were enhanced in neonates exposed in
utero to 20 ppm benzene. Some of the mice exposed to 10 ppm prenatally were re-exposed to 10 ppm as
adults. Their hematopoietic progenitor cell numbers were depressed compared with controls exposed for
the first time as adults. No tests were conducted on the dams after benzene exposure.
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In a follow-up study, pregnant Swiss Webster mice were exposed 6 hours/day on Gd 615 to 5, 10, or
20 ppm benzene (Keller and Snyder 1988). The results indicated that 16-day fetuses, when checked for
erythrocyte and leukocyte counts, hemoglobin analysis, and the proliferating pool of differentiating
hematopoietic cells, had no noteworthy change at any of the exposure levels. In contrast, 2-day neonates
exposed in utero to all concentrations of benzene exhibited a reduced number of circulating erythroid
precursor cells and, at 20 ppm, had increased numbers of hepatic hematopoietic blast cells and
granulopoietic precursor cells accompanied by decreased numbers of erythropoietic precursor cells.
Six-week-old adult mice exposed in utero to 20 ppm of benzene had a similar pattern of enhanced
granulopoiesis. However, this effect was not clearly evident in 6-week-old adult mice exposed in utero to
5 or 10 ppm.
The results of inhalation studies conducted in experimental animals have been fairly consistent across
species. It has been suggested that benzene fetotoxicity in animals is a function of maternal toxicity
because the joint occurrence of a decrease in fetal weight and an increase in skeletal variants usually
occurs when there is a decrease in maternal weight (Tatrai et al. 1980b). However, the mechanism
underlying developmental toxicity has not been fully elucidated, and there are few data on the effect of
benzene on maternal food consumption and on blood levels of benzene and its metabolites in the dams
and their fetuses. There are apparently none of the usual fetotoxic findings after exposure in utero to low
concentrations of benzene (10 ppm) (Coate et al. 1984; Kuna and Kapp 1981). As stated above, there is
evidence for persistent hematopoietic anomalies in animals exposed in utero to benzene at 20 ppm (Keller
and Snyder 1988). They may also exist at lower concentrations, but adequate testing has not been
performed.
The highest NOAEL value and all reliable LOAEL values for developmental effects in each species
following acute exposure are recorded in Table 3-1 and plotted in Figure 3-1.
3.2.1.7 Cancer
Epidemiological studies and case reports provide clear evidence of a causal relationship between
occupational exposure to benzene and benzene-containing solvents and the occurrence of acute
nonlymphocytic leukemia (ANLL), particularly the myeloid cell type (acute myelogenous leukemia,
AML) (EPA 1985b, 1998; Hayes et al. 1997; IARC 1982, 1987; IRIS 2007; Rinsky et al. 1987, 2002; Yin
et al. 1996a, 1996b). Some of the studies also provide suggestive evidence of associations between
benzene exposure and non-Hodgkins lymphoma (NHL) and multiple myeloma (Hayes et al. 1997;
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Rinsky et al. 1987). The epidemiological studies are generally limited by confounding chemical
exposures and methodological problems, including inadequate or lack of exposure monitoring and low
statistical power (due to small numbers of cases), but a consistent excess risk of leukemia across studies
indicates that benzene is the causal factor. Many of the earlier studies are additionally limited by a lack of
information on leukemia cell types other than AML, because leukemia used to be considered a single
diagnostic category for epidemiological purposes, due in part to historical nomenclature, small numbers
of deaths by cell type, and unavailability of cell-type-specific rates for comparison.
Two series of studies on workers exposed to benzene in Ohio (the Pliofilm study) (e.g., Rinsky et al.
1981, 1987, 2002) and China (the NCI/CAPM study) (e.g., Hayes et al. 1997; Yin et al. 1996a, 1996b)
have yielded particularly strong data on the leukemogenic potential of benzene and are summarized
below. The studies of the Pliofilm cohort provide the best set of data for evaluating human cancer risks
from benzene exposure because, in comparison to other published studies, the Pliofilm workers had the
fewest reported co-exposures to other potentially carcinogenic substances and experienced a greater range
of estimated exposures to benzene (EPA 1998). The NCI/CAPM Chinese study is one of the largest of its
type ever undertaken and evaluated many thousands of benzene-exposed workers, enabling detection of
significantly elevated risks at unusually low levels of exposure.
A number of studies were performed on a cohort of workers exposed to benzene in three rubber
hydrochloride (Pliofilm) manufacturing plants in Ohio (Collins et al. 1997; Finkelstein 2000; Infante
1978; Infante et al. 1977; Ireland et al. 1997; Paxton et al. 1994a, 1994b; Rinsky et al. 1981, 1987, 2002;
Schnatter et al. 1996a; Wong 1995). Analyses using various expansions and updates of the cohort,
statistical methods, and exposure estimates have consistently shown a significant relationship between
exposure to benzene and excess mortality from leukemia. In the report by Rinsky et al. (1987), a cohort
of 1,165 white males employed between 1940 and 1965 and followed through 1981 experienced
increased mortality from all leukemias (9 observed versus 2.7 expected; standardized mortality ratio
[SMR]=3.37, 95% CI 1.546.41) and multiple myeloma (4 observed versus 1 expected; SMR=4.09, 95%
CI 1.1010.47). Death rates for age-matched U.S. white males during the same calendar period were
used for comparison with death rates observed in the cohort. Assessment after an additional 15 years of
follow-up showed declines in the SMRs for both leukemias (2.56, 95% CI 1.434.22) and multiple
myeloma (2.12, 95% CI 0.694.96), suggesting that the excess risks diminished with time since exposure
(Rinsky et al. 2002). Exposures in the most recent 10 years were most strongly associated with leukemia
risk, and there was no significant relation between leukemia death and benzene exposures received more
than 20 years previously (Finkelstein 2000). AML accounted for most of the increased leukemia
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(SMR=5.03, 95% CI 1.8410.97) (Schnatter et al. 1996a; Wong 1995). The risk of all leukemias and
AML, but not multiple myeloma, significantly increased with increasing cumulative exposure above
200 ppm-years (Rinsky et al. 1987, 2002; Wong 1995). Analysis of 4,417 workers from one of the
Pliofilm plants showed no clear evidence that the risks of all leukemias, ANLL, multiple myeloma, or
other lymphohematopoietic cancers increased with increasing exposure at lower levels of cumulative
exposure (172 ppm-years), and the number of peak exposures over 100 ppm for 40 days was a better
predictor of risk for all leukemias and multiple myeloma (Collins et al. 2003; Ireland et al. 1997). No
clear associations between exposure to benzene and mortality from NHL or Hodgkins disease were
reported (Collins et al. 2003; Ireland et al. 1997; Rinsky et al. 2002).
A collaborative study between the National Cancer Institute and the Chinese Academy of Preventive
Medicine (NCI/CAPM) evaluated incidence rates (occurrence of disease and cause of death) for
lymphohematopoietic malignancies and other hematologic disorders in a cohort of 74,828 benzeneexposed and 35,805 nonexposed workers employed in 672 factories in 12 cities in China (Hayes et al.
1996, 1997, 2001; Linet et al. 1996). The joint NCI/CAPM study is an expansion of earlier studies
performed by CAPM alone (Yin et al. 1987a, 1987b, 1987c, 1989). The workers were employed from
19721987, followed for an average of nearly 12 years, and worked in various job categories using
benzene as a solvent for paints, varnishes, glues, coatings, and other products. The derivation of the
cohort from many different factories and job types suggests that the members were concurrently exposed
to many other chemicals. Findings in the exposed workers included significantly increased relative risk
(RR) for all hematologic neoplasms (RR=2.6, 95% CI 1.44.7), all leukemias (RR=2.5, 95% CI 1.25.1),
ANLL (RR=3.0, 95% CI 1.08.9), and combined ANLL and precursor myelodysplastic syndromes
(ANLL/MDS; RR=4.1, 95% CI 1.411.6) (Hayes et al. 1997). Increases were also observed for other
(unspecified) leukemias (RR=2.0, 95% CI=0.75.4), but the results were not statistically significant.
Analysis by level of average benzene exposure (<10, 1024, and 25 ppm) and cumulative exposure
(<40, 4099, and 100 ppm-years) indicated that the risk for all hematologic neoplasms was significantly
increased at <10 ppm (RR=2.2, 95% CI 1.14.2) and <40 ppm-years (RR=2.2, 95% CI 1.14.5),
respectively. The risks for all leukemias, ANLL, and ANLL/MDS were increased at exposure levels of
1024 ppm (average) and 4099 ppm-years (cumulative). The elevated risks showed weak tendencies to
increase with increasing average and cumulative levels of exposure. Analysis by duration of exposure
(<5, 59, or 10 years) did not show increased risk with increasing exposure duration. Analysis by
occupational group (coatings, rubber, chemical, shoe, other/mixed) showed that the increased risks for
ANLL and ANLL/MDS were consistent across the spectrum of industries studied, implying that the
associations were due to the common exposure to benzene rather than other industry-specific exposures.
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The results of a study of shoe factory workers in Italy are similar to those of the Pliofilm and NCI/CAPM
studies in showing that the risk of leukemia increases with increasing exposure to benzene (Costantini et
al. 2003; Paci et al. 1989). The cohort was followed from 1950 to 1999 and consisted of 891 men and
796 women who were exposed to estimated benzene concentrations ranging from 0 to 92 ppm, and had
mean cumulative exposures of 71.8 and 43.4 ppm-years, respectively (exposure durations not reported).
Leukemia risk was significantly increased in both sexes in the highest of four exposure categories and
most apparent in the men. For cumulative exposures of <40, 4099, 100199, and >200 ppm-years, the
leukemia SMR values for the men were 1.4 (95% CI 0.25.0), 3.7 (95% CI 0.120.6), 3.0 (95% CI 0.4
10.9), and 7.0 (95% CI 1.918.0), respectively. Leukemia subtypes were not evaluated. The findings are
consistent with earlier epidemiologic studies and case reports showing increased incidences of leukemia
in shoe factory and rotogravure plant workers exposed to high benzene levels during its use as a solvent
(Aksoy et al. 1974, 1987; IARC 1982; Vigliani and Forni 1976).
No significant increases in leukemia were found in chemical industry workers (Bloemen et al. 2004; Bond
et al. 1986; Ott et al. 1978) or petroleum industry workers (Lewis et al. 1997; Raabe and Wong 1996;
Rushton and Romaniuk 1997; Schnatter et al. 1993, 1996b, 1996c; Tsai et al. 1983) exposed to lower
levels of benzene. Cause-specific mortality was determined in a prospective study of 2,266 chemical
workers who were exposed to benzene in various Dow Chemical Company manufacturing processes
between 1938 and 1970 (Bloemen et al. 2004). The workers were followed from 19401996 and had an
average duration of exposure, intensity of exposure, and cumulative exposure of 4.8 years, 9.6 ppm, and
39.7 ppm-years, respectively. There were no significant increases in risk for any lymphohematopoietic
malignancies, including all leukemias, ANLL, chronic lymphatic leukemia (CLL), NHL, and multiple
myeloma. A previous investigation of this cohort found a significantly increased risk of myelogenous
leukemias (SMR=4.44, 95% CI not reported) (Bond et al. 1986; Ott et al. 1978), but this result is less
reliable than those of the current study because it is based on a much smaller number of deaths.
A meta-analysis was conducted on 19 cohorts of petroleum workers in the United States and the United
Kingdom that were pooled into a single database for cell-type-specific leukemia analysis (Raabe and
Wong 1996). The combined cohort consisted of 208,741 workers, mainly refinery employees who
contributed more than 4.6 million person-years of observation. Benzene exposures were mainly from
handling gasoline and the estimated mean and cumulative exposures for the most exposed jobs were
<1 ppm and <45 ppm-years, respectively. No increased risks were found for mortality from AML,
chronic myeloid leukemia (CML), acute lymphocytic leukemia (ALL), or CLL. Analyses limited to
BENZENE
101
3. HEALTH EFFECTS
studies of refinery workers or studies with at least 15 years of follow-up yielded similar results. The
negative results of this meta-analysis are consistent with those of case-control studies of the common
leukemia cell types in petroleum workers (Rushton and Romaniuk 1997; Schnatter et al. 1993, 1996b,
1996c). A case-control study of hairy cell leukemia, a rare B-lymphoid chronic leukemia, found no
association between reported exposure to benzene and the risk of this cancer in workers from various
occupations with generally low levels of exposure (Clavel et al. 1996).
A possible association between occupational exposure to benzene and NHL is suggested by results of the
most recent analysis (Hayes et al. 1997) of the cohort from the NCI/CAPM Chinese study summarized
above. The relative risk for mortality from NHL in the whole cohort was 3.0 (95% CI 0.910.5), an
increase that was not statistically significant. However, the risk for NHL did significantly increase at the
highest level and duration of benzene exposure. For exposure to average concentrations of <10, 1024,
and 25 ppm, the RR values were 2.7 (95% CI 0.710.6), 1.7 (95% CI 0.310.2), and 4.7 (95% CI 1.2
18.1), respectively (P for trend=0.04). For cumulative exposures of <40, 4099, and 100 ppm-years, the
RR values were 3.3 (95% CI 0.813.1), 1.1 (95% CI 0.111.1), and 3.5 (95% CI 0.913.2), respectively
(P for trend=0.02). Additionally, the risk for NHL significantly increased with increasing duration of
exposure among workers exposed for <5 years (RR=0.7, 95% CI 0.17.2), 59 years (RR=3.3, 95% CI
0.714.7), and >10 years (RR=4.2, 95% CI 1.115.9) (P for trend=0.01).
Although the results of the NCI/CAPM Chinese study (Hayes et al. 1997) indicated a possible association
between exposure to benzene and NHL, other cohort mortality studies found no significant increases in
NHL mortality. These studies include the entire Pliofilm cohort (Rinsky et al. 2002); a total of
4,417 workers from one of the three Pliofilm plants assessed for cumulative exposures of 0, <1, 16, and
>6 ppm-years (Collins et al. 2003); a cohort of 2,266 chemical manufacturing workers exposed to a
cumulative benzene level of 39.7 ppm-years (Bloemen et al. 2004); and a meta-analysis of 26 cohorts of
petroleum workers from the United States and five other countries with expected low exposures to
benzene (Wong and Raabe 2000). Furthermore, case-control studies provide no indications of an
association between benzene exposure and risk of NHL (Schnatter et al. 1996b; Wong and Raabe 2000),
and the adequacy of the data from the NCI/CAPM Chinese study (Hayes et al. 1997) has been questioned
(Dosemeci et al. 1994; Wong 1999; Wong and Raabe 2000).
Other evidence suggests a possible association between occupational exposure to benzene and increased
risk of multiple myeloma. Mortality from multiple myeloma was initially elevated in the Pliofilm cohort,
but appeared to diminish with increasing duration of observation. Based on an evaluation of 1,165 white
BENZENE
102
3. HEALTH EFFECTS
male workers followed through 1981, Rinsky et al. (1987) found an SMR for multiple myeloma of
4.09 (95% CI 1.1010.47), an increase that was statistically significant. Assessment of the cohort after an
additional 6 and 15 years of follow-up yielded lower SMRs of 2.91 (95% CI 0.797.45) (Wong 1995) and
2.12 (95% CI 0.694.96) (Rinsky et al. 2002), respectively, risks that were no longer statistically
significant. The risk of multiple myeloma did not increase with increasing level of cumulative exposure
to benzene or with duration of employment, but this finding is particularly limited by the small number of
deaths. No increases in mortality from multiple myeloma were found in other cohort mortality studies,
including the NCI/CAPM study of Chinese workers (no observed deaths) (Yin et al. 1996a, 1996b), the
Bloemen et al. (2004) study of chemical workers (SMR=0.72, 95% CI 0.152.10), and the Paci et al.
(1989) study of shoe factory workers (no observed deaths in males; SMR=1.11 in females, 95% CI not
calculated) summarized above. Additionally, mortality from multiple myeloma was not increased in a
meta-analysis of 22 other cohorts of petroleum workers with potential exposure to benzene or benzenecontaining petroleum products (Wong and Raabe 1997). The combined cohort consisted of
250,816 workers (from the United States, Canada, the United Kingdom, and Australia) who were
observed over a period of 55 years from 1937 to 1991. The overall SMR for multiple myeloma was
0.93 (95% CI=0.811.07), and analyses by type of facility/industrial process (refinery, distribution, and
crude oil production and pipeline workers) and duration of observation showed no differences in risk.
The individual cohorts used in the meta-analysis also had no increased risk of multiple myeloma.
As summarized above, one of the early assessments of the Pliofilm cohort found an increased risk of
mortality from multiple myeloma (Rinsky et al. 1987). The implications of this finding are unclear
because the risk declined to non-significant levels in subsequent follow-ups (Rinsky et al. 2002; Wong
1995), and was not supported by the findings of other cohort mortality studies (Bloemen et al. 2004; Paci
et al. 1989; Wong and Raabe 2000; Yin et al. 1996a, 1996b). Additionally, population-based and
hospital-based case-control studies indicate that benzene exposure is not likely to be causally related to
the risk of mulitple myeloma (Bezabeh et al. 1996; Heineman et al. 1992; Linet et al. 1987; Schnatter et
al. 1996b; Sonoda et al. 2001; Wong and Raabe 1997). A meta-analysis of case-control studies found no
significant association between occupational exposure to benzene and benzene-containing products and
risk of multiple myeloma from sources categorized as benzene and/or organic solvents (summary odds
ratio [OR]=0.74, 95% CI 0.600.90), petroleum (summary OR=1.11, 95% CI 0.961.28), or petroleum
products (summary OR=1.08, 95% CI 0.891.33) (Sonoda et al. 2001).
BENZENE
103
3. HEALTH EFFECTS
Studies in animals provide supporting evidence for the carcinogenicity of inhaled benzene. As
summarized below, inhalation exposure to benzene induced tumors at multiple sites in rats and mice, with
a tendency towards induction of lymphomas in mice.
Carcinomas of the Zymbal gland and oral cavity were clearly increased in Sprague-Dawley rats that were
exposed to 200300 ppm benzene for 47 hours/day, 5 days/week for up to 104 weeks (Maltoni et
al. 1982a, 1982b, 1983, 1985, 1989). The Zymbal gland is a specialized sebaceous gland and a site for
benzene-induced tumors. Marginal increases in nasal cavity carcinomas, mammary tumors, and
hepatomas were also observed. The significance of these findings is unclear because statistical analyses
were not performed and there were no clearly increased incidences of Zymbal gland carcinoma or tumors
at other sites in Sprague-Dawley rats that were similarly exposed to 100 or 300 ppm benzene for
6 hours/day, 5 days/week for life (Snyder et al. 1978b, 1984).
Benzene was carcinogenic in mice exposed to 100 or 300 ppm benzene for 6 hours/day, 5 days/week for
16 weeks and observed for 18 months or life (Cronkite 1986; Cronkite et al. 1984, 1985, 1989; Farris et
al. 1993). These exposures consistently induced a variety of tumors, including leukemia (mainly thymic
lymphoma) and Zymbal gland and ovarian tumors in C57BL/6 mice, and myelogenous leukemias,
malignant lymphomas, and Zymbal gland and lung tumors in CBA/Ca mice. Incidences of lymphocytic
lymphoma with thymic involvement were significantly increased in C57BL/6 mice similarly exposed to
300 ppm benzene for life (Snyder et al. 1980). Intermittent lifetime exposure to 300 ppm benzene
(6 hours/day, 5 days/week on every third week) was more tumorigenic (in the Zymbal gland and lungs) to
CD-1 and C57BL/6 mice than short-term exposure to 1,200 ppm (6 hours/day, 5 days/week for 10 weeks)
followed by lifetime observation, although neither of these exposures induced leukemia (Snyder et al.
1988).
The cancer effect levels (CELs) for each species and duration category are recorded in Table 3-1 and
plotted in Figure 3-1.
carcinogen. The Department of Health and Human Services (NTP 2005) determined that benzene is a
known carcinogen based on human evidence showing a causal relationship between exposure to benzene
and cancer. IARC (1987, 2004, 2007) classified benzene in Group 1 (carcinogenic to humans) based on
sufficient evidence in both humans and animals. EPA (IRIS 2007) classified benzene in Category A
(known human carcinogen) based on convincing evidence in humans supported by evidence from animal
BENZENE
104
3. HEALTH EFFECTS
studies. Under EPAs most recent guidelines for carcinogen risk assessment, benzene is characterized as
a known human carcinogen for all routes of exposure based on convincing human evidence as well as
supporting evidence from animal studies (IRIS 2007). Based on the Rinsky et al. (1981, 1987) human
leukemia data, EPA derived a range of inhalation unit risk values of 2.2x10-67.8x10-6 (g/m3)-1 for
benzene (IRIS 2007). For risks ranging from 1x10-4 to 1x10-7, the corresponding air concentrations for
lifetime exposure range from 13.045.0 g/m3 (414 ppb) to 0.0130.045 g/m3 (0.0040.014 ppb),
respectively. These risk levels are presented in Figure 3-1.
3.2.2
Oral Exposure
3.2.2.1 Death
Individual case reports of death from acute oral exposure to benzene have appeared in the literature since
the early 1900s. The benzene concentrations encountered by the victims were often not known.
However, lethal oral doses for humans have been estimated at 10 mL (8.8 g or 125 mg/kg for a 70-kg
person) (Thienes and Haley 1972). Lethality in humans has been attributed to respiratory arrest, central
nervous system depression, or cardiac collapse (Greenburg 1926). Accidental ingestion and/or attempted
suicide with lethal oral doses of benzene have produced the following signs and symptoms: staggering
gait; vomiting; shallow and rapid pulse; somnolence; and loss of consciousness, followed by delirium,
pneumonitis, collapse, and then central nervous system depression, coma, and death (Thienes and Haley
1972). Ingestion of lethal doses may also result in visual disturbances and/or feelings of excitement and
euphoria, which may quite suddenly change to weariness, fatigue, sleepiness, convulsion, coma, and
death (Von Oettingen 1940).
Animal lethality data indicate that benzene is of low toxicity following acute oral exposure (O'Bryan and
Ross 1988). Oral LD50 values for rats ranged from 930 to 5,600 mg/kg; the values varied with age and
strain of the animals (Cornish and Ryan 1965; Wolf et al. 1956). Male Sprague-Dawley rats were given
various doses of benzene to determine the LD50 (Cornish and Ryan 1965). The LD50 for nonfasted rats
was found to be 930 mg/kg. In 24-hour fasted rats, the LD50 was 810 mg/kg. No increase in mortality
was reported in Fischer 344 rats or B6C3F1 mice treated with 600 mg/kg/day for up to 17 weeks (Huff et
al. 1989; NTP 1986).
Sprague-Dawley rats (3035 males, 3035 females) were exposed to benzene by ingestion (stomach
tube), in olive oil, at 0, 50, or 250 mg/kg/day for 45 days weekly for 52 weeks and then kept under
supervision until spontaneous death (Maltoni et al. 1983). Exposure to 50 mg/kg/day benzene after
BENZENE
105
3. HEALTH EFFECTS
52 weeks resulted in deaths in 9 of 30 male (same as controls) and 2 of 30 female rats. At 250 mg/kg/day
exposure, 13 of 35 males and 9 of 35 females died. For rats receiving only olive oil, 9 of 30 males and
0 of 30 females died. In a companion study, Sprague-Dawley rats were exposed to 500 mg/kg/day
benzene by ingestion (stomach tube), in olive oil, 45 days/week for 92 weeks, and then kept under
observation until spontaneous death (Maltoni et al. 1983). Mortality rates were the same as the controls.
In a chronic-duration oral study conducted by the NTP (1986), increased mortality was observed in male
Fischer 344 rats exposed to 200 mg/kg/day benzene in corn oil, and in female Fischer 344 rats exposed to
50 mg/kg/day benzene. B6C3F1 mice given 100 mg/kg/day also had increased mortality compared to
control mice.
The LD50 values and all reliable LOAEL values for death in each species following acute and chronic
exposure are recorded in Table 3-2 and plotted in Figure 3-2.
No studies were located regarding respiratory, cardiovascular, musculoskeletal, hepatic, renal, endocrine,
ocular, metabolic, or body weight effects in humans. Human and animal data pertaining to other systemic
effects are presented below.
The highest NOAEL value and all reliable LOAEL values for systemic effects in each species and
Respiratory Effects.
Male and female Fischer 344 rats and B6C3F1 mice were given oral doses of 0,
25, 50, 100, 200, 400, or 600 mg/kg/day benzene in corn oil for 120 days (NTP 1986). No histopathological lesions were observed in lungs, trachea, or mainstream bronchi. After chronic-duration exposure
to 50, 100, or 200 mg/kg/day (male rats) or 25, 50, or 100 mg/kg/day (female rats, male and female mice),
no histopathological lesions were observed in trachea, lungs, or mainstream bronchi in rats (NTP 1986).
In mice, a significantly increased incidence of alveolar hyperplasia was observed at 50 and
100 mg/kg/day in females and at 100 mg/kg/day in males.
Cardiovascular Effects.
No histopathological lesions were observed in cardiac tissue from male and
female Fischer 344 rats or B6C3F1 mice given oral doses of 0, 25, 50, 100, 200, 400, or 600 mg/kg/day
benzene in corn oil for 120 days (NTP 1986). After chronic-duration exposure to 200 mg/kg/day (male
Table 3-2 Levels of Significant Exposure to Benzene - Oral
BENZENE
a
Key to Species
Figure (Strain)
Exposure/
Duration/
Frequency
(Route)
LOAEL
NOAEL
System
Less Serious
(mg/kg/day)
(mg/kg/day)
Serious
Reference
(mg/kg/day)
Chemical Form
Comments
ACUTE EXPOSURE
Death
1
Human
once
126
(death)
Thienes and Haley 1972
126
338
Rat
(SpragueDawley)
once
(G)
930 M (LD50)
Cornish and Ryan 1965
930
121
Rat
(Wistar)
once
(GO)
5600 M (LD50)
Wolf et al. 1956
5600
162
Gd 6-15
daily
(G)
Renal
Exxon 1986
1000 F
1000
709
Dermal
50 F (alopecia of hindlimbs
and trunk)
50
Bd Wt
500 F
3. HEALTH EFFECTS
Systemic
Rat
4
(SpragueDawley)
1000 F (body weight decreased

11%)
500
1000
Other
50 F
250 F (decreased food

consumption)
50
250
Rat
(CF)
1-3 d
Hepatic
Pawar and Mungikar 1975
1402 M (increased liver weight,

biochemical changes)
1402
138
Neurological
Human
6
once
126
(muscular incoordination, Thienes and Haley 1972

unconsciousness)
126
350
106
Rat
(SpragueDawley)
BENZENE
a
Key to Species
Figure (Strain)
Exposure/
Duration/
Frequency
(Route)
(continued)
LOAEL
NOAEL
System
Less Serious
(mg/kg/day)
1d
(mg/kg/day)
Reference
Chemical Form
1870 M (tremors)
88 M (slight CNS depression)

88
Serious
(mg/kg/day)
Comments
Cornish and Ryan 1965
1870
122
Rat
(SpragueDawley)
once
(G)
Kanada et al. 1994
950 M (altered neurotransmitter

concentrations)
950
752
Gd 6-15
daily
(G)
1000 F
Exxon 1986
1000 F
Exxon 1986
1000
229
Developmental
Rat
10
(SpragueDawley)
Gd 6-15
daily
(G)
1000
230
11
Mouse
(ICR/SIM)
Gd 8-12
(GO)
3. HEALTH EFFECTS
Reproductive
Rat
9
(SpragueDawley)
Seidenberg et al. 1986
1300 F (decreased pup weight

on neonatal days 1-3)
1300
266
Death
12
Rat
(SpragueDawley)
52 wk
4-5 d/wk
1 x/d
(GO)
250 F (9/35 died)
250
234
107
Systemic
Rat
13
(SpragueDawley)
52 wk
4-5 d/wk
1 x/d
(GO)
BENZENE
a
Key to Species
Figure (Strain)
Exposure/
Duration/
Frequency
(Route)
(continued)
LOAEL
NOAEL
System
Less Serious
(mg/kg/day)
Bd Wt
(mg/kg/day)
250
50
50
(body weight decreased

19%)
Serious
Reference
(mg/kg/day)
Chemical Form
Comments
250
660
14
Rat
(F-344/N)
<1 yr
5 d/wk
(GO)
Hemato
50 M (lymphocytopenia and
leukocytopenia)
50
NTP 1986
25 F (lymphocytopenia and
leukocytopenia)
Bd Wt
100
200 M (body weight decreased

11% or more in 25
weeks)
100
200
3. HEALTH EFFECTS
25
450
108
15
Rat
(F-344/N)
60-120 d
5 d/wk
(GO)
BENZENE
a
Key to Species
Figure (Strain)
Exposure/
Duration/
Frequency
(Route)
(continued)
LOAEL
NOAEL
System
Less Serious
(mg/kg/day)
Resp
(mg/kg/day)
Serious
Reference
(mg/kg/day)
Chemical Form
Comments
NTP 1986
600
600
702
Cardio
600
600
Gastro
600
600
Hemato
200
200
(dose-related leukopenia
at 60 days)
(dose-related leukopenia
at 120 days)
200
(body weight decreased

14% in males and 16% in
females)
3. HEALTH EFFECTS
25
25
Musc/skel
600
600
Hepatic
600
600
Renal
600
600
Endocr
600
600
Bd Wt
200
16
Rat
6 wk
(Fischer- 344) 5 d/wk
(GO)
Hepatic
Taningher et al. 1995
400 M
400
736
Bd Wt
400 M
400
17
Rat
(Wistar)
6 mo
5 d/wk
(GO)
Hemato
50 F (leukopenia,
erythrocytopenia)
1F
1
Wolf et al. 1956
50
163
109
18
Mouse
(C57BL/6)
28 d
(W)
BENZENE
a
Key to Species
Figure (Strain)
Exposure/
Duration/
Frequency
(Route)
(continued)
LOAEL
NOAEL
System
Less Serious
(mg/kg/day)
Bd Wt
(mg/kg/day)
Serious
Reference
(mg/kg/day)
Chemical Form
Comments
Fan 1992
1000 M
1000
714
19
Mouse
(CD-1)
4 wk
ad lib
(W)
Hemato
8 M (erythrocytopenia,
increased mean
corpuscular volume;
leukopenia)
Hsieh et al. 1988b
31.5 M (decreased erythrocyte,

leukocyte counts)
Hsieh et al. 1990
166
Mouse
(CD-1)
4 wk
(W)
Hemato
31.5
431
Hepatic
31.5 M
31.5
Renal
31.5 M
31.5
Bd Wt
31.5 M
3. HEALTH EFFECTS
20
31.5
21
Mouse
(B6C3F1)
<1 yr
5 d/wk
(GO)
Hemato
25
(lymphocytopenia)
NTP 1986
25
451
Bd Wt
100
100
110
22
Mouse
(B6C3F1)
60-120 d
5 d/wk
(GO)
BENZENE
a
Key to Species
Figure (Strain)
Exposure/
Duration/
Frequency
(Route)
(continued)
LOAEL
NOAEL
System
Less Serious
(mg/kg/day)
Resp
(mg/kg/day)
Serious
Reference
(mg/kg/day)
Chemical Form
Comments
NTP 1986
600
600
707
Cardio
600
600
Gastro
600
600
Hemato
200 F
200
50 M (dose-related leukopenia)
50
25
25
600
3. HEALTH EFFECTS
Musc/skel
400 F (dose-related leukopenia)

400
600
Hepatic
600
600
Renal
600
600
Endocr
600
600
Bd Wt
600
600
23
Mouse
(B6C3F1)
30 d
ad lib
(W)
Hemato
195 F (decreased leukocytes)
12 F
12
350 F (decreased hemoglobin,

hematocrit, leukocytes,
MCV, and MCH)
195
Shell 1992
350
231
Hepatic
350 F
350
Renal
350 F
350
Bd Wt
350 F
350
Other
12 F (decreased fluid intake)

12
111
BENZENE
a
Key to Species
Figure (Strain)
Exposure/
Duration/
Frequency
(Route)
(continued)
LOAEL
NOAEL
System
Less Serious
(mg/kg/day)
Immuno/ Lymphoret
Rat
60-120 d
24
5 d/wk
(F-344/N)
(GO)
(mg/kg/day)
200
Serious
Reference
(mg/kg/day)
Chemical Form
Comments
Huff et al. 1989; NTP 1986
(lymphopenia at 60 days,
lymphoid depletion in
B-cell of the spleen)
200
25 F dose-related
lymphopenia at 120
days)
25
703
25
6 mo
5 d/wk
(GO)
1F
1
50 F (leukopenia)
Wolf et al. 1956
27 M (decreased number of
splenocytes and IL-2
production)
Fan 1992
50
161
26
Mouse
(C57BL/6)
28 d
(W)
27
3. HEALTH EFFECTS
Rat
(Wistar)
502
27
Mouse
(CD-1)
4 wk
ad lib
(W)
8 M (leukopenia and
lymphopenia; enhanced
splenic lymphocyte
proliferation)
Hsieh et al. 1988b
340
28
Mouse
(CD-1)
4 wk
(W)
Hsieh et al. 1990

31.5 M (reduction in thymus
mass; suppresion of both
B- and T-cell
mitogeneses;
suppressed IL-2
secretions; leukopenia,
lymphopenia)
31.5
432
112
29
Mouse
(CD-1)
BENZENE
a
Key to Species
Figure (Strain)
Exposure/
Duration/
Frequency
(Route)
(continued)
LOAEL
NOAEL
System
Less Serious
(mg/kg/day)
(mg/kg/day)
4 wk
ad lib
(W)
Serious
Reference
(mg/kg/day)
Chemical Form
40 M (elevated corticosterone
levels; T-lymphocyte
suppression)
Comments
Hsieh et al. 1991
40
898
30
Mouse
(B6C3F1)
60-120 d
5 d/wk
(GO)
25 M
50 M (dose-related
lymphopenia)
25
200 F
200
50
400 F
400
31
Mouse
(B6C3F1)
30 d
ad lib
(W)
12 F (decreased leukocytes)
Shell 1992
12
232
Neurological
Rat
32
(F-344/N)
60-120 d
5 d/wk
(GO)
NTP 1986
600
600
3. HEALTH EFFECTS
708
704
33
Mouse
(CD-1)
4 wk
ad lib
(W)
8 M (fluctuation of
neurotransmitter levels)
Hsieh et al. 1988a
165
34
Mouse
(CD-1)
4 wk
(W)
31.5 M (decreased NE, DA,

5-HT)
Hsieh et al. 1990
31.5
886
35
Mouse
(CD-1)
4 wk
ad lib
(W)
40 M (increased hypothalmic
NE and VMA)
8M
8
Hsieh et al. 1991
40
113
443
36
Mouse
(B6C3F1)
60-120 d
5 d/wk
(GO)
BENZENE
a
Key to Species
Figure (Strain)
Exposure/
Duration/
Frequency
(Route)
(continued)
LOAEL
NOAEL
System
Less Serious
(mg/kg/day)
400
200
200
(mg/kg/day)
Serious
Reference
(mg/kg/day)
Chemical Form
Comments
NTP 1986
(intermittent tremors)
400
709
37
Mouse
(B6C3F1)
30 d
ad lib
(W)
Shell 1992
350 F (decreased brain weight)
195 F
195
350
233
Reproductive
Rat
38
(F344/N)
600
NTP 1986
600
NTP 1986
600
705
39
Mouse
(B6C3F1)
17 wk
5 d/wk
(GO)
600
710
Cancer
40
Rat
(SpragueDawley)
52 wk
4-5 d/wk
1 x/d
(GO)
50 F (CEL: Zymbal gland

carcinoma in 2/30; oral
cavity carcinoma)
50
Maltoni et al. 1989
3. HEALTH EFFECTS
17 wk
5 d/wk
(GO)
50
237
41
Rat
(SpragueDawley)
52 wk
4-5 d/wk
1 x/d
(GO)
(CEL: Zymbal gland

carcinoma)
50
250
(CEL: nasal cavity

carcinoma, fore-stomach
liver angiosarcoma;
Zymbal gland carcinoma)
250
146
114
42
Mouse
(RF/J)
BENZENE
a
Key to Species
Figure (Strain)
Exposure/
Duration/
Frequency
(Route)
(continued)
LOAEL
NOAEL
System
Less Serious
(mg/kg/day)
(mg/kg/day)
52 wk
4-5 d/wk
(GO)
Serious
Reference
(mg/kg/day)
Chemical Form
500
(CEL: mammary
pulmonary leukemias)
Comments
Maltoni et al. 1989
500
141
CHRONIC EXPOSURE
Death
43
Rat
(F-344/N)
2 yr
5 d/wk
(GO)
200 M (30/50 died)

200
NTP 1986
50 F (14/50 died)
50
711
Mouse
(B6C3F1)
2 yr
5 d/wk
(GO)
100
(41/50 males died, 35/50 NTP 1986

females died)
100
717
Systemic
Human
45
6.1 yr (avg)
(Occup)
Hemato
0.29
0.29
(reduced WBC and

platelet counts,
approximately 7-18%
lower than control
values)
Lan et al. 2004a
Route-to-route
extrapolation from the
reported LOAEL of
0.57 ppm for
occupational exposure
was used by ATSDR to
estimate equivalent
oral dose.
3. HEALTH EFFECTS
44
1035
115
46
Rat
(F-344/N)
2 yr
5 d/wk
(GO)
BENZENE
a
Key to Species
Figure (Strain)
Exposure/
Duration/
Frequency
(Route)
(continued)
LOAEL
NOAEL
System
Less Serious
(mg/kg/day)
Resp
(mg/kg/day)
Reference
Chemical Form
Comments
200 M
200
Serious
(mg/kg/day)
100 F
100
712
Cardio
200 M
200
100 F
100
Gastro
100
200
Hemato
50 M (lymphocytopenia and
leukocytopenia)
50
25 F (lymphocytopenia and
leukocytopenia)
3. HEALTH EFFECTS
200 M (hyperkeratosis and

acanthosis in
nonglandular
forestomach)
100
25
Musc/skel
200 M
200
100 F
100
Hepatic
200 M
200
100 F
100
Renal
200 M
200
100 F
100
Endocr
200 M
200
100 F
100
Dermal
200 M
200
Ocular
200 M
200
100 F
100
116
100 F
100
47
Rat
(F-344/N)
2 yr
5 d/wk
(GO)
BENZENE
a
Key to Species
Figure (Strain)
Exposure/
Duration/
Frequency
(Route)
(continued)
LOAEL
NOAEL
System
Less Serious
(mg/kg/day)
Bd Wt
(mg/kg/day)
Serious
Reference
(mg/kg/day)
Chemical Form
Comments
200 M (body weights decreased Huff et al. 1989; NTP 1986

23% in 103 weeks)
100 M
100
200
712
48
92 wk
4-5 d/wk
1 x/d
(GO)
Hemato
500
500
236
Bd Wt
500
500
(decreased body weights

in 92 weeks)
(decreased RBCs and

WBCs after 84 weeks)
3. HEALTH EFFECTS
Rat
(SpragueDawley)
117
49
Mouse
(B6C3F1)
2 yr
5 d/wk
(GO)
BENZENE
a
Key to Species
Figure (Strain)
Exposure/
Duration/
Frequency
(Route)
(continued)
LOAEL
NOAEL
System
Less Serious
(mg/kg/day)
Resp
100 M (alveolar hyperplasia)
50 M
50
100
25 F
25
(mg/kg/day)
Serious
Reference
(mg/kg/day)
Chemical Form
Comments
NTP 1986
50 F
50
718
Cardio
100
100
Gastro
25
(epithelial hyperplasia
and hyperkeratosis of
forestomach)
25
(lymphocytopenia;
increased frequency of
micronucleated
normochromatic
peripheral erythrocytes)
25
(hyperplasia of adrenal
gland and harderian
gland)
25
25
Musc/skel
100
100
Hepatic
100
3. HEALTH EFFECTS
Hemato
100
Renal
100
100
Endocr
25
Dermal
100
100
Bd Wt
50
100
50
(mean body weight

decreased 10% in 47
weeks to 19% in 103
weeks in males;
decreased 14-15% in
week 99-103 in females)
100
118
BENZENE
a
Key to Species
Figure (Strain)
Exposure/
Duration/
Frequency
(Route)
(continued)
LOAEL
NOAEL
System
Less Serious
(mg/kg/day)
Immuno/ Lymphoret
Rat
2 yr
50
5 d/wk
(F-344/N)
(GO)
(mg/kg/day)
50 M (lymphoid depletion of
spleen and thymus)
50
Serious
Reference
(mg/kg/day)
Chemical Form
Comments
25 F (lymphoid depletion of
spleen and thymus)
25
713
51
92 wk
4-5 d/wk
1 x/d
(GO)
500
(decreased WBCs after

84 weeks)
25
(lymphopenia,
hematopoietic
hyperplasia in the bone
marrow, splenic
hematopoiesis)
500
905
52
Mouse
(B6C3F1)
2 yr
5 d/wk
(GO)
3. HEALTH EFFECTS
Rat
(SpragueDawley)
25
719
Neurological
Rat
53
(F-344/N)
2 yr
5 d/wk
(GO)
200 M
200
NTP 1986
100 F
100
714
54
Mouse
(B6C3F1)
2 yr
5 d/wk
(GO)
100
NTP 1986
100
720
119
Reproductive
Rat
55
(F-344/N)
2 yr
5 d/wk
(GO)
BENZENE
a
Key to Species
Figure (Strain)
Exposure/
Duration/
Frequency
(Route)
(continued)
LOAEL
NOAEL
System
Less Serious
(mg/kg/day)
(mg/kg/day)
Reference
Chemical Form
100 F (endometrial polyps)
200 M
200
Serious
(mg/kg/day)
Comments
NTP 1986
100
50 F
50
715
56
Mouse
(B6C3F1)
2 yr
5 d/wk
(GO)
25
NTP 1986
(preputial gland
hyperplasia in males;
ovarian hyperplasia and
senile atrophy in
females)
57
Rat
(F-344/N)
2 yr
5 d/wk
(GO)
50 M (CEL: squamous cell

papillomas and
carcinomas of the oral
cavity)
50
3. HEALTH EFFECTS
25
721
Cancer
25 F (CEL: Zymbal gland

carcinomas)
25
955
58
Rat
(SpragueDawley)
92 wk
4-5 d/wk
1 x/d
(GO)
500
(CEL: Zymbal gland

carcinoma; oral and
nasal cavity carcinoma;
angiosarcoma of the
liver)
500
238
120
59
Rat
(Wistar)
BENZENE
a
Key to Species
Figure (Strain)
Exposure/
Duration/
Frequency
(Route)
(continued)
LOAEL
NOAEL
System
(mg/kg/day)
Less Serious
(mg/kg/day)
104 wk
4-5 d/wk
1 x/d
(GO)
Serious
Reference
(mg/kg/day)
Chemical Form
500
(CEL: Zymbal gland and

oral and nasal cavity
carcinoma;
angiosarcoma of the
liver)
Comments
Maltoni et al. 1989
500
140
60
Rat
(SpragueDawley)
104 wk
5 d/wk
(GO)
50 M (CEL: Zymbal gland

carcinoma)
Maltoni et al. 1989
25
(CEL: harderian gland

adenoma, lymphoma in
males; CEL: lymphoma
in females)
500
(CEL: mammary tumors,

lung tumors, Zymbal
gland)
Maltoni et al. 1989
50
61
Mouse
(B6C3F1)
2 yr
5 d/wk
(GO)
25
956
62
Mouse
(Swiss)
78 wk
4-5 d/wk
1 x/d
(GO)
3. HEALTH EFFECTS
167
500
143
a The number corresponds to entries in Figure 3-2.

b Differences in levels of health effects and cancer effects between male and females are not indicated in Figure 3-2. Where such differences exist, only the levels of effect for the
most sensitive gender are presented.
c Study results used to derive a chronic-duration oral minimal risk level (MRL) of 0.0005 mg/kg/day based on route-to-route extrapolation, as described in detail in Chapter 2 and
Appendix A. Benchmark dose (BMD) analysis was performed on B-lymphocyte counts to select a point of departure, which was adjusted for intermittent exposure. An equivalent oral
dose was estimated based on route-to-route extrapolation to determine a point of departure for deriving a chronic-duration oral MRL for benzene, which was divided by an uncertainty
factor of 30 (10 for human variability and 3 for uncertainty in route-to-route extrapolation).
121
ad lib = ad libitum; Bd Wt = body weight; CEL = cancer effect level; CNS = central nervous system; d = day(s); DA = dopamine; F = female; (F) = feed; (G) = gavage; Gd = gestational
day; (GO) = gavage in oil; Hemato = hematological; 5-HT = 5-hydroxystryptamine; LD50 = lethal dose, 50% kill; LOAEL = lowest-observed-adverse-effect level; M = male; MCH =
mean corpuscular hemoglobin; MCHC = mean corpuscular hemoglobin concentration; MCV = mean corpuscular volume; mo = month(s); NE = norepinephrine; NOAEL =
no-observed-adverse-effect level; NS = not specified; RBC = red blood cells; VMA = vanillin mandelic acid; (W) = water; WBC = white blood cells; wk = week(s); yr = year(s); x =
times
Figure 3-2 Levels of Significant Exposure to Benzene - Oral

BENZENE
Acute (14 days)

Systemic
mg/kg/day
ath
De
tic
pa
He
na
Re
al
rm
De
ht
dy
Bo
eig
al
ur
he
Ot
Ne
tiv
ic
log
pr
Re
c
du
tal
en
m
lop
ve
De
10000
3r
7r
5r
11m
4r
2r
4r
8r
9r
3. HEALTH EFFECTS
1000
10r
4r
4r
100
7r
4r
4r
10
-Humans
k-Monkey
m-Mouse
h-Rabbit
a-Sheep
f-Ferret
n-Mink
j-Pigeon
o-Other
e-Gerbil
s-Hamster
g-Guinea Pig

NOAEL - Animals

NOAEL - Humans
LD50/LC50
Minimal Risk Level
for effects
other than
Cancer
122
c-Cat
d-Dog
r-Rat
p-Pig
q-Cow
Figure 3-2 Levels of Significant Exposure to Benzene - Oral (Continued)

BENZENE

Systemic
mg/kg/day
ath
c
as
a
pir
De
Re
ula
y
tor
v
dio
i
tro
Ca
al
al
tin
Ga
s
nte
e
sk
ato
lo
cu
He
al
let
ic
log
Mu
c
ati
He
Re
ht
eig
ne
al
ri
oc
En
yW
Bo
10000
1000
18m
22m
15r
22m
15r
22m
15r
22m
22m
22m
15r
22m
23m
16r
23m
15r
23m
100
22m
15r
22m
23m
13r
21m
22m
20m
21m
22m
17r
14r
15r
13r
20m
20m
20m
3. HEALTH EFFECTS
23m
12r
15r
23m
10
19m
17r
0.1
-Humans
k-Monkey
m-Mouse
h-Rabbit
a-Sheep
f-Ferret
n-Mink
j-Pigeon
o-Other
e-Gerbil
s-Hamster
g-Guinea Pig

NOAEL - Animals

NOAEL - Humans
LD50/LC50
Minimal Risk Level
for effects
other than
Cancer
123
c-Cat
d-Dog
r-Rat
p-Pig
q-Cow
Systemic
mg/kg/day
e
yW
d
Bo
ho
t
igh
p
ym
he
Ot
g
olo
ve
cti
u
Ne
mm
ica
/L
o
un
BENZENE
u
od
r*
ce
p
Re
n
Ca
10000
1000
32r
14r
100
15r
24r
36m
37m
36m
37m
39m
38r
42m
3. HEALTH EFFECTS
16r
41r
14r
26m
23m
10
30m
29m
28m
30m
25r
24r
40r
41r
31m
27m
34m
35m
33m
35m
25r
0.1
-Humans
k-Monkey
m-Mouse
h-Rabbit
a-Sheep
f-Ferret
n-Mink
j-Pigeon
o-Other
e-Gerbil
s-Hamster
g-Guinea Pig

NOAEL - Animals

NOAEL - Humans
LD50/LC50
Minimal Risk Level
for effects
other than
Cancer
124
c-Cat
d-Dog
r-Rat
p-Pig
q-Cow
BENZENE
Chronic (365 days)
Systemic
mg/kg/day
ath
De
ry
ato
ir
sp
Re
al
va
io
rd
Ca
la
cu
te
oin
tr
as
al
n
sti
a
gic
lo
to
ma
He
sk
ulo
c
us
t
ele
ti
pa
l
na
He
Re
rin
c
do
En
ma
r
De
lar
u
Oc
yW
d
Bo
ho
t
igh
/L
no
u
mm
p
ym
1000
48r
100
44m
43r
46r
49m
49m
49m
46r
46r
46r
49m
48r
49m
49m
46r
49m
46r
46r
49m
46r
46r
49m
46r
46r
49m
47r
49m
49m
47r
52m
10
3. HEALTH EFFECTS
1
45
0.1
0.01
0.001
0.0001
1E-5
1E-6
-Humans
k-Monkey
m-Mouse
h-Rabbit
a-Sheep
f-Ferret
n-Mink
j-Pigeon
o-Other
e-Gerbil
s-Hamster
g-Guinea Pig

NOAEL - Animals

NOAEL - Humans
LD50/LC50
Minimal Risk Level
for effects
other than
Cancer
125
c-Cat
d-Dog
r-Rat
p-Pig
q-Cow
ho
mg/kg/day
p
ym
mm
ica
/L
o
un
BENZENE
Chronic (365 days)
g
olo
u
Ne
ve
cti
u
od
p
Re
r*
ce
n
Ca
1000
51r
100
54m
50r
53r
62m
58r
57r
61m
57r
59r
55r
56m
60r
10
3. HEALTH EFFECTS
0.1
0.01
10-4
0.001
Estimated
Upper-Bound
10-5
0.0001
Human Cancer
Risk Levels
10-6
1E-5
1E-6
-Humans
k-Monkey
m-Mouse
h-Rabbit
a-Sheep
f-Ferret
n-Mink
j-Pigeon
o-Other
e-Gerbil
s-Hamster
g-Guinea Pig

NOAEL - Animals

NOAEL - Humans
LD50/LC50
Minimal Risk Level
for effects
other than
Cancer
126
c-Cat
d-Dog
r-Rat
p-Pig
q-Cow
10-7
BENZENE
127
3. HEALTH EFFECTS
rats) or 100 mg/kg/day (female rats, male and female mice) no histopathological lesions were observed
in the heart (NTP 1986).
Gastrointestinal Effects.
A man swallowed an unspecified amount of benzene and survived, but
developed an intense toxic gastritis and later pyloric stenosis (Greenburg 1926).
No histopathological lesions were observed in esophageal and stomach tissue or in the small intestine and
colon from male and female Fischer 344 rats or B6C3F1 mice given oral doses of 0, 25, 50, 100, 200, 400,
or 600 mg/kg/day benzene in corn oil for 120 days (NTP 1986). After chronic-duration exposure to 50
200 mg/kg/day (male rats) or 25100 mg/kg/day (female rats, male and female mice), male rats exhibited
hyperkeratosis and acanthosis in the nonglandular forestomach at 200 mg/kg/day, and mice exhibited
epithelial hyperplasia and hyperkeratosis in the forestomach at 25 mg/kg/day (NTP 1986).
Prior to 1913, benzene was used as a treatment for leukemia. Benzene was
given in gelatin capsules starting with 43 mg/kg/day and increasing to 71 mg/kg/day for unspecified
durations (Selling 1916). Leukemia patients showed a great reduction in leukocyte count and multiple
hemorrhages with advanced anemia. However, it is difficult to determine which effects were due to the
leukemia and which were due to the benzene treatment.
Intermediate-duration studies in animals have revealed decreases in numbers of erythrocytes and
leukocytes following exposure to benzene. Male and female Fischer 344 rats and B6C3F1 mice were
given oral doses of 0, 25, 50, 100, 200, 400, and 600 mg/kg/day benzene in corn oil for 120 days. Doserelated leukopenia and lymphopenia were observed at 200 and 600 mg/kg/day for both male and female
rats killed on day 60, and at all doses in female rats killed on day 120. Dose-related leukopenia and
lymphopenia were observed in male mice at 50 mg/kg/day and female mice at 400 mg/kg/day for
120 days, but not for 60 days. Mice exposed to 8 mg/kg/day in the drinking water for 4 weeks had
decreased numbers of erythrocytes, increased mean corpuscular volumes, and decreased numbers of
lymphocytes (Hsieh et al. 1988b, 1990). Female B6C3F1 mice were exposed to 0, 12, 195, or
350 mg/kg/day benzene in drinking water for 30 days (Shell 1992). Decreased hemoglobin, hematocrit,
leukocytes, MCV, and MCH were observed at 350 mg/kg/day. Decreased leukocytes also occurred at
195 mg/kg/day. Decreased leukocytes also occurred at 195 mg/kg/day. Rats gavaged with 50 mg/kg/day
of benzene 5 days/week for 6 months also had decreased numbers of erythrocytes and leukocytes (Wolf et
al. 1956). One chronic-duration study showed that gavage doses of 25 mg/kg/day resulted in leukopenia
and/or lymphocytopenia in both rats and mice, both at the interim sacrifices at 318 months, and at the
BENZENE
128
3. HEALTH EFFECTS
end of 2 years (Huff et al. 1989; NTP 1986). Increased frequency of micronucleated normochromatic
peripheral erythrocytes was observed in mice at 25 mg/kg/day after 2 years. Sprague-Dawley rats were
exposed to 500 mg/kg benzene by ingestion (stomach tube), in olive oil, once daily, 45 days/week for
92 weeks, and then kept under observation until spontaneous death (Maltoni et al. 1983). Decreased
erythrocytes and leukocytes were observed after 84 weeks.
Musculoskeletal Effects.
No histopathological lesions were observed in femoral tissue from male
and female Fischer 344 rats or B6C3F1 mice given oral doses of 0, 25, 50, 100, 200, 400, or
600 mg/kg/day benzene in corn oil for 120 days, or in the sternebrae, femur, or vertebrae from
Fischer 344 rats and mice exposed to 50200 mg/kg/day (male rats) or 25100 mg/kg/day (female rats,
male and female mice) for 2 years (NTP 1986).
Hepatic Effects.
Acute oral administration of 1,402 mg/kg/day benzene for 3 days induced hepatic
changes in rats evidenced by increased liver weight, decreased protein in the postmitochondrial
supernatant fractions (9,000 times specific gravity), and changes in hepatic drug metabolism and lipid
peroxidation (Pawar and Mungikar 1975). The initiation-promotion-progression (IPP) model for the
induction of malignant neoplasms in the liver was evaluated for benzene in male and female SpragueDawley rats (Dragan et al. 1993). Initiation was begun in 5-day-old rats with administration of a single
intraperitoneal injection of diethylnitrosamine during the time when the liver is undergoing rapid growth.
Promotion began at 6 months of age with phenobarbital in the feed, and continued into young adulthood.
Partial hepatectomy was performed, and at the height of the regenerative proliferation phase following the
hepatectomy, benzene (1 g/kg) was administered by gavage; phenobarbital treatment was maintained after
the administration of benzene. A slight increase in the incidence of altered hepatic foci was observed
after initiation and promotion. Few hepatic foci were observed in the livers of male Fischer 344 rats
treated by gavage with 400 mg/kg/day benzene in corn oil for 5 days/week for 6 weeks (Taningher et al.
1995). No histopathological non-neoplastic lesions were observed in hepatic tissue from male and female
Fischer 344 rats given oral doses of 0, 25, 50, 100, 200, 400, or 600 mg/kg/day benzene in corn oil for
120 days or in male rats exposed to 50200 mg/kg/day and female rats exposed to 25100 mg/kg/day for
2 years (NTP 1986).
Female B6C3F1 mice were exposed to 0, 12, 195, or 350 mg/kg/day benzene in drinking water for
30 days (Shell 1992). No adverse liver effects, as evidenced by gross necropsy, liver weight
determination, and serum levels of hepatic enzymes, were observed. Oral administration of
31.5 mg/kg/day benzene continuously in drinking water for 4 weeks did not affect liver weight in CD-1
BENZENE
129
3. HEALTH EFFECTS
mice (Hsieh et al. 1990). No histopathological non-neoplastic lesions effects were observed in hepatic
tissue from male and female B6C3F1 mice given oral doses of 0, 25, 50, 100, 200, 400, or 600 mg/kg/day
benzene in corn oil for 120 days, or in male and female mice exposed to 25100 mg/kg/day for 2 years
(NTP 1986).
Renal Effects.
Female Sprague-Dawley rats were dosed by gavage with 0, 50, 250, 500, or
1,000 mg/kg/day benzene on Gd 615 and killed on Gd 20 (Exxon 1986). No adverse effects were noted
in the kidneys based on gross necropsy. No adverse effects based on histological examination were
observed on renal tissue or the urinary bladder from male and female Fischer 344 rats given oral doses of
0, 25, 50, 100, 200, 400, or 600 mg/kg/day benzene in corn oil for 120 days or in male rats exposed to
50200 mg/kg/day and female rats exposed to 25100 mg/kg/day for 2 years (NTP 1986). Female
B6C3F1 mice were exposed to 0, 12, 195, or 350 mg/kg benzene in drinking water for 30 days (Shell
1992). No adverse effects were observed in the kidneys, based on kidney weights, gross examination, and
blood urea nitrogen and creatinine determinations. Oral administration of 31.5 mg/kg/day benzene
continuously in drinking water for 4 weeks did not affect kidney weight in CD-1 mice (Hsieh et al. 1990).
No adverse effect based on histological examination was observed on renal tissue or the urinary bladder
from male and female B6C3F1 mice given oral doses of 0, 25, 50, 100, 200, 400, or 600 mg/kg/day
benzene in corn oil for 120 days, or in male and female mice exposed to 25100 mg/kg/day for 2 years
(NTP 1986).
Endocrine Effects.
No histopathological lesions were observed in salivary, thyroid, parathyroid,
pancreas, adrenal, or pituitary glands from male and female Fischer 344 rats or B6C3F1 mice given oral
doses of 0, 25, 50, 100, 200, 400, or 600 mg/kg/day benzene in corn oil for 120 days (NTP 1986). In the
companion chronic-duration oral study, male Fischer 344 rats were exposed to 25, 50, 100, or
200 mg/kg/day benzene, while female rats received 25, 50, or 100 mg/kg/day benzene. Hyperplasia of
the Zymbal gland was increased in low-dose males and in mid-dose females. In the adrenal gland,
hyperplasia was observed in both sexes (males: 27 and 4% at 50 and 200 mg/kg, respectively; females:
34% at 25 mg/kg). In the thyroid gland, incidences of C-cell hyperplasia were 14, 26, 15, and 15% in
males treated with 0, 50, 100, and 200 mg/kg, respectively. Analysis of the pituitary gland showed
incidence of hyperplasia in males treated with 0, 50, 100, and 200 mg/kg at 6, 16, 20, and 10%,
respectively; in females treated with 0, 25, 50, and 100 mg/kg at 11, 20, 10, and 14%, respectively. None
of the increased incidences of hyperplasia in these glands were considered to be treatment-related by
NTP. The non-dose-related increase of hyperplasia of the Zymbal gland could represent a progression to
the neoplasms (see Section 3.2.2.7). In mice, Zymbal gland lesions showed epithelial hyperplasia in
BENZENE
130
3. HEALTH EFFECTS
males (0, 9, 30, and 26%) and in females (2, 3, 5, and 19%) exposed to 0, 25, 50, or 100 mg/kg,
respectively. Hyperplasia of the adrenal cortex occurred at incidences of 4, 67, 29, and 9% in males and
10, 43, 68, and 13% in females, respectively. Hyperplasia of the harderian gland occurred at incidences
of 0, 11, 22, and 15% in males and 13, 23, 22, and 21% in females, respectively (NTP 1986).
Dermal Effects.
A case of accidental poisoning in which the patient survived but developed an odd
skin condition consisting of swelling and edema has been reported (Greenburg 1926).
Female Sprague-Dawley rats were dosed by gavage with 0, 50, 250, 500, or 1,000 mg/kg/day benzene
daily on Gd 615 and killed on Gd 20 (Exxon 1986). Alopecia of the hind limbs and trunk was noted in
all dose groups.
No histopathological lesions were observed in the skin of male and female Fischer 344 rats and B6C3F1
mice after chronic oral exposure to 50200 mg/kg/day (male rats) or 25100 mg/kg/day (female rats and
male and female mice) (NTP 1986).
Ocular Effects.
No histopathological lesions were noted in the eyes of male and female Fischer 344
rats and B6C3F1 mice after chronic-duration oral exposure to 50200 mg/kg/day (male rats) or 25
100 mg/kg/day (female rats and male and female mice) (NTP 1986).
Body Weight Effects.
No significant change in body weight was observed in male Fischer 344 rats
treated by gavage with 400 mg/kg/day benzene in corn oil for 5 days/week for 6 weeks (Taningher et al.
1995). Body weight was unaffected in male and female Fischer 344 rats given oral doses of 0, 25, 50, or
100 mg/kg/day benzene in corn oil for 120 days (NTP 1986). However, animals receiving 200, 400, or
600 mg/kg/day benzene exhibited a 1422% decrease in body weight after 120 days. Female SpragueDawley rats were dosed by gavage with 0, 50, 250, 500, or 1,000 mg/kg benzene daily on Gd 615 and
killed on Gd 20 (Exxon 1986). Maternal body weight decreased 11% at the high dose. C57BL/6 male
mice were given benzene at concentration levels of 200 and 1,000 mg/L (assumed benzene intake of
27 and 154 mg/kg/day) in drinking water for 28 days (Fan 1992). Control groups were given untreated
tap water. Groups of mice were killed on day 7, 14, 21, and 28 of administration, and on day 7, 14, 21,
and 28 after the last administration of benzene at 200 mg/L. There was no effect of treatment on body
weight. Female B6C3F1 mice were exposed to 0, 12, 195, or 350 mg/kg/day benzene in drinking water
for 30 days (Shell 1992). There was no significant effect on body weight at the highest treatment level.
Oral administration of 31.5 mg/kg/day benzene continuously in drinking water for 4 weeks did not affect
BENZENE
131
3. HEALTH EFFECTS
body weight in CD-1 mice (Hsieh et al. 1990). No adverse effect was observed on body weight of male
and female B6C3F1 mice given oral doses of 0, 25, 50, 100, 200, 400, or 600 mg/kg/day benzene in corn
oil for 120 days (NTP 1986).
Sprague-Dawley rats were exposed to benzene by gavage in olive oil, at 0, 50, or 250 mg/kg/day body
weight for 45 days/week for 52 weeks, and then kept under supervision until spontaneous death (Maltoni
et al. 1983, 1985). A 19% decrease in body weight was reported in animals exposed to 250 mg/kg
benzene for 52 weeks. In a companion study, Sprague-Dawley rats were exposed to 500 mg/kg benzene
by ingestion (stomach tube), in olive oil, once daily, 45 days/week for 92 weeks, and then kept under
observation until spontaneous death (Maltoni et al. 1983). Decreased body weight was observed after
92 weeks. In a chronic-duration oral study, male Fischer 344 rats exhibited a decrease in body weight of
11% or more after 25 weeks exposure to doses of 200 mg/kg/day benzene in corn oil (NTP 1986).
Females rats and male and female B6C3F1 mice in the same study exposed to doses up to 100 mg/kg/day
benzene did not show any change in body weight after 12 months of exposure, or after 2 years exposure
(female rats). Male and female mice exhibited body weight effects after chronic exposure. In male mice
given 100 mg/kg, mean body weights decreased from 10% after 47 weeks to 19% in 103 weeks of
exposure relative to controls. In female mice given 100 mg/kg, mean body weights decreased 1415% in
weeks 99103 of exposure (NTP 1986).
Other Systemic Effects.
C57BL/6 male mice were given benzene at concentration levels of 200 and
1,000 mg/L (assumed benzene intake of 27 and 154 mg/kg/day) in drinking water for 28 days (Fan 1992).
Control groups were given untreated tap water. Groups of mice were killed on day 7, 14, 21, and 28 of
administration, and on day 7, 14, 21, and 28 after the last administration of benzene at 200 mg/L. There
was no effect of treatment on food or water consumption. Female Sprague-Dawley rats were dosed by
gavage with 0, 50, 250, 500, or 1,000 mg/kg/day on Gd 615 and killed on Gd 20 (Exxon 1986).
Decreased feed consumption was noted at doses of 250 mg/kg and above, and body weight decreased
11% at the high dose. Female B6C3F1 mice were exposed to 0, 12, 195, or 350 mg/kg/day benzene in
drinking water for 30 days (Shell 1992). Decreased fluid consumption was observed at >12 mg/kg.
No studies were located regarding immunological effects in humans after oral exposure to benzene.
BENZENE
132
3. HEALTH EFFECTS
Oral administration of benzene to CD-1 mice produced an immunotoxic effect on both the humoral and
cellular immune responses (Hsieh et al. 1988b). Exposure to benzene at 8, 40, or 180 mg/kg/day for
4 weeks caused a significant dose-response reduction of total peripheral blood leukocytes and
erythrocytes. Lymphocytes, but not neutrophils or other leukocytes, were decreased in number. Splenic
lymphocyte proliferative response to B- and T-cell mitogens was biphasicenhanced in the 8 mg/kg/day
dosage group and depressed in the 40 and 180 mg/kg/day dosage groups. Cell-mediated immunity
evaluated in mixed-lymphocyte reaction and in the 51Cr-release assay showed a similar biphasic response.
Antibody production was significantly suppressed in mice dosed at 40 and 180 mg/kg/day. The results
indicate that administration of 40 mg/kg/day benzene has an immunosuppressive effect evident in
decreased immune functions evaluated in in vitro assays for cell-mediated immunity and antibody
production. A dose-related decrease in spleen weight was observed, which was significant only in the
180 mg/kg/day group.
C57BL/6 male mice were given benzene at concentration levels of 200 and 1,000 mg/L (assumed benzene
intake of 27 and 154 mg/kg/day) in drinking water for 28 days (Fan 1992). Control groups were given
untreated tap water. Mice were sacrificed on days 7, 14, 21, and 28 of administration. Selected mice of
the 27 mg/kg/day dose group were sacrificed on postexposure days 7, 14, and 21 in order to assess the
postexposure time course of the benzene toxicity. At 27 mg/kg/day, a decreased number of splenocytes
was observed on day 21 and 28 of exposure. At 154 mg/kg/day, spleen cell numbers decreased
significantly as a function of time in mice treated for 14, 21, and 28 days. Benzene treatment for 3 weeks
raised natural killer (NK) cell activity significantly at both doses. However, after another week of
benzene treatment, NK cell activity resumed to control levels. Significant depression of interleukin-2(IL-2) production was detected in both levels for 28 days. The NK cell activity showed normal levels on
day 7, 14, and 21 after the last administration of benzene exposure for 28 days at 27 mg/kg/day.
IL-2 production decreased significantly on day 7 and 14 after cessation of benzene administration, but
recovered with time (43, 71, and 79% of control on day 7, 14, and 21, respectively). Spleen cell number
decreased significantly on the 7th day, but recovered on day 14 and 21. Female B6C3F1 mice were
exposed to 0, 12, 195, or 350 mg/kg/day benzene in drinking water for 30 days (Shell 1992). Decreased
leukocytes were observed at 12 mg/kg/day, and decreased spleen cell number was observed at
195 mg/kg/day.
In the NTP-sponsored intermediate-duration oral study using Fischer 344 rats and B6C3F1 mice, doserelated leukopenia and lymphopenia were observed for both male and female Fischer 344 rats at 200 and
600 mg/kg/day killed on day 60, and at all doses in female rats killed on day 120 (NTP 1986). Decreased
BENZENE
133
3. HEALTH EFFECTS
leukocytes were observed in male and female rats exposed for 60 days to 200 and 600 mg/kg/day
benzene. Lymphoid depletion in the B-cells of the spleen was observed in animals exposed to
200 mg/kg/day (3 of 5 males, 4 of 5 females) and 600 mg/kg/day (5 of 5 males, 5 of 5 females) benzene
for 60 days and in animals that received 600 mg/kg/day (10 of 10 males, 10 of 10 females) benzene for
120 days. At 600 mg/kg/day benzene exposure, increased extramedullary hematopoiesis was observed in
the spleen of 4 of 5 male and 3 of 5 female rats. Dose-related leukopenia and lymphopenia were observed
for both male and female mice exposed for 120 days, but not for 60 days. Leukocytes and lymphocytes
were significantly decreased in male mice exposed for 120 days to 50, 100, 200, 400, and 600 mg/kg/day
benzene. At 120 days of exposure, leukocytes were significantly decreased in female mice at
600 mg/kg/day and lymphocytes at 400 and 600 mg/kg/day. Histological examination revealed no
adverse effects in mandibular lymph node or the thymus for either rats or mice (Huff et al. 1989; NTP
1986). Rats exposed to benzene at 50 and 100 mg/kg/day for 6 months had significant leukopenia (Wolf
et al. 1956).
Leukopenia and lymphopenia were observed in mice at 31.5 mg/kg/day after 4 weeks of oral exposure
(Hsieh et al. 1990). Reduction in thymus mass, suppression of B- and T-cell mitogenesis, and suppressed
IL-2 release were also noted. Similar results were noted at 40 mg/kg/day (Hsieh et al. 1991). Oral
administration of benzene to B6C3F1 mice and Fischer 344 rats at doses of 50200 mg/kg/day (male rats)
or 25100 mg/kg/day (female rats and male and female mice), 5 days/week for 103 weeks resulted in
significant leukocytopenia and lymphocytopenia in both species (Huff et al. 1989; NTP 1986). In the
thymus, lymphoid depletion was observed at 0, 10, 20, or 28% in male rats treated with 0, 50, 100, or
200 mg/kg/day, respectively. Increased incidences of lymphoid depletion of the spleen were observed in
male rats treated with 0 (0%), 50 (40%), 100 (17%), and 200 (49%) mg/kg/day and in female rats treated
with 0 (0%), 25 (22%), 50 (16%), and 100 (20%) mg/kg/day. In mice, an increased incidence of
hematopoietic hyperplasia was observed in the bone marrow of dosed animals in both sexes. Splenic
hematopoiesis was increased in dosed animals of both sexes of mice (Huff et al. 1989; NTP 1986).
Maltoni et al. (1983, 1985) observed decreased leukocytes in Sprague-Dawley rats dosed with
500 mg/kg/day benzene for 84 weeks or more.
The highest NOAEL values and all reliable LOAEL values for immunologic effects in each species and
BENZENE
134
3. HEALTH EFFECTS

In humans, symptoms of central nervous system toxicity (including euphoria, vertigo, muscular
incoordination, and unconsciousness) have been reported following one-time ingestion of benzene at
125 mg/kg (Thienes and Haley 1972).
Neurochemical profiles were conducted on rats after oral exposure to benzene (Kanada et al. 1994).
Sprague-Dawley rats received a single dose of 950 mg/kg benzene by gavage and were sacrificed 2 hours
after treatment. The control group received nothing. Brains were dissected into small-brain areas and
stored until analysis. Acetylcholine, 3,4-dihydroxyphenylalanine (DOPA), dopamine, 3,4-dihydroxyphenylacetic acid (DOPAC), homovanillic acid (HVA), norepinephrine, 3-methoxy-4-hydroxyphenylglycol (MHPG), serotonin, and 5-hydroxyindoleacetic acid (5HIAA) contents in the small-brain
regions were measured. Results showed that benzene decreased acetylcholine content of rat
hippocampus. DOPA and norepinephrine content decreased in the rat midbrain. Dopamine, serotonin
and 5HIAA content increased in the rat midbrain. Dopamine, DOPAC, norepinephrine, and 5HIAA
content increased and serotonin content decreased in the rat hypothalmus after oral administration of
benzene. Increased dopamine, HVA, MHPG, and serotonin content of rat medulla oblongata was
observed. Decreased norepinephrine and 5HIAA content of rat medulla oblongata by benzene treatment
was observed.
Oral exposure to benzene induced both synthesis and catabolism of monoamine neurotransmitters in
CD-1 mice (Hsieh et al. 1988a). Mice given 8, 40, or 180 mg/kg/day of benzene for 4 weeks in drinking
water exhibited changes in the levels of norepinephrine, dopamine, serotonin, and catecholamine
metabolites in several brain regions but no treatment-related behavioral changes. Similar results were
seen at 31.540 mg/kg/day (Hsieh et al. 1990, 1991). Because of the lack of association with behavioral
changes, the effects on the neurotransmitters cannot be adequately assessed. Female B6C3F1 mice were
exposed to 0, 12, 195, or 350 mg/kg/day benzene in drinking water for 30 days (Shell 1992). Decreased
brain weight was observed at 350 mg/kg/day. Sprague-Dawley rats given one oral dose of 88 mg/kg
benzene exhibited slight central nervous system depression, whereas at 1,870 mg/kg/day, tremors were
observed (Cornish and Ryan 1965). Histological examination of the brain revealed no treatment-related
lesions after gavage treatment of male and female Fischer 344 rats and B6C3F1 mice with doses up to
600 mg/kg/day for 120 days (NTP 1986). In the same experiment, B6C3F1 mice exhibited tremors
intermittently at doses of 400 mg/kg/day, which were more pronounced in males during the last 3 weeks
of the study. No adverse effects based on histological examination of brain or spinal cord were observed
BENZENE
135
3. HEALTH EFFECTS
in male and female Fischer 344 rats and B6C3F1 mice after chronic oral exposure to 50200 mg/kg/day
(male rats) or 25100 mg/kg/day (female rats and male and female mice) (NTP 1986).
The highest NOAEL values and all reliable LOAEL values for neurological effects in each species and
No studies were located regarding reproductive effects in humans after oral exposure to benzene.
Female Sprague-Dawley rats were dosed by gavage with 0, 50, 250, 500, or 1,000 mg/kg benzene daily
on Gd 615 and killed on Gd 20 (Exxon 1986). No adverse effects were noted on reproductive
competency. No histological changes were reported in the prostate, testes, ovaries, mammary gland, or
uterus of male and female Fischer 344 rats and B6C3F1 mice dosed by gavage with up to 600 mg/kg/day
benzene for 17 weeks (NTP 1986). In male and female Fischer 344 rats and B6C3F1 mice after chronic
oral exposure to 50200 mg/kg/day (male rats) or 25100 mg/kg/day (female rats and male and female
mice), endometrial stromal polyps occurred with a significant positive trend in female rats (NTP 1986).
The incidence in high dose group (14/50) was significantly greater than that in the control (7/50). In
mice, analysis of preputial gland lesions in male mice dosed at 0, 25, 50, or 100 mg/kg showed increased
incidences of focal, diffuse or epithelial hyperplasia (5, 65, 31, and 3%, respectively). The lower
incidences of hyperplasia in the higher dose groups were probably due to the progression of the preputial
gland lesions to neoplasias (see Section 3.2.2.7). Various non-neoplastic and neoplastic ovarian lesions
were observed in dosed female mice, including epithelial hyperplasia and senile atrophy (NTP 1986).
The NOAEL and LOAEL values for reproductive effects in rats and mice are recorded in Table 3-2 and
plotted in Figure 3-2.
No studies were located regarding developmental effects in humans after oral exposure to benzene.
Benzene was embryotoxic as evidenced by reduced pup body weights when mice were administered
1,300 mg/kg/day of benzene by gavage on Gd 812 (Seidenberg et al. 1986). No maternal toxicity was
observed. Female Sprague-Dawley rats were dosed by gavage with 0, 50, 250, 500, or 1,000 mg/kg
BENZENE
136
3. HEALTH EFFECTS
benzene daily on Gd 615 and killed on Gd 20 (Exxon 1986). No adverse effects were noted on
morphological development.
The NOAEL value for rats and the LOAEL value for mice for developmental effects following acute oral
exposure are recorded in Table 3-2 and plotted in Figure 3-2.
3.2.2.7 Cancer
Essentially no information was located regarding the oral carcinogenicity of benzene in humans.
Lymphatic and hematopoietic cancers were increased in vehicle maintenance workers who occasionally
siphoned gasoline by mouth (Hunting et al. 1995), but the skin and lungs were the main routes of
exposure (see Section 3.2.3.7).
Benzene has been shown to be a multiple site carcinogen by the oral route in animals (Huff et al. 1989;
Maltoni et al. 1983, 1985, 1989; NTP 1986). In bioassays conducted by the NTP, benzene in corn oil was
administered groups of 50 F344/N rats and 50 B6C3F1 of each sex by gavage on 5 days/week for
103 weeks (Huff et al. 1989; NTP 1986). The male rats were exposed to dose levels of 0, 50, 100, or
200 mg/kg/day, and female rats and mice of both sexes were exposed to 0, 25, 50, or 100 mg/kg/day. In
the rats, benzene caused significantly increased incidences of Zymbal gland carcinomas in males at
100 mg/kg/day and females at 25 mg/kg/day, oral cavity squamous cell papillomas and carcinomas in
males at 50 mg/kg/day and females at 25 mg/kg/day, and skin squamous cell papillomas and
carcinomas in males at 200 mg/kg/day. In the mice, benzene mainly caused significantly increased
incidences of malignant lymphomas in both sexes at 25 mg/kg/day, Zymbal gland carcinomas in males
at 50 mg/kg/day and females at 100 mg/kg/day, lung alveolar/bronchiolar adenomas and carcinomas in
males at 100 mg/kg/day and females at 50 mg/kg/day, Harderian gland adenomas in males at
25 mg/kg/day, preputial gland squamous cell carcinomas in males at 50 mg/kg/day, and mammary
gland carcinomas in females at 50 mg/kg/day. NTP (1986) concluded that there was clear evidence of
carcinogenicity of benzene in male and female F344/N rats and B6C3F1 mice under the conditions of
these studies.
Maltoni et al. (1983, 1985, 1989) assessed carcinogenicity in groups of 3050 rats and mice of each sex
that were exposed to benzene in olive oil by gavage on 45 days/week for up to 104 weeks and observed
for life. Sprague-Dawley rats were exposed to 0, 50, or 250 mg/kg/day for 52 weeks or 0 or
500 mg/kg/day for 104 weeks; effects included increased incidences of Zymbal gland carcinomas in
BENZENE
137
3. HEALTH EFFECTS
females at 50 mg/kg/day, oral cavity carcinomas in females at 250 mg/kg/day and both sexes at
500 mg/kg/day, forestomach carcinomas in females at 500 mg/kg/day, nasal cavity and skin carcinomas
in males at 500 mg/kg/day, and liver angiosarcomas in both sexes at 500 mg/kg/day. Wistar rats were
exposed to 0 or 500 mg/kg/day for 104 weeks; effects included increased incidences of Zymbal gland,
nasal cavity, and oral cavity carcinomas. Swiss mice were exposed to 0 or 500 mg/kg/day for 78 weeks;
effects included increased incidences Zymbal gland carcinomas in males, mammary carcinomas in
females, and lung adenomas in both sexes. RF/J mice were exposed to 0 or 500 mg/kg/day for 52 weeks;
effects included increased incidences of mammary carcinomas in females and lung adenomas in both
sexes.
As discussed in Section 3.2.1.7, there is a consensus that benzene is a human carcinogen (IARC 1987,
2004, 2007; IRIS 2007; NTP 2005). This conclusion is based on sufficient inhalation data in humans
supported by animal evidence, including the oral studies summarized above. The human cancer induced
by inhalation exposure to benzene is predominantly acute nonlymphocytic leukemia, whereas benzene is
a multiple site carcinogen in animals by both the inhalation and oral routes. Due to the lack of oral
carcinogenicity data in humans, as well as the lack of a well-demonstrated and reproducible animal model
for leukemia from benzene exposure, EPA extrapolated an oral slope factor from the inhalation unit risk
range (IRIS 2007). The oral slope factor ranges from 1.5x10-2 to 5.5x10-2 (mg/kg/day)-1, and for cancer
risks of 1x10-41x10-7, the corresponding dose levels range from 6.7x10-31.8x10-3 to 6.7x10-61.8x10-6
mg/kg/day, respectively. These risk levels are presented in Figure 3-1.
3.2.3
Dermal Exposure
3.2.3.1 Death
No studies were located regarding deaths in animals after dermal exposure to benzene.
A cohort of 338 men was investigated as to causes of death among employees of the fleet maintenance
division of Washington DC's Department of Public Works (Hunting et al. 1995). This mortality study
was undertaken because of three cases of leukemia among car and mobile equipments mechanics.
Preliminary evaluation showed that the garage mechanics regularly used gasoline to clean parts and wash
their hands; these workers also experienced dermal and inhalation exposure to gasoline during
maintenance of vehicles. The men were employed for at least 1 year between January 1, 1977, and
December 31, 1989. Cause-specific SMRs were calculated. Increased risk of death was found in some
categories.
BENZENE
138
3. HEALTH EFFECTS

No studies were located regarding respiratory, cardiovascular, gastrointestinal, musculoskeletal, hepatic,
renal, endocrine, or body weight effects in humans or animals after dermal exposure to benzene.
Available data pertaining to hematological, dermal, and ocular effects are presented below. All reliable
LOAEL values for systemic effects in humans and rabbits for acute- and chronic-duration dermal
exposure are recorded in Table 3-3.
Tondel et al. (1995) reported a case of myelofibrosis that was diagnosed in a
46-year-old man who had worked from 1962 to 1979 as a gasoline station attendant. Although the
exposure was primarily by inhalation, it is likely that dermal exposure also occurred.
Dermal Effects.
In humans, benzene is a skin irritant. By defatting the keratin layer, it may cause
erythema, vesiculation, and dry and scaly dermatitis (Sandmeyer 1981). Acute fatal exposure to benzene
vapors caused second degree burns on the face, trunk, and limbs of the victims (Avis and Hutton 1993).
Fifteen male workers were exposed to benzene vapors (>60 ppm) over several days during the removal of
residual fuel from shipyard fuel tanks (Midzenski et al. 1992). Exposures to benzene range from 1 day to
3 weeks (mean of 5 days), 2.58 hours/day (mean of 5.5 hours). Workers with more than 2 days
(16 hours) exposure reported mucous membrane irritation (80%), and skin irritation (13%) after exposure
to the vapor.
Benzene was slightly irritating to the skin of rabbits (Wolf et al. 1956). The skin showed moderate
erythema, edema, and moderate necrosis following application 1 time/day for 4 weeks.
Ocular Effects.
Solvent workers who were exposed to 33 (men) or 59 (women) ppm benzene
exhibited eye irritation while being exposed to the vapors (Yin et al. 1987b).
A transient increase in lacrimation was observed in male rats exposed to 10300 ppm benzene for
6 hours/day, 5 days/week (Shell 1980). Moderate conjunctival irritation and transient corneal damage
were observed in rabbits subsequent to placement of 2 drops of benzene onto the eyeball (Wolf et al.
1956).
Table 3-3 Levels of Significant Exposure to Benzene - Dermal
BENZENE
Species
(Strain)
Exposure/
Duration/
Frequency
(Route)
LOAEL
Reference
System
NOAEL
Less Serious
Serious
Chemical Form
Comments
ACUTE EXPOSURE
Systemic
Human
1-21 d
2.5-8 hr/d
Dermal
60 M
ppm
921
Wolf et al 1956
164
Shell 1980
930
Yin et al. 1987b
952
ppm
921
Rabbit
(NS)
(mucous membrane and

skin irritation)
once
Ocular
2
Unknown
164
(moderate conjunctival
irritations; light corneal
injury)
Unknown
Systemic
Rat
(CD)
10 wk
5 d/wk
6 hr/d
Ocular
1M
ppm
ppm
10 M
ppm
(lacrimation)
33 M
ppm
(eye irritation)
59 F
ppm
(eye irritation)
ppm
930
CHRONIC EXPOSURE
Systemic
Human
>1 yr
(occup)
Ocular
952
3. HEALTH EFFECTS
ppm
ppm
d = day(s); F = female; hr = hour(s); LOAEL = lowest-observed-adverse-effect level; M = male; NOAEL = no-observed-adverse-effect level; NS = not specified; occup = occupational;
yr = year
139
BENZENE
140
3. HEALTH EFFECTS

No studies were located regarding immunological and lymphoreticular effects in humans or animals after
dermal exposure to benzene.
Tondel et al. (1995) reported the case of a gasoline station attendant who had worked from 1962 to 1979
and who developed myelofibrosis. The patient described symptoms of fatigue for 3 weeks and night
sweats, among other symptoms. Although the exposure was primarily by inhalation, it is probable that
dermal exposure also occurred.
No studies were located regarding neurological effects in animals after dermal exposure to benzene.
No studies were located regarding reproductive effects in humans or animals after dermal exposure to
benzene.
No studies were located regarding developmental effects in humans or animals after dermal exposure to
benzene.
3.2.3.7 Cancer
A cohort mortality study was conducted to estimate the relative risk of hematological cancer among
335 male vehicle maintenance workers who were employed for at least 1 year between 1977 and 1989
and followed through 1991 (Hunting et al. 1995). The workers were car and mobile equipment
mechanics who regularly used gasoline to clean parts and wash their hands. Exposure to gasoline also
occurred via inhalation, and some of the workers occasionally siphoned gasoline by mouth. Since most of
the cohort consisted of non-white (race data were missing for 9.5% of the cohort) vehicle maintenance
workers who lived within the District of Columbia, death rates for age- and calendar-matched non-white
males in the District of Columbia were used to calculate expected deaths for the cohort. Three deaths due
to lymphatic and hematopoietic cancer were observed, yielding SMRs of 3.63 (95% CI 0.7510.63) in the
whole cohort and 4.22 (95% CI 0.8712.34) in a subgroup of 297 workers with the highest potential for
BENZENE
141
3. HEALTH EFFECTS
exposure. Neither of these SMRs were significant, although analysis by cancer subtype showed a
significantly elevated risk for leukemia and aleukemia in the high exposure subgroup (two cases,
SMR=9.26, 95% CI 1.1233.43). Aleukemia refers to leukemias in which the blood has normal or nearnormal white blood cell counts, but few numbers of young leukocytes. Two additional cases of leukemia
were identified among workers who were not included in the cohort (one died after the end of the followup period and another was still alive). Mortalities from causes other than lymphatic/hematopoietic cancer
were not significantly increased.
Application of benzene to the skin of animals has not produced evidence of carcinogenicity, although
most studies were inadequate for evaluation. As summarized by IARC (1982, 1987), many dermal
carcinogenicity studies of chemicals other than benzene used benzene as a vehicle, and treated large
numbers of control animals (mice) with benzene alone. None of these studies indicated that benzene
induced skin tumors; however, all possible tumor sites usually were not examined.
3.3
GENOTOXICITY
The genotoxic effects of benzene have been studied extensively. The in vivo and in vitro data are
summarized in Tables 3-4 and 3-5, respectively. In chronically-exposed humans, benzene and/or its
metabolites primarily cause chromosomal aberrations (Andreoli et al. 1997; Bogadi-are et al. 1997; Ding
et al. 1983; Forni and Moreo 1967, 1969; Forni et al. 1971a; Hartwich et al. 1969; Hedli et al. 1991;
Karacic et al. 1995; Kauba et al. 2000; Major et al. 1992, 1994; Picciano 1979; Popp et al. 1992;
Rothman et al. 1995; Sardas et al. 1994; Sasiadek et al. 1989; Sellyei and Kelemen 1971; Smith et al.
1998; Sul et al. 2002; Tompa et al. 1994; Tough and Court Brown 1965; Tough et al. 1970; Trkel and
Egeli 1994; Van den Berghe et al. 1979; Yardley-Jones et al. 1990; Zhang et al. 1998b, 1999).
Chromosomal aberrations in humans are frequently demonstrated in peripheral blood lymphocytes and
bone marrow. Although inhalation, oral, and dermal routes are all potential pathways of exposure
relevant to humans, available in vivo human data are usually drawn from occupational settings in which
inhalation and dermal exposure routes are most prevalent. In most of these studies, chromosome
abnormalities were detected in workers exposed to high concentrations of benzene, sufficient to produce
blood dyscrasias. However, Qu et al. (2003a, 2003b) noted a concentration-related increase in
chromosomal aberrations across a wide range of exposure concentrations, including workers with
relatively low-level benzene exposure. Limitations of many of the occupational studies include lack of
accurate exposure data, possible coexposure to other chemicals, and lack of appropriate control groups.
BENZENE
142
3. HEALTH EFFECTS
Table 3-4. Genotoxicity of Benzene In Vivo

Species (test system)
End point
Result Reference
DNA synthesis
Hellmr and Bolcsfoldi 1992a
Sex-linked recessive lethal

Recombination
Recombination
Heritable translocation
Kale and Baum 1983
Mouse (bone marrow)
Chromosomal aberrations
Mouse (bone marrow)

Mouse (bone marrow)
Mouse (spleen lymphocytes)
+
(+)
+
Mouse (lymphoid cells, myeloid

cells)
Rat (bone marrow)
Giver et al. 2001; Shelby and

Witt 1995; Siou et al. 1981
Meyne and Legator 1980
Tice et al. 1980; 1982
Au et al. 1991; Rithidech et al.
1987
Giver et al. 2001
Rat (bone marrow)
+b
Rat (bone marrow)

Chinese hamster (bone marrow)
Rabbit (bone marrow)
Human (occupational
exposure/lymphocytes)
+
+
+
Human (occupational
Human (occupational
Mouse (bone marrow)
(+)
Micronuclei
Mouse (bone marrow PCEs)

Moused (bone marrow PCEs)
Micronuclei
Micronuclei
+c
+e
Prokaryotic cells:
Escherichia coli (host mediated
DNA repair)
Invertebrate animal cells:
Drosophila melanogaster
Spermatocytes
Spermatogonia
Spermatocytes
Mammalian cells:
Fujie et al. 1990; Hoechst

1977; Philip and Jensen 1970;
Styles and Richardson 1984
Anderson and Richardson
1981b
Hoechst 1977
Siou et al. 1981
Kissling and Speck 1972; 1973
Bogadi-are et al. 1997; Ding
et al. 1983; Forni et al. 1971a;
Kauba et al. 2000; Picciano
1979; Sasiadek 1992; Sasiadek
and Jagielski 1990; Sasiadek et
al. 1989; Smith et al. 1998;
Tompa et al. 1994; Tough and
Court Brown 1965; Tough et al.
1970; Zhang et al. 1998a, 1999
Yardley-Jones et al. 1990
Bogadi-are et al. 1997;
Jablonick et al. 1987
Shelby and Witt 1995; Shelby

et al. 1993
Ciranni et al. 1988
Suzuki et al. 1989
BENZENE
143
3. HEALTH EFFECTS

End point
Result Reference
Micronuclei

Mouse (bone marrow NCEs)
Mouse (bone marrow NCEs)
Mouse (pregnant/bone marrow
PCEs)
Mouse (peripheral blood)
Micronuclei
Micronuclei
Micronuclei
Micronuclei
Micronuclei
+a
+f
+
+f
(+)
Micronuclei
Mouse (peripheral blood PCEs) Micronuclei

Mouse (peripheral blood PCEs) Micronuclei
Mouse (peripheral blood NCEs) Micronuclei
+
+c,g
+
Mouse (peripheral blood NCEs)

Mouse (lung fibroblasts)
Mouse (fetus/liver cells)
Rat (lymphocytes)
Chinese hamster (bone marrow)
Human (lymphocytes)
Human (occupational
Human (occupational
Mouse (bone marrow)
Mouse (pregnant/bone marrow)
Mouse (lymphocytes)
Mouse (fetus/liver cells)
Rat (lymphocytes)
Human (occupational
Human (occupational
Micronuclei
Micronuclei
Micronuclei
Micronuclei
Micronuclei
Micronuclei
Micronuclei
+c,f
+
+
+
+
+
+
Micronuclei
Sister chromatid exchange

+
+
+
+
+
+
Au et al. 1990; Barale et al.

1985; Chen et al. 1994; Diaz et
al. 1980; Erexson et al. 1986;
Farris et al. 1996; Harper et al.
1984; Hite et al. 1980; Siou et
al. 1981; Toft et al. 1982
Meyne and Legator 1980
Eastmond et al. 2001
Farris et al. 1996
Eastmond et al. 2001
Ciranni et al. 1988
Hayashi et al. 1992; Healy et
al. 2001
Farris et al. 1996
Luke et al. 1988a
Barale et al. 1985; Choy et al.
1985; Farris et al. 1996;
Rithidech et al. 1988
Luke et al. 1988a
Ranaldi et al. 1998
Ciranni et al. 1988
Erexson et al. 1986
Siou et al. 1981
Robertson et al. 1991
Liu et al. 1996
Pitarque et al. 1996; Surralls
et al. 1997
Tice et al. 1980; 1982
Sharma et al. 1985
Erexson et al. 1986
Sharma et al. 1985
Erexson et al. 1986
Popp et al. 1992
Mouse (spleen lymphocytes)

Mutations
Mouse (lung tissue)
Mutations
Mouse embryo (premelanocytes) Mutations (deletions)
+
+
+
Kauba et al. 2000; Pitarque et

al. 1997; Seiji et al. 1990;
Yardley-Jones et al. 1988
Ward et al. 1992
Mullin et al. 1998
Schiestl et al. 1997
Human (bone marrow)
Rothman et al. 1995
Mutations (geneduplicating)
BENZENE
144
3. HEALTH EFFECTS

End point
Result Reference
Rothman et al. 1995
Mouse (bone marrow)
Mutations (geneinactivating)
DNA adducts
Mouse (white blood cells)

Mouse (liver)
DNA adducts
DNA adducts
+
+
Rat (bone marrow)
DNA adducts
Arfellini et al. 1985; Creek et al.

1997; Lvay et al. 1996; Pathak
et al. 1995; Turteltaub and
Mani 2003
Lvay et al. 1996
1997; Mani et al. 1999;
Turteltaub and Mani 2003
1997; Lvay et al. 1996; Pathak
et al. 1995; Turteltaub and
Mani 2003
Rat (liver)
DNA adducts
Mouse (peripheral blood

lymphocytes)
Rat (lymphocytes, bone marrow,

spleen, liver)
Human (occupational
Mouse (peripheral blood
lymphocytes, bone marrow)
Human (occupational
Rat (bone marrow)

Human (occupational
Mouse (bone marrow)

Rabbit (bone marrow)
Mouse (bone marrow)
Rat (liver mitochondria)
DNA strand breaks

1997; Lutz and Schlatter 1977;
Mani et al. 1999; Mazzullo et al.
1989; Turteltaub and Mani
2003
Tuo et al. 1996
DNA strand breaks
Lee et al. 2005
DNA strand breaks
DNA damage
Andreoli et al. 1997; Nilsson et
al. 1996; Sul et al. 2002
Chang et al. 2005
DNA repair efficiency
Hallberg et al. 1996
DNA oxidative damage

DNA oxidative damage
+
+
Kolachana et al. 1993
Liu et al. 1996
DNA synthesis inhibition

DNA synthesis inhibition
RNA synthesis inhibition
RNA synthesis inhibition
+
+
+
+
Lee et al. 1988
Kissling and Speck 1972
Kissling and Speck 1972
Kalf et al. 1982
Human (bone marrow)
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3. HEALTH EFFECTS

Mouse (spermatogonia)
End point
Result Reference
Sperm head abnormality
Topham 1980
This result was observed following both oral and intraperitoneal exposure.
This result was observed following both inhalation and intraperitoneal exposure.
c
Males affected to a significantly greater degree than females.
d
Two strains of mouse were tested; Ms/Ae and CD-1. The result applies to both strains.
e
This result was observed following both oral and intraperitoneal exposure; however, oral exposure produced the
greater effect.
f
Increase in micronuclei was exposure duration-dependent.
g
Increase in micronuclei was exposure duration-independent.
+ = Positive result; = negative result; (+) = weakly positive result; DNA = deoxyribonucleic acid;
NCEs = normochromatic erythrocytes; PCEs = polychromatic erythrocytes; RNA = ribonucleic acid
BENZENE
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3. HEALTH EFFECTS
Table 3-5. Genotoxicity of Benzene In Vitro

Prokaryotic organisms:
Salmonella typhimurium
(Ames test)
S. typhimurium (histidine
reversion)
S. typhimurium (azaquanine
reversion)
Bacillus subtilis (histidine
reversion)
Escherichia coli (DNA
polymerase 1/cell-free DNA
synthetic system)
E. coli (host meditated DNA

repair)
Plasmid DNA X-174 RF I
Eukaryotic organisms:
Fungi:
Aspergillus nidulans
(methionine supressors)
Mammalian cells:
Mouse (L5178Y cells/TK
test)
Chinese hamster (ovary

cell culture)
Human (lymphocyte cell
culture)
culture)
Human (lymphoblastoid
culture)
cell culture)
Human (whole blood cells)

cell culture)
culture)
culture)
Rabbit (bone marrow
mitoplasts)
End point
Result
With
Without
activation activation
Reference
Gene mutation
De Flora et al. 1984
Gene mutation
Glatt et al. 1989
Gene mutation
No data
Gene mutation
Kaden et al. 1979;

Seixas et al. 1982
Tannoka 1977
DNA synthesis
inhibition
No data
Lee et al. 1988
DNA synthesis
No data
No data
DNA degradation
No data
Hellmr and Bolcsfoldi

1992b
Li et al. 1995
Gene mutation
No data
Crebelli et al. 1986
Gene mutation
Oberly et al. 1984
Chromosomal
aberrations
Chromosomal
aberrations
Chromosomal
aberrations
Intrachromosomal
recombination
Micronuclei
Gulati et al. 1989
No data
No data
No data
Eastmond et al. 1994;
Morimoto 1976
Gerner-Smidt and
Friedrich 1978
Aubrecht et al. 1995
Douglas et al. 1985
No data
Zarani et al. 1999
Douglas et al. 1985;
Gulati et al. 1989
Morimoto 1983
No data
No data
Micronuclei
Sister chromatid
exchange
Sister chromatid
exchange
Sister chromatid
exchange
DNA adducts
Gerner-Smidt and
Friedrich 1978
Rushmore et al. 1984
BENZENE
147
3. HEALTH EFFECTS
End point
Result
With
Without
Rat (liver mitoplasts)
Calf thymus DNA
Human (bone marrow)
DNA adducts
DNA adducts
DNA adducts
No data
No data
No data
+
+
+
Human (leukemia cells)
DNA adducts
No data
No data
+
+a
No data
No data
Pellack-Walker and
Blumer 1986
Kolachana et al. 1993
No data
Hallberg et al. 1996
No data
Dees and Travis 1994
No data
(+)
Glauert et al. 1985
No data
Probst and Hill 1985;

Williams et al. 1985
Barrett 1985
No data
Lee et al. 1988
(+)
Lee et al. 1989
No data
Lee et al. 1988
No data
Painter and Howard

1982
Post et al. 1985
Rat (hepatocytes)
DNA breaks
DNA breaks
cell culture)

DNA breaks
cell culture)
Chinese hamster (V79 cell

DNA breaks
culture)
Mouse (L5178Y cell
DNA breaks
culture)
Human (leukemia cells)

DNA oxidative
damage
DNA repair
culture)
Rat liver epithelial cells

DNA hyperphos
phorylatioin
Rat (hepatocyte culture)
Unscheduled DNA
synthesis
Rat (hepatocyte culture)
Unscheduled DNA
synthesis
Human (HeLa S3 cells)
Unscheduled DNA
synthesis
Mouse (bone marrow cell DNA
synthesis
culture)
inhibition
Mouse (bone marrow cell DNA
synthesis
culture)
inhibition
Calf (thymus DNA
DNA synthesis
polymerase /cell-free
inhibition
DNA synthetic system)
Human (HeLa cells)
DNA synthesis
inhibition
Mouse (spleen
RNA synthesis
lymphocytes)
inhibition
Reference
Rushmore et al. 1984
Chenna et al. 1995
Bodell et al. 1993;
Lvay and Bodell 1992
Bodell et al. 1993;
Lvay and Bodell 1992
Bradley 1985
Douglas et al. 1985
Lakhanisky and
Hendricks 1985
Swenberg et al. 1976
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3. HEALTH EFFECTS

Rat (liver mitoplasts)
Cat, rabbit (bone marrow
mitoplasts)
a
End point
RNA synthesis
inhibition
RNA synthesis
inhibition
Result
With
Without
Reference
No data
Kalf et al. 1982
No data
Kalf et al. 1982
Benzene's effect on DNA breaks was reduced when metabolic activators were used.
= negative results; + = positive results; (+) = weakly positive result; DNA = deoxyribonucleic acid;
RNA = ribonucleic acid
BENZENE
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3. HEALTH EFFECTS
Chromosomal aberrations observed in workers chronically exposed to benzene include hypo- and
hyperdiploidy, deletions, breaks, and gaps. For example, analysis of peripheral lymphocytes of workers
exposed to benzene vapors at a mean concentration of 30 ppm revealed significant increases in
monosomy of chromosomes 5, 7, and 8 (but not 1), and tri- and/or tetrasomy of chromosomes 1, 5, 7, and
8 (Zhang et al. 1998b, 1999). In another series of epidemiological studies in workers chronically exposed
to benzene, nonrandom effects were apparent in chromosomes 1, 2, 4, and 9; nonrandom breaks in
chromosomes 2, 4, and 9 were twice as prevalent in benzene-exposed workers versus controls; and
chromosomes 1 and 2 were nearly twice as prone to gaps (Sasiadek and Jagielski 1990; Sasiadek et al.
1989). Twenty-one people with hematological signs of chronic benzene poisoning exhibited significantly
more chromosomal abnormalities than controls (Ding et al. 1983). A significant increase in dicentric
chromosomes and unstable aberrations was noted in 36 female workers exposed to benzene in a shoe
factory for up to 32 years (Kauba et al. 2000). Significant increases in hyperploidy of chromosomes 8
and 21 and translocations between chromosomes 8 and 21 were observed in workers exposed to benzene
vapors at a mean TWA of 31 ppm (Smith et al. 1998).
DNA repair efficiency was evaluated in blood lymphocytes collected from exposed or unexposed workers
in a petrochemical plant (Hallberg et al. 1996). Plasmids (pCMVCAT) were irradiated with UV light
(254 nm) to induce thymidine dimers and were then transfected into blood lymphocytes from workers.
Transfected plasmids that were repaired in the lymphocytes would express the chloramphenicol
actetyltransferase reporter gene (CAT) product whereas unrepaired plasmids would not. Lymphocytes
from exposed or unexposed workers did not show significant differences in their ability to repair lightdamaged DNA; however, the authors suggest that the sample population was too small to detect any
differences given the large individual variations in repair capacity (Hallberg et al. 1996).
Oxidative DNA damage was assessed in workers (n=87) exposed to benzene by conducting
measurements of 8-hydroxy-2-deoxyguanosine (8-OHdG) in peripheral blood lymphocytes (Liu et al.
1996). 8-OHdG is formed by hydroxy radical (OH) addition at the C-8 position of deoxyguanosine
(Kasai and Nishimura 1986) and appears to be a biomarker of oxidative DNA damage (Liu et al. 1996).
The exposure to benzene was classified as low, medium, or high (mean benzene levels of 2.46, 103, or
424 mg/m3, respectively [corresponding to 0.78, 32.2, or 133 ppm]). Levels of 8-OHdG in exposed
workers rose in a concentration-related manner although the increases were significant only in the
medium and high exposure groups. Toluene, also detected in the workplace air, did not alter levels of
8-OHdG. Formation of micronuclei, a measure of DNA damage, increased in a concentration-related
manner to levels that were significantly higher than those in controls. The levels of urinary trans,trans-
BENZENE
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3. HEALTH EFFECTS
muconic acid, a biomarker of benzene exposure, were well correlated with levels of 8-OHdG which, in
turn, were well correlated with levels of lymphocyte micronuclei. The findings of Liu et al. (1996) are
supported by the results of a study by Nilsson et al. (1996) in which concentration-related increased
urinary levels of 8-OHdG were reported in male workers (n=30) at a gasoline station. Breathing zone
benzene concentrations ranged from 0.003 to 0.6 ppm (mean 0.13 ppm). Significant concentrationrelated increases in DNA single-strand breaks were also noted in these workers. Collectively, the results
of Liu et al. (1996) and Nilsson et al. (1996) provide suggestive evidence that benzene metabolites may
induce reactive oxygen species, which could result in oxidative DNA damage and formation of
hydroxylated bases such as 8-OHdG (Liu et al. 1996).
Rothman et al. (1995) used the glycophorin A (GPA) gene loss mutation assay to assess the nature of
DNA damage in workers heavily exposed to benzene. The GPA assay measures the frequency of variant
cells that have lost the expression of the M form of the GPA gene in the peripheral blood of heterozygous
(MN) subjects. The variant cells possess the NN phenotype (double-copy expression of the N allele and
no expression of M) or the N phenotype (single-copy expression of the N allele and no expression of
M). The NN variants are thought to arise from mitotic recombination, chromosome loss, and
reduplication, or gene conversion. The N variants appear to be associated with point mutations,
deletions, or gene inactivation. Rothman et al. (1995) demonstrated a significant increase in the
frequency of NN cells in benzene-exposed workers (compared with unexposed control subjects) in the
absence of a significant effect on the frequency of N cells. These results suggest that benzene induces
gene-duplicating, but not gene-inactivating, mutations at the GPA locus in benzene-exposed humans.
Sister chromatid exchange was not found to be a significant effect of benzene exposure in humans
(Kauba et al. 2000; Seiji et al. 1990; Yardley-Jones et al. 1988); however, the selection of control
subjects in the studies by Seiji et al. (1990) and Yardley-Jones et al. (1988) was poor. Refer to Table 3-4
for a further summary of these results.
In vivo animal studies provide convincing evidence of benzene's genotoxicity (Table 3-4). Furthermore,
the finding that male mice are more sensitive than females to benzene-induced chromosomal damage is
consistent among reports (Armstrong and Galloway 1993; Barale et al. 1985; Choy et al. 1985; Ciranni et
al. 1988; Hatakeyama et al. 1992; Meyne and Legator 1980; Siou et al. 1981). Consistently positive
findings for chromosomal aberrations in bone marrow and lymphocytes in animals support the human
case reports and epidemiological studies in which chromosomal damage was linked to benzene exposure.
Micronucleus assays are popular methods for crudely analyzing DNA damage in animals. Positive results
BENZENE
151
3. HEALTH EFFECTS
were observed in all studies testing for increased micronuclei frequencies. One of these micronucleus
assays (Luke et al. 1988a) investigated the effects of different inhalation exposure durations on
polychromatic erythrocytes (PCEs) and normochromatic erythrocytes (NCEs) in peripheral blood. PCEs
are newly formed erythrocytes that contain mRNA and as a result, exhibit staining when RNA staining
reagents are used. NCEs are mature erythrocytes that lack mRNA and are not stained under the same
conditions. The researchers found that PCEs are good indicators of recent and acute exposure, while
NCEs are good indicators of accumulated long-term exposure (Luke et al. 1988a). Although no human
studies were located that reported increased sister chromatid exchange in exposed individuals, increases
in sister chromatid exchange were reported in mice and rats (Erexson et al. 1986; Sharma et al. 1985; Tice
et al. 1980, 1982). In vivo, trans,trans-muconaldehyde (a metabolite of benzene) has also been shown to
induce highly significant increases in sister chromatid exchanges in mice (Witz et al. 1990a). In addition
to oral and inhalation routes, many researchers tested subcutaneous and intraperitoneal routes as well; the
results for these alternate routes of exposure were largely positive for chromosomal aberrations in bone
marrow (Anderson and Richardson 1981; Kissling and Speck 1972, 1973; Kolachana et al. 1993; Meyne
and Legator 1980; Philip and Jensen 1970), micronuclei in bone marrow (Diaz et al. 1980), and sister
chromatid exchange in mouse fetus liver cells (Sharma et al. 1985). Binding of benzene and/or its
metabolites to DNA, RNA, and proteins has been consistently observed in rats and mice (Arfellini et al.
1985; Creek et al. 1997; Lvay et al. 1996; Mani et al. 1999; Mazullo et al. 1989; Turteltaub and Mani
2003). Arfellini et al. (1985) noted that binding to RNA and proteins was more prevalent than binding to
DNA. Lvay et al. (1996) observed dose- and time-dependent formation of two DNA adducts in white
blood cells and bone marrow cells of mice administered benzene for 7 days via intraperitoneal injection.
Turteltaub and Mani (2003) consistently observed DNA and protein binding in mice at intraperitoneal
doses as low as 5 g/kg body weight. One study using intraperitoneal injections reported a dosedependent increase in sperm head abnormalities in mice exposed to 0.5 or 0.6 mL benzene/kg/day
(Topham 1980). The author views sperm head abnormality as a possible indication of heritable
mutations. However, from this study alone, it cannot be determined if benzene causes such transmissible
genetic mutations. For these and other results from animal in vivo studies, see Table 3-4.
Molecular mechanisms of benzene-induced genotoxicity have been studied to some extent in laboratory
animals. Schiestl et al. (1997) noted benzene-induced reversion of the mouse pink-eye unstable mutation
in premelanocytes of mouse embryos following intraperitoneal injection of the chemical into pregnant
dams; the reversions resulted from genomic DNA deletions. Results of Chen et al. (1994) and Eastmond
et al. (2001) indicate that benzene-induced micronuclei in mouse bone marrow erythrocytes are formed
predominantly from chromosome breakage, but also from aneuploidy.
BENZENE
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3. HEALTH EFFECTS
In vitro studies strongly imply that benzene's genotoxicity is derived primarily from its metabolites.
Positive results were obtained for gene mutation in Salmonella typhimurium (Glatt et al. 1989; Kaden et
al. 1979; Seixas et al. 1982) and sister chromatid exchange in human lymphocyte cell culture (Morimoto
1983) only when exogenous metabolic activators of benzene were used. Similarly, endogenous metabolic
activation was required for effects to be seen on DNA synthesis in rat hepatocyte culture (Glauert et al.
1985), DNA adduct formation in rat liver mitoplasts (Rushmore et al. 1984), and RNA synthesis in rat
liver mitoplasts and in rabbit and cat bone marrow mitoplasts (Kalf et al. 1982). Endogenous activation
occurs naturally by enzymes already within the cells. Exogenous activation requires the addition of
enzymes to cellular preparations. In one study using benzene and 13 possible metabolites, Glatt et al.
(1989) found that trans-1,2-dihydrodiol (with metabolic activators) and the diol epoxides (with or without
metabolic activators) produced histidine reversion in S. typhimurium. The same researchers also
investigated the genotoxicity of these 13 proposed metabolites in V79 Chinese hamster cells; anti-diol
epoxide, syn-diol epoxide, 1,2,3-trihydroxybenzene, 1,2,4-trihydroxybenzene, quinone, hydroquinone,
catechol, phenol, and 1,2-dihydrodiol were found to produce genotoxic effects ranging from sister
chromatid exchange and micronuclei increases to gene mutations (Glatt et al. 1989). Similar studies with
trans,trans-muconaldehyde showed that this metabolite is strongly mutagenic in V79 cells and weakly
mutagenic in bacteria (Glatt and Witz 1990). Chang et al. (1994) showed that muconaldehyde and its
aldehyde metabolites 6-hydroxy-trans,trans-2,4-hexadienal and 6-oxo-trans,trans-hexadienoic acid were
mutagenic in V79 cells.
Several studies revealed a possible connection between certain benzene metabolites and DNA damage via
the formation of oxygen radicals. Lewis et al. (1988) found that both 1,2,4-benzenetriol and
hydroquinone produce DNA strand breaks, 1,2,4-benzenetriol to a greater degree than hydroquinone.
Consistent with these findings was the observation that 1,2,4-benzenetriol generates a greater
concentration of oxygen radicals than hydroquinone. The study concluded that 1,2,4-benzenetriol
damages DNA by producing oxygen radicals, while hydroquinone probably exerts its genotoxic effects by
some other mechanism (Lewis et al. 1988). However, results of Li et al. (1995b) indicate that much of
the benzene-mediated DNA damage may result from the generation of reactive oxygen species via a
copper-redox cycling mechanism involved in the oxidation of the benzene metabolite, hydroquinone.
Increased recombination, which can lead to adverse genetic changes, was observed in Chinese hamster
ovary cells exposed to phenol, catechol, or benzoquinone (Winn 2003). Benzene had no effect on
recombination. The observed increases in recombination were abolished by the addition of catalase to the
cells, suggesting that the effect of the benzene metabolites was elicited by oxidative stress (Winn 2003).
BENZENE
153
3. HEALTH EFFECTS
Oxidative damage (manifested as DNA strand breaks) was observed in HL60 cells treated with
1,4-benzoquinone or 1,4-hydroquinone (Hiraku and Kawanishi 1996). 1,4-Benzoquinone and
hydroquinone induced DNA strand breaks in Chinese hamster ovary cells while phenol, catechol,
1,2,4-benzenetriol, trans,trans-muconic acid, and S-phenyl mercapturic acid did not (Sze et al. 1996). No
synergism between hydroquinone and the other metabolites was observed.
Phenol, catechol, hydroquinone, and benzene induced morphological transformation and gene mutations
in Syrian hamster embryo cells (Tsutsui et al. 1997). Chromosomal aberrations, sister chromatid
exchange, and unscheduled DNA synthesis were increased by the benzene metabolites while aneuploidy
was observed in cells treated with benzene or catechol (Tsutsui et al. 1997). There was no significant
increase (p>0.005) in DNA fragmentation in human respiratory epithelial cells exposed to an atmosphere
of 5 mg benzene/m3 for 8 hours, although there was evidence of an inflammatory response (Gosepath et
al. 2003). Chromosomal breaks and hyperdiploidy were observed in human lymphocytes after exposure
to hydroquinone in vitro (Eastmond et al. 1994). Aneusomy of chromosomes 7 and 8 were observed in
human umbilical cord blood cells treated with hydroquinone (2, 10, or 50 M) for 72 hours (Smith et al.
2000). Monosomy 7 and trisomy 8 are two common clonal aberrations observed in myeloid leukemias
(Smith et al. 2000). Hydroquinone (2649 M) also induced monosomy of chromosomes 5, 7, and 8 in a
human lymphoblast cell line (Stillman et al. 1997). Hydroquinone (10100 M) and 1,2,4-benzenetriol
(1050 M) significantly increased monosomy 5 and 7 in human lymphocytes and long-arm deletions in
chromosomes 5 and 7 (Zhang et al. 1998b). Benzene metabolites have been shown to form DNA adducts
in human bone marrow and HL-60 cells (Bodell et al. 1993; Lvay and Bodell 1992). Zhang et al. (1993)
showed that 1,2,4-benzenetriol increased the frequency of micronuclei formation in human lymphocytes
in culture, and in HL60 cells in a dose-related manner. An increase in the level of oxidative damage to
DNA was also noted in HL60 cells in culture. Extracts from human cells have been shown to have repair
activity toward benzoquinone-DNA adducts in vitro (Chenna et al. 1995). Benzene (15 mM) did not
elicit micronuclei formation in whole blood cells treated for 48 hours with or without metabolic activation
with 10% S9 rat liver fraction for 2 hours (Zarani et al. 1999). The assay was conducted with blood
collected from four subjects. Chen and Eastmond (1995) showed that benzene metabolites can adversely
affect human topoisomerases, enzymes involved in DNA replication and repair. No effect of any
metabolite was seen on human topoisomerase I or for topoisomerase II for hydroquinone, phenol,
2,2'-biphenol, 4,4'-biphenol, and catechol at concentrations as high as 500 M. 1,4-Benzoquinone and
1,2,4-benzenetriol inhibited human topoisomerase II in vitro, at 500 and 250 M without bioactivation.
However, following bioactivation, phenol and 2,2'-biphenol showed inhibitory effects at doses as low as
50 M, whereas 4,4'-biphenol inhibited topoisomerase II at concentrations of 10 M. More recently,
BENZENE
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3. HEALTH EFFECTS
Eastmond et al. (2001) demonstrated decreased activity of topoisomerase II activity in nucleated bone
marrow cells of mice administered benzene by oral gavage for subchronic durations.
Available in vitro data suggest that benzene itself is genotoxic. Two studies reported that benzene
produced DNA breaks in Chinese hamster ovary cells independent of metabolic activators (Douglas et al.
1985; Lakhanisky and Hendrickx 1985). In a study by Aubrecht et al. (1995), benzene was shown to
induce intrachromosomal recombination in human lymphoblastoid cell culture. Therefore, benzene
appears to have some genotoxic capabilities of its own, but its metabolites seem to be the primary
genotoxins in systems in which normal metabolism is occurring. Refer to Table 3-5 for the results of
these and other in vitro studies.
In summary, chromosome aberrations have been found consistently in bone marrow cells of persons
occupationally exposed to benzene. The conclusion, based on human epidemiological studies, that
benzene is a human clastogen is well supported by in vivo animal studies and in vitro cell cultures and
subcellular studies. Virtually all studies that looked for effects at the chromosomal level were positive
when the ability to metabolize benzene was present. These experimental results are consistent with the
chromosomal damage seen in exposed humans. The leukemia observed in some benzene-exposed
persons may result from the appearance of a clone of chromosomally abnormal cells in the bone marrow.
With respect to genetic effects, no safe human exposure level can be determined from available
epidemiological data. Significant increases in sister chromatid exchanges were produced in bone marrow
cells and lymphocytes of animals. The significance of sister chromatid exchanges is unknown, but their
production by a chemical is generally considered to indicate a genotoxic potential. Exposures generally
occur via inhalation, and based on animal studies, effects following oral exposure may be greater than
effects following inhalation exposure to comparable levels of benzene. Data presented in this section and
elsewhere in this profile (Section 3.4) show that benzene metabolites are the genotoxic entities. It is
possible that each metabolite causes a different genotoxic effect. Differences in metabolic capability are
probably responsible for some of the variations in response to benzene seen in different test systems.
3.4
TOXICOKINETICS
The toxicokinetics of benzene has been extensively studied. Inhalation exposure is probably the major
route of human exposure to benzene, although oral and dermal exposure are also important. Benzene is
readily absorbed following inhalation or oral exposure. Although benzene is also readily absorbed from
the skin, a significant amount of a dermal application evaporates from the skin surface. Absorbed
BENZENE
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3. HEALTH EFFECTS
benzene is rapidly distributed throughout the body and tends to accumulate in fatty tissues. The liver
serves an important function in benzene metabolism, which results in the production of several reactive
metabolites. Although it is widely accepted that benzene toxicity is dependent upon metabolism, no
single benzene metabolite has been found to be the major source of benzene hematopoietic and
leukemogenic effects. At low exposure levels, benzene is rapidly metabolized and excreted
predominantly as conjugated urinary metabolites. At higher exposure levels, metabolic pathways appear
to become saturated and a large portion of an absorbed dose of benzene is excreted as parent compound in
exhaled air. Benzene metabolism appears to be qualitatively similar among humans and various
laboratory animal species. However, there are quantitative differences in the relative amounts of benzene
metabolites.
3.4.1
Absorption
3.4.1.1 Inhalation Exposure

Inhalation exposure is probably the major route of human exposure to benzene, and numerous studies of
absorption of benzene after inhalation exposure in different settings have been conducted (Ashley et al.
1994; Avis and Hutton 1993; Boogaard and van Sittert 1995; Brunnemann et al. 1989; Byrd et al. 1990;
Etzel and Ashley 1994; Fustinoni et al. 1995; Ghittori et al. 1995; Gordian and Guay 1995; Hajimiragha
et al. 1989; Hanzlick 1995; Karacic et al. 1995; Kok and Ong 1994; Lagorio et al. 1994a; Laitinen et al.
1994; Lauwerys et al. 1994; Lindstrom et al. 1994; Mannino et al. 1995; Nomiyama and Nomiyama
1974a; Ong et al. 1994, 1995; Pekari et al. 1992; Popp et al. 1994; Rauscher et al. 1994; Rothman et al.
1995; Ruppert et al. 1995; Scherer et al. 1995; Shamy et al. 1994; Srbova et al. 1950; Yu and Weisel
1996). Existing evidence indicates that benzene is rapidly absorbed by humans following inhalation
exposure. Results from a study of 23 subjects who inhaled 47110 ppm benzene for 23 hours showed
that absorption was highest in the first few minutes of exposure, but decreased rapidly thereafter (Srbova
et al. 1950). In the first 5 minutes of exposure, absorption was 7080%, but by 1 hour, it was reduced to
approximately 50% (range, 2060%). Respiratory uptake (the amount of benzene absorbed from the
lungs following inhalation of the vapors) in six volunteers including males and females exposed to 52
62 ppm benzene for 4 hours was determined to be approximately 47%(Nomiyama and Nomiyama 1974a).
In a similar study, three healthy nonsmoking volunteers were exposed to benzene at levels of 1.6 or
9.4 ppm for 4 hours (Pekari et al. 1992). The amount of benzene absorbed was estimated from the
difference between the concentration inhaled and the concentration exhaled. Estimates were 48% for the
high dose and 52% for the low dose, supporting the evidence of Nomiyama and Nomiyama (1974a). Yu
and Weisel (1996) measured the uptake of benzene by three female subjects exposed to benzene in smoke
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generated by burning cigarettes, which resulted in airborne benzene concentrations in the range of 3269
ppm. Average absorption for exposure periods of 30 or 120 minutes was 64% and did not appear to be
influenced by exposure duration.
Studies of occupational exposure to benzene suggest that absorption occurs both by inhalation and
dermally in many workplace settings. In a study conducted in 1992 in Finland, car mechanics exposure
to benzene was evaluated (Laitinen et al. 1994). Different work phases were measured at five Finnish
garages. Blood samples from car mechanics (eight nonsmokers) were taken 39 hours after exposure to
benzene. The results were approximated to the time point of 16 hours after exposure. Fourteen air
samples were taken from the breathing zone and five stationary samples were collected from the middle
of the garage for background concentration levels. The average background concentration (stationary
samples) of gasoline vapors was 67 cm3/m3 (22 ppm) and the concentration of benzene was under the
detection limit of 0.2 cm3/m3 (0.1 ppm). The concentrations of benzene in the breathing zone varied from
the detection limit of 0.2 cm3/m3 to 1.3 cm3/m3 (0.10.4 ppm) for unleaded gasoline and from the
detection limit to 3.7 cm3/m3 (1.2 ppm) for leaded gasoline. The highest benzene exposure level (2.4
3.7 cm3/m3 or 0.81.2 ppm) was measured when changing the filter to the fuel pump. The mechanics
worked without protective gloves, and the risk of contamination and penetration through the skin was
significant. During carburetor renewal and gathering, benzene concentrations were 0.51.1 cm3/m3 (0.2
0.3 ppm). During changing of the fuel filter to electronic fuel-injection system, benzene concentration
ranged from 0.9 to 3.4 cm3/m3 (0.31.1 ppm). The approximated benzene concentrations in blood
corresponding to the time point of 16 hours after the exposure showed much higher levels of exposure
than could be expected according to the corresponding air measurements (8-hour TWA). The comparison
of expected benzene concentrations in blood, if no dermal exposure were present, to the levels at the time
point of 16 hours after the exposure showed that the dermal route must be the source of about 68% of
exposure (range 1.188.2%). Two of eight workers had minimal exposure through the skin (01.1%).
The other six workers showed high dermal exposure (79.4%).
Exposure to benzene-contaminated water can also provide an opportunity for both inhalation and dermal
absorption. In a series of experiments conducted in a single-family residence from June 11 to 13, 1991,
exposure to benzene through contaminated residential water was monitored (Lindstrom et al. 1994). The
residential water was contaminated with benzene and other hydrocarbons in 1986. Periodic testing
conducted from 1986 to 1991 showed benzene concentrations ranging from 33 to 673 g/L (ppb). The
experiment involved an individual taking a 20-minute shower with the bathroom door closed, followed by
5 minutes for drying and dressing; then the bathroom door was opened and this individual was allowed to
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leave the house. Integrated 60- and 240-minute whole-air samples were collected from the bathroom, an
adjacent bedroom, living room, and in ambient air. Glass, gas-tight syringe grab samples were
simultaneously collected from the shower, bathroom, bedroom, and living room at 0, 10, 18, 20, 25, 25.5,
and 30 minutes. Two members of the monitoring team were measured for 6 hours using personal Tenax
gas GC monitors. For the first 30 minutes of each experiment, one member was based in the bathroom
and the other in the living room. Benzene concentrations in the shower head ranged from 185 to
367 g/L (ppb), while drain level samples ranged from below the detectable limit (0.6 g/L or ppb) to
198 g/L (ppb). Analysis of the syringe samples suggested a pulse of benzene moving from the shower
stall to the rest of the house over approximately 60 minutes. Peak levels of benzene measured 758
1,670 g/m3 (235518 ppb) in the shower stall at 1820 minutes, 366498 g/m3 (113154 ppb) in the
bathroom at 1025 minutes, 81146 g/m3 (2545 ppb) in the bedroom at 25.530 minutes, and 40
62 g/m3 (1219 ppb) in the living room at 3670 minutes. The individual who took the 20-minute
shower had estimated inhalation doses of 79.6, 105, and 103 g (mean=95.9 g) for the 3 consecutive
sampling days. These doses were estimated by taking the products of the concentration of benzene in
water, the minute ventilation rate, the duration of exposure, and a 70% benzene absorption factor. This
was 2.14.9 times higher than corresponding 20-minute bathroom exposures. Adding the average dose
absorbed in the bathroom during the 5.5 minutes following the shower (using the overall 2025 minutes
mean syringe level of 318 g/m3 [99 ppb]) gave a total average shower-related inhalation dose of 113 g.
An average dermal dose of 168 g was estimated for the 20-minute shower by multiplying the average
concentration of benzene in water by the surface area of the male volunteer, an exposure factor of 75%
body surface area exposed, a dermal permeability constant for benzene of 0.11 cm/hour, an exposure
duration of 0.33 hours, and a unit conversion factor of 11/1,000 cm2. The total benzene dose resulting
from the shower was estimated to be approximately 281 g (40% via inhalation and 60% via dermal),
suggesting a higher potential exposure to benzene via dermal contact from the water than through
vaporization and inhalation. This exposure was 23.5 times higher than the mean 6-hour inhalation dose
received by the sampling members. The estimated inhalation and dermal doses reported by Lindstrom et
al. (1994) have not been validated by others and are therefore of questionable value for quantitative
analysis.
Additional evidence of benzene absorption following inhalation exposure comes from data on cigarette
smokers. Benzene levels were significantly higher in the venous blood of 14 smokers (median level of
493 ng/L) than in a control group of 13 nonsmokers (median level of 190 ng/L) (Hajimiragha et al. 1989).
Cigarette smoke is known to contain benzene (Brunnemann et al. 1989; Byrd et al. 1990), and the subjects
had no known exposure to other sources of benzene (Hajimiragha et al. 1989). Kok and Ong (1994)
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report blood and urine levels of benzene as 110.9 and 116.4 ng/L, respectively for nonsmokers, and
328.8 and 405.4 ng/L, respectively for smokers. The National Association of Medical Examiners
Pediatric Toxicology (PedTox) Registry reported blood benzene concentrations ranging from 0.2 to
4.9 mg/L in eight children who died in fires and were dead at the scene, indicating absorption of benzene
from burning materials (Hanzlick 1995). Blood benzene levels taken from U.S. engineers (Group I) and
firefighters (Group II) working at burning oil wells in Kuwait were compared to blood benzene levels
from non-exposed U.S. citizens (Etzel and Ashley 1994). The median concentrations of benzene in whole
blood from Groups I, II, and U.S. reference group were 0.035 g/L (range ND0.055 g/L), 0.18 g/L
(range 0.0631.1 g/L), and 0.066 g/L (range ND0.54 g/L), respectively. The median concentration
in group II was generally higher than the median concentrations in group I or the reference group.
Statistically significant higher concentrations of benzene (p<0.0001) were found in group II smokers than
in Group II nonsmokers.
Animal data confirm that benzene is rapidly absorbed through the lungs. Inhalation studies with
laboratory dogs indicate that distribution of benzene throughout the animal's body is rapid, with tissue
values dependent on blood supply. A linear relationship existed between the concentration of benzene in
air (2001,300 ppm) and the equilibrium concentration in blood (Schrenk et al. 1941). At these
exposures, the concentrations of benzene in the blood of dogs exposed to benzene reached a steady state
within 30 minutes.
In rodents, the extent of uptake increased linearly with concentration for exposures up to 200 ppm. At
concentrations of >200 ppm, zero-order kinetics were observed (i.e., uptake became nonlinear, indicating
saturation of the metabolic capacity). The percentage of inhaled benzene that was absorbed and retained
during a 6-hour exposure period decreased from 33 to 15% in rats and from 50 to 10% in mice as the
exposure concentration was increased from about 10 to 1,000 ppm (Sabourin et al. 1987). When rats and
mice were exposed to approximately 300 ppm, mice had greater uptake than rats. Mice and rats had
different absorption characteristics; the cumulative inhaled dose in mice was greater than that in rats
(Eutermoser et al. 1986; Sabourin et al. 1987). Purebred Duroc-Jersey pigs were exposed to 0, 20, 100,
and 500 ppm benzene vapors 6 hours/day, 5 days/week for 3 weeks (Dow 1992). The average
concentration of phenol in the urine increased linearly with dose.
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3.4.1.2 Oral Exposure

Although definitive scientific data are not available on oral absorption of benzene in humans, case studies
of accidental or intentional poisoning indicate that benzene is absorbed by the oral route (Thienes and
Haley 1972).
Benzene appears to be efficiently absorbed following oral dosing in animals. Oral absorption of benzene
was first demonstrated by Parke and Williams (1953a). After radiolabeled (14C) benzene was
administered orally to rabbits (340500 mg/kg), the total radioactivity eliminated in exhaled air and urine
accounted for approximately 90% of the administered dose, indicating that at least this much of the
administered dose was absorbed. Studies in rats and mice showed that gastrointestinal absorption was
greater than 97% in both species when the animals were administered benzene by gavage (in corn oil) at
doses of 0.5150 mg/kg/day (Sabourin et al. 1987). In many animal studies, benzene is administered
orally in oil to insure predictable solubility and dose concentration control. This is unlike the predicted
human oral exposure, which is likely to be in drinking water. There are a number of studies in which
benzene has been administered to animals in the drinking water, which more closely resembles predicted
human oral exposure (Lindstrom et al. 1994). Although no information was located regarding the extent
of oral absorption of benzene in aqueous solutions, it is reasonable to assume that oral absorption from
water solutions would be nearly 100%.
The bioavailability of pure as opposed to soil-adsorbed benzene was conducted in adult male rats (Turkall
et al. 1988). Animals were gavaged with an aqueous suspension of benzene alone, or adsorbed to clay or
sandy soil. Plasma concentration, half-life, tissue distribution, respiratory excretion, and urinary
excretion were monitored. Peak plasma concentration of radioactivity was increased in the presence of
either soil as opposed to benzene alone, while sandy soil also decreased the time to peak plasma
concentration as opposed to benzene alone. Soil increased the area under the plasma radioactivity-time
curve as opposed to benzene alone, a difference that was significant with clay soil. The half-life in
plasma was not affected by soil.
3.4.1.3 Dermal Exposure
Studies conducted in vivo in humans and in vitro using human skin indicates that benzene can be
absorbed dermally. The movement of a substance through the skin to the blood occurs by passive
diffusion and has been described mathematically by Fick's law. However, this is an oversimplification of
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the process of skin absorption; various factors (e.g., interaction of benzene with molecules within the
skin) affect the transport of the solvent through the skin (Lodn 1986).
In vivo experiments on four volunteers, to whom 0.0026 mg/cm2 of 14C-benzene was applied to forearm
skin, indicated that approximately 0.05% of the applied dose was absorbed (Franz 1984). Absorption was
rapid, with more than 80% of the total excretion of the absorbed dose occurring in the first 8 hours after
application. Calculations were based on urinary excretion data and no correction was made for the
amount of benzene that evaporated from the applied site before absorption occurred. In addition, the
percentage of absorbed dose excreted in urine that was used in the calculation was based only on data
from rhesus monkeys and may not be accurate for humans. In another study, 3543 cm2 of the forearm
was exposed to approximately 0.06 g/cm2 of liquid benzene for 1.252 hours (Hanke et al. 1961). The
absorption was estimated from the amount of phenol eliminated in the urine. The absorption rate of liquid
benzene by the skin (under the conditions of complete saturation) was calculated to be low, approximately
0.4 mg/cm2/hour. The absorption due to vapors in the same experiment was negligible. The results
indicate that dermal absorption of liquid benzene is of concern, while dermal absorption from vapor
exposure may not be of concern because of the low concentration of benzene in vapor form at the point of
contact with the skin. No signs of acute intoxication due to liquid benzene dermally absorbed were noted.
These results confirm that benzene can be absorbed through skin. However, non-benzene-derived phenol
in the urine was not accounted for.
Studies of occupational exposure to benzene suggest that absorption occurs both by inhalation and
dermally in many workplace settings. In a study conducted in 1992 in Finland, car mechanics exposure
to benzene was evaluated (Laitinen et al. 1994). Different work phases were measured at five Finnish
garages. Blood samples from car mechanics (eight nonsmokers) were taken 39 hours after exposure to
benzene. The results were approximated to the time point of 16 hours after exposure. Fourteen air
samples were taken from the breathing zone and five stationary samples were collected from the middle
of the garage for background concentration levels. The mechanics worked without protective gloves, and
the risk of contamination and penetration through the skin was significant. The approximated benzene
concentrations in blood corresponding to the time point of 16 hours after the exposure showed much
higher levels of exposure than could be expected according to the corresponding air measurements
(8 hour TWA). The comparison of expected benzene concentrations in blood, if no dermal exposure was
present, to the levels at the time point of 16 hours after the exposure showed that the dermal route must be
the source of about 68% of exposure (range 1.188.2%). Two of eight workers had minimal exposure
through the skin (01.1%). The other six workers showed high dermal exposure (79.4%).
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Exposure to benzene-contaminated water can also provide an opportunity for both inhalation and dermal
absorption. In a series of experiments conducted in a single-family residence from June 11 to 13, 1991,
exposure to benzene through contaminated residential water was monitored (Lindstrom et al. 1994). The
residential water was contaminated with benzene and other hydrocarbons in 1986. Exposure was
monitored for a person taking a 20-minute shower and for people in other parts of the house during and
after the shower. An average dermal dose of 168 g was estimated for a 20-minute shower using this
water. The total benzene dose resulting from the shower was estimated to be approximately 281 g (40%
via inhalation and 60% via dermal), suggesting a higher potential exposure to benzene via dermal contact
from the water than through vaporization and inhalation (see Section 3.4.1.1 for a more detailed
discussion). This exposure was 23.5 times higher than the mean 6-hour inhalation dose received by the
sampling team members in other parts of the house. The estimated inhalation and dermal doses reported
by Lindstrom et al. (1994) have not been validated by others and are therefore of questionable value for
quantitative analysis.
In vitro experiments using human skin support the fact that benzene can be absorbed dermally. An
experiment on the permeability of excised human skin with regard to benzene (specific activity
99.8 mCi/mmol; total volume of applied benzene not reported) resulted in the absorption of 0.17 mg/cm2
after 0.5 hours and 1.92 mg/cm2 after 13.5 hours (Lodn 1986). Following application of 5, 120, 270, and
520 L/cm2 of benzene to human skin, total absorption was found to be 0.01, 0.24, 0.56, and 0.9 L/cm2,
respectively. Thus, the total amount absorbed appears to increase linearly with dose. The study author
indicated that evaporation of benzene did not exceed 5%. When exposure time (i.e., the time to complete
evaporation) at each dose was measured and plotted as the ordinate of absorption, total absorption was
found to increase linearly with exposure time. The percentage of the applied dose absorbed at each
concentration was constant at about 0.2% (Franz 1984).
Using results from an in vitro study, it was estimated that an adult working in ambient air containing
10 ppm benzene would absorb 7.5 L/hour from inhalation and 1.5 L/hour from whole-body (2 m2)
dermal exposure (Blank and McAuliffe 1985). It was also estimated that 100 cm2 of smooth and bare
skin in contact with gasoline containing 5% benzene would absorb 7.0 L/hour. Diffusion through the
stratum corneum was considered the most likely rate-limiting step for dermal absorption because of
benzene's low water solubility (Blank and McAuliffe 1985).
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Based on an observational study of workers in a tire factory, it was estimated that a worker exposed to
benzene as a result of direct skin contact with petroleum naphtha containing 0.5% benzene could absorb
48 mg of benzene per day through intact skin (Susten et al. 1985). This amount absorbed was compared
with an estimated 14 mg of benzene absorbed as a result of inhalation of 1 ppm for an 8-hour day. The
estimate for dermal absorption is theoretical since in many facilities the concentration of benzene in
rubber solvents such as petroleum naphtha is less than 0.5% and may be as low as 0.09%.
Benzene is also absorbed dermally by animals. In Rhesus monkeys, minipigs, and hairless mice, dermal
absorption was <1% following a single direct (unoccluded) application of liquid benzene (Franz 1984;
Maibach and Anjo 1981; Susten et al. 1985). As with humans, absorption appeared to be rapid, with the
highest urinary excretion of the absorbed dose observed in the first 8 hours following exposure (Franz
1984). Multiple applications, as well as application to stripped skin, resulted in greater skin penetration
(Maibach and Anjo 1981). The percentage of absorption of the applied dose of benzene in each of these
animals was approximately 23-fold higher than that of humans.
Data indicate that soil adsorption decreases the dermal bioavailability of benzene. A study in which male
rats were treated dermally with 0.004 mg/cm2 14C-benzene, with or without 1 g of clay or sandy soil,
reported benzene absorption half-lives of 3.1, 3.6, and 4.4 hours for pure benzene, sandy soil, and clay
soil, respectively (Skowronski et al. 1988).
Benzene in air was rapidly absorbed through the skin of hairless mice that were attached to respirators to
avoid pulmonary uptake of the benzene vapors (Tsuruta 1989). The rate of absorption of benzene through
the skin increased linearly with dose. The skin absorption rate for 200 ppm was 4.11 nmol/cm2/hour
(0.31 g/cm2/hour); at 1,000 ppm, the rate was 24.2 nmol/cm2/hour (1.89 g/cm2/hour), and at 3,000 ppm,
the rate was 75.5 nmol/cm2/hour (5.90 g/cm2/hour). The skin absorption coefficient was 0.619 cm/hour.
McDougal et al. (1990) estimated permeability constants of 0.15 and 0.08 cm/hour for rat and human
skin, respectively, based on the appearance of benzene in the blood of rats dermally exposed to benzene
vapors at a concentration of 40,000 ppm for 4 hours. A physiologically based pharmacodynamic (PBPK)
model was used to estimate the permeability of the vapor in rat and human skin. These results indicate
that dermal absorption of benzene may be greater in rats than humans. Therefore, results in rats may
provide a conservative estimate of dermal absorption of benzene in humans.
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In an in vitro experiment using Fischer 344 rat skin, the partition coefficient for skin:air was determined
for benzene at 203 ppm (Mattie et al. 1994). The partition coefficient of a chemical in skin is an indicator
of the capacity of the skin to retain the chemical, and may reflect the rate at which a chemical is absorbed
through the skin and enters the circulation. Results indicated a partition coefficient of 35, with an
equilibration time of 4 hours. The skin:air partition coefficient is necessary for developing the dermal
compartment of a PBPK model.
Based on data for skin absorption of benzene vapors in mice and occupational exposure data, Tsuruta
(1989) estimated the ratio of skin absorption rate to pulmonary uptake for humans exposed to benzene to
be 0.037. Dermal absorption could account for a relatively higher percentage of total benzene uptake in
occupational settings where personnel, using respirators but not protective clothing, are exposed to high
concentrations of benzene vapor.
3.4.2
Distribution

Information on the distribution of benzene in humans comes primarily from case studies. The data
suggest that benzene is distributed throughout the body following absorption into blood. Since benzene is
lipophilic, a high distribution to fatty tissue might be expected. Following inhalation exposure to
benzene, the chemical has been detected in the biological fluids and tissues of the subjects (Pekari et al.
1992; Tauber 1970; Winek and Collom 1971; Winek et al. 1967). Fluid and tissue levels of benzene have
been reported in cases of both accidental and intentional lethal exposures. Levels of 0.38 mg% in blood
(mg% = mg per 100 mL of blood or mg per 100 g of tissue), 1.38 mg% in the brain, and 0.26 mg% in the
liver were reported in a worker who died from exposure to very high air concentrations of the chemical
(Tauber 1970). An autopsy (time after death not indicated) performed on a youth who died while sniffing
reagent-grade benzene revealed benzene concentrations of 2.0 mg% in blood, 3.9 mg% in brain, 1.6 mg%
in liver, 1.9 mg% in kidney, 1 mg% in stomach, 1.1 mg% in bile, 2.23 mg% in abdominal fat, and
0.06 mg% in urine (Winek and Collom 1971). Benzene crosses the human placenta and is present in the
cord blood in amounts equal to or greater than those in maternal blood (Dowty et al. 1976). Benzene is
expected to readily bind to plasma proteins (Travis and Bowers 1989). Furthermore, benzene metabolites
have been found to form covalent adducts with proteins from blood in humans (Bechtold et al. 1992b;
Rappaport et al. 2002a, 2002b; Yeowell-OConnell et al. 1998) and mice (McDonald et al. 1994). The
relatively widespread distribution of benzene and its metabolites to other tissues and organs indicates that
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protein adduct formation in the blood does not adversely affect distribution, although no confirming
studies were located.
Results from animal studies indicate that absorbed benzene is distributed among several compartments.
The parent compound is preferentially stored in the fat, although the relative uptake in tissues also
appears to be dependent on the perfusion rate of tissues by blood.
Following a 10-minute inhalation exposure of pregnant mice to 2,000 ppm benzene, parent compound and
its metabolites were found to be present in lipid-rich tissues, such as brain and fat, and in well-perfused
tissues, such as liver and kidney. Benzene was also found in the placenta and fetuses immediately
following inhalation of benzene (Ghantous and Danielsson 1986). During inhalation exposure of rats to
500 ppm, benzene levels reached a steady-state concentration within 4 hours in blood (11.5 g/mL),
6 hours in fat (164.4 g/g), and less than 2 hours in bone marrow (37.0 g/g) (Rickert et al. 1979).
Benzene was also distributed to the kidney, lung, liver, brain, and spleen. The benzene metabolites
phenol, catechol, and hydroquinone were detected in blood and bone marrow following 6 hours of
exposure to benzene, with levels in bone marrow exceeding the respective levels in blood. The levels of
phenol in blood and bone marrow decreased much more rapidly after exposure ceased than did those of
catechol or hydroquinone, suggesting the possibility of accumulation of the latter two compounds.
Benzene was rapidly distributed throughout the bodies of dogs exposed via inhalation to concentrations of
800 ppm for up to 8 hours/day for 822 days (Schrenk et al. 1941). Fat, bone marrow, and urine
contained about 20 times the concentration of benzene in blood; benzene levels in muscles and organs
were 13 times that in blood; and erythrocytes contained about twice the amount of benzene found in
plasma. During inhalation exposure of rats to 1,000 ppm (2 hours/day, for 12 weeks), benzene was stored
longer (and eliminated more slowly) in female and male rats with higher body fat content than in leaner
animals (Sato et al. 1975).
Benzene was detected in the liver, lung, and blood of rats and mice examined immediately following a
6-hour exposure to benzene vapors at a concentration of 50 ppm (Sabourin et al. 1988). Sabourin and
coworkers (Sabourin et al. 1987, 1988) also examined effects exposure concentration, exposure rate, and
route of administration on the comparative metabolism of benzene in rats and mice. Results of these
studies are summarized in Section 3.4.3 (Metabolism).
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No studies were located regarding distribution in humans after oral exposure to benzene.
In Sprague-Dawley rats administered a single dose of 0.15, 1.5, 15, 150, or 500 mg/kg of 14C-benzene by
gavage, benzene was rapidly absorbed and distributed to various organs and tissues within 1 hour of
administration (Low et al. 1989). One hour after rats were dosed with 0.15 or 1.5 mg/kg of benzene,
tissue distribution of benzene was highest in liver and kidney, intermediate in blood, and lowest in the
Zymbal gland, nasal cavity tissue, and mammary gland. At higher doses, beginning with 15 mg/kg,
benzene disproportionately increased in the mammary glands and bone marrow. Bone marrow and
adipose tissue proved to be depots of benzene at the higher dose levels. The highest tissue concentrations
of benzenes metabolite hydroquinone 1 hour after administration of 15 mg/kg of benzene were in the
liver, kidney, and blood, while the highest concentrations of the metabolite phenol were in the oral cavity,
nasal cavity, and kidney. The major tissue sites of benzenes conjugated metabolites were blood, bone
marrow, oral cavity, kidney, and liver for phenyl sulfate and hydroquinone glucuronide; muconic acid
was also found in these sites. Additionally, the Zymbal gland and nasal cavity were depots for phenyl
glucuronide, another conjugated metabolite of benzene. The Zymbal gland is a specialized sebaceous
gland and a site for benzene-induced tumors. Therefore, it is reasonable to expect that lipophilic
chemicals like benzene would partition readily into this gland. However, benzene did not accumulate in
the Zymbal gland; within 24 hours after administration, radiolabel derived from 14C-benzene in the
Zymbal gland constituted less than 0.0001% of the administered dose.
The bioavailability of pure as opposed to soil-adsorbed benzene was conducted in adult male rats (Turkall
et al. 1988). Animals were gavaged with an aqueous suspension of 14C-benzene alone, or adsorbed to
clay or sandy soil. Two hours after exposure, stomach tissue contained the highest amount of
radioactivity, followed by fat in all treatment groups. No differences in tissue distribution patterns were
detected for the three treatments.
No studies were located regarding distribution in humans after dermal exposure to benzene.
A study of male rats treated dermally with 0.004 mg/cm2 of 14C-benzene, with and without 1 g of clay or
sandy soil, revealed soil-related differences in tissue distribution following treatment. The 14C activity
(expressed as a percentage of initial dose per g of tissue) 48 hours after treatment with soil-adsorbed
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benzene was greatest in the treated skin (0.0590.119%), followed by the kidney (0.024%) and liver
(0.0130.015%), in both soil groups. In the pure benzene group, the kidney contained the largest amount
of radioactivity (0.026%), followed by the liver (0.013%) and treated skin (0.11%) (Skowronski et al.
1988). In all three groups, <0.01% of the radioactivity was found in the following tissues: duodenum,
fat, bone marrow, esophagus, pancreas, lung, heart, spleen, blood, brain, thymus, thyroid, adrenal, testes,
untreated skin, and remaining carcass.
3.4.3
Metabolism
Although the metabolism of benzene has been studied extensively, the steps leading to benzene toxicity
are not yet fully understood. It is generally understood that both cancer and noncancer effects are caused
by one or more reactive metabolites of benzene. Available data indicate that metabolites produced in the
liver are carried to the bone marrow where benzene toxicity is expressed. Benzene metabolism may
occur, at least in part, in the bone marrow. Benzene metabolism has been demonstrated in isolated
perfused rabbit lung preparations (Powley and Carlson 2002). As discussed in detail in Section 3.5,
available evidence suggests that multiple benzene metabolites may collectively be responsible for the
expression of benzene toxicity.
Data regarding metabolism of benzene in humans are derived primarily from studies using inhalation
exposures. Benzene is excreted both unchanged via the lungs and as metabolites (but also as parent
compound in small amounts) in the urine. The rate and percentage of excretion via the lungs are
dependent on exposure dose and route. Qualitatively, the metabolism and elimination of benzene appear
to be similar in humans and laboratory animals, but no directly comparable studies are available
(Henderson et al. 1989; Sabourin et al. 1988).
The metabolic scheme shown in Figure 3-3 is based on results of numerous mechanistic studies of
benzene metabolism (see Henderson et al. 1989; Huff et al. 1989; and Ross 1996, 2000 for
comprehensive reviews of benzene metabolism). The first step is the cytochrome P-450 2E1 (CYP2E1)
catalyzed oxidation of benzene to form benzene oxide (Lindstrom et al. 1997), which is in equilibrium
with its oxepin (Vogel and Gnther 1967). Several pathways are involved in the metabolism of benzene
oxide. The predominant pathway involves nonenzymatic rearrangement to form phenol (Jerina et al.
1968), the major initial product of benzene metabolism (Parke and Williams 1953a). Phenol is oxidized
in the presence of CYP2E1 to catechol or hydroquinone, which are oxidized via myeloperoxidase (MPO)
to the reactive metabolites 1,2- and 1,4-benzoquinone, respectively (Nebert et al. 2002). The reverse
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Figure 3-3. Metabolic Pathways for Benzene

Benzene
dihydrodiol
OH
Benzene
Catechol
OH
DHDD
OH
OH
H3C
NH
OH
CYP2E1
EH
O
S
ADH
ALDH
FE/OH
CYP2E1
OH
O
GSH
S-Phenylmercapturic
acid
O
Benzene
oxide
OH
trans,trans-Muconaldehyde
trans,trans-Muconic acid
Benzene
oxepin
Rearrangement
Hydroquinone
Phenol
OH
Catechol
OH
OH
CYP2E1
CYP2E1
HO
NQO1
OH
CYP2E1
CYP2E1
MPO
MPO
NQO1
1,2,4-Benzenetriol
OH
O
O
1,4-Benzoquinone
HO
OH
O
1,2-Benzoquinone
ADH = alcohol dehydrogenase; ALDH = aldehyde dehydrogenase; CYP2E1 = cytochrome P-450 2E1;
DHDD = dihydrodiol dehydrogenase; EH = epoxide hydrolase; GSH = glutathione; MPO = myeloperoxidase;
NQ01 = NAD(P)H:quinone oxidoreductase
Source: adapted from Nebert et al. 2002; Ross 2000
BENZENE
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reaction (reduction of 1,2- and 1,4-benzoquinone to catechol and hydroquinone, respectively) is catalyzed
by NAD(P)H:quinone oxidoreductase (NQ01) (Nebert et al. 2002). Both catechol and hydroquinone may
be converted to the reactive metabolite 1,2,4-benzenetriol via CYP2E1 catalysis. Alternatively, benzene
oxide may undergo epoxide hydrolase-catalyzed conversion to benzene dihydrodiol and subsequent
dihydrodiol dehydrogenase-catalyzed conversion to catechol (Nebert et al. 2002; Snyder et al. 1993a,
1993b). Each of the phenolic metabolites of benzene (phenol, catechol, hydroquinone, and
1,2,4-benzenetriol) can undergo sulfonic or glucuronic conjugation (Nebert et al. 2002; Schrenk and Bock
1990); the conjugates of phenol and hydroquinone are major urinary metabolites of benzene (Sabourin et
al. 1989a; Wells and Nerland 1991). Other pathways of benzene oxide metabolism include: (1) reaction
with glutathione (GSH) to form S-phenylmercapturic acid (Nebert et al. 2002; Sabourin et al. 1988;
Schafer et al. 1993; Schlosser et al. 1993; Schrenk et al. 1992; van Sittert et al. 1993), and (2) ironcatalyzed ring-opening conversion to trans,trans-muconic acid, presumably via the reactive trans,transmuconaldehyde intermediate (Bleasdale et al. 1996; Nebert et al. 2002; Ross 2000; Witz et al. 1990b,
1990c, 1996).
Results of several studies provide strong evidence for the involvement of CYP2E1 in the oxidation of
benzene. For example, no signs of benzene-induced toxicity were observed in transgenic CYP2E1
knockout mice (that do not express hepatic CYP2E1 activity) following exposure to benzene vapors
(200 ppm, 6 hours/day for 5 days) that caused severe genotoxicity and cytotoxicity in wild-type mice
(Valentine et al. 1996a, 1996b). Pretreatment of mice with CYP inhibitors (toluene, propylene glycol,
-diethyl amino ethyl diphenyl propyl acetate hydrogen chloride [SKF-525A]) has been demonstrated to
reduce both benzene metabolite formation (Andrews et al. 1977; Gill et al. 1979; Ikeda et al. 1972; Tuo et
al. 1996) and resulting genotoxicity (DNA damage as assessed by the alkaline comet assay) in mice (Tuo
et al. 1996). Pretreatment with CYP inducers (3-methylcholanthrene and -naphthoflavone) increased
both benzene metabolism and benzene clastogenicity (Gad-El-Karim et al. 1986). Immunoinhibition
studies in rat and rabbit hepatic microsomes provide additional support to the major role of CYP2E in
benzene metabolism (Johansson and Ingelman-Sundberg 1988; Koop and Laethem 1992).
Occupationally exposed workers with a phenotype corresponding to rapid CYP2E1 metabolism were
more susceptible to benzene hematotoxicity than workers not expressing this phenotype (Rothman et al.
1997). In vitro studies using human liver microsomes demonstrate a positive correlation between
benzene metabolism and CYP2E1 activity (Nedelcheva et al. 1999; Seaton et al. 1994). Although
CYP2E1 appears to be the major catalyzing agent in initial benzene metabolism, other CYPs, such as
CYP2B1 and CYP2F2, may also be involved (Gut et al. 1996a, 1996b; Powley and Carlson 2000, 2001;
Sheets and Carlson 2004; Sheets et al. 2004; Snyder et al. 1993a, 1993b).
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CYPs involved in benzene metabolism are found in all tissues. However, the predominant repository is
the liver, which is considered to be the primary site of benzene metabolism. By demonstrating that partial
hepatectomy diminished both the rate of metabolism of benzene and its toxicity in rats exposed to
benzene via subcutaneous injection, Sammett et al. (1979) provided suggestive evidence that one or more
benzene metabolites formed in the liver are necessary for toxicity. In vitro studies have demonstrated that
pulmonary microsomes of humans and laboratory animals are capable of metabolizing benzene, which
appears to be catalyzed by both CYP2E1 and CYP2F2 (Powley and Carlson 1999, 2000; Sheets et al.
2004). There is some indication that CYP2E1-catalyzed benzene metabolism may also occur in bone
marrow, a major target tissue of benzene toxicity. Andrews et al. (1979) demonstrated that rabbit bone
marrow is capable of metabolizing benzene. Schnier et al. (1989) subsequently found that rabbit bone
marrow contains CYP2E1. Irons et al. (1980) demonstrated that benzene metabolism by rat bone marrow
(in situ) was complete and independent of metabolism by the liver, with concentrations of phenol greater
than catechol and hydroquinone. Although the total metabolism by bone marrow was limited (total
metabolites present were 25% of those in blood), the concentration of metabolites in the bone marrow
exceeded that in the blood. Similar studies have been conducted in mice (Ganousis et al. 1992).
Fibroblasts had elevated levels of glutathione-S-transferase activity relative to macrophages, whereas
macrophages had higher levels of UDP-glucuronyltransferase and peroxidase activity. These data suggest
that cell-specific metabolism of benzene in the marrow may contribute to the toxicity of benzene in this
tissue compartment. In addition, comparison of the detoxifying activities of rat and mouse bone marrow
stromal cells indicates that rats have higher levels of glutathione and quinone reductase, which are known
to play critical roles in modulating hydroquinone-induced toxicity; this suggests a metabolic basis for the
observed increased susceptibility of mice to benzene-induced hematotoxicity (Zhu et al. 1995). Bernauer
et al. (1999, 2000) recently noted the presence of CYP2E1 in bone marrow samples of mice (several
strains), rats, rabbits, and humans. However, although Irons et al. (1980) demonstrated that the isolated
perfused rat femur was capable of metabolizing a very small amount of benzene (approximately 0.0002%
of 14C-benzene was recovered as metabolites), neither benzene oxide nor phenol were detected in a test of
benzene metabolism using microsomal preparations of bone marrow from rats (Lindstrom et al. 1999),
indicating that bone marrow is not a likely source of initial metabolic oxidation for benzene. No studies
were located the potential for human bone marrow tissue to metabolize benzene.
Mouse liver microsomes and cytosol have been shown to catalyze ring opening in the presence of
nicotinamide adenine dinucleotide phosphate (NADPH) in vitro, producing trans,trans-muconaldehyde, a
six-carbon diene dialdehyde also referred to as muconic dialdehyde (Goon et al. 1993; Latriano et al.
BENZENE
170
3. HEALTH EFFECTS
1986), a known hematotoxin (Witz et al. 1985) and toxic metabolite of benzene (Henderson et al. 1989).
Metabolism of benzene and trans,trans-muconaldehyde in the isolated perfused rat liver indicated that
benzene was metabolized to muconic acid, a ring-opened metabolite of benzene (Grotz et al. 1994).
Trans,trans-muconaldehyde was metabolized to muconic acid and three other metabolites. These studies
indicate that ring-opening of benzene occurs in the liver. Other recent literature identifies the following
metabolites after incubation of benzene with mouse liver microsomes: phenol, hydroquinone,
trans,trans-muconaldehyde, 6-oxo-trans,trans-2,4-hexadienoic acid, 6-hydroxy-trans,trans-2,4-hexadienal, and 6-hydroxy-trans,trans-2,4-hexadienoic acid (Zhang et al. 1995a). -Hydroxymuconaldehyde,
a new metabolite, was also identified. Additional work by Zhang et al. (1995b) suggests that cis,cismuconaldehyde is formed first, followed by cis,trans-muconaldehyde, and finally converted to
trans,trans-muconaldehyde. Muconic dialdehyde has been shown to be metabolized in vivo in mice to
muconic acid (Witz et al. 1990c). These data suggest that muconic dialdehyde is the precursor of
muconic acid in animals exposed to benzene. Small amounts of muconic acid were found in the urine of
rabbits and mice that received oral doses of 14C-benzene (Gad-El-Karim et al. 1985; Parke and Williams
1953a). The percentage of this metabolite formed varied with the administered benzene dose and was
quite high at low doses (17.6% of 0.5 mg/kg benzene administered to C57BL/6 mice) (Witz et al. 1990c).
Other studies in animals support these results (Brondeau et al. 1992; Ducos et al. 1990; McMahon and
Birnbaum 1991; Sabourin et al. 1989a; Schad et al. 1992). This pathway also appears to be active in
humans (Bechtold and Henderson 1993; Ducos et al. 1990, 1992; Lee et al. 1993; Melikian et al. 1993,
1994). For instance, urine samples from male and female smokers and nonsmokers were obtained from
subjects who applied for life insurance (Melikian et al. 1994). Samples from pregnant women were
obtained during 735 weeks of pregnancy. Questionnaires were filled out on smoking history and
occupation. The levels of muconic acid and cotinine (a biomarker for cigarette smoking) in the urine for
the groups of pregnant and nonpregnant smokers and nonsmokers were compared with previously
reported data in male smokers. Results showed the mean levels of muconic acid in the groups of male,
female-nonpregnant, and female-pregnant smokers were 3.6-, 4.8-, and 4.5-fold higher than the mean
concentration of this acid in the nonsmoking groups. The differences in the mean muconic acid
concentrations between smoking and nonsmoking groups were significant in male, and nonpregnant and
pregnant female smokers. Mean concentrations of muconic acid levels in nonpregnant female smokers
are similar to that of male smokers. Mean concentrations of muconic acid in groups of 42 male smokers
and 53 female smokers were 0.220.03 and 0.240.02 mg/g creatinine, or 0.130.06 and
0.130.07 mg/mg cotinine, respectively. Mean concentrations of muconic acid in groups of 63 pregnant
and 53 nonpregnant female smokers were 0.270.04 and 0.240.02 mg/g creatinine, or 0.240.06 and
0.130.07 mg/mg cotinine, respectively. Because of its relative importance in benzene toxicity,
BENZENE
171
3. HEALTH EFFECTS
additional modeling studies, including molecular orbital studies, have been conducted to further describe
how trans,trans-muconaldehyde is transformed to muconic acid (Bock et al. 1994).
Kenyon et al. (1995) compared their urinary profile of metabolites in B6C3F1 mice after oral dosing with
phenol with the results of Sabourin et al. (1989a) who administered a comparable oral dose of benzene to
B6C3F1 mice. The analysis of Kenyon et al. (1995) indicated that phenol administration resulted in lower
urinary levels of hydroquinone glucuronide, and higher levels of phenol sulfate and phenol glucuronide
compared to benzene administration. Kenyon et al. (1995) hypothesized that the differences in the
urinary metabolite profiles between phenol and benzene after oral dosing were due to zonal differences in
the distribution of metabolizing enzymes within the liver. Conjugating enzymes are more concentrated in
the periportal area of the liver, the first region to absorb the compound, whereas oxidizing enzymes are
more concentrated in the pericentral region of the liver. Based on this hypothesis, during an initial pass
through the liver after oral administration, phenol would have a greater opportunity to be conjugated as it
was absorbed from the gastrointestinal tract into the periportal region of the liver, thus resulting in less
free phenol being delivered into the pericentral region of the liver to be oxidized. With less free phenol
available for oxidation, less hydroquinone would be produced, relative to conjugated phenol metabolites.
In contrast, benzene must be oxidized before it can be conjugated. Therefore, metabolism of benzene
would be minimal in the periportal region of the liver, with most of the benzene reaching the pericentral
region to be oxidized to hydroquinone. Based on this scheme, the authors suggest that benzene
administration would result in more free phenol being delivered to oxidizing enzymes in the pericentral
region of the liver than administration of phenol itself (Kenyon et al. 1995).
Benzene has been found to stimulate its own metabolism, thereby increasing the rate of toxic metabolite
formation. Pretreatment of mice, rats, and rabbits subcutaneously with benzene increased benzene
metabolism in vitro without increasing CYP2E1 concentrations (Arinc et al. 1991; Gonasun et al. 1973;
Saito et al. 1973). In contrast, there was no significant effect on the metabolism of benzene when
Fischer 344 rats and B6C3F1 mice, pretreated with repeated inhalation exposure to 600 ppm of benzene,
were again exposed to 600 ppm benzene (Sabourin et al. 1990). The rate of benzene metabolism can be
altered by pretreatment with various compounds. Benzene is a preferential substrate of CYP2E1, which
also metabolizes alcohol and aniline. CYP2E1 can be induced by these substrates and is associated with
the generation of hydroxyl radicals, probably via futile cycling of the cytochrome (Chepiga et al. 1991;
Parke 1989; Snyder et al. 1993a, 1993b). It is possible that hydroxy radical formation by CYP2E1 may
play a role in the benzene ring-opening pathway, leading to the formation of trans,trans-muconaldehyde.
Phenol, hydroquinone, benzoquinone, and catechol have also been shown to induce CYPs in human
BENZENE
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3. HEALTH EFFECTS
hematopoietic stem cells (Henschler and Glatt 1995). Therefore, exposure to chemicals that stimulate the
activity of this enzyme system prior to exposure to benzene could increase the rate of benzene
metabolism.
Both NADPH-linked and ascorbate-induced lipid peroxidation activities induced in vitro were lowered
5.5 and 26%, respectively, in rats following oral administration of 1,400 mg/kg/day of benzene for 3 days,
followed by intraperitoneal injection of phenobarbital. These results suggest that benzene alters hepatic
drug metabolism and lipid peroxidation. The decrease in lipid peroxidation could be due to the
antioxidant property of the metabolites (Pawar and Mungikar 1975).
The ultimate disposition and metabolic fate of benzene depends on animal species, dose, and route of
exposure. The dose of benzene affects both the total metabolism and the concentrations of individual
metabolites formed. In mice, the percentage of hydroquinone glucuronide decreased as the dose
increased. In both rats and mice, the percentage of muconic acid decreased as the dose increased. The
shift in metabolism may affect the dose-response relationship for toxicity, and has been observed in all
animal species studies thus far (Sabourin et al. 1989a, 1992; Witz et al. 1990b, 1990c). The effect of
species differences in metabolism of inhaled benzene was evidenced by the fact that mice have a higher
minute volume per kg body weight than rats (1.5 times higher). This caused the blood concentration of
benzene to reach equilibrium more quickly in mice than in rats, but the steady-state level in blood was not
influenced (Sabourin et al. 1987). Species differences in benzene metabolism following oral exposure
were elucidated in rats and mice administered benzene by gavage at doses of 0.5150 mg/kg/day
(Sabourin et al. 1987). At doses below 15 mg/kg, >90% of the benzene was metabolized, while at doses
above 15 mg/kg, an increasing percentage of orally administered benzene was exhaled unmetabolized.
Total metabolites per unit body weight were equal in rats and mice at doses up to 50 mg/kg/day.
However, total metabolites in mice did not increase at higher doses, suggesting saturation of metabolic
pathways (Sabourin et al. 1987).
The integrated dose to a tissue over a 14-hour period (6-hour exposure, 8 hours following exposure) was
calculated for benzene metabolites in rats and mice that were exposed to 50 ppm of radiolabeled (3H)
benzene (Sabourin et al. 1988). The major metabolic products in rats were detoxification products that
were marked by phenyl conjugates. In contrast, mice had substantial quantities of the markers for
toxification pathways (muconic acid, hydroquinone glucuronide, and hydroquinone sulfate) in their
tissues. Muconic acid and hydroquinone glucuronide were also detected in mouse bone marrow. These
results may explain why mice are more susceptible to benzene-induced toxicity than rats.
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3. HEALTH EFFECTS
In a study by Orzechowski et al. (1995), hepatocytes from adult male Wistar rats and NMRI mice were
incubated for 1 hour with 0.5 mM 14C-benzene, and the supernatant analyzed for metabolites. Formation
of sulfate conjugates of benzene, hydroquinone, and 1,2,4-benzenetriol was also studied in a separate
experiment. Mouse hepatocytes produced two metabolites (1,2,4-trihydroxybenzene sulfate and hydroquinone sulfate) that were not found in rat hepatocyte incubations. These sulfate metabolites were found
in incubations including benzene, or the metabolites themselves, hydroquinone and 1,2,4-benzenetriol.
Mouse hepatocytes were almost three times more effective in metabolizing benzene, compared to rat
hepatocytes. This difference was accounted for in the formation of hydroquinone, hydroquinone sulfate,
and 1,2,4-trihydroxybenzene sulfate. These in vitro experiments indicate there are both quantitative and
qualitative differences in rodent metabolism of benzene.
Data produced in vitro by mouse and rat liver microsomes also indicate species differences in benzene
metabolism (Schlosser et al. 1993). Quantitation of metabolites from the microsomal metabolism of
benzene indicated that after 45 minutes, mouse liver microsomes from male B6C3F1 mice had converted
20% of the benzene to phenol, 31% to hydroquinone, and 2% to catechol. In contrast, rat liver
microsomes from male Fischer 344 rats converted 23% to phenol, 8% to hydroquinone, and 0.5% to
catechol. Mouse liver microsomes continued to produce hydroquinone and catechol for 90 minutes,
whereas rat liver microsomes had ceased production of these metabolites by 90 minutes. Muconic acid
production by mouse liver microsomes was <0.04 and <0.2% from phenol and benzene, respectively,
after 90 minutes.
There are quantitative differences in the benzene metabolites produced by different species (Sabourin et
al. 1988). Fischer 344 rats exposed to 50 ppm benzene had undetectable amounts of phenol, catechol, and
hydroquinone in the liver, lungs, and blood. The major water-soluble metabolites were muconic acid,
phenyl sulfate, prephenyl mercapturic acid, and an unknown. The unknown was present in amounts equal
to the amounts of phenyl sulfate in the liver; phenyl sulfate and the unknown were the major metabolites
in the liver. B6C3F1 mice exposed to 50 ppm benzene had detectable levels of phenol and hydroquinone
in the liver, lungs, and blood; catechol was detectable only in the liver and not in the lungs or blood. As
in the rat, the unknown was present in amounts equal to the amounts of phenyl sulfate in the liver. Mice
had more muconic acid in the liver, which indicates a greater risk for them from trans,trans-muconaldehyde (Sabourin et al. 1988).
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The effect of dose rate on benzene metabolism was studied in Fischer 344 rats and B6C3F1 mice that had
either long inhalation exposures to low concentrations or short exposures to high concentrations of
benzene (Sabourin et al. 1989a, 1989b). Inhalation occurred at 1 of 3 exposure regimens, all having the
same integral amount of benzene: 600 ppm benzene for 0.5 hour, 150 ppm for 2 hours, or 50 ppm for
6 hours. Results indicated no dose-rate effect in rats. In mice, however, the fast exposure rate
(0.5 hour times 600 ppm) produced less muconic acid in the blood, liver, and lungs. In the blood and
lungs, less hydroquinone glucuronide and more prephenyl mercapturic acid were produced at the higher
exposure rates. At the highest benzene exposure concentrations or fastest benzene exposure rate in mice,
there was a reduction in the ratios of muconic acid and hydroquinone glucuronide to the metabolite
phenylsulfate. Furthermore, with increased dose rate or increased exposure concentration, mice tended to
shift a greater portion of their benzene metabolism toward detoxification pathways. Likewise, the
detoxification pathways for benzene appear to be low-affinity, high-capacity pathways, whereas pathways
leading to the putative toxic metabolites appear to be high-affinity, low-capacity systems (Henderson et
al. 1989). Accordingly, if the exposure dose regimen, via inhalation, extends beyond the range of linear
metabolism rates of benzene (200 ppm by inhalation) (Sabourin et al. 1989b), then the fraction of toxic
metabolites formed relative to the amount administered will be reduced. Bois and Paxman (1992) used a
PBPK model to assess effects of dose rate on the disposition of benzene metabolites. Simulations were
performed for rats exposed either for 15 minutes to 32 ppm or for 8 hours to 1 ppm (equivalent 8-hour
TWAs). The amount of metabolites (hydroquinone, catechol, and muconaldehyde) formed was 20%
higher after the 15-minute exposure at the higher level than after the 8-hour exposure at the lower level.
Differences between the model predictions (Bois and Paxman 1992) and the empirical data of Sabourin et
al. (1989a, 1989b) may be related, at least in part, to the higher benzene exposure levels (50, 150, and
600 ppm) used by Sabourin and coworkers.
A number of investigators have suggested that covalent binding of benzene metabolites to cellular
macromolecules is related to benzene's mechanism of toxicity, although the relationship between adduct
formation and toxicity is not clear. Benzene metabolites have been found to form covalent adducts with
proteins from blood in humans (Bechtold et al. 1992b). Benzene metabolites form covalent adducts with
nucleic acids and proteins in rats and mice (Norpoth et al. 1988; Rappaport et al. 1996); covalently bind to
proteins in mouse or rat liver, bone marrow, kidney, spleen, blood, and muscle in vivo (Bechtold and
Henderson 1993; Bechtold et al. 1992a, 1992b; Creek et al. 1997; Longacre et al. 1981a, 1981b; Sun et
al. 1990); bind to proteins in perfused bone marrow preparations (Irons et al. 1980) and in rat and mouse
liver DNA in vivo (Creek et al. 1997; Lutz and Schlatter 1977); and bind to DNA in rabbit and rat bone
marrow mitochondria in vitro (Rushmore et al. 1984). Exposure-related increases in blood levels of
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3. HEALTH EFFECTS
albumin adducts of benzene oxide and 1,4-benzoquinone were noted among workers occupationally
exposed to benzene air concentrations ranging from 0.07 to 46.6 ppm (Rappaport et al. 2002a, 2002b).
Several reactive metabolites of benzene have been proposed as agents of benzene hematotoxic and
leukemogenic effects. These metabolites include benzene oxide, reactive products of the phenol pathway
(catechol, hydroquinone, and 1,4-benzoquinone), and trans,trans-muconaldehyde. See Section 3.5.2 for a
discussion of mechanisms of benzene toxicity.
3.4.4
Elimination and Excretion

Available human data indicate that following inhalation exposure to benzene, the major route for
elimination of unmetabolized benzene is via exhalation. Absorbed benzene is also excreted in humans via
metabolism to phenol and muconic acid followed by urinary excretion of conjugated derivatives (sulfates
and glucuronides). In six male and female volunteers exposed to 5262 ppm benzene for 4 hours,
respiratory excretion (the amount of absorbed benzene excreted via the lungs) was approximately 17%;
no gender-related differences were observed (Nomiyama and Nomiyama 1974a, 1974b). Results from a
study of 23 subjects who inhaled 47110 ppm benzene for 23 hours showed that 16.441.6% of the
retained benzene was excreted by the lungs within 57 hours (Srbova et al. 1950). The rate of excretion
of benzene was the greatest during the first hour. The study also showed that only 0.070.2% of the
retained benzene was excreted in the urine. Other studies suggest that benzene in the urine may be a
useful biomarker of occupational exposure (Ghittori et al. 1993). Results of a study involving a single
human experimental subject exposed to concentrations of benzene of 6.4 and 99 ppm for 8 hours and
1 hour, respectively, suggested that excretion of benzene in breath has three phases and could possibly
have four phases. The initial phase is rapid and is followed by two (or three) slower phases (Sherwood
1988). The initial phase with a high exposure concentration (99 ppm) and a short-term exposure duration
(1 hour) had a more rapid excretion rate (half-life=42 minutes) and a greater percentage of the total dose
excreted (17%) than did the initial phase with a low exposure concentration (6.4 ppm) and longer
exposure duration (8 hours) (half-life=1.2 hours, percentage of total dose excreted=9.3%). Subsequent
phases showed an increase in the half-lives. These results also showed that urinary excretion of phenol
conjugate was biphasic, with an initial rapid excretion phase, followed by a slower excretion phase. A
greater proportion of the total dose was excreted in urine than in breath (Sherwood 1988). The urinary
excretion of phenol in workers was measured following a 7-hour workshift exposure to 1200 ppm
benzene. A correlation of 0.881 between exposure level and urinary phenol excretion was found (Inoue et
al. 1986). Urine samples were collected from randomly chosen subjects not exposed to known sources of
BENZENE
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3. HEALTH EFFECTS
benzene, from subjects exposed to sidestream cigarette smoke, or from supermarket workers presumed
exposed to benzene from polyvinyl chloride (PVC) meat packing wrap (Bartczak et al. 1994). Samples
were analyzed for identification of muconic acid. Muconic acid concentrations of 8550 ng/mL were
found in all urine samples. Kok and Ong (1994) report blood and urine levels of benzene as 110.9 and
116.4 ng/L, respectively, in nonsmokers, and 328.8 and 405.4 ng/L, respectively in smokers. A
significant correlation was found between benzene levels in blood and benzene levels in urine. Similar
results were found for filling station attendants in Italy (Lagorio et al. 1994b).
Popp et al. (1994) reported a mean blood benzene level in car mechanics of 3.3 g/L. Urinary muconic
acid and S-phenylmercapturic acid levels increased during the work shift, and were well correlated with
the blood levels and the benzene air levels, which reached a maximum of 13 mg/m3.
As discussed in Section 3.4.3, the mean urinary levels of muconic in groups of male, female-nonpregnant,
and female-pregnant smokers were 3.6-, 4.8-, and 4.5-fold higher than the mean concentration of this acid
in the nonsmoking groups (Melikian et al. 1994). The differences in the mean muconic acid
concentrations between smoking and nonsmoking groups were significant in male (p=0.001), and
nonpregnant (p=0.001) and pregnant female smokers (p=0.002). Mean concentrations of muconic acid
levels in nonpregnant female smokers are similar to that of male smokers. Mean concentrations of
muconic acid in groups of 42 male smokers and 53 female smokers were 0.220.03 and 0.240.02 mg/g
creatinine, or 0.130.06 and 0.130.07 mg/mg cotinine, respectively. Mean concentrations of muconic
acid in groups of 63 pregnant and 53 nonpregnant female smokers were 0.270.04 and 0.240.02 mg/g
creatinine, or 0.240.06 and 0.130.07 mg/mg cotinine, respectively. Mean concentrations of urinary
cotinine in pregnant smokers were significantly lower than in the group of nonpregnant female smokers
(1.130.12 mg/g creatinine compared to 1.820.14 mg/g creatinine). Benzene levels ranging from 0.01 to
0.18 g/kg have been detected in samples of human breast milk (Fabietti et al. 2004).
Animal data show that exhalation is the main route for excretion of unmetabolized benzene and that
metabolized benzene is excreted primarily in urine. Only a small amount of an absorbed dose is
eliminated in feces. A biphasic pattern of excretion of unmetabolized benzene in expired air was
observed in rats exposed to 500 ppm for 6 hours, with half-times for expiration of 0.7 hour for the rapid
phase and 13.1 hours for the slow phase (Rickert et al. 1979). The half-life for the slow phase of benzene
elimination suggests the accumulation of benzene. The major route of excretion following a 6-hour noseonly inhalation exposure of rats and mice to various concentrations of 14C-benzene appeared to be
dependent on the inhaled concentration (Sabourin et al. 1987). At similar exposures to vapor
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3. HEALTH EFFECTS
concentrations of 101,000 ppm, the mice received 150200% of the equivalent dose in rats on a per kg
body weight basis. At all concentrations, fecal excretion accounted for <3.5% of the radioactivity for rats
and <9% for mice. At lower exposure concentrations (i.e., 13130 ppm in rats and 11130 ppm in mice),
<6% of the radioactivity was excreted in expired air. At the highest exposure concentrations (rats,
870 ppm; mice, 990 ppm), both rats and mice exhaled a significant amount of unmetabolized benzene
(48 and 14%, respectively) following termination of the exposure. The majority of the benzeneassociated radioactivity that was not exhaled was found in the urine and in the carcass 56 hours after the
end of exposure to these high concentrations. The radioactivity in the carcass was associated with the pelt
of the animals. The authors assumed that this was due to contamination of the pelt with urine, since the
inhalation exposure had been nose-only. Further investigation confirmed that the radioactivity was
associated with the fur of the animals. Accordingly, the percentage of the total radioactivity excreted by
these animals (urine and urine-contaminated pelt) that was not exhaled or associated with feces was 47
92% for rats and 8094% for mice. At exposures of 260 ppm in rats, 8592% of the radioactivity was
excreted as urinary metabolites, while at exposures of 130 ppm in mice, 8894% of the radioactivity was
excreted as urinary metabolites. The total urinary metabolite formation was 537% higher in mice than in
rats at all doses. This may be explained by the greater amount of benzene inhaled by mice per kg of body
weight (Sabourin et al. 1987). Purebred Duroc-Jersey pigs were exposed to 0, 20, 100, and 500 ppm
benzene vapors 6 hours/day, 5 days/week for 3 weeks (Dow 1992). The average concentration of phenol
in the urine increased linearly with dose.
No studies were located regarding excretion in humans after oral exposure to benzene. Data on excretion
of benzene or its metabolites in human breast milk after oral exposure were not found.
Radiolabeled benzene (340 mg/kg) was administered by oral intubation to rabbits; 43% of the label was
recovered as exhaled unmetabolized benzene and 1.5% was recovered as carbon dioxide (Parke and
Williams 1953a). Urinary excretion accounted for about 33% of the dose. The isolated urinary
metabolites were mainly in the form of conjugated phenols. Phenol was the major metabolite accounting
for about 23% of the dose or about 70% of the benzene metabolized and excreted in the urine. The other
phenols excreted (percentage of dose) were hydroquinone (4.8%), catechol (2.2%), and trihydroxybenzene (0.3%). L-Phenyl-N-acetyl cysteine accounted for 0.5% of the dose. Muconic acid accounted
for 1.3%; the rest of the radioactivity (510%) remained in the tissues or was excreted in the feces (Parke
and Williams 1953a).
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Mice received a single oral dose of either 10 or 200 mg/kg radiolabeled benzene (McMahon and
Birnbaum 1991). Radioactivity was monitored in urine, feces, and breath. At the low dose, urinary
excretion was the major route of elimination. Hydroquinone glucuronide, phenylsulfate, and muconic
acid were the major metabolites at this dose, accounting for 40, 28, and 15% of the dose, respectively. At
200 mg/kg, urinary excretion decreased to account for 4247% of the administered dose, while
respiratory excretion of volatile components increased to 4656% of the administered dose. Fecal
elimination was minor and relatively constant over both doses, accounting for 0.53% of the dose.
The effect of dose on the excretion of radioactivity, including benzene and metabolites, following oral
administration of 14C-benzene (0.5300 mg/kg) has been studied in rats and mice (Sabourin et al. 1987).
At doses of <15 mg/kg for 1 day, 90% of the administered dose was excreted in the urine of both species.
There was a linear relationship for the excretion of urinary metabolites up to 15 mg/kg; above that level,
there was an increased amount of 14C eliminated in the expired air. Mice and rats excreted equal amounts
up to 50 mg/kg; above this level, metabolism apparently became saturated in mice. In rats, 50% of the
150 mg/kg dose of 14C was eliminated in the expired air; in mice, 69% of the 150 mg/kg dose of 14C was
eliminated in expired air (Sabourin et al. 1987). The label recovered during exhalation was largely in the
form of unmetabolized benzene, suggesting that saturation of the metabolic pathways had occurred. Dose
also affected the metabolite profile in the urine. At low doses, a greater fraction of the benzene was
converted to putative toxic metabolites than at high doses, as reflected in urinary metabolites.
Mathews et al. (1998) reported similar results following oral (gavage) administration of 14C-benzene to
rats, mice, and hamsters in single doses from as low as 0.2 mg/kg and up to 100 mg/kg. For example,
>95% of a 0.5 mg/kg dose was recovered in the urine of rats; a small amount (3%) was recovered in
expired air. At benzene doses of 10 and 100 mg/kg, elimination in the breath rose to 9 and 50%,
respectively, indicating the likely saturation of benzene metabolism. Excretion in the feces was minimal
at all dose levels. Similar results were noted for mice and hamsters. Both dose and species differences
were noted in the composition of urinary metabolites. Phenyl sulfate was the major metabolite in rat
urine at all dose levels, accounting for 6473% of urinary radioactivity. Phenyl sulfate (2432%) and
hydroquinone glucuronide (2729%) were the predominant urinary metabolites in mice. At a dose of
0.1 mg/kg, mice produced a considerably higher proportion of muconic acid than rats (15 versus 7%). In
hamsters, hydroquinone glucuronide (2429%) and muconic acid (1931%) were the primary urinary
metabolites. Two additional metabolites (1,2,4-trihydroxybenzene and catechol sulfate) were recovered
from the urine of hamsters, but not rats or mice.
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Limited data on excretion of benzene after dermal exposure in humans were found. Four human male
subjects were given a dermal application of 0.0024 mg/cm2 14C benzene (Franz 1984). A mean of
0.023% (range, 0.0060.054%) of the applied radiolabel was recovered in the urine over a 36-hour period.
Urinary excretion of the radiolabel was greatest in the first two hours following skin application. More
than 80% of the total excretion occurred in the first 8 hours. In another study, 3543 cm2 of the forearm
were exposed to approximately 0.06 g/cm2 of liquid benzene for 1.252 hours (Hanke et al. 1961). The
absorption was estimated from the amount of phenol eliminated in the urine. The absorption rate of liquid
benzene by the skin (under the conditions of complete saturation) was calculated to be low, approximately
0.4 mg/cm2/hour. The absorption due to vapors in the same experiment was negligible. Although there
was a large variability in the physiological values, the amount of excreted phenol was 8.014.7 mg during
the 24-hour period after exposure. It is estimated that approximately 30% of dermally absorbed benzene
is eliminated in the form of phenol in the urine.
Data on excretion of benzene or its metabolites in human breast milk after dermal exposure were not
found.
Monkeys and minipigs were exposed dermally to 0.00260.0036 mg/cm2 of 14C-benzene (Franz 1984).
After application, the urine samples were collected over the next 24 days at 5-hour intervals. The rate of
excretion was highest in the first two collection periods. The total urinary excretion of radioactivity was
found to be higher in monkeys than in minipigs with the same exposure. Mean excretion in monkeys was
0.065% (range, 0.0330.135%) of the applied dose compared to 0.042% (range, 0.0300.054%) in
minipigs.
Results of a study in which male rats were dermally treated with 0.004 mg/cm2 of 14C-benzene, with or
without 1g of clay or sandy soil, showed that for all treatment groups, the major routes of excretion were
the urine and, to a lesser extent, the expired air (Skowronski et al. 1988). The highest amount of
radioactivity in urine appeared in the first 1224 hours after treatment (58.8, 31.3, and 25.1% of the
absorbed dose, respectively, for pure benzene, sandy soiladsorbed benzene, and clay soil-adsorbed
benzene). In the group treated with pure benzene, 86.2% of the absorbed dose was excreted in the urine.
Sandy soil and clay soil significantly decreased urinary excretion to 64.0 and 45.4%, respectively, of the
absorbed dose during the same time period. Rats receiving pure benzene excreted 12.8% of the absorbed
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dose in expired air within 48 hours. Only 5.9% of the radioactivity was collected in expired air 48 hours
after treatment with sandy soiladsorbed benzene, while experiments with clay soiladsorbed benzene
revealed that 10.1% of the radioactivity was located in expired air. Less than 1% of the absorbed dose
was expired as 14CO2 in all groups. The 14C activity in the feces was small (<0.5% of the applied
radioactivity) in all groups 48 hours after treatment. Phenol was the major urinary metabolite detected in
the 012-hour urine samples of all treatment groups. The percentage of total urinary radioactivity
associated with phenol was 37.7% for benzene alone, 44.2% for benzene adsorbed to sandy soil, and
45.5% for benzene adsorbed to clay soil. Smaller quantities of hydroquinone, catechol, and benzenetriol
were also detected (Skowronski et al. 1988).
3.4.4.4 Other Routes of Exposure
The metabolic fate of benzene can be altered in fasted animals. In nonfasted rats that received an
intraperitoneal injection of 88 mg of benzene, the major metabolites present in urine were total conjugated
phenols (1419% of dose), glucuronides (34% of dose), and free phenol (23% of dose). However, in
rats fasted for 24 hours preceding the same exposure, glucuronide conjugation increased markedly (18
21% of dose) (Cornish and Ryan 1965). Free phenol excretion (810% of dose) was also increased in
fasted, benzene-treated rats. There was no apparent increase in total conjugated phenol excretion in
fasted rats given benzene.
When 14C-benzene (0.5 and 150 mg/kg) was injected intraperitoneally into rats and mice, most of the
14
C-benzene and 14C-metabolites were excreted in the urine and in the expired air. A smaller amount of
14
C-benzene was found in the feces due to biliary excretion (Sabourin et al. 1987). Monkeys were dosed
intraperitoneally with 5500 mg/kg radiolabeled benzene, and urinary metabolites were examined
(Sabourin et al. 1992). The proportion of radioactivity excreted in the urine decreased with increasing
dose, whereas as the dose increased, more benzene was exhaled unchanged. This indicated saturation of
benzene metabolism at higher doses. Phenyl sulfate was the major urinary metabolite. Hydroquinone
conjugates and muconic acid in the urine decreased as the dose increased. When C57BL/6 mice and
DBA/2 mice were given benzene subcutaneously in single doses (440, 880, or 2,200 mg/kg) for 1 day, or
multiple doses (880 mg/kg) 2 times daily for 3 days, no strain differences were observed in the total
amount of urinary ring-hydroxylated metabolites (Longacre et al. 1981a). Although each strain excreted
phenol, catechol, and hydroquinone, differences in the relative amounts of these metabolites were noted.
The more sensitive DBA/2 mice excreted more phenol but less hydroquinone than the more resistant
C57BL/6 mice, while both strains excreted similar amounts of catechol. DBA/2 mice excreted more
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phenyl glucuronide but less sulfate conjugate. Both strains excreted similar amounts of phenyl
mercapturic acid (Longacre et al. 1981a).
3.4.5
Physiologically Based Pharmacokinetic (PBPK)/Pharmacodynamic (PD) Models
Physiologically based pharmacokinetic (PBPK) models use mathematical descriptions of the uptake and
disposition of chemical substances to quantitatively describe the relationships among critical biological
processes (Krishnan et al. 1994). PBPK models are also called biologically based tissue dosimetry
models. PBPK models are increasingly used in risk assessments, primarily to predict the concentration of
potentially toxic moieties of a chemical that will be delivered to any given target tissue following various
combinations of route, dose level, and test species (Clewell and Andersen 1985). Physiologically based
pharmacodynamic (PBPD) models use mathematical descriptions of the dose-response function to
quantitatively describe the relationship between target tissue dose and toxic end points.
PBPK/PD models refine our understanding of complex quantitative dose behaviors by helping to
delineate and characterize the relationships between: (1) the external/exposure concentration and target
tissue dose of the toxic moiety, and (2) the target tissue dose and observed responses (Andersen and
Krishnan 1994; Andersen et al. 1987). These models are biologically and mechanistically based and can
be used to extrapolate the pharmacokinetic behavior of chemical substances from high to low dose, from
route to route, between species, and between subpopulations within a species. The biological basis of
PBPK models results in more meaningful extrapolations than those generated with the more conventional
use of uncertainty factors.
The PBPK model for a chemical substance is developed in four interconnected steps: (1) model
representation, (2) model parameterization, (3) model simulation, and (4) model validation (Krishnan and
Andersen 1994). In the early 1990s, validated PBPK models were developed for a number of
toxicologically important chemical substances, both volatile and nonvolatile (Krishnan and Andersen
1994; Leung 1993). PBPK models for a particular substance require estimates of the chemical substancespecific physicochemical parameters, and species-specific physiological and biological parameters. The
numerical estimates of these model parameters are incorporated within a set of differential and algebraic
equations that describe the pharmacokinetic processes. Solving these differential and algebraic equations
provides the predictions of tissue dose. Computers then provide process simulations based on these
solutions.
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The structure and mathematical expressions used in PBPK models significantly simplify the true
complexities of biological systems. If the uptake and disposition of the chemical substance(s) are
adequately described, however, this simplification is desirable because data are often unavailable for
many biological processes. A simplified scheme reduces the magnitude of cumulative uncertainty. The
adequacy of the model is, therefore, of great importance, and model validation is essential to the use of
PBPK models in risk assessment.
PBPK models improve the pharmacokinetic extrapolations used in risk assessments that identify the
maximal (i.e., the safe) levels for human exposure to chemical substances (Andersen and Krishnan 1994).
PBPK models provide a scientifically sound means to predict the target tissue dose of chemicals in
humans who are exposed to environmental levels (for example, levels that might occur at hazardous waste
sites) based on the results of studies where doses were higher or were administered in different species.
Figure 3-4 shows a conceptualized representation of a PBPK model.
If PBPK models for benzene exist, the overall results and individual models are discussed in this section
in terms of their use in risk assessment, tissue dosimetry, and dose, route, and species extrapolations.
Several PBPK models have been developed that simulate the disposition of benzene in humans (Bois et
al. 1996; Brown et al. 1998; Fisher et al. 1997; Medinsky et al. 1989c; Sinclair et al. 1999; Travis et al.
1990), mice (Cole et al. 2001; Medinsky et al. 1989a, 1989b; Sun et al. 1990; Travis et al. 1990), and rats
(Bois et al. 1991a; Medinsky et al. 1989a, 1989b; Sun et al. 1990; Travis et al. 1990). A comparative
summary of the models is provided in Table 3-6. All of the models have the same general structure
(Figure 3-5). Most of the models simulate inhalation and oral exposures; one model provides a
simulation of dermal absorption (Sinclair et al. 1999). Physiological parameters and partition coefficients
for simulating benzene biokinetics of human females were reported for the Brown et al. (1998) and Fisher
et al. (1997) models. Flow-limited exchange of benzene between blood and tissues is assumed in all
models, with excretion of benzene in exhaled air and, in one case, to breast milk (Fisher et al. 1997). All
models include simulations of blood, fat, liver, lung, and lumped compartments representing other
slowly-perfused tissues (e.g., skeletal muscle) and rapidly-perfused tissues (e.g., kidneys, other viscera).
Simulation of bone marrow, the primary target for benzene toxicity, is included in the models reported by
Bois et al. (1991a, 1996), Sinclair et al. (1999), and Travis et al. (1990).
Simulations of metabolism in the various models vary in complexity. In the simplest representation,
metabolic elimination of benzene is simulated as a single capacity-limited process, represented with
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Figure 3-4. Conceptual Representation of a Physiologically Based
Pharmacokinetic (PBPK) Model for a
Hypothetical Chemical Substance
In h a le d c h e m ic a l
E x h a le d c h e m ic a l
In g e s tio n
Lungs
L iv e r
V max
Km
V
E
N
O
U
S
GI
T ra c t
F at
S lo w ly
p e rfu s e d
tis s u e s
R ic h ly
p e rfu s e d
tis s u e s
B
L
O
O
D
F eces
K id n e y
A
R
T
E
R
I
A
L
B
L
O
O
D
U rin e
S k in
C h e m ic a ls
c o n ta c tin g s k in
Note: This is a conceptual representation of a physiologically based pharmacokinetic (PBPK) model for a
hypothetical chemical substance. The chemical substance is shown to be absorbed via the skin, by inhalation, or by
ingestion, metabolized in the liver, and excreted in the urine or by exhalation.
Source: adapted from Krishnan and Andersen 1994
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Figure 3-5. General Structure of Physiologically Based Pharmacokinetic Models

of Benzene*
Air
Alveolar Space
Air
Lung Blood
Fat
Bone Marrow
Metabolites
Rapidly-perfused
Slowly-perfused
Urine
Mammary
Milk
Liver
Gastrointestinal
Tract
Metabolites
*Tissues shown with dashed lines are not simulated in all models. Flow-limited exchange of benzene between blood
and tissues is assumed. Metabolism is simulated to varying degrees of complexity (see Table 3-6 for model
comparison).
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Table 3-6. Summary Comparison of Physiologically Based Pharmacokinetic
Models for Benzene
Simulated areas
Reference
Excretion
Absorption
Speciesa pathwaysb Tissuesc Metabolic pathwaysd pathwayse Comment
Bois et al. R
1991a
IH, OR
BL, BM,
FA, LI,
LU, RP,
SP
BM, LI: BZBO(c)
EH: BZ
BOBG(c)
UR: PH
BOPH(f)
BOGSH(c)
BGDI(c)
Simulates metabolic
pathways in bone
marrow, and phenol
conjugation in lung
and gastrointestinal
tract
PHHQ(c)
PHCA(c)
Bois et al. H
1996
IH
Brown et H (m,f)
al. 1998
IH
Cole et
al. 2001
IH, OR
BL, BM,
FA, LU,
LI, RP,
SP
BL, FA,
LI, LU,
RP, SP
BL, FA,
LI, LU,
RP, SP
BM,
PHPHCO(c)
LU,
LU, GI:
EH: BZ
BM, LI: BZMtot(c)
LI:
PHXendPH(z) UR: Mtot

PH
EH: BZ
BZMtot(f)
LI:
BZBO(c)
EH: BZ
BOPH(f)
UR: CA
LI:
BOPMA(f)
MA
BOMA(f)
PHCO
PHHQ(c)
PMA
PHPHCO(c)
HQCO
PHCA(c)
THB
Simulates metabolic
pathways in bone
marrow, and endogenous production
of phenolic meta
bolites
Simulates males or
females
All metabolism is
assigned to the liver
CATHB(c)
HQHQCO(c)
Fisher et H
al. 1997
IH
Medinsky H, M, R
et al.
1989a,
1989b,
1989c
IH, OR
FA, LU,
LI, RP,
SP, MI
FA, LI,
LU, RP,
SP
LI:
BZMtot(c)
EH: BZ
MI: BZ
LI:
BZBO(c)
EH: BZ
BOPHCO(c)
BOPMA(c)
BOHQCO(c)
BOMA(c)
Simulates transfer of
benzene to breast
milk
All metabolism is
assigned to the liver
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Table 3-6. Summary Comparison of Physiologically Based Pharmacokinetic
Models for Benzene
Simulated areas
Reference
Excretion
Absorption
Speciesa pathwaysb Tissuesc Metabolic pathwaysd pathwayse Comment
Sinclair et H
al. 1999
IH, OR, DE BL, BM,

LI, LU,
MU, RP
Sun et al. M, R
1990
IH, OR
BM, LI: BZMtot(c)
BL, FA, LI:

LI, LU,
RBC, RP,
SP
BZBO(c)
EH: BZ
UR: Mtot
PH
EH: BZ
Simulates dermal
exposure and
absorption
EH: BZ
Total metabolism of
benzene in the bone
marrow and liver
BOPHCO(c)
BOPMA(c)
BOHQCO(c)
Simulates formation
of hemoglobin
adducts in red blood
cells derived from
benzene oxide
BOMA(c)
RBC:
Travis et H, M, R
al. 1990
IH, OR
BL, BM,
FA, LI,
LU, MU,
RP
BOHBA(c,f)
BM, LI: BZMtot(c)
Species simulated: H=human; M=mouse; R=rat; m=male; f=female

Absorption pathways simulated: IH=inhalation; OR=oral; DE=dermal
c
Tissues simulated: BL=blood; BM=bone marrow; FA=fat; LI=liver; LU=lung; MU=muscle; RBC=red blood cells;
RP=other rapidly-perfused tissues; SP=other slowly-perfused tissues
d
Metabolic pathways simulated: BZ=benzene; BD=benzene diols; BO=benzene oxide; BG=Benzene glycol;
CA=catechol; HBA=hemoglobin adduct; HQ=hydroquinone; HQCO=ydroqinione conjugates; MA=muconic acid;
Mtot=total metabolites; PH=phenol; PHCO=phenol conjugates; PMA=phenylmercapturic acid;
THB=trihydroxybenzene; PHXend=endogenous phenolic metabolites; (c)=capacity-limited; (f)=first-order; (z)= zeroorder
e
Excretion pathways simulated: EH=exhalation; MI=breast milk; UR=urine
b
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Michaelis-Menten function of benzene concentration in tissue (Bois et al. 1996; Brown et al. 1998; Fisher
et al. 1997; Sinclair et al. 1999; Travis et al. 1990). In the more complex representations, the major
pathways of metabolism of benzene, including conjugation reactions, are simulated as capacity-limited or
first-order processes (Bois et al. 1991a; Cole et al. 2001; Medinsky et al. 1989a, 1989b, 1989c; Sun et al.
1990). In most of the models, all metabolic pathways are attributed to the liver; however, four of the
models include simulations of metabolism in bone marrow (Bois et al. 1991a, 1996; Sinclair et al. 1999;
Travis et al. 1990), and one model includes simulations of the formation of sulfate and glucuronide
conjugates of phenol in the gastrointestinal and respiratory tracts (Bois et al. 1991a). The Sun et al.
(1990) model includes a simulation of the formation of hemoglobin adducts derived from benzene oxide.
In models that simulate the disposition of the metabolites, metabolites are assumed to be excreted in urine
either at a rate equal to their formation (Cole et al. 2001), or in accordance with a first-order excretion rate
constant (Bois et al. 1991a, 1996; Sinclair et al. 1999); the difference being, in the latter, the mass balance
for formation and excretion of metabolites is simulated, allowing predictions of metabolite levels in
tissues. All of the models use typical parameters and values for species-specific blood flows and tissue
volumes.
Brief summaries of the models presented in Table 3-6 are provided below, with emphasis on unique
features that are applicable to risk assessment.
Medinsky et al. 1989a, 1989b, 1989c
Description of the Model.
The Medinsky et al. (1989a, 1989b, 1989c) model simulates absorption
and disposition of benzene in the human, mouse, and rat. Tissues simulated include the blood, bone
marrow, fat, liver, lung, other slowly-perfused tissues, and other rapidly-perfused tissues. Gastrointestinal
absorption of benzene is simulated as a first-order process; absorption and excretion of benzene in the
lung are assumed to be flow-limited. Exchange of benzene between blood and tissues is assumed to be
flow-limited. The model simulates capacity-limited (i.e., Michaelis-Menten) metabolism of benzene to
benzene oxide as a function of the concentration of benzene in liver. Conversion of benzene oxide to
phenol conjugates, phenylmercapturic acid, hydroquinone conjugates, and muconic acid are simulated as
parallel, capacity-limited reactions in liver. The model simulates rates of formation of metabolites, but
not the disposition (e.g., excretion) of metabolites. Metabolism parameter values (Vmax, Km) for the
mouse and rat models were estimated by optimization of the model to observations of total metabolites
formed in mice and rats exposed by inhalation or oral routes to benzene (Medinsky et al. 1989b; Sabourin
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et al. 1987). Human metabolism parameter values were derived from allometric scaling of the values for
mice (Medinsky et al. 1989c).
Risk Assessment.
The model has been used to predict the amounts of benzene metabolites formed in
rats and mice after inhalation or oral exposures (Medinsky et al. 1989a, 1989b). For inhalation
concentrations up to 1,000 ppm, mice were predicted to metabolize at least 23 times more benzene than
rats. For oral doses >50 mg/kg, rats were predicted to metabolize more benzene on a kg-body weight
basis than mice. The model also predicts different metabolite profiles in the two species: mice were
predicted to produce primarily hydroquinone glucuronide and muconic acid, metabolites linked to toxic
effects, whereas rats were predicted to produce primarily phenyl sulfate, a detoxification product. These
predictions agree with experimental data and provide a framework for understanding the greater
sensitivity of the mouse to benzene toxicity.
Validation of the Model.
The model was calibrated with data from Sabourin et al. (1987). Bois et al.
(1991b) compared predictions made to observations of benzene exhaled by rats following exposures to
490 ppm benzene, reported by Rickert et al. (1979), as well as the data from which the model was
calibrated (Sabourin et al. 1987). In general, the model tended to overestimate observations to which it
was not specifically fitted.
Target Tissues.
The model simulates amounts and concentrations of benzene in blood, liver, fat, and
lumped compartments for other rapidly-perfused and slowly-perfused tissues as well as amounts of
metabolites formed. It does not simulate concentrations of metabolites in these tissues. It does not
simulate bone marrow, a target of benzene metabolites.
Species Extrapolation.
The model has been applied to simulations of mice, rats, and humans
(Medinsky et al. 1989a, 1989b, 1989c).

High-low Dose Extrapolation.
The model has been evaluated for simulating inhalation exposures in
rodents ranging from 1 to 1,000 ppm and oral gavage doses ranging from 0.1 to 300 mg/kg (Bois et al.
1991b; Medinsky et al. 1989a, 1989b, 1989c).
Interroute Extrapolation.
The model simulates inhalation and oral exposures and has been applied
to predicting internal dose metrics (e.g., amounts of metabolites formed) resulting from exposures by
these routes (Medinsky et al. 1989a, 1989b, 1989c).
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Strengths and Limitations.
Strengths of the model are that it simulates disposition of inhaled and
ingested (single dose) benzene, including rates and amounts of major metabolites formed in mice, rats,
and humans. Limitations include: (1) the model has not been evaluated for multiple exposures; (2) the
model attributes all metabolism to the liver; (3) the model does not simulate the fate of metabolites
formed and, therefore, cannot be used to predict concentrations of metabolites (e.g., muconaldehyde) in
tissues; and (4) the model does not simulate bone marrow, a major target tissue for benzene metabolites.
Sun et al. 1990
The Sun et al. (1990) model is an extension of the mouse and rat models
developed by Medinsky et al. (1989a, 1989b, 1989c). The Sun et al. (1990) model includes a simulation
of the formation of hemoglobin adducts derived from benzene oxide. Adduct formation is represented as
the sum of capacity-limited and first-order functions of the concentration of benzene oxide in the liver.
Parameter values were estimated by optimization to measurements of hemoglobin adduct formations in
rats and mice exposed to single oral gavage doses of benzene (Sun et al. 1990).
Risk Assessment.
The model has been applied to predicting the levels of hemoglobin adducts in
mice and rats following inhalation or oral exposures to benzene. This approach could be potentially
useful for predicting exposure levels that correspond to measured hemoglobin adduct levels, for use of
adducts as an exposure biomarker.
The model was calibrated against measurements of hemoglobin adduct
formation in mice and rats that received single oral gavage doses of benzene ranging from 0.008 to
800 mg/kg (Sun et al. 1990). The model was evaluated by comparing predictions to observations of
amounts of hemoglobin adducts formed in mice and rats exposed to benzene vapor concentrations of 5,
50, or 600 ppm for 6 hours (Sabourin et al. 1989a).
Target Tissues.
The model predicts hemoglobin adduct formation after oral and inhalation exposure
(Sun et al. 1990).

The model has been applied to simulations for mice and rats.
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The model was calibrated with observations made in mice and rats
exposed to single gavage doses ranging from 0.1 to 10,000 mol/kg (0.008800 mg/kg), and evaluated
for predicting observations in mice and rats exposed by inhalation to 600 ppm benzene.
The Sun model examined two routes of exposure, oral and inhalation.
The model was found to be useful in predicting the concentrations of hemoglobin adducts in blood in
rodents after oral and inhalation exposure.
Strengths of the model are that it extends the Medinsky et al. (1989a,
1989b, 1989c) models to simulate hemoglobin adduct formation secondary to formation of benzene
oxide. A limitation of the adduct model is that it simulates production of adducts as a function of benzene
oxide concentration in liver and does not consider other potential pathways of adduct formation through
hydroquinone, phenol, or muconaldehyde.
Travis et al. 1990
The Travis et al. (1990) model simulates the absorption and disposition
of benzene in the human, mouse, and rat. Tissues simulated include the blood, bone marrow, fat, liver,
lung, other slowly-perfused tissues, and rapidly-perfused tissues. Gastrointestinal absorption of benzene
is simulated as a first-order process. Absorption and excretion of benzene in the lung are assumed to be
flow-limited, as are exchanges of benzene between blood and tissues. The model simulates capacitylimited (i.e., Michaelis-Menten) metabolic elimination of benzene as a function of the concentration of
benzene in bone marrow and liver. The model simulates rates of metabolic elimination of benzene, but
not the rates of formation of specific metabolites or their disposition (e.g., excretion). For the purpose of
comparing model predictions to observations, 80% of the total metabolite formed in 24 hours (and
excreted in urine) was assumed to be phenol. Metabolism parameter values (Vmax, Km) were estimated by
optimization of the model to observations of total metabolites formed (i.e., excreted in urine) in humans,
mice, and rats exposed to benzene by inhalation or oral routes to benzene. The Vmax for metabolism in
bone marrow in humans was assumed to be 4% of that of liver, consistent with optimized values for
rodents.
Risk Assessment.
This model has been used to predict the amounts of benzene in expired air,
concentrations of benzene in blood, and total amount of benzene metabolized following inhalation
exposures to humans and inhalation, intraperitoneal, oral gavage, or subcutaneous exposures in mice or
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rats (Travis et al. 1990). Cox (1996) applied the model to derive internal dose-response relationships for
benzene in humans.
The model was evaluated by comparing predictions with observations
made in mouse and rat inhalation studies (Rickert et al. 1979; Sabourin et al. 1987; Sato et al. 1975;
Snyder et al. 1981); mouse oral gavage studies (Sabourin et al. 1987); mouse subcutaneous injection
studies (Andrews et al. 1977); and rat intraperitoneal injection studies (Sato and Nakajima 1979).
Predictions of benzene in expired air and/or blood concentrations were also compared to observations
made in humans who inhaled concentrations ranging from 5 to 100 ppm benzene (5 ppm: Berlin et al.
1980; Sherwood 1972; 2557 ppm: Sato et al. 1975; Sherwood 1972; Nomiyama and Nomiyama 1974a,
1974b; 99100 ppm: Sherwood 1972; Teisinger and Fiserova-Bergerova 1955). Further evaluations of
predictions of benzene in workers are reported in Sinclair et al. (1999) and Sherwood and Sinclair (1999),
who compared model predictions with observations of benzene in exhaled breath and urinary excretion of
phenol in workers who were exposed to benzene at concentrations ranging from 1 to 1,100 ppm.
Target Tissues.
The model simulates amounts and concentrations of benzene in blood, bone marrow
(a target tissue), liver, fat, and lumped compartments for other rapidly-perfused and slowly-perfused
tissues; and amounts of metabolites formed in liver and bone marrow. It does not simulate concentrations
of metabolites in these tissues.
The model has been applied to simulations for mice, rats, and humans
(Sherwood and Sinclair 1999; Sinclair et al. 1999; Travis et al. 1990).
humans ranging from 1 to 1,110 ppm (Sherwood and Sinclair 1999; Sinclair et al. 1999; Travis et al.
1990). Evaluations of predictions in rodents included observations made during inhalation exposures that
ranged from 11 to 1,000 ppm and oral gavage doses that ranged from 0.5 to 300 mg/kg.
to predicting internal dose metrics (e.g., benzene concentration in blood, amount benzene metabolized)
resulting from exposures by these routes (Travis et al. 1990).
Strengths of the model are that it simulates (1) disposition of inhaled
and ingested (single dose) benzene in mice, rats, and humans; and (2) concentrations of benzene, and
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rates and amount of benzene metabolized in bone marrow, a target tissue for benzene metabolites.
Limitations of the model include: (1) the model simulates metabolic elimination of benzene, but not the
rates of formation of major metabolites; and (2) the model does not simulate fate of metabolites formed
and, therefore, cannot be used to predict concentrations of metabolites in tissues.
Fisher et al. 1997
The Fisher et al. (1997) model extends the model reported by Travis et al.
(1990) to include a simulation of lactational transfer of benzene to breast milk in humans. Other tissues
simulated include blood, fat, liver, lung, other slowly-perfused tissues, and rapidly-perfused tissues.
Absorption and excretion of benzene in the lung and exchange of benzene between blood and tissues are
assumed to be flow-limited, as is excretion of benzene in breast milk. The lactational transfer model
includes simulations of breast milk production and loss from nursing; the latter is represented as a firstorder process. Estimates of blood:air and blood:milk partition coefficients during lactation (from which
the milk:blood partition coefficient could be calculated) were measured in nine lactating subjects (Fisher
et al. 1997). The model simulates capacity-limited (i.e., Michaelis-Menten) metabolism of metabolic
elimination of benzene as a function of the concentration of benzene in liver. Rates of formation of
specific metabolites and their disposition (e.g., excretion) are not simulated. Metabolism parameter
values (Km,Vmax) and tissue:blood partition coefficients were derived from Travis et al. (1990).
Risk Assessment.
This model has been used to predict benzene concentrations in breast milk and
lactational transfers to breast feeding infants (Fisher et al. 1997). Exposures to the threshold limit value
(TLV) (10 ppm, 8 hours/day, 5 days/week) were predicted to yield 0.053 mg of benzene in breast milk
per 24 hours. This approach has potential applicability to assessing lactational exposures to infants
resulting from maternal exposures.
The lactation model was evaluated (Fisher et al. 1997) by comparing
predictions for perchloroethylene (not benzene) with those predicted by a perchloroethylene model
developed by Schreiber (1993). Other components of the biokinetics model were derived from the Travis
et al. (1990) model, which has undergone evaluations against data obtained from studies in humans.
Target Tissues.
The model simulates concentrations of benzene in blood, breast milk, liver, fat, and
lumped compartments for other rapidly-perfused and slowly-perfused tissues as well as rates of metabolic
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elimination of benzene. It does not simulate concentrations of metabolites in these tissues and does not
simulate metabolism in bone marrow, a major target of benzene metabolites.
The model has been applied to simulations for humans (Fisher et al. 1997).
The lactational model has not been evaluated for simulating
inhalation exposures to benzene in humans; therefore, applicability to high-low dose extrapolations

cannot be assessed.
The model was developed to simulate inhalation exposures. Extrapolation
to other routes (e.g., oral, dermal) would require the extension of the model to include simulations of
absorption from these routes.
Strengths of the model are that it simulates the disposition of inhaled
benzene in females during lactation, including transfers of benzene to breast milk and nursing infants;
concentrations of benzene in blood and tissues; and rates of metabolic elimination of benzene
metabolized. Limitations of the model include that the model does not simulate rates of formation of
major metabolites and that the model does not simulate kinetics of uptake or metabolism of benzene in
bone marrow, a major target of benzene toxicity.
Sinclair et al. 1999
The Sinclair et al. (1999) model is an extension of the human model
developed by Travis et al. (1990) to include a simulation of first-order urinary excretion of total
metabolites and phenol, and dermal absorption of benzene.
Risk Assessment.
The model has been applied to predicting the levels of benzene in exhaled air and
phenol in urine in workers exposed to benzene (Sherwood and Sinclair 1999; Sinclair et al. 1999).
The model was evaluated against measurements of benzene in exhaled
breath and urinary excretion of phenol in workers who were exposed to benzene at concentrations ranging
from 1 to 1,100 ppm (Sherwood and Sinclair 1999; Sinclair et al. 1999).
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Target Tissues.
The model simulates amounts and concentrations of benzene in blood, bone marrow
(a target tissue), liver, fat, and lumped compartments for other rapidly-perfused and slowly-perfused
tissues; rates of metabolic elimination of benzene in liver and bone marrow; and excretion of total
metabolites formed and phenol.
The model has been applied to simulations for humans (Sinclair et al. 1999).
The model was evaluated against observations of benzene in exhaled
breath and urinary excretion of phenol in workers who were exposed to benzene at concentrations ranging
from 1 to 1,100 ppm (Sherwood and Sinclair 1999; Sinclair et al. 1999).
The model simulates inhalation, oral, and dermal exposures.

Strengths of the model are that it extends the Travis et al. (1990) model
to include simulation of dermal absorption of benzene.

Bois et al. 1991a
The Bois et al. (1991a) model simulates absorption and disposition of
benzene and the benzene metabolite, phenol, in the rat. Tissues simulated include the blood, bone
marrow, fat, liver, lung, other slowly-perfused tissues, and other rapidly-perfused tissues. Gastrointestinal
absorption of benzene and phenol are simulated as a first-order function for dose. Absorption and
excretion of benzene in the lung are assumed to be flow-limited as are exchanges of benzene and phenol
between blood and tissues. Excretion of phenol is simulated as a first-order transfer to urine. The model
simulates capacity-limited (i.e., Michaelis-Menten) and first-order metabolism of benzene and
metabolites in bone marrow, liver, gastrointestinal tract, and respiratory tract (see Table 3-6). All
pathways are assumed to be capacity-limited reactions, except for the spontaneous hydrolysis of benzene
oxide to form phenol, which is simulated as a first-order process. The model simulates rates of formation
of metabolites and first-order excretion of phenol; however, disposition (e.g., excretion) of other
metabolites is not simulated. Parameter values, including metabolism parameter values, were optimized
to a reference set of observations of metabolites formed in rats exposed by inhalation or to single gavage
doses of benzene (see below).
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Risk Assessment.
This model has been used to predict amounts of benzene and phenol metabolites
formed in rats during oral gavage exposures to benzene equivalent to those administered in NTP (1986)
and to inhalation exposures equivalent to the OSHA PEL (Bois and Paxman 1992; Bois et al. 1991a).
Model simulations indicate that dose rate may be an important factor in benzene toxicity. For example,
when the model was applied to simulations for rats exposed either for 15 minutes to a benzene vapor
concentration of 32 ppm or for 8 hours to 1 ppm (equivalent 8-hour TWAs), the amount of metabolites
(hydroquinone, catechol, and muconaldehyde) formed was 20% higher after the 15-minute exposure at
the higher level than after the 8-hour exposure at the lower level (Bois and Paxman 1992). These
metabolites have been identified as being important in the genesis of bone marrow toxicity after benzene
exposure (Eastmond et al. 1987). These types of analyses, if extended to humans, would be applicable to
evaluations of the adequacy of short-term exposure limits.
The model was calibrated (Bois and Paxman 1992; Bois et al. 1991a) with
observations made in rats exposed to single oral gavage doses of benzene, or to inhalation exposures of
13870 ppm (Sabourin et al. 1987, 1989b), in rats administered single parenteral doses of phenol
(Cassidy and Houston 1984), and in in vitro metabolism studies (Sawahata and Neal 1983). Further
evaluations against data not used in the calibration were not reported.
Target Tissues.
The model simulates amounts and concentrations of benzene and phenol in bone
marrow, a target tissue for benzene metabolites, as well as in blood, liver, fat, and lumped compartments
for other rapidly-perfused and slowly-perfused tissues. The model also simulates amounts of specific
metabolites formed and urinary excretion of the major urinary metabolite, phenol. It does not simulate
concentrations of metabolites, other than phenol, in these tissues.
The model has been applied to simulations for rats. A human model has
been developed that implements a scaled-down version of the rat metabolism model (see Bois et al.
1996).
rats ranging from 13 to 870 ppm and oral gavage doses ranging from 15 to 300 mg/kg.
these routes.
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ingested benzene (and phenol), including rates and amounts of most of the major metabolites formed in
rats. Limitations include: (1) the model has not been evaluated for multiple exposures; (2) although the
model simulates the fate of benzene and phenol, it does not simulate the fate of other metabolites formed
and, therefore, cannot be used to predict concentrations of these metabolites in tissues; and (3) the model,
as configured in Bois et al. (1991a), does not simulate benzene disposition in humans.
Bois et al. 1996
The Bois et al. (1996) model simulates inhalation absorption and
disposition of benzene in humans. Tissues simulated include the blood, bone marrow, fat, liver, lung,
other slowly-perfused tissues, and other rapidly-perfused tissues. Absorption and excretion of benzene in
the lung are assumed to be flow-limited as are exchanges of benzene between blood and tissues. The
model simulates metabolic elimination of benzene as a single capacity-limited (i.e., Michaelis-Menten)
reaction, occurring in bone marrow and liver. Endogenous formation of phenolic metabolites is also
simulated as a zero-order process occurring in liver. The model simulates first-order excretion of total
metabolites and the phenol fraction (approximately 80% of total). Parameter values (physiological and
chemical) were estimated by Bayesian optimization techniques (Markov Chain Monte Carlo analysis)
using reference observations of benzene concentration in blood and urinary excretion of phenol in human
subjects who were exposed to benzene in air (Pekari et al. 1992).
Risk Assessment.
The model has been used to predict rates and amounts of benzene metabolized in
human populations (Bois et al. 1996). The population model (population geometric means and standard
deviations of parameter values) was derived using Markov Chain Monte Carlo analysis with observations
from three human subjects serving as the reference data for inter-individual variability (from Pekari et al.
1992). The population model predicts probability distributions of model outputs (for example, rates or
amounts of benzene metabolized for a given exposure). This approach could be used to evaluate
uncertainty factors in risk assessments intended to account for uncertainties in our understanding of
benzene pharmacokinetics variability.
The model was calibrated with observations of benzene concentrations in
blood and urinary phenol levels, made in three human subjects who were exposed to 1.7 or 10 ppm
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benzene for 4 hours (Pekari et al. 1992). Further evaluations against data not used in the calibration have
not been reported.
Target Tissues.
The model simulates amounts and concentrations of benzene in bone marrow, a
target of benzene toxicity, as well as blood, liver, fat, and lumped compartments for other rapidlyperfused and slowly-perfused tissues. Amounts of total metabolites formed and excreted are simulated;
however, the model does not simulate concentrations of metabolites in these tissues.
The model has been applied to simulations for humans (Bois et al. 1996).
human subjects ranging from 1.7 to 10 ppm (Bois et al. 1996).

The model simulates inhalation exposures. Extrapolation to other routes
(e.g., oral, dermal) would require the extension of the model to include simulations of absorption from
these routes.
Strengths of the model are that it simulates disposition of inhaled
benzene and rates of total metabolism in humans. Limitations include that the model has not been
evaluated for multiple exposures and that the model simulates total metabolism of benzene, and not the
rates of formation of the major metabolites of benzene of toxicological interest.
Brown et al. 1998
The Brown et al. (1998) model simulates inhalation absorption and
disposition of benzene in humans. Tissues simulated include the blood, fat, liver, lung, other slowlyperfused tissues, and other rapidly-perfused tissues. Absorption and excretion of benzene in the lung and
exchange of benzene between blood and tissues are assumed to be flow-limited. The model simulates
capacity-limited (i.e., Michaelis-Menten) metabolic elimination of benzene as a function of the
concentration of benzene in liver. Rates of formation of specific metabolites, or their disposition (e.g.,
excretion), are not simulated. For the purpose of comparing model predictions to observations, 80% of
the total metabolites formed and excreted in urine (i.e., amount of benzene eliminated by metabolism) in
24 hours was assumed to be phenol. The Km parameter for metabolism was derived from Travis et al.
1990; the Vmax was estimated by optimization of the model to observations of blood concentrations of
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benzene and benzene in exhaled breath of female and male subjects who were exposed to 25 ppm
benzene for 2 hours (Sato et al. 1975). Partition coefficients for males and females were derived from
vial equilibrium studies conducted on blood and/or tissues from males and females (Fisher et al. 1997;
Paterson and Mackay 1989).
Risk Assessment.
This model has been used to predict the benzene concentrations in blood and
amounts of benzene metabolized in females and males who experience the same inhalation exposure
scenarios. Females were predicted to metabolize 2326% more benzene than similarly-exposed males.
This difference was attributed, in part, to a higher blood:air partition coefficient for benzene in females.
The model was calibrated by comparing predictions of blood
concentrations of benzene and benzene in exhaled breath of female and male subjects who were exposed
to 25 ppm benzene for 2 hours (Brown et al. 1998; Sato et al. 1975). Further evaluations against data not
used in the calibration were not been reported.
Target Tissues.
The model simulates concentrations of benzene in blood, liver, fat, and lumped
compartments for other rapidly-perfused and slowly-perfused tissues as well as rates of metabolic
elimination of benzene. It does not simulate concentrations of metabolites in these tissues and does not
simulate metabolism in bone marrow, a major target of benzene metabolites.
The model has been applied to simulations for humans.
humans. Evaluations of predictions included observations made during inhalation exposures to 25 ppm
(Brown et al. 1998; Sato et al. 1975).
The model simulates inhalation and has been applied to predicting internal
dose metrics (e.g., benzene concentration in blood, amount benzene metabolized) resulting from
exposures by this route (Brown et al. 1998). Extrapolation to other routes (e.g., oral, dermal) would
require the extension of the model to include simulations of absorption from these routes.
Strengths of the model are that it simulates disposition of inhaled
benzene in female and male humans as well as the concentrations of benzene and rates of metabolic
elimination of benzene metabolized. Limitations of the model include: (1) the model does not simulate
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rates of formation of major benzene metabolites; (2) the model does not simulate fate of metabolites
formed and, therefore, cannot be used to predict concentrations of metabolites in tissues; and (3) the
model does not simulate kinetics of uptake or metabolism of benzene in bone marrow, a major target of
benzene toxicity.
Cole et al. 2001
The Cole et al. (2001) model simulates absorption and disposition of
benzene in the mouse. Tissues simulated include the blood, fat, liver, lung, other slowly-perfused tissues,
and other rapidly-perfused tissues. Gastrointestinal absorption of benzene is simulated as a first-order
process. Absorption and excretion of benzene in the lung are assumed to be flow-limited as are
exchanges of benzene between blood and tissues. The model simulates capacity-limited (i.e., MichaelisMenten) and first-order metabolism of benzene and metabolites in liver (see Table 3-6). Capacity-limited
reactions in bone marrow and liver include benzene to benzene oxide, phenol to hydroquinone, phenol to
catechol, catechol to trihydroxybenzene, and conjugation of phenol and hydroquinone. First-order
reactions in liver include conversion of benzene oxide to phenol, muconic acid, and phenylmercapturic
acid. The model simulates rates of formation of metabolites, tissue distribution of benzene oxide, phenol,
and hydroquinone; and first-order excretion of metabolites in urine. Capacity-limited metabolism
parameter values were estimated from in vitro studies of mouse liver (Lovern et al. 1999; Nedelcheva et
al. 1999; Seaton et al. 1995); first-order parameters were estimated by optimization of model output to
observations of metabolites formed in mice exposed by inhalation or to single gavage doses (Kenyon et
al. 1995; Mathews et al. 1998; Sabourin et al. 1988). Blood:tissue partition coefficients for benzene and
metabolites were derived from Medinsky et al. (1989a) or estimated based on the n-octanol-water
partition coefficient (Poulin and Krishnan 1995).
Risk Assessment.
This model has been used to predict amounts of benzene exhaled and amounts of
benzene metabolites produced in mice during inhalation exposures or following oral gavage exposures to
benzene (Cole et al. 2001).
The model was calibrated with observations made in mice exposed to
single oral gavage doses of benzene, or to inhalation exposures (Cole et al. 2001; Kenyon et al. 1995;
Mathews et al. 1998; Sabourin et al. 1988). Further evaluations against data not used in the calibration
have not been reported.
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Target Tissues.
The model simulates amounts and concentrations of benzene in blood, liver, fat, and
lumped compartments for other rapidly perfused and slowly perfused tissues; rates of formation of
metabolites; tissue distribution of benzene oxide, phenol, and hydroquinone; and first-order excretion of
metabolites in urine. It does not simulate concentrations of metabolites in bone marrow, a target tissue
for benzene metabolites.
The model has been applied to simulations for mice (Cole et al. 2001).
mice (50 ppm) and oral gavage doses ranging from 0.1 to 100 mg/kg (Cole et al. 2001; Kenyon et al.
1995; Mathews et al. 1998; Sabourin et al. 1988).
these routes (Cole et al. 2001).
ingested benzene, including rates and amounts of major metabolites. Most of the metabolism parameter
values were derived empirically from in vitro studies, rather than by model optimization. Limitations
include: (1) the model has not been evaluated for multiple exposures; (2) the model does not simulate the
metabolism of benzene in bone marrow, a major target of benzene toxicity; (3) the model, as configured
in Cole et al. (2001), does not simulate benzene disposition in humans.
3.5
3.5.1
MECHANISMS OF ACTION
Pharmacokinetic Mechanisms
Benzene is readily absorbed via all natural routes of exposure (inhalation, oral, and dermal) and
distributed throughout the body via the blood. Based on physical properties such as slight water
solubility, high lipid solubility, and nonpolarity, benzene is expected to enter the blood via passive
diffusion from gut, lungs, and skin. Benzene is expected to readily bind to plasma proteins (Travis and
Bowers 1989). Being moderately lipophilic, benzene tends to accumulate in fatty tissues. However,
benzene metabolism is relatively rapid and required for hematopoietic and leukemogenic effects to be
expressed. Multiple reactive metabolites appear to be involved in benzene toxicity. As discussed in
detail in Section 3.5.2, potential candidates include benzene oxide, phenolic metabolites (phenol,
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catechol, hydroquinone, 1,2,4-benzenetriol, and 1,2- and 1,4-benzoquinone), and trans,trans-muconaldehyde. Both human and animal data demonstrate the importance of CYP2E1 in benzene metabolism (see
Section 3.4.3 for a detailed discussion). Metabolism is assumed to take place primarily in the liver, with
some secondary metabolism in the bone marrow, the site of characteristic benzene toxicity. Processes
involved in transport of hepatic metabolites of benzene to the critical toxicity target (bone marrow) are
not known, although some degree of covalent binding of reactive benzene metabolites to blood proteins is
expected. At relatively low exposure levels, urinary excretion of conjugated benzene derivatives
represents the major excretory pathway for benzene. Biliary excretion represents a minor excretory
pathway.
3.5.2
Mechanisms of Toxicity
Numerous mechanistic studies have been conducted in an effort to elucidate mechanisms of benzeneinduced hematotoxic and leukemogenic effects, widely recognized as the most critical effects of benzene
exposure. Conversely, benzene-induced effects on reproduction, development, and the nervous system
have not been studied in sufficient detail to assess mechanisms of toxicity for these end points. The
database of information for benzene-induced hematotoxic and leukemogenic effects has been reviewed
extensively (e.g., Bird et al. 2005; Irons 2000; Morgan and Alvares 2005; Ross 1996, 2000, 2005;
Schnatter et al. 2005; Smith 1996a, 1996b; Snyder 2000a, 2000b, 2002; Snyder and Hedli 1996; Snyder
and Kalf 1994). It is generally believed that reactive hepatic metabolites of benzene are transported to the
major toxicity target (bone marrow). Additional metabolism likely occurs in bone marrow. Phenolic
metabolites (phenol, hydroquinone, catechol, 1,2,4-benzenetriol, and 1,2- and 1,4-benzoquinone) appear
to play a major role in benzene toxicity. Smith (1996a, 1999b) noted that the phenolic metabolites can be
metabolized by bone marrow peroxidases, such as myeloperoxidase (MPO), to highly reactive
semiquinone radicals and quinones that stimulate the production of reactive oxygen species. These steps
lead to damage to tubulin, histone proteins, topoisomerase II, other DNA associated proteins, and DNA
itself (clastogenic effects such as strand breakage, mitotic recombination, chromosome translocations, and
aneuploidy). Damage to stem or early progenitor cells would be expressed as hematopoietic and
leukemogenic effects.
Results of several mechanistic studies demonstrate that benzene hematotoxicity is dependent upon
metabolism (see Section 3.4.3 for a detailed discussion). Inhibition of benzene metabolism reduced its
toxicity (Andrews et al. 1977). Partial hepatectomy decreased both benzene metabolism and toxicity
(Sammett et al. 1979). Pretreatment with inducers of metabolism increased both benzene metabolism and
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toxicity (Gad-El-Karim et al. 1985, 1986). Inhibition of CYPs (enzymes that catalyze oxidation pathways
in benzene metabolism) reduced benzene-induced genotoxicity (Tuo et al. 1996). Mice lacking CYP2E1
or microsomal epoxide hydrolase expression were not susceptible to benzene levels known to cause
myelotoxicity and cytotoxicity in wild type mice (Bauer et al. 2003; Valentine et al. 1996a, 1996b).
Occupationally-exposed workers with a phenotype corresponding to rapid CYP2E1 metabolism were
more susceptible to benzene toxicity than those expressing slow CYP2E1 metabolism (Rothman et al.
1997). The enzyme NAD(P)H:quinone oxidoreductase (NQ01), which maintains quinones in reduced
form where they are more readily conjugated and excreted (Nebert et al. 2002), is another example of the
importance of metabolism in benzene hematotoxicity. Between 22% (Caucasian) and 45% (Asian) of the
population is homozygous for an NQ01 allele whereby NQ01 production is negligible. Rothman et al.
(1997) found that workers homozygous for an NQ01 allele whereby NQ01 production is negligible (wild
type) exhibited a 2.4-fold increased risk for benzene hematotoxicity than workers with the normal
genotype. Greater than 7-fold increased risk of benzene hematotoxicity was noted in workers who
expressed both rapid CYP2E1 metabolism and the NQ01 wild type (Rothman et al. 1997).
Benzene metabolism involves the production of reactive metabolites that may act directly on cellular
macromolecules (proteins and DNA). No single metabolite has been implicated; effects are probably due
to many metabolites, which include benzene oxide, reactive products of the phenol pathway (catechol,
hydroquinone, and 1,4-benzoquinone), and trans,trans-muconaldehyde. Evidence that benzene oxide
may play a role in benzene toxicity includes findings that benzene oxide is a product of oxidative benzene
metabolism in mouse, rat, and human liver microsomes (Lovern et al. 1997), benzene oxide can be
released from the liver into the blood (Lindstrom et al. 1997), benzene oxide-protein adducts have been
found in the blood and bone marrow of mice exposed to benzene (McDonald et al. 1994), and benzene
oxide hemoglobin and albumin adducts have been detected in the blood of workers exposed to benzene
(Rappaport et al. 2002a, 2002b; Yeowell-OConnell et al. 1998).
Urinary trans,trans-muconic acid has been detected in humans and animals following benzene exposure,
although its purported reactive precursor (trans,trans-muconaldehyde; see Figure 3-3) has not been
detected in vivo. The highly reactive trans,trans-muconaldehyde, which has been found in mouse hepatic
microsomes (Latriano et al. 1986), can undergo reductive and oxidative metabolism (Goon et al. 1992)
and has been shown to be hematotoxic (Witz et al. 1985). A small amount (<0.05%) of parenterallyadministered trans,trans-muconaldehyde to mice reached the bone marrow (Zhang et al. 1997). Rivedal
and Witz (2005) found trans,trans-muconaldehyde to be a strong inhibitor of gap junction intercellular
communication in rat liver epithelial cells.
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Phenolics (phenol, catechol, hydroquinone; 1,2,4-benzenetriol; 1,2- and 1,4-benzoquinone) are major
metabolites of benzene that have been shown to persist in bone marrow following inhalation exposure
(Rickert et al. 1979; Sabourin et al. 1988). Hydroquinone induces chromosomal damage in lymphocytes
in vitro in a manner similar to that observed in lymphocytes of benzene-exposed workers (Eastmond et al.
1994; Stillman et al. 1997; Zhang et al. 1998b). Kolachana et al. (1993) demonstrated that both
hydroquinone and 1,2,4-benzenetriol cause oxidative damage to DNA in mouse bone marrow (in vivo)
and human myeloid cells (in vitro). Glutathione adducts of 1,4-benzoquinone have been shown to be
hematotoxic in bone marrow of mice exposed to benzene (Bratton et al. 1997). Additional evidence that
phenolic metabolites may play an important role in benzene toxicity includes the finding that MPO, an
enzyme found in high concentration in bone marrow (Bainton et al. 1971), catalyzes the oxidation of
polyphenols to reactive quinones, semiquinones, and oxygen radicals (Nebert et al. 2002). This can lead
to strand breaks and inhibition of topoisomerases and microtubule assembly, which could result in
chromosome damage (Chen and Eastmond 1995; Eastmond et al. 2001; Irons and Neptune 1980; Smith
1996a, 1996b). Eastmond et al. (2005) demonstrated that hydroquinone can be readily activated to a
potent topoisomerase II inhibitor in the presence of human MPO and H2O2 and that partial inhibition
occurs at hydroquinone concentrations as low as 50 nM. Irons and Neptune (1980) suggested that
benzene-derived hydroquinone may inhibit cell replication by covalently binding to tubulin, a protein
essential for spindle formation in mitosis.
Benzene-induced effects on DNA have been studied in some detail. Schwartz et al. (1985) demonstrated
that benzene metabolites inhibit mitochondrial DNA polymerase. Hydroquinone has been shown to
inhibit ribonucleotide reductase, a key step in DNA synthesis (Li et al. 1997, 1998). Rushmore et al.
(1984) found that benzene metabolites covalently bind to mitochondrial DNA and inhibit RNA synthesis.
Benzene metabolites bound to hepatic DNA have been observed in animals following inhalation exposure
to radiolabeled benzene (Lutz and Schlatter 1977). However, the reported levels of DNA adduct
formation appear to be low. For example, Creek et al. (1997) observed adducts to both protein and DNA
in the range of nanograms per kilogram in mice given radiolabeled benzene.
Benzene-induced DNA damage may result from the oxidation of DNA by reactive oxygen species that
are produced during benzene metabolism. Both 1,4-benzoquinone and hydroquinone are known to
increase superoxide, nitric oxide, and hydrogen peroxide in HL-60 cells (Rao and Snyder 1995). Chen et
al. (2004) observed nitric oxide-derived benzene metabolites, namely nitrobenzene, nitrophenyl, and
nitrophenol isomers in the bone marrow of mice 1 hour following intraperitoneal injection of a 400 mg/kg
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dose of benzene. These nitro metabolites were either not detected in other tissues or were present in
much smaller concentrations, indicating that they were most likely produced in bone marrow. Brunmark
and Cadenas (1988) described a metabolic pathway from hydroquinone leading to the production of
glutathionyl-benzenetriol, which can undergo autoxidation leading to superoxide formation. Rao (1996)
suggested that benzene-induced DNA damage is mediated by the release of free iron in the bone marrow
(probably by polyphenolic metabolites), followed by the chelation of iron by hydroquinone or
benzenetriol to yield a reactive oxygen-generating species such as superoxide, which causes oxidative
damage to DNA.
The expression of benzene toxicity may involve multiple benzene metabolites. All of the known
unconjugated metabolites of benzene, with the exception of phenol and 1,2,4-benzenetriol, have been
shown to decrease erythropoiesis (Snyder and Hedli 1996). In mice, the combination of phenol and
hydroquinone resulted in exacerbated loss of bone marrow cellularity (Eastmond et al. 1987), increased
peroxidatic activation of hydroquinone (Subrahmanyam et al. 1989, 1990), and increased DNA damage
(Lvay and Bodell 1992; Marrazzini et al. 1994). Combinations of either phenol and hydroquinone, or
phenol and catechol, were more hematotoxic than any of the metabolites given alone (Guy et al. 1991).
The combination of hydroquinone and muconaldehyde was the most potent in inhibiting erythropoiesis
(Snyder et al. 1989). Catechol was found to stimulate the peroxidase-mediated activation of
hydroquinone and produced a synergistic genotoxic effect in lymphocytes (Robertson et al. 1991).
3.5.3
Animal-to-Human Extrapolations
Pathways of benzene metabolism are generally similar among various rodent and nonhuman primate
species. However, species differences exist regarding capacity to metabolize benzene and relative
proportions of various benzene metabolites formed.
Species differences exist in absorption and retention of benzene. For example, following 6-hour
exposures to low concentrations (710 ppm) of benzene vapors, mice retained 20% of the inhaled
benzene, whereas rats and monkeys retained only 34% (Sabourin et al. 1987, 1992). Mice exhibit a
greater overall capacity to metabolize benzene, compared to rats. An Inhalation exposure to 925 ppm
resulted in an internal dose of 152 mg/kg in mice, approximately 15% of which was excreted as parent
compound, and an internal dose of 116 mg/kg in rats, approximately 50% of which was excreted
unchanged (Henderson et al. 1992; Sabourin et al. 1987).
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The proportions of benzene metabolites produced depend on both species and exposure concentration.
Hydroquinones and muconic acid (potential sources of benzene toxicity) were detected in much higher
concentrations in the blood, liver, lung, and bone marrow of mice than rats, following a 6-hour inhalation
exposure to benzene at a concentration of 50 ppm (Sabourin et al. 1988). It is generally understood that
metabolic profiles of benzene in mice and humans are more similar than those of humans and rats.
Sabourin et al. (1989a) noted increased production of detoxification metabolites (phenylglucuronide and
prephenylmercapturic acid) and decreased production of potentially toxic metabolites (hydroquinones and
muconic acid) in both mice and rats exposed to benzene at much higher concentrations (600 ppm in air or
200 mg/kg orally), which indicates that extrapolation of toxicological results from studies using high
exposure concentrations to low exposure scenarios may result in an underestimation of risk.
Recent PBPK models have tried to address benzene metabolism in an effort to derive animal-to-human
extrapolations (Bois et al. 1991a, 1996; Cole et al. 2001; Medinsky 1995; Medinsky et al. 1989a, 1989b,
1989c; Travis et al. 1990). Each model described a multicompartmental model that attempted to relate
the generation of metabolites to end points of benzene toxicity. The generation of hydroquinone and
muconaldehyde in the liver, with further metabolism in the bone marrow, has been addressed as well as
the available data allow. However, the models are not sufficiently refined to allow them to accurately
predict human metabolism.
3.6
TOXICITIES MEDIATED THROUGH THE NEUROENDOCRINE AXIS
Recently, attention has focused on the potential hazardous effects of certain chemicals on the endocrine
system because of the ability of these chemicals to mimic or block endogenous hormones. Chemicals
with this type of activity are most commonly referred to as endocrine disruptors. However, appropriate
terminology to describe such effects remains controversial. The terminology endocrine disruptors,
initially used by Thomas and Colborn (1992), was also used in 1996 when Congress mandated the EPA to
develop a screening program for ...certain substances [which] may have an effect produced by a
naturally occurring estrogen, or other such endocrine effect[s].... To meet this mandate, EPA convened a
panel called the Endocrine Disruptors Screening and Testing Advisory Committee (EDSTAC), and in
1998, the EDSTAC completed its deliberations and made recommendations to EPA concerning endocrine
disruptors. In 1999, the National Academy of Sciences released a report that referred to these same types
of chemicals as hormonally active agents. The terminology endocrine modulators has also been used to
convey the fact that effects caused by such chemicals may not necessarily be adverse. Many scientists
agree that chemicals with the ability to disrupt or modulate the endocrine system are a potential threat to
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the health of humans, aquatic animals, and wildlife. However, others think that endocrine-active
chemicals do not pose a significant health risk, particularly in view of the fact that hormone mimics exist
in the natural environment. Examples of natural hormone mimics are the isoflavinoid phytoestrogens
(Adlercreutz 1995; Livingston 1978; Mayr et al. 1992). These chemicals are derived from plants and are
similar in structure and action to endogenous estrogen. Although the public health significance and
descriptive terminology of substances capable of affecting the endocrine system remains controversial,
scientists agree that these chemicals may affect the synthesis, secretion, transport, binding, action, or
elimination of natural hormones in the body responsible for maintaining homeostasis, reproduction,
development, and/or behavior (EPA 1997). Stated differently, such compounds may cause toxicities that
are mediated through the neuroendocrine axis. As a result, these chemicals may play a role in altering,
for example, metabolic, sexual, immune, and neurobehavioral function. Such chemicals are also thought
to be involved in inducing breast, testicular, and prostate cancers, as well as endometriosis (Berger 1994;
Giwercman et al. 1993; Hoel et al. 1992).
No information was located to indicate that benzene may adversely affect the endocrine system.
3.7
CHILDRENS SUSCEPTIBILITY
This section discusses potential health effects from exposures during the period from conception to
maturity at 18 years of age in humans, when all biological systems will have fully developed. Potential
effects on offspring resulting from exposures of parental germ cells are considered, as well as any indirect
effects on the fetus and neonate resulting from maternal exposure during gestation and lactation.
Relevant animal and in vitro models are also discussed.
Children are not small adults. They differ from adults in their exposures and may differ in their
susceptibility to hazardous chemicals. Childrens unique physiology and behavior can influence the
extent of their exposure. Exposures of children are discussed in Section 6.6, Exposures of Children.
Children sometimes differ from adults in their susceptibility to hazardous chemicals, but whether there is
a difference depends on the chemical (Guzelian et al. 1992; NRC 1993). Children may be more or less
susceptible than adults to health effects, and the relationship may change with developmental age
(Guzelian et al. 1992; NRC 1993). Vulnerability often depends on developmental stage. There are
critical periods of structural and functional development during both prenatal and postnatal life, and a
particular structure or function will be most sensitive to disruption during its critical period(s). Damage
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may not be evident until a later stage of development. There are often differences in pharmacokinetics
and metabolism between children and adults. For example, absorption may be different in neonates
because of the immaturity of their gastrointestinal tract and their larger skin surface area in proportion to
body weight (Morselli et al. 1980; NRC 1993); the gastrointestinal absorption of lead is greatest in infants
and young children (Ziegler et al. 1978). Distribution of xenobiotics may be different; for example,
infants have a larger proportion of their bodies as extracellular water, and their brains and livers are
proportionately larger (Altman and Dittmer 1974; Fomon 1966; Fomon et al. 1982; Owen and Brozek
1966; Widdowson and Dickerson 1964). The infant also has an immature blood-brain barrier (Adinolfi
1985; Johanson 1980) and probably an immature blood-testis barrier (Setchell and Waites 1975). Many
xenobiotic metabolizing enzymes have distinctive developmental patterns. At various stages of growth
and development, levels of particular enzymes may be higher or lower than those of adults, and
sometimes unique enzymes may exist at particular developmental stages (Komori et al. 1990; Leeder and
Kearns 1997; NRC 1993; Vieira et al. 1996). Whether differences in xenobiotic metabolism make the
child more or less susceptible also depends on whether the relevant enzymes are involved in activation of
the parent compound to its toxic form or in detoxification. There may also be differences in excretion,
particularly in newborns who all have a low glomerular filtration rate and have not developed efficient
tubular secretion and resorption capacities (Altman and Dittmer 1974; NRC 1993; West et al. 1948).
Children and adults may differ in their capacity to repair damage from chemical insults. Children also
have a longer remaining lifetime in which to express damage from chemicals; this potential is particularly
relevant to cancer.
Certain characteristics of the developing human may increase exposure or susceptibility, whereas others
may decrease susceptibility to the same chemical. For example, although infants breathe more air per
kilogram of body weight than adults breathe, this difference might be somewhat counterbalanced by their
alveoli being less developed, which results in a disproportionately smaller surface area for alveolar
absorption (NRC 1993).
No clear evidence of age-related differences in susceptibility to benzene toxicity was located. Benzene
crosses the placenta and can be found in cord blood at concentrations that equal or exceed those of
maternal blood (Dowty et al. 1976). Nursing infants can be exposed to benzene in the breast milk
(Fabietti et al. 2004). Limited animal studies indicate that in utero exposure to benzene results in
hematological changes similar to those observed in animals exposed only as adults (Corti and Snyder
1996; Keller and Snyder 1986, 1988). There is some indication that parental occupational exposure to
benzene may play a role in childhood leukemia (Buckley et al. 1989; McKinney et al. 1991; Shaw et al.
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1984; Shu et al. 1988). However, none of these studies indicate whether children may be at greater risk
than adults for benzene toxicity. Results of Infante-Rivard et al. (2005) indicate that maternal exposure to
benzene during pregnancy or from 2 years before pregnancy up to birth does not result in increased risk of
childhood acute lymphoblastic leukemia (ALL), a frequent form of childhood cancer (Infante-Rivard et
al. 2005). However, no information was located regarding the risk of childhood AML, the form of
leukemia most frequently associated with exposure to benzene.
Children could potentially be at increased risk for significant benzene exposure via the inhalation route
based on higher activity levels and ventilation rates than adults. However, no information was located to
indicate that children are at increased risk for benzene toxicity. Age-related differences in benzene
metabolism could potentially affect susceptibility. Results of one human study indicate that CYP2E1, a
major enzyme involved in benzene metabolism, is not present in the fetus, but appears in rapidly
increasing concentrations during early postnatal development (Vieira et al. 1996). This suggests that
fetuses and neonates may be at decreased risk of benzene toxicity due to a reduced metabolic capacity.
No information was located regarding potential age-related differences in pharmacodymanic processes
such as benzene-target interactions in the hematopoietic system.
3.8
BIOMARKERS OF EXPOSURE AND EFFECT
Biomarkers are broadly defined as indicators signaling events in biologic systems or samples. They have
been classified as markers of exposure, markers of effect, and markers of susceptibility (NAS/NRC
1989).
Due to a nascent understanding of the use and interpretation of biomarkers, implementation of biomarkers
as tools of exposure in the general population is very limited. A biomarker of exposure is a xenobiotic
substance or its metabolite(s) or the product of an interaction between a xenobiotic agent and some target
molecule(s) or cell(s) that is measured within a compartment of an organism (NAS/NRC 1989). The
preferred biomarkers of exposure are generally the substance itself, substance-specific metabolites in
readily obtainable body fluid(s), or excreta. However, several factors can confound the use and
interpretation of biomarkers of exposure. The body burden of a substance may be the result of exposures
from more than one source. The substance being measured may be a metabolite of another xenobiotic
substance (e.g., high urinary levels of phenol can result from exposure to several different aromatic
compounds). Depending on the properties of the substance (e.g., biologic half-life) and environmental
conditions (e.g., duration and route of exposure), the substance and all of its metabolites may have left the
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body by the time samples can be taken. It may be difficult to identify individuals exposed to hazardous
substances that are commonly found in body tissues and fluids (e.g., essential mineral nutrients such as
copper, zinc, and selenium). Biomarkers of exposure to benzene are discussed in Section 3.8.1.
Biomarkers of effect are defined as any measurable biochemical, physiologic, or other alteration within an
organism that, depending on magnitude, can be recognized as an established or potential health
impairment or disease (NAS/NRC 1989). This definition encompasses biochemical or cellular signals of
tissue dysfunction (e.g., increased liver enzyme activity or pathologic changes in female genital epithelial
cells), as well as physiologic signs of dysfunction such as increased blood pressure or decreased lung
capacity. Note that these markers are not often substance specific. They also may not be directly
adverse, but can indicate potential health impairment (e.g., DNA adducts). Biomarkers of effects caused
by benzene are discussed in Section 3.8.2.
A biomarker of susceptibility is an indicator of an inherent or acquired limitation of an organism's ability
to respond to the challenge of exposure to a specific xenobiotic substance. It can be an intrinsic genetic or
other characteristic or a preexisting disease that results in an increase in absorbed dose, a decrease in the
biologically effective dose, or a target tissue response. If biomarkers of susceptibility exist, they are
discussed in Section 3.10, Populations That Are Unusually Susceptible.
3.8.1
Biomarkers Used to Identify or Quantify Exposure to Benzene
Several biomarkers of exposure to benzene have been reported in the literature. Unmetabolized benzene
can be detected in the expired air and urine of humans exposed to benzene vapors (Farmer et al. 2005;
Fustinoni et al. 2005; Ghittori et al. 1993; Nomiyama and Nomiyama 1974a, 1974b; Sherwood 1988;
Srbova et al. 1950; Waidyanatha et al. 2001). Urinary phenol measurements have routinely been used for
monitoring occupational exposure to benzene (OSHA 1987), and urinary phenol levels appear to be
correlated with exposure levels (Astier 1992; Inoue et al. 1986, 1988b; Karacic et al. 1987; Pagnotto et al.
1961; Pekari et al. 1992).
Urinary trans,trans-muconic acid has been widely studied as a biomarker of exposure to benzene
(Boogaard and van Sittert 1995, 1996; Ducos et al. 1990, 1992; Inoue et al. 1989b, 2000; Lee et al. 1993;
Melikian et al. 1993, 1994; Pezzagno et al. 1999; Popp et al. 1994; Qu et al. 2005; Rothman et al. 1998;
Ruppert et al. 1997; Sanguinetti et al. 2001; van Sittert et al. 1993; Weaver et al. 2000). Urinary
S-phenylmercapturic acid levels have also been correlated with occupational exposure to benzene
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(Boogaard and van Sittert 1995, 1996; Farmer et al. 2005; Inoue et al. 2000; Jongeneelen et al. 1987;
Popp et al. 1994; Qu et al. 2005). Significant exposure-response trends for urinary trans,trans-muconic
acid and S-phenylmercapturic acid levels have been demonstrated in occupationally-exposed subjects at
exposure levels of 1 ppm (Qu et al. 2005). The American Conference of Governmental Industrial
Hygienists (ACGIH) has established 25 g S-phenylmercapturic acid/g creatinine in the urine and 500 g
trans,trans-muconic acid/g creatinine in the urine as Biological Exposure Indices (BEIs) for benzene
exposure in the workplace (ACGIH 2006). The BEI is primarily an index of exposure and not a level at
which health effects might occur from exposure to benzene. Positive correlations have been made
between benzene workplace air levels and urinary catechol and hydroquinone in exposed workers (Inoue
et al. 1988a, 1988b; Rothman et al. 1998).
Hemoglobin and albumin adducts of the benzene metabolites, benzene oxide and 1,4-benzoquinone, have
been used as biomarkers of exposure to benzene (Bechtold and Henderson 1993; Bechtold et al. 1992a,
1992b; Smith and Rothman 2000; Yeowell-OConnell et al. 1998, 2001). Furthermore, DNA adducts
with benzene metabolites have been found after benzene exposure (Hedli et al. 1991; Lutz and Schlatter
1977; Reddy et al. 1989).
Ong et al. (1995) evaluated various biomarkers of benzene exposure for their relationship with
environmental benzene levels. Muconic acid in the urine correlated best with environmental benzene
concentrations. Urinary hydroquinone levels were the most accurate biomarker of exposure for the
phenolic metabolites of benzene, followed by phenol and catechol. No correlation was found between
environmental benzene levels and unmetabolized benzene in the urine, although other studies suggest that
benzene in the urine may be a useful biomarker of occupational exposure (Ghittori et al. 1993).
The biomarkers discussed in the preceding paragraphs appear to be adequate indicators of exposure to
benzene at relatively high occupational exposure levels, and may serve as biomarkers in acute exposure
scenarios involving relatively high levels of benzene. However, some of these biomarkers do not appear
to be reliable indicators of environmental exposure to benzene (concentrations below the common
industrial standard of 1 ppm TWA). For example, results from data collected on 152 chemical workers
showed a linear relationship between the concentration of benzene in breathing zone air (when greater
than 10 ppm) and urinary concentrations of catechol and hydroquinone (Inoue et al. 1988a). Workers
who had an average work-site exposure of 10 ppm benzene showed no significant differences in the
concentration of urinary catechol or hydroquinone when compared to a group of unexposed subjects. In a
study of pharmacy workers exposed to benzene levels measured in the parts per billion (ppb) range, there
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was no significant difference in urinary levels of trans,trans-muconic acid between subjects exposed to
1.5 ppb and those exposed to 2.5 ppb (Sanguinetti et al. 2001). Recent reports indicate that urinary
benzene may serve as the most sensitive biomarker of exposure to benzene concentrations well below
1 ppm (Farmer et al. 2005; Fustinoni et al. 2005).
Several additional factors must be taken into account when assessing the reliability of biomarkers of
exposure to benzene. High and variable background levels of phenol and its metabolites result from
ingestion of vegetables, exposure to other aromatic compounds, ingestion of ethanol, and inhalation of
cigarette smoke (Nakajima et al. 1987). Relatively high urinary phenol levels (542 mg/L) have been
found in persons with no known exposure to benzene (NIOSH 1974). Although muconic acid is used as a
marker for benzene exposure, muconic acid in the urine can also result from ingestion of sorbic acid, a
common food preservative (Ducos et al. 1990). Inoue et al. (1989b) suggested that individual urinary
trans,trans-muconic acid content was not a useful index of benzene exposure due to large variations in
measured individual background urinary trans,trans-muconic acid values.
In summary, several benzene metabolites may serve as biomarkers of exposure to benzene. Urinary
benzene appears to be the most sensitive biomarker for low-level exposure to benzene. Refer to
Tables 7-1 and 7-2 for information regarding analytical methods for determining benzene in biological
samples and benzene metabolites in urine.
3.8.2
Biomarkers Used to Characterize Effects Caused by Benzene
In addition to using levels of benzene and benzene metabolites for monitoring purposes, various
biological indices might also be helpful in characterizing the effects of exposure to benzene. As with
monitoring for benzene exposure, monitoring for effects may best be accomplished through the use of a
series of biomarkers with correlation of the results. Decreases in erythrocyte and leukocyte counts have
been used as an indicator of high occupational exposures. Monitoring of benzene workers has included
monthly blood counts, with workers being removed from areas of high benzene exposure when leukocyte
counts fell below 4,000/mm3 or erythrocyte counts fell below 4,000,000/mm3 (ITII 1975; OSHA 1987).
Hayes (1992) indicates that benzene-related leukopenia, commonly thought of as an intermediate end
point in the process of developing benzene-related leukemia, is considered a biomarker of benzene
poisoning in China, not necessarily related to leukemia. Leukocyte alkaline phosphatase (LAP) activity
was increased in benzene workers exposed to about 31 ppm for a chronic time period (Songnian et al.
1982). Increased LAP activity is an indicator of myelofibrosis and is associated with both decreased
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white blood cell counts and changes in bone marrow activity. The change in LAP activity could be used
in the diagnosis of benzene poisoning since it was more sensitive than the change in the leukocyte count,
although it is not a biomarker that is specific for benzene. Additionally, it seems reasonable that
chromosomal aberrations in bone marrow and peripheral blood lymphocytes and sister chromatid
exchanges could be used to monitor for benzene effects (Eastmond et al. 1994; Van Sittert and de Jong
1985). Benzene metabolites have also been found to form adducts with DNA (Chenna et al. 1995; Lutz
and Schlatter 1977; Norpoth et al. 1988; Rushmore et al. 1984; Snyder et al. 1987).
Exposure to benzene causes toxic effects in the bone marrow via its metabolites and possibly by benzene
via solvent effects (Eastmond et al. 1987; Gad-El-Karim et al. 1985; Hedli et al. 1991; Irons et al. 1980).
Therefore, it is possible that hematological tests could be used as markers of hematotoxicity. To date,
surveillance and early diagnosis of benzene hematotoxicity rely primarily on the complete blood count,
including hemoglobin, hematocrit, erythrocyte count, leukocyte count, and differential and platelet
counts. In effect, complete blood counts and marrow exams should be good for early detection of
preleukemic lesions. Additionally, cytogenetic tests of marrow cells are being used. Workers exposed to
benzene in the air have shown elevated levels of delta-aminolevulinic acid in erythrocytes and elevated
coproporphyrin in the urine (Kahn and Muzyka 1973). These may be biomarkers for disruption of
porphyrin synthesis and may be early indicators of adverse hematological effects. These effects are not
specific for benzene. Hedli et al. (1991) observed that in rats, benzene metabolite-DNA adducts were
observed in the bone marrow at doses that did not affect bone marrow cellularity, and suggested that
monitoring of the bone marrow DNA adducts might be a sensitive bioassay of genotoxic effects of
benzene exposure. Work by other researchers also suggests that monitoring DNA adducts or products of
DNA damage might be useful (Bodell et al. 1993; Chen et al. 1994; Lagorio et al. 1994a; Reddy et al.
1989).
3.9
INTERACTIONS WITH OTHER CHEMICALS
Studies have been conducted on the interaction of benzene with other chemicals, both in vivo and in the
environment. Benzene metabolism is complex, and various xenobiotics can induce or inhibit specific
routes of detoxification and/or activation in addition to altering the rate of benzene metabolism and
clearance from the body. Toluene, Aroclor 1254, phenobarbital, acetone, and ethanol are known to alter
the metabolism and toxicity of benzene. Interactions reported in in vivo studies occurred at relatively
high benzene exposure levels, which would not likely be encountered near hazardous waste sites.
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Ethanol and benzene induce the formation of the hepatic cytochrome P-450 isoenzyme, CYP2E1, in
rabbits and rats (Gut et al. 1993; Johansson and Ingelman-Sundberg 1988), although benzene derivatives,
such as toluene and xylene, can inhibit the enzymatic activity of the isozyme (Koop and Laethem 1992).
Ethanol enhances both the metabolism (in vitro) and the toxicity (in vivo) of benzene in animals (Baarson
et al. 1982; Nakajima et al. 1985). Administration of ethanol (5 or 15% in drinking water, 4 days/week
for 13 weeks) to mice exposed to benzene vapors at a concentration of 300 ppm, 6 hours/day,
5 days/week for 13 weeks) resulted in greater severity of benzene-induced hematological effects (anemia,
lymphocytopenia, bone marrow aplasia, transient increases in normoblasts and peripheral blood atypia)
relative to benzene-exposed mice not given ethanol (Baarson et al. 1982). The modulating effects of
benzene were dose dependent. The enhancement of the hematotoxic effects of benzene by ethanol may
be of particular concern for benzene-exposed workers who consume alcohol (Nakajima et al. 1985),
although the interactions demonstrated in the mice occurred at much higher benzene exposure
concentrations than would likely be experienced in workplace air. Benzene can interfere with the
disappearance of ethanol from the body. Accordingly, increased central nervous system disturbances
(e.g., depression) may occur following concurrent exposure to high levels of benzene and ethanol.
Other chemicals that induce specific isoenzymes of cytochrome P-450 can increase the rate of benzene
metabolism and may alter metabolism pathways favoring one over another. Ikeda and Ohtsuji (1971)
presented evidence that benzene hydroxylation was stimulated when rats were pretreated with
phenobarbital and then exposed to 1,000 ppm of benzene vapor for 8 hours/day for 2 weeks.
Additionally, phenobarbital pretreatment increased the rate of metabolism by 40% in rats and 70% in
mice (Pawar and Mungikar 1975). In contrast, rats exposed to phenobarbital showed no effects on the
metabolism of micromolar amounts (35112.8 mol) of benzene in vitro (Nakajima et al. 1985).
Co-administration of toluene inhibited the biotransformation of benzene to phenol in rats (Ikeda et al.
1972; Inoue et al. 1988b). This was due to competitive inhibition of the oxidation mechanisms involved
in the metabolism of benzene. Phenobarbital pretreatment of the rats alleviated the suppressive effect of
toluene on benzene hydroxylation by the induction of oxidative activities in the liver. This effect has
been observed in other studies in rats (Purcell et al. 1990).
Mathematical models of benzene and phenol metabolism suggest that the inhibition by benzene of phenol
metabolism, and by phenol on benzene metabolism, occurs through competition for a common reaction
site, which can also bind catechol and hydroquinone (Schlosser et al. 1993). Flavonoids have been shown
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to inhibit phenol hydroxylase or increase phenol hydroxylase activity in a dose-dependent manner,

dependent on the oxidation potential of the flavonoid (Hendrickson et al. 1994).
SKF-525A and carbon monoxide are classic inhibitors of cytochrome P-450s. The binding between
P-450 and carbon monoxide or SKF-525A is coordinate covalent. Carbon monoxide inhibits all
cytochrome P-450 isoenzymes since it binds to the heme component of cytochrome P-450, whereas
SKF-525A inhibits specific types. SKF-525A inhibited benzene metabolism in the rat (Ikeda et al. 1972).
Injection of 80 mg/kg of SKF-525A in rats resulted in a depression of phenol excretion. It also prolonged
phenol excretion and interfered in the conversion of benzene to glucuronides and free phenols. Carbon
monoxide, aniline, aminopyrine, cytochrome C, and metyrapone inhibited benzene metabolism in vitro by
mouse liver microsomes (Gonasun et al. 1973).
3.10 POPULATIONS THAT ARE UNUSUALLY SUSCEPTIBLE
A susceptible population will exhibit a different or enhanced response to benzene than will most persons
exposed to the same level of benzene in the environment. Reasons may include genetic makeup, age,
health and nutritional status, and exposure to other toxic substances (e.g., cigarette smoke). These
parameters result in reduced detoxification or excretion of benzene, or compromised function of organs
affected by benzene. Populations who are at greater risk due to their unusually high exposure to benzene
are discussed in Section 6.7, Populations with Potentially High Exposures.
Variability in human susceptibility to benzene toxicity may be related, at least in part, to genetic
polymorphisms associated with metabolic processes. As discussed in Section 3.4.3, the flavoenzyme,
NAD(P)H:quinone oxidoreductase (NQ01), catalyzes the reduction of 1,2- and 1,4-benzoquinone
(reactive metabolites of benzene) to catechol and hydroquinone, respectively (Nebert et al. 2002), thus
protecting cells from oxidative damage by preventing redox cycling. The NQ01*1 (wild type) allele
codes for normal NQ01 enzyme and activity. An NQ01*2 allele encodes a nonsynonymous mutation that
has negligible NQ01 activity. Approximately 5% of Caucasians and African Americans, 15% of
Mexican-Americans, and 20% of Asians are homozygous for the NQ01*2 allele (Kelsey et al. 1997;
Smith and Zhang 1998). Rothman et al. (1997) demonstrated that workers expressing negligible NQ01
activity were at increased risk of benzene poisoning (Rothman et al. 1997). In the same group of workers,
those expressing rapid CYP2E1 activity were also at increased risk of benzene poisoning. Those workers
with polymorphisms for both negligible NQ01 activity and rapid CYP2E1 activity exhibited greater than
7-fold increased risk of benzene poisoning than workers not expressing these polymorphisms. These
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results indicate that individuals expressing rapid CYP2E1 activity may also be at increased risk for
benzene toxicity.
Polymorphisms have been described for many of the glutathione genes (Cotton et al. 2000; Hengstler et
al. 1998; Strange and Fryer 1999), the myeloperoxidase gene (Williams 2001), and the epoxide hydrolase
gene (Omiecinski et al. 2000), which are known to be involved in benzene metabolism. However, the
potential involvement of such polymorphisms in benzene toxicity have not been demonstrated in
benzene-exposed workers.
Individuals with medical conditions that include reduced bone marrow function or decreased blood
factors would be at increased risk for benzene toxicity. Treatments for certain medical conditions might
result in decreases in particular blood factors, which could lead to increased susceptibility to benzene
poisoning.
Ethanol can increase the severity of benzene-induced anemia, lymphocytopenia, and reduction in bone
marrow cellularity, and produce transient increases in normoblasts in the peripheral blood and atypical
cellular morphology (Baarson et al. 1982). The enhancement of the hematotoxic effects of benzene by
ethanol is of particular concern for benzene-exposed workers who consume alcohol (Nakajima et al.
1985). Accordingly, increased central nervous system disturbances (e.g., depression) may be expected
following concurrent exposure to benzene and ethanol.
Gender-related differences in susceptibility to benzene toxicity have been observed in animals. For
example, Kenyon et al. (1998) exposed male and female mice to benzene vapor concentrations of 100 or
600 ppm and found increased benzene metabolism and associated genotoxicity in males, relative to
females. Brown et al. (1998) used a PBPK modeling approach to assess potential gender-related
differences in susceptibility to benzene. Their results suggest that women exhibit a higher blood/air
partition coefficient and maximum velocity of benzene metabolism than men, and that women metabolize
23-26% more benzene than men under similar exposure scenarios. However, gender-related differences
in susceptibility among benzene-exposed workers were not located in available reports.
At early stages of human development, metabolic pathways may not be fully functional, which might
result in a lower level of susceptibility. Young children might experience increased susceptibility to
benzene by inhalation due to increased breathing rates and potential for increased absorption, relative to
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adults. However, no definitive human or animal data were located regarding age-related differences in
susceptibility to benzene.
3.11 METHODS FOR REDUCING TOXIC EFFECTS
This section will describe clinical practice and research concerning methods for reducing toxic effects of
exposure to benzene. However, because some of the treatments discussed may be experimental and
unproven, this section should not be used as a guide for treatment of exposures to benzene. When
specific exposures have occurred, poison control centers and medical toxicologists should be consulted
for medical advice. The following texts provide specific information about treatment following exposures
to benzene:
Goldfrank LR, Flomenbaum NE, Lewin NA, et al., eds. 1998. Goldfranks toxicologic emergencies. 6th
ed. Stamford: Appleton & Lange, 1384-1398.
Haddad LM, Shannon MW, Winchester JF. 1998. Clinical management of poisoning and drug overdose.
3rd ed. Philadelphia, PA: W.B Saunders Company, 940-941.
3.11.1 Reducing Peak Absorption Following Exposure
Human exposure to benzene can occur by inhalation, oral, or dermal routes. General recommendations
for reducing absorption of benzene following acute high-level inhalation exposure have included moving
the patient to fresh air and monitoring for respiratory distress (Bronstein and Currance 1988; Haddad et
al. 1998; HSDB 2007; Kunisaki and Augenstein 1994; Stutz and Janusz 1988). The administration of
100% humidified supplemental oxygen with assisted ventilation, when required, has also been suggested
(Bronstein and Currance 1988; Haddad et al. 1998; Kunisaki and Augenstein 1994; Stutz and Janusz
1988). In the case of eye exposure, irrigation with copious amounts of water or saline has been
recommended (Bronstein and Currance 1988; Haddad et al. 1998; HSDB 2007; Stutz and Janusz 1988).
The removal of contaminated clothing and a thorough washing of exposed areas with soap and water have
also been recommended (Bronstein and Currance 1988; Ellenhorn and Barceloux 1988; Haddad et al.
1998; Kunisaki and Augenstein 1994; Stutz and Janusz 1988). Some sources suggest the administration
of water or milk to the victim after ingestion of benzene (Stutz and Janusz 1988). Emesis may be
indicated following recent substantial ingestion of benzene (Ellenhorn and Barceloux 1988); however,
other sources do not recommend the use of emetics because of the risk of aspiration pneumonitis,
especially once the patient loses consciousness (Bronstein and Currance 1988; Haddad et al. 1998). Some
sources recommend gastric lavage if indicated (Haddad et al. 1998; Kunisaki and Augenstein 1994).
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While lavage may be useful, induction of emesis should be regarded with great caution because the rapid
onset of central nervous system depression may lead to aspiration. Although of unproven value,
administration of a charcoal slurry, aqueous or mixed with saline cathartic or sorbitol, has also been
suggested as a way to stimulate fecal excretion of the chemical before it is completely absorbed by the
body (HSDB 2007; Kunisaki and Augenstein 1994; Stutz and Janusz 1988). Diazepam and phenytoin
may be helpful in controlling seizures (Bronstein and Currance 1988; HSDB 2007; Stutz and Janusz
1988). Administration of epinephrine or other catecholamines has not been recommended because of the
possibility of myocardial sensitization and subsequent arrhythmia (Haddad et al. 1998; HSDB 2007;
Nahum and Hoff 1934).
3.11.2 Reducing Body Burden
Following absorption into the blood, benzene is rapidly distributed throughout the body. Since benzene is
lipophilic, it is preferentially distributed to lipid-rich tissues. The initial stage of benzene metabolism is
the formation of benzene oxide via P-450 mixed-function oxidases. Detoxification pathways generally
involve the formation of glutathione conjugates of benzene oxide and glucuronide or sulfate conjugates of
phenol or its subsequent metabolites, catechol, hydroquinone, and trihydroxybenzene. Other metabolites
of benzene also have known toxic effects. Exhalation is the main route for excretion of unmetabolized
benzene, while metabolized benzene is excreted primarily in the urine. Studies in humans and animals
indicate that both exhalation and urinary excretion occur in several phases, with half-lives of minutes
to hours. Hence, benzene and its metabolites have relatively short half-lives in the body, and while some
of these metabolites are clearly toxic, accumulation of substantial body burdens are not expected.
No methods are currently used for reducing the body burden of benzene. It is possible that methods could
be developed to enhance the detoxification and elimination pathways, such as ensuring sufficient
glutathione stores in the body by the administration of N-acetyl-L-cysteine.
3.11.3 Interfering with the Mechanism of Action for Toxic Effects
Administration of indomethacin, a nonsteroidal anti-inflammatory drug, has been shown to prevent
benzene-induced myelotoxicity in mice and the accompanying increase in prostaglandin E in the bone
marrow (Kalf et al. 1989; Renz and Kalf 1991). Co-administration of indomethacin also prevented an
increase in the number of micronucleated polychromatic erythrocytes in peripheral blood. The authors
suggest that these results suggest a role for prostaglandin synthetase in benzene-induced myelotoxic and
genotoxicity, and a way to interfere with that process with substances such as indomethacin.
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Prostaglandins have been shown to inhibit hematopoiesis (Kalf et al. 1989). Additionally, prostaglandin
synthetase could be involved in the oxidation of phenol and/or hydroquinone to toxic metabolites (Kalf et
al. 1989).
The use of indomethacin to block benzene toxicity has led to data that indicate that myelotoxicity may
involve the destruction of stromal macrophages that produce IL-1, a cytokine essential for hematopoiesis
(Renz and Kalf 1991). External administration of recombinant IL-1 to mice prior to benzene
administration prevents the myelotoxicity, presumably by providing a source of the cytokine (Renz and
Kalf 1991). Further indication that IL-1 is affected by benzene exposure comes from the work of
Carbonnelle et al. (1995), who showed that exposure of human monocytes to micromolar amounts of
hydroquinone for 2 hours resulted in significantly decreased secretion of IL-1 and IL-1 at
concentrations of 5 M and above. RNA and protein synthesis were also inhibited. Additional research
in this area indicates that tumor necrosis factor may provide protection against the inhibitory effects of
hydroquinone on human hematopoietic progenitor cells (Colinas et al. 1995). The research of Shankar et
al. (1993) determined that pretreatment with Protein A, a glycoprotein that acts as a multipotent
immunostimulant, modulated the toxicity of benzene. Groups of six female albino rats (Swiss Wistar)
were injected intraperitoneally with 1.0 mL/kg/body weight (879 mg/kg) benzene once daily for
3 consecutive days. Another group (six per group) was administered intravenously with 60 g/kg Protein
A twice weekly for 2 weeks and then injected with 879 mg/kg benzene intraperitoneally once daily for
3 consecutive days. Controls were injected with normal saline. All of the animals were killed 24 hours
after receiving the last benzene injection. Blood was collected from the jugular vein for enumeration of
total leukocyte counts. Routine autopsy was performed on all animals and the liver, thymus, spleen, and
kidney organs were collected for organ weights. In benzene-only treated animals, there was a significant
decrease in the total leukocyte counts in the peripheral blood as well as a significant decrease in the
number of lymphocytes, a decrease in the gross organ weights of thymus and spleen, a significant
increase in the iron content and lipid peroxidation of the liver and bone marrow, and an increase in low
molecular weight iron in the bone marrow. Pretreatment with Protein A prevented these parameters from
changing.
3.12 ADEQUACY OF THE DATABASE
Section 104(I)(5) of CERCLA, as amended, directs the Administrator of ATSDR (in consultation with the
Administrator of EPA and agencies and programs of the Public Health Service) to assess whether
adequate information on the health effects of benzene is available. Where adequate information is not
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available, ATSDR, in conjunction with the National Toxicology Program (NTP), is required to assure the
initiation of a program of research designed to determine the health effects (and techniques for developing
methods to determine such health effects) of benzene.
The following categories of possible data needs have been identified by a joint team of scientists from
ATSDR, NTP, and EPA. They are defined as substance-specific informational needs that if met would
reduce the uncertainties of human health assessment. This definition should not be interpreted to mean
that all data needs discussed in this section must be filled. In the future, the identified data needs will be
evaluated and prioritized, and a substance-specific research agenda will be proposed.
3.12.1 Existing Information on Health Effects of Benzene
The existing data on health effects of inhalation, oral, and dermal exposure of humans and animals to
benzene are summarized in Figure 3-6. The purpose of this figure is to illustrate the existing information
concerning the health effects of benzene. Each dot in the figure indicates that one or more studies provide
information associated with that particular effect. The dot does not necessarily imply anything about the
quality of the study or studies, nor should missing information in this figure be interpreted as a data
need. A data need, as defined in ATSDRs Decision Guide for Identifying Substance-Specific Data
Needs Related to Toxicological Profiles (Agency for Toxic Substances and Disease Registry 1989), is
substance-specific information necessary to conduct comprehensive public health assessments.
Generally, ATSDR defines a data gap more broadly as any substance-specific information missing from
the scientific literature.
Virtually all of the information regarding health effects in humans comes from studies of workers
exposed to benzene-containing solvents and/or adhesives. Exposures to benzene occurred at rotogravure
printing shops; at shoe, rubber, and raincoat manufacturing plants; and during chemical manufacturing
processes. Case reports and cohort studies describe both acute and chronic health effects. The
predominant route of exposure in these studies is inhalation. Dermal contact is also suspected as a
possible route of exposure in these studies.
As seen in Figure 3-6, inhalation information for humans is available regarding death; acute-,
intermediate-, and chronic-duration systemic effects; immunologic, neurologic, reproductive,
developmental, and genotoxic effects; and cancer. However, as mentioned above, human exposure to
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Figure 3-6. Existing Information on Health Effects of Benzene
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benzene in specific work environments probably occurs not only by inhalation, but also by the dermal
route. Limited information is available regarding direct skin contact with benzene by humans.
Additionally, oral studies in humans are limited to isolated case reports of death and acute-duration
systemic effects subsequent to accidental or intentional ingestion of benzene, although one study (Hunting
et al. 1995) described effects in vehicle maintenance workers who siphoned gasoline by mouth. There is
limited information on effects of dermal exposure of humans to benzene, including death, acute-duration
effects, and cancer.
Inhalation and oral studies in animals provide data on death; systemic effects after acute-, intermediate-,
and chronic-duration exposure; and immunologic, neurologic, reproductive, developmental, genotoxic,
and cancer effects. Furthermore, data exist regarding acute- and intermediate-duration systemic effects
and cancer in animals after dermal exposure to benzene.
3.12.2 Identification of Data Needs
Acute-Duration Exposure.
There are reports on the health effects resulting from acute exposure of
humans and animals to benzene via the inhalation, oral, and dermal routes. The primary target
organs for acute exposure are the hematopoietic system, nervous system, and immune system. Acute
effects on the nervous system and immune system are discussed below under Neurotoxicity and
Immunotoxicity. Information is also available for levels that cause death in humans (e.g., Cronin 1924;
Flury 1928; Greenburg 1926; Tauber 1970; Thienes and Haley 1972) and in animals (e.g., Cornish and
Ryan 1965; Drew and Fouts 1974; Smyth et al. 1962) following inhalation and oral exposures.
No acute human or animal data on hematological effects from oral or dermal exposure are available.
However, there are acute inhalation data that characterize the effects of benzene on the hematological
system in humans and animals. Data regarding effects on the human hematological system following
acute inhalation exposure to benzene are scant, but indicate leukopenia, anemia, and thrombocytopenia
after more than 2 days of occupational exposure to more than 60 ppm benzene (Midzenski et al. 1992).
Data for hematological effects in animals after acute-duration inhalation exposure are more extensive.
Changes in peripheral erythrocytes (Chertkov et al. 1992; Cronkite et al. 1985; Rozen et al. 1984; Ward et
al. 1985), in peripheral leukocytes (Aoyama et al. 1986; Chertkov et al. 1992; Gill et al. 1980; Green et al.
1981b; Li et al. 1986; Ward et al. 1985; Wells and Nerland 1991), and in bone marrow cells (Chertkov et
al. 1992; Corti and Snyder 1996; Cronkite et al. 1989; Dempster and Snyder 1991; Gill et al. 1980; Green
et al. 1981b; Neun et al. 1992; Toft et al. 1982) were seen in rats and mice. An acute-duration inhalation
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MRL of 0.009 ppm was determined based on the LOAEL for immunologic effects in the mouse (Rozen et
al. 1984) (discussed below under Immunotoxicity). No acute-duration oral studies were suitable for
deriving MRLs. Additional studies that include dose-response information on hematological effects
following acute oral exposure could be designed to provide information that could be useful in deriving
an acute-duration oral MRL for benzene. Such studies could be designed to serve as validation for
existing PBPK models. Acute dermal exposure at levels that are likely to be found in the environment
and at hazardous waste sites is not likely to cause adverse health effects.
Intermediate-Duration Exposure.
There is sufficient information in humans and animals to
identify the hematopoietic, nervous, and immunological systems as targets for benzene toxicity. The
effects on the nervous system and immune system are discussed in the sections below titled Neurotoxicity
and Immunotoxicity. Data on adverse hematological effects in humans are available following
intermediate-duration exposures to benzene in the workplace (Aksoy and Erdem 1978; Aksoy et al.
1972). However, the exposure levels and durations were not well defined and, therefore, could not be
used to calculate an MRL. Studies in rats and mice following inhalation exposure can be used to define
NOAELs and LOAELs for hematological and immunological effects (e.g., Baarson et al. 1984; Cronkite
et al. 1982, 1985, 1989; Dow 1992; Farris et al. 1993, 1997a, 1997b; Green et al. 1981a, 1981b; Luke et
al. 1988b; Plappert et al. 1994a, 1994b; Rosenthal and Snyder 1987; Seidel et al. 1989; Snyder et al.
1978a, 1980; Toft et al. 1982; Vacha et al. 1990; Ward et al. 1985; Wolf et al. 1956). An intermediateduration inhalation MRL of 0.006 ppm was derived, based on a LOAEL for immunologic effects in the
mouse (Rosenthal and Snyder 1987) (discussed below under Immunotoxicity). Data on hematological
effects from oral exposures in animals are also available (Hsieh et al. 1988b, 1990, 1991; NTP 1986;
Shell 1992; Wolf et al. 1956), but no intermediate-duration oral MRL could be derived because the
threshold for hematological (and immunological) effects could not be identified. Additional studies that
include dose-response information on hematological effects following intermediate-duration oral
exposure could be designed to provide information that could be useful in deriving an intermediateduration oral MRL for benzene. Such studies could be designed to serve as validation for existing PBPK
models. Intermediate-duration dermal exposure at levels that are likely to be found in the environment
and at hazardous waste sites is not likely to cause adverse health effects.
Chronic-Duration Exposure and Cancer.
The primary target for adverse systemic effects of
benzene following chronic exposure is the hematological system. Hematotoxicity was reported in studies
of humans chronically exposed to benzene in the workplace air (Aksoy and Erdem 1978; Aksoy et al.
1971, 1972, 1974, 1987; Cody et al. 1993; Dosemeci et al. 1996; Doskin 1971; Erf and Rhoads 1939;
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Goldwater 1941; Greenburg et al. 1939; Kipen et al. 1989; Lan et al. 2004a, 2004b; Li et al. 1994; Qu et
al. 2002, 2003a, 2003b; Rothman et al. 1996a, 1996b; Townsend et al. 1978; Ward et al. 1996; Yin et al.
1987c). Several studies of occupational inhalation exposure to benzene did not find clinically-defined
hematological effects (Collins et al. 1991, 1997; Tsai et al. 1983, 2004), but they had a high degree of
uncertainty regarding estimations of benzene exposure levels. The study of workers of shoe
manufacturing industries in Tianjin, China (Lan et al. 2004a, 2004b) identified the lowest LOAEL for
hematotoxicity and was selected as the principal study for deriving a chronic-duration inhalation MRL of
0.003 ppm for benzene. Chronic-duration animal studies are available for hematological effects
following inhalation exposure and provide support to the human data (Snyder et al. 1978a, 1980, 1982,
1984). No human data are available to evaluate hematological effects following oral exposure. Although
chronic duration oral animal studies are available for hematological effects (Huff et al. 1989; Maltoni et
al. 1983, 1985; NTP 1986), the most extensive study (NTP 1986) did not conclusively define a NOAEL
or less serious LOAEL for end points that could be used to derive an MRL. However, a chronic-duration
oral MRL of 0.0005 mg/kg/day was derived for benzene based on route-to-route extrapolation of the same
results (Lan et al. 2004a, 2004b) that served as the basis for the chronic-duration inhalation MRL.
Additional chronic-duration oral animal data could be designed to provide support to the chronic-duration
oral MRL and to assist in defining threshold levels for populations living near hazardous waste sites.
Dermal data for humans and animals were not available. However, chronic-duration dermal exposure at
levels that are likely to be found in the environment and at hazardous waste sites is not likely to cause
adverse health effects.
carcinogen based on sufficient data in humans supported by animal evidence (IARC 2004; IRIS 2007;
NTP 2005). Epidemiological studies and case reports provide clear evidence of a causal relationship
between occupational exposure to benzene and the occurrence of acute myelogenous leukemia (AML)
(Hayes et al. 1997; IARC 1982, 1987; EPA 1995, 1998; IRIS 2007; Rinsky et al. 1987, 2002; Yin et al.
1996a, 1996b), as well as is suggestive evidence of associations between benzene and non-Hodgkins
lymphoma (NHL) and multiple myeloma (Hayes et al. 1997; Rinsky et al. 1987). Studies of workers in
Ohio (the Pliofilm study) (e.g., Rinsky et al. 1981, 1987; 2002) and China (the NCI/CAPM study) (e.g.,
Hayes et al. 1997; Yin et al. 1996a, 1996b) provide the strongest data on the leukemogenic potential of
benzene, including exposure-response information, and data from the Pliofilm study was used as the basis
of inhalation and oral cancer risk values for benzene (EPA 1998; IRIS 2007). Additional studies on these
and other cohorts could better characterize exposure level and exposure duration relationships for benzene
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and leukemia, particularly at low levels of exposure, and clarify the potential of benzene to induce NHL
and multiple myeloma.
Benzene is a multiple site carcinogen in rats and mice following inhalation exposure (Cronkite 1986;
Cronkite et al. 1984, 1985, 1989; Farris et al. 1993; Maltoni et al. 1982a, 1982b, 1983, 1985, 1989;
Snyder et al. 1980) and oral exposure (Huff et al. 1989; Maltoni et al. 1983, 1985, 1989; NTP 1986),
inducing lymphomas and other neoplasms in numerous tissues not affected in humans. Although
contributing to the weight of evidence for carcinogenicity, the animal studies do not identify a suitable
model for leukemia in humans. An appropriate animal model would help to provide a better
understanding of how benzene causes cancer, particularly the mechanism of benzene leukemogenesis.
The exact mechanism of benzene carcinogenicity is not known, but it has been postulated that some
benzene metabolites are capable of forming adducts with DNA and are responsible for reduced immune
function which could potentially lead to cancer. The clastogenic properties of benzene may play a role in
its carcinogenicity. DNA adduct formation could occur with both inhalation and oral exposures (Ding et
al. 1983; Sasiadek et al. 1989). Questions that need to be answered with regard to the mechanism of
benzene carcinogenesis include how benzene metabolites produce greater-than-additive effects,
determination of the critical target genes, whether aplastic anemia is essential to the development of
leukemia, and determination of the role of cytokines and growth factor pathways in benzene toxicity.
Genotoxicity.
Evidence for the genotoxicity of benzene in humans comes from studies of chronically-
exposed populations (Andreoli et al. 1997; Bogadi-are et al. 1997; Ding et al. 1983; Forni and Moreo
1967, 1969; Forni et al. 1971a; Hallberg et al. 1996; Hartwich et al. 1969; Hedli et al. 1991; Karacic et al.
1995; Kauba et al. 2000; Liu et al. 1996; Major et al. 1992, 1994; Nilsson et al. 1996; Picciano 1979;
Pitarque et al. 1996, 1997; Popp et al. 1992; Qu et al. 2003a, 2003b; Rothman et al. 1995; Sardas et al.
1994; Sasiadek et al. 1989; Sellyei and Kelemen 1971; Smith et al. 1998; Sul et al. 2002; Tompa et al.
1994; Tough and Court Brown 1965; Tough et al. 1970; Trkel and Egeli 1994; Van den Berghe et al.
1979; Yardley-Jones et al. 1990; Zhang et al. 1998b, 1999). These exposures have occurred primarily via
inhalation, although some dermal exposure cannot be ruled out. In spite of the lack of accurate exposure
data, exposure to multiple chemicals, and often inappropriate control groups, the association between
benzene exposure and the appearance of structural and numerical chromosome aberrations in human
lymphocytes suggests that benzene can be considered a human clastogen. Benzene-induced cytogenetic
effects, including chromosome and chromatid aberrations, sister chromatid exchanges, and micronuclei,
have been consistently found in in vivo animal studies (Anderson and Richardson 1981; Au et al. 1991;
Chen et al. 1994; Chang et al. 2005; Eastmond et al. 2001; Erexson et al. 1986; Farris et al. 1996; Fujie et
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al. 1992; Healy et al. 2001; Lee et al. 2005; Kolachana et al. 1993; Ranaldi et al. 1998; Siou et al. 1981;
Toft et al. 1982; Ward et al. 1992). Binding of benzene and/or its metabolites to DNA, RNA, and
proteins has been consistently observed in rats and mice (Arfellini et al. 1985; Creek et al. 1997; Lvay et
al. 1996; Mani et al. 1999; Mazullo et al. 1989; Turteltaub and Mani 2003). Inhalation exposure of mice
has yielded identifiable benzene-derived hemoglobin adducts (Sabourin et al. 1990). Ward et al. (1992)
have shown that intermediate-duration exposure of mice to benzene by inhalation at levels below the
current PEL may induce gene mutations in the lymphocytes.
Standard short-term in vitro assays indicate that benzene's genotoxicity is derived primarily from its
metabolites, although benzene has been shown to produce DNA breaks in Chinese hamster ovary cells
independent of metabolic activators (Douglas et al. 1985; Eastmond et al. 1994; Lakhanisky and
Hendrickx 1985; Zhang et al.1993). There is good evidence that benzene affects cell cycle progression,
RNA and DNA synthesis, as well as DNA binding (Forni and Moreo 1967, 1969; Hartwich et al. 1969;
Sellyei and Kelemen 1971; Van den Berghe et al. 1979). Chen and Eastmond (1995) showed that
benzene metabolites can adversely affect human topoisomerases (enzymes involved in DNA replication
and repair); however, evidence exists that some repair of DNA binding with benzene metabolites occurs
in human cells (Chenna et al. 1995).
The genotoxicity of benzene has been extensively studied and demonstrated in both humans and
laboratory animals. Although some information is available regarding possible mechanisms of benzene
genotoxicity and carcinogenicity, additional mechanistic studies would be helpful to more completely
characterize mechanisms responsible for these effects.
Reproductive Toxicity.
Reproductive data are available on women occupationally exposed to
benzene (Mukhametova and Vozovaya 1972; Vara and Kinnunen 1946). The data suggest spontaneous
abortions, menstrual disturbances, and ovarian atrophy. These studies are limited by the difficulty in
identifying appropriate controls, problems in controlling for concomitant exposures to other chemicals,
and inadequate follow-up. Only one study was found that described the reproductive effects on men
exposed to benzene (Stucker et al. 1994). There are some data available from inhalation studies on
reproductive effects of benzene in animals. Although there are data on adverse gonadal effects (e.g.,
atrophy/degeneration, decrease in spermatozoa, moderate increases in abnormal sperm forms) (Ward et
al. 1985; Wolf et al. 1956), data on reproductive outcomes are either negative (Coate et al. 1984; Green et
al. 1978; Kuna et al. 1992; Murray et al. 1979; Tatrai et al. 1980a) or inconclusive or conflicting (i.e.,
number of live fetuses, incidences of pregnancies) (Murray et al. 1979; Ungvary and Tatrai 1985).
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Gofmekler (1968) showed infertility in female rats after intermediate-duration inhalation exposure to
210 ppm, but the results are poorly reported. Thus, they provide only suggestive evidence that benzene
may have an adverse effect on reproductive outcomes. No data were located on reproductive effects
following oral exposure to benzene in humans. Negative effects on reproductive outcome have been
reported in one oral study in animals (Exxon 1986). NTP (1986) reported neoplastic changes in the
reproductive tissues of female rats and male and female mice after chronic oral exposure. Given the
paucity of available data across all exposure routes, it would be useful to have additional 90-day studies
conducted by the oral and inhalation routes that assess reproductive organs histologically or
cytogenetically. Although dermal contact is not likely to be the most relevant route of exposure for
humans at hazardous waste sites, data on dermal exposure would also be useful. If the results of the
suggested inhalation or oral studies indicate reproductive toxicity, multigeneration or continuous breeding
studies for oral and inhalation exposures would help clarify the potential for benzene to cause
reproductive effects in humans.
Developmental Toxicity.
Based on epidemiological studies at hazardous waste sites, an increased
susceptibility to benzene of pregnant women or their offspring has not been demonstrated (Budnick et al.
1984; Goldman et al. 1985; Heath 1983; Olsen 1983). However, these studies have several limitations
that make it impossible to assess the effect of benzene on the human fetus. For example, the few studies
that do exist are limited by a lack of control incidences for end points, problems in identifying exposed
populations, a lack of data on exposure levels, and/or exposure to multiple substances (Budnick et al.
1984; Forni et al. 1971a; Funes-Cravioto et al. 1977; Goldman et al. 1985; Heath 1983; Olsen 1983). In
the occupational setting, however, there may be stronger evidence of increased susceptibility to benzene
of pregnant women and/or their offspring (Forni et al. 1971a; Funes-Cravioto et al. 1977). For example,
severe pancytopenia and increased chromosomal aberrations occurred in a pregnant worker exposed to
benzene (levels not reported) during the entire pregnancy. She gave birth to a healthy boy (Forni et al.
1971b). On the other hand, increased frequency of chromatid and isochromatid breaks and sister
chromatid exchanges was found in lymphocytes from 14 children of female workers exposed to benzene
(levels not reported) and other organic solvents during pregnancy (Funes-Cravioto et al. 1977).
Irrespective of the hematological effects reported in the pregnant worker or the 14 children of female
workers, the lack of complete and detailed medical records and the lack of follow-up limit the
significance of effects with regard to post exposure morbidity.
A number of investigations have evaluated the developmental/maternal toxicity of benzene in animals
following inhalation exposures. In these investigations, fetotoxicity was evidenced by reduced fetal
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weight and/or minor skeletal variations at concentrations 47 ppm (Coate et al. 1984; Green et al. 1978;
Kimmel and Wilson 1973; Kuna and Kapp 1981; Murray et al. 1979; Tatrai et al. 1980a, 1980b; Ungvary
and Tatrai 1985). However, persistently altered fetal hematopoiesis occurred in mice at 20 ppm (Keller
and Snyder 1986, 1988). Additional information designed to assess the hematopoietic system of the
developing fetus (human or animal) following low-level in utero exposures to benzene is needed. Oral
data are limited to two animal studies in which benzene was shown to reduce pup body weight when mice
were administered a high single oral dose (Seidenberg et al. 1986) or have no effect after gestational
exposure (Exxon 1986). Additional oral studies designed to assess the developmental effects (human or
animal) of low-level exposures to benzene would be useful. No data are available on the developmental
toxicity of benzene following dermal exposure. Although these data would be useful, the most likely
routes of exposure for humans at hazardous waste sites are the inhalation and oral routes.
Immunotoxicity.
The immune system is known to be a target for benzene toxicity. The effects do not
appear to be route- or species-specific. The evidence for immunotoxicity in humans comes from workers
exposed by inhalation of intermediate and chronic durations. A series of studies demonstrated decreases
in numbers of circulating leukocytes (Aksoy et al. 1971, 1972, 1974), but levels of exposure were not
well documented. In other studies, alterations in human serum immunoglobulins were observed, but there
was concomitant exposure to xylene and toluene (Lange et al. 1973a, 1973b). There is a need to test for
subtle alterations in the immune system and immune competence in workers with intermediate- and
chronic-duration exposure to benzene. Animal studies support the findings of immune dysfunction and
indicate that additional parameters of the immune system are affected by exposure to benzene in the air
for acute (Aoyama 1986; Chertkov et al. 1992; Cronkite 1986; Cronkite et al. 1982, 1985, 1989; Gill et al.
1980; Green et al. 1981a; Li et al. 1986; Neun et al. 1992; Rozen et al. 1984; Rosenthal and Snyder 1985;
Toft et al. 1982; Ward et al. 1985; Wells and Nerland 1991), intermediate (Baarson et al. 1984; Cronkite
et al. 1982, 1985, 1989; Dow 1992; Gill et al. 1980; Green et al. 1981a, 1981b; Li et al. 1992; Plappert et
al. 1994b; Rosenthal and Snyder 1987; Rozen and Snyder 1985; Seidel et al. 1990; Snyder et al. 1980;
Stoner et al. 1981; Ward et al. 1985; Wolf et al. 1956; Songnian et al. 1982), and chronic (Snyder et al.
1980, 1984, 1988) periods. An acute-duration inhalation study found depressions of proliferative
responses of bone-marrow-derived B-cells and splenic T-cells in mice at 10 ppm (Rozen et al. 1984), and
an acute-duration inhalation MRL has been derived based on this study. An intermediate-duration
inhalation study found delayed splenic lymphocyte reaction to foreign antigens evaluated by in vitro
mixed lymphocyte culture following exposure of mice to benzene vapors at a concentration of 10 ppm
(Rosenthal and Snyder 1987), and an intermediate-duration inhalation MRL has been derived based on
this study. No human data on the oral route were available, but animal data showed immunological
BENZENE
228
3. HEALTH EFFECTS
effects after intermediate (Fan 1992; Hsieh et al. 1988b, 1990, 1991; Shell 1992; Wolf et al. 1956) and
chronic exposures (Huff et al. 1989; Maltoni et al. 1983, 1985; NTP 1986) via the oral route. Since the
immune system is known to be a target organ, it is important to have information on subtle alterations in
immune competence in people exposed to benzene in the air and in the drinking water. A decrease in
cell-mediated immune functions, including alloantigen response and cytotoxicity, was reported in mice
following intermediate inhalation exposure to 100 ppm of benzene (Rosenthal and Snyder 1987). This
impaired cell-mediated immune function was also apparent in other in vivo studies (Stoner et al. 1981).
Mice exposed to 100 ppm of benzene for a total of 100 days had reduced tumor resistance when
challenged with syngeneic tumor cells and developed tumors that were lethal (Rosenthal and Snyder
1987). Further animal studies would also be useful in defining more NOAEL and LOAEL values. No
data are available that document immunotoxicity in humans or animals exposed by dermal application.
Dermal sensitization tests may also provide useful data on the likelihood of an allergic response in
humans, since skin contact may occur in the workplace and at hazardous waste sites.
Neurotoxicity.
In humans, the nervous system is a target of benzene toxicity following both inhalation
and oral exposures. No data are available that demonstrate neurologic effects in humans or animals
exposed dermally. There are sufficient data to suggest that it is the central nervous system which is
affected following acute exposures. Neurological symptoms reported in humans following acute oral and
inhalation exposures are similar and include drowsiness, dizziness, headache, vertigo, tremor, delirium,
and loss of consciousness (Cronin 1924; Flury 1928; Greenburg 1926; Midzenski et al. 1992; Tauber
1970; Thienes and Haley 1972). Acute- and intermediate-duration inhalation and oral animal studies
provide supportive evidence that benzene effects the central nervous system (Carpenter et al. 1944;
Cornish and Ryan 1965; Evans et al. 1981; Frantik et al. 1994). Effects observed include narcosis,
hyperactivity, tremors, tonic-clonic convulsions, decreased evoked electrical activity in the brain, and
slight nervous system depression. Behavioral changes were found in mice after 1 week of exposure to
300 ppm, and decreased grip strength was found after three exposures to 1,000 ppm (Dempster et
al. 1984). An intermediate-duration inhalation study found increased rapid response in mice at 0.78 ppm
(Li et al. 1992), but this study was limited due to apparent discrepancies between reported and actual
benzene exposure levels, since neurological effects reported in other animal studies occurred only at
much higher exposure levels. Acute-duration oral exposures of 950 mg/kg benzene altered neurotransmitter levels in the brains of rats (Kanada et al. 1994). Intermediate-duration oral exposures of
8 mg/kg/day resulted in changes in the levels of monoamine transmitters in the brain without treatmentrelated behavioral changes (Hsieh et al. 1988a).
BENZENE
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3. HEALTH EFFECTS
One chronic-duration occupational study reported neurological abnormalities of the peripheral nervous
system (global atrophy of lower extremities and distal neuropathy of upper extremities) in four of six
benzene-exposed patients with aplastic anemia (Baslo and Aksoy 1982). Additional studies are needed to
verify the peripheral nervous system effects that might occur following chronic exposures to low doses of
benzene.
Although there are sufficient data to indicate that the nervous system is a target of benzene toxicity, the
neurotoxicity of benzene has not been extensively studied. Additional studies in animals are needed to
identify the thresholds of neurotoxicity following acute-, intermediate-, and chronic-duration inhalation
and oral exposures.
Epidemiological and Human Dosimetry Studies.
A large segment of the U.S. population is
exposed to benzene. This exposure occurs primarily as a result of benzene emitted to the air from tobacco
smoke, gasoline stations, and automobile exhaust. Benzene has been found in about a third of the NPL
hazardous waste sites. The magnitude of exposure is greater for those occupationally exposed.
The predominance of available epidemiological data comes from occupational studies and associated
analysis (Aksoy and Erdem 1978; Aksoy et al. 1971, 1972, 1974; Baslo and Aksoy 1982; Ciccone et al.
1993; Cody et al. 1993; Dosemeci et al. 1994, 1996; Doskin 1971; EPA 1986, 1995, 1998; Erf and
Rhoads 1939; Goldwater 1941; Greenburg et al. 1939; Hayes et al. 1997; IARC 1982, 1987; Infante 1978;
Infante et al. 1977; IRIS 2007; Kipen et al. 1989; Lan et al. 2004a, 2004b; Li et al. 1994; Ott et al. 1978;
Paustenbach et al. 1992; Paxton et al. 1994a, 1994b; Qu et al. 2002, 2003a, 2003b; Rinsky et al. 1981,
1987, 2002; Rothman et al. 1996a, 1996b; Townsend et al. 1978; Travis et al. 1994; Utterback and Rinsky
1995; Ward et al. 1996; Yin et al. 1987c, 1989, 1994, 1996a, 1996b). While these studies provide data on
hematological and neurotoxic effects and evidence of carcinogenicity, they only offer information on the
effects of inhalation exposure. Retrospective and prospective studies of populations that have been
identified as being exposed to contaminated groundwater or drinking water could provide information on
long-term health effects from oral exposures. The Benzene Subregistry Baseline Technical Report of the
National Exposure Registry contains information on 1,143 persons who had documented exposure to
benzene in their drinking water and were exposed for at least 30 days (Agency for Toxic Substances and
Disease Registry 2001). No causal relationship has been proposed for health conditions identified in the
base subregistry (Agency for Toxic Substances and Disease Registry 1995) or continued follow-up of the
population (Agency for Toxic Substances and Disease Registry 2001).
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3. HEALTH EFFECTS
Few well-conducted epidemiological studies of exposed populations exist. An epidemiological study of a

population with chronic exposure to two NPL hazardous waste sites illustrates the problems inherent in
assessing adverse health effects from waste site exposure (Dayal et al. 1995). Benzene was only 1 of
20 compounds or groups of compounds identified as being chemicals of concern in the waste site. The
authors note that assessment of long-term exposure to a dumpsite is problematic because of a lack of
history of materials deposited, the complexity of the waste components, the interaction of the components
over time, and the emphasis on statistically significant versus biologically significant effects. The study
had certain limitations, including lifestyle characteristics, the use of mailed-in responses, subjective
symptom-reports without independent confirmation, and various problems with exposure estimation. A
relationship was detected between exposed individuals and neurological symptoms, including learning
difficulties, nervousness, numbness in fingers or toes, sleeping difficulties, unusual fatigue, and decreased
sense of smell. Chest pains, irregular heartbeat, skin rashes, and detection of a peculiar odor or taste also
occurred more often in the exposed population. However, no effort was made to relate the symptoms to
particular chemicals in the waste site. Clearly, more studies of exposed populations surrounding waste
sites would be warranted. Other studies include analyses of workers in China (Dosemeci et al. 1994,
1996; Lan et al. 2004a, 2004b; Qu et al. 2002, 2003a, 2003b; Rothman et al. 1996a, 1996b; Yin et al.
1987c, 1994), reanalysis of data from the Pliofilm cohort (Paxton et al. 1994a, 1994b; Utterback and
Rinsky 1995; Ward et al. 1996), and assessment of clinical manifestations of benzene hematotoxicity in
U.S. industries with potential for benzene exposure (Collins et al. 1991, 1997; Tsai et al. 1983, 2004). A
publication by Lioy and Pellizzari (1995) outlines an appropriate framework for the National Human
Exposure Assessment Survey, using benzene as a candidate compound. Other articles also address
population-based exposure models and measurements (MacIntosh et al. 1995; Pellizzari et al. 1995).
Analysis of benzene dosimetry in animal studies suggests that the metabolic pathways leading to the
production of the putative toxic metabolites appear to be low-capacity, high-affinity pathways that are
saturated at relatively low-exposure concentrations (Henderson et al. 1992). The authors suggest that this
could also be true for humans. Additional dosimetry studies are necessary to further define the role of
toxic metabolites in the development of benzene-related leukemia.
Studies of DNA damage have primarily been performed on workers with preexisting hematological
effects and cancer (Ding et al. 1983; Forni et al. 1971a; Picciano 1979; Yardley-Jones et al. 1988) or as
biomonitoring of benzene exposure (Andreoli et al. 1997; Bogadi-are et al. 1997; Hallberg et al. 1996;
Kauba et al. 2000; Liu et al. 1996; Nilsson et al. 1996; Pitarque et al. 1996; Sul et al. 2002; Surralls et
al. 1997; Zhang et al. 1999). Additional data on benzene-induced DNA damage could be useful in
BENZENE
231
3. HEALTH EFFECTS
monitoring possible adverse effects in individuals living near hazardous waste sites if more quantitative
dose-response data were available for clastogenicity.
Studies to determine whether benzene causes solid tumors and hematological malignancies other than
acute nonlymphocytic leukemia in humans have been suggested as a data need (Goldstein and Warren
1993). These data could be obtained by revisiting already established cohorts of exposed individuals, or
by examining new cohorts.
Biomarkers of Exposure and Effect.
Exposure. There is no clinical disease state unique to benzene toxicity. However, the effects on the
hematopoietic and immune systems are well recognized, and analytical methodologies exist for
monitoring benzene levels in expired breath and blood (Brugnone et al. 1989, 1992; DeLeon and Antoine
1985; Pellizzari et al. 1988). Several biomarkers of exposure to benzene exist, including unmetabolized
benzene in expired air and urine (Farmer et al. 2005; Fustinoni et al. 2005; Ghittori et al. 1993;
Nomiyama and Nomiyama 1974a, 1974b; Sherwood 1988; Srbova et al. 1950; Waidyanatha et al. 2001),
urinary phenol levels (Astier 1992; Inoue et al. 1986, 1988b; Jongeneelen et al. 1987; Karacic et al. 1987;
Pagnotto et al. 1961; Pekari et al. 1992), and urinary trans,trans-muconic acid and S-phenylmercapturic
acid levels (Boogaard and van Sittert 1995, 1996; Ducos et al. 1990, 1992; Farmer et al. 2005; Inoue et al.
1989b, 2000; Lee et al. 1993; Melikian et al. 1993, 1994; Pezzagno et al. 1999; Popp et al. 1994; Qu et al.
2005; Rothman et al. 1998; Ruppert et al. 1997; Sanguinetti et al. 2001; van Sittert et al. 1993; Weaver et
al. 2000). Estimates of benzene exposure can be made by comparing the ratio of inorganic to organic
sulfates in the urine (Hammond and Hermann 1960). However, urinary sulfate levels are variable, and
they have not been used to identify exposure levels of benzene associated with minimal toxic effect.
Furthermore, benzene-derived adducts with hemoglobin and albumin may be useful as biomarkers
(Bechtold et al. 1992a, 1992b; Bechtold and Henderson 1993; Farmer et al. 2005; Fustinoni et al. 2005;
Ghittori et al. 1993; Hedli et al. 1991; Lutz and Schlatter 1977; Nomiyama and Nomiyama 1974a, 1974b;
Pekari et al. 1992; Reddy et al. 1989; Schad et al. 1992; Sherwood 1988; Smith and Rothman 2000;
Srbova et al. 1950; Waidyanatha et al. 2001; Yeowell-OConnell et al. 1998, 2001).
Phenol measurements have routinely been used for monitoring occupational exposures, and there is
evidence that urinary phenol levels can be correlated with exposure levels (Astier 1992; Inoue et al. 1986;
Pekari et al. 1992). Correlating urinary phenol with benzene exposure is complicated by high and
variable background levels in nonexposed persons. The data suggest that variations in urinary phenol will
BENZENE
232
3. HEALTH EFFECTS
obscure phenol formed from low levels of benzene (Ong and Lee 1994; Perbellini et al. 1988). For
exposures to 1 ppm or less, the current workplace standard, urinary phenol is not an adequate assay to
determine the extent of benzene exposure. In addition, urinary excretion of phenol is lowered by
coexposure to toluene (Inoue et al. 1988b). In many exposure situations (e.g., occupational settings and
hazardous waste sites), toluene and other chemicals are present and could interfere with the metabolism
and elimination of benzene. Studies that attempt to identify another biomarker that is specific to benzene,
such as S-phenylmercapturic acid, the proposed urinary BEI (ACGIH 1996b), may be helpful for medical
surveillance.
The metabolism of benzene to ring-opened compounds (e.g., muconic acid) may be pertinent to the
development of a biomarker of benzene exposure and effect (Ducos et al. 1990, 1992; Inoue et al. 1989b;
Lee et al. 1993; Melikian et al. 1993; Ong and Lee 1994; Witz et al. 1990b). However, the dose-response
curve for metabolism of benzene to urinary muconic acid in humans needs to be determined. If urinary
muconic acid increases in humans at low benzene exposures, as it does in mice, then it may be a valid
biomarker of benzene exposure. Excretion of urinary muconic acid in humans has also been shown to be
lowered by coexposure to toluene (Inoue et al. 1989b). The effect of coexposure to toluene and other
chemicals on muconic acid formation needs to be assessed in order to determine the usefulness of
muconic acid as a biomarker of benzene exposure.
Breath levels of benzene have been used as a measure of exposure (Brugnone et al. 1989; Money and
Gray 1989; Nomiyama and Nomiyama 1974a, 1974b; Ong and Lee 1994; Pekari et al. 1992). However,
the amount of benzene lost in expired air will vary not only with the dose, duration, and time from
exposure, but also with the extent of metabolism in the body. Benzene levels in blood have been
measured. However, blood levels rapidly decrease after exposure (Brugnone et al. 1989; DeLeon and
Antoine 1985; Schrenk et al. 1941).
Benzene metabolites also form DNA adducts (Popp et al. 1992). The use of hemoglobin adducts formed
from benzene metabolites as a biomarker of benzene exposure has been developed by Sun et al. (1990).
Bechtold and colleagues have conducted further studies using this model (Bechtold et al. 1992a; Bechtold
and Henderson 1993). Further refinement of this model would be useful to quantitate cumulative lowlevel exposures to benzene. Development of specific markers of exposure to use in epidemiological
studies to clarify the shape of the dose-response curve in the low-dose region has been suggested as a data
need (Goldstein and Warren 1993). Recent reports indicate that urinary benzene may serve as a sensitive
BENZENE
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3. HEALTH EFFECTS
biomarker of exposure to benzene concentrations well below 1 ppm (Farmer et al. 2005; Fustinoni et al.
2005).
Effect. Monitoring of benzene workers has included monthly blood counts. Toxic effects occur in the
bone marrow and arise either from benzene via a solvent effect, or via its metabolites (Gad-El-Karim et
al. 1985; Irons et al. 1980). Hematological tests could be used as markers of hematotoxicity, but medical
laboratories lack tests specific to benzene hematotoxicity. The tests cannot be relied upon to find
preclinical disease but can identify the subtle changes that are early indicators of effects. Furthermore,
these markers of effect may not be useful for long periods following cessation of exposure, nor do they
distinguish between acute and chronic exposures. As stated above, additional studies are needed to define
the role of benzene-related leukopenia in the disease process initiated by benzene exposure, to determine
if it is a biomarker of effect, or an intermediate end point in the development of leukemia (Hayes 1992).
Absorption, Distribution, Metabolism, and Excretion.
Data from both humans and animals
consistently indicate that benzene is rapidly absorbed through the lungs (Eutermoser et al. 1986;
Nomiyama and Nomiyama 1974a; Sabourin et al. 1987; Schrenk et al. 1941; Srbova et al. 1950; Yu and
Weisel 1996). Although experimentally-acquired data are not available on oral absorption of benzene in
humans, case reports of accidental or intentional poisoning suggest that benzene is rapidly absorbed from
the gastrointestinal tract (Thienes and Haley 1972). The efficient absorption of oral doses in animals is
well documented (Cornish and Ryan 1965; Parke and Williams 1953a; Sabourin et al. 1987). Benzene
can be absorbed through the skin, but the rate of absorption is much lower than that for inhalation
(Maibach and Anjo 1981; Susten et al. 1985; Tsuruta 1989). Following absorption into the body, benzene
is widely distributed to tissues, with the relative uptake dependent on the perfusion of the tissue by blood,
and the total potential uptake dependent on fat content and metabolism (Sato et al. 1975; Tauber 1970).
There is no evidence to suggest that the route of administration has any substantial effect on the
subsequent metabolism of benzene, either in humans or animals. Benzene is metabolized primarily in the
liver and to a lesser extent, in the bone marrow. Benzene is a preferential substrate of CYP2E1, which
also metabolizes alcohol. The induction of CYP2E1 by benzene (and some of its metabolites) with
subsequent generation of reactive metabolites, oxygen radicals, circulating lipid peroxides, and hydroxyl
radicals could be associated with hematopoietic toxicity and carcinogenicity of benzene (Irons 2000;
Parke 1989; Ross 1996, 2000; Smith 1996a, 1996b; Snyder 2000a, 2000b, 2002; Snyder and Hedli 1996;
Snyder and Kalf 1994). CYP2E1 is not confined to the liver, but has also been detected in bone marrow.
Andrews et al. (1979) demonstrated that rabbit bone marrow is capable of metabolizing benzene. Schnier
BENZENE
234
3. HEALTH EFFECTS
et al. (1989) subsequently found that rabbit bone marrow contains CYP2E1. Irons et al. (1980)
demonstrated that benzene metabolism by rat bone marrow (in situ) was complete and independent of
metabolism by the liver, with concentrations of phenol greater than catechol and hydroquinone. Although
the total metabolism by bone marrow was limited (total metabolites present were 25% of those in blood),
the concentration of metabolites in the bone marrow exceeded that in the blood. Similar studies have
been conducted in mice (Ganousis et al. 1992). Benzene metabolism in bone marrow is not well
understood; additional data regarding the initial oxidation step and the comparatively low levels of
CYP2E1 activity in bone marrow would be useful in identifying the mechanisms of benzene's
hematotoxicity. This aspect of metabolism may have implications for long-term exposures, which could
be explored in chronic exposure studies. The intermediary metabolites of benzene are responsible for
many of the toxic effects observed (Eastmond et al. 1987; Gad-El-Karim et al. 1985). Biotransformation
is believed to be essential for benzene-induced bone marrow damage.
However, there is disagreement as to whether benzene is activated in the marrow, activated elsewhere and
transported to the marrow, or metabolized in the liver and the metabolites activated in the marrow.
Further studies on the metabolism of benzene would help define its mechanism of action. Additionally,
more information is needed on the pathways of metabolism in humans, the chemical nature of the toxic
metabolites, and the mechanism of toxicity. Recently published data comparing urinary metabolite
profiles of orally administered benzene and phenol in mice suggest that zonal differences in metabolism
in the liver may be responsible for relative differences in the production of hydroquinone, thus explaining
the higher toxicity observed after benzene administration compared with phenol administration (Kenyon
et al. 1995). Additional work in this area would aid in further understanding the kinetic determinants of
benzene toxicity. Ethanol and dietary factors such as food deprivation and carbohydrate restriction
enhance the hematotoxic effects of benzene. Therefore, more information regarding differences in
metabolic pattern according to sex, age, nutritional status, and species, and correlation to differences in
health effects would be useful.
Humans and animals both excrete inhaled benzene via expiration. Additionally, benzene metabolites are
excreted primarily in the urine in both humans and animals. No studies in humans exist for excretion of
oral doses of benzene. Studies in several animal species indicate that the route of excretion of benzene
and/or its metabolites is a function of exposure level and the saturation of metabolic systems (Henderson
et al. 1989). Data regarding excretion following dermal exposure in humans are limited. However, the
major route of excretion in both humans and animals following dermal exposure is the urine.
BENZENE
235
3. HEALTH EFFECTS
Comparative Toxicokinetics.
Qualitatively, absorption, distribution, metabolism, and excretion
appear to be similar in humans and laboratory animals. However, quantitative variations in the
absorption, distribution, metabolism, and excretion of benzene have been observed with respect to
exposure routes, sex, nutritional status, and species. Further studies that focus on these differences and
their implications for human health would be useful. Additionally, in vitro studies using human tissue
and further research into PBPK modeling in animals would contribute significantly to the understanding
of the kinetics of benzene and would aid in the development of pharmacokinetic models of exposure in
humans. These topics are being addressed in ongoing studies (see Section 3.12.3).
Methods for Reducing Toxic Effects.
Development of methods and practices that are specific for
benzene is needed for reducing peak absorption, body burden and for interfering with the mechanism of
action following benzene exposures. Since benzene metabolites are thought to play the major role in the
toxicity and carcinogenicity, more information is needed about their covalent binding to nucleic acids and
cellular macromolecules. This information would help the development of methods for possible
prevention of benzene-induced toxicity. Related lines of investigation include the use of non-steroidal
anti-inflammatory drugs to block prostaglandin and prostaglandin synthetase-mediated activity after
benzene exposure, and the role of IL-1 cytokine activity in preventing depression of hematopoiesis.
Childrens Susceptibility.
Data needs relating to both prenatal and childhood exposures, and
developmental effects expressed either prenatally or during childhood, are discussed in detail in the
Developmental Toxicity subsection above.
No clear evidence of age-related differences in susceptibility to benzene toxicity was located. Benzene
crosses the placenta (Dowty et al. 1976). Nursing infants can be exposed to benzene in the breast milk
(Fabietti et al. 2004). Limited animal studies indicate that in utero exposure to benzene results in
hematological changes similar to those observed in animals exposed only as adults (Corti and Snyder
1996; Keller and Snyder 1986, 1988). There is some indication that parental occupational exposure to
benzene may play a role in childhood leukemia (Buckley et al. 1989; McKinney et al. 1991; Shaw et al.
1984; Shu et al. 1988). However, none of these studies indicate whether children may be at greater risk
than adults for benzene toxicity. Children could potentially be at increased risk for benzene toxicity via
the inhalation exposure route based on higher activity levels and ventilation rates than adults. Age-related
differences in benzene metabolism could potentially affect susceptibility. Well-designed animal studies
should be performed to adequately assess the potential for age-related increased susceptibility to benzene.
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3. HEALTH EFFECTS
Child health data needs relating to exposure are discussed in Section 6.8.1, Identification of Data Needs:
Exposures of Children.
3.12.3 Ongoing Studies
Ongoing studies pertaining to benzene have been identified and are shown in Table 3-7 (FEDRIP 2005).
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3. HEALTH EFFECTS
Table 3-7. Ongoing Studies on the Health Effects of Benzene

Investigator Affiliation
Research description
Conaway CC University of California

Lawrence Livermore
National Laboratory
French JE
National Institute of
Environmental Health
Sciences
French JE
Sciences
Sabri MI
Oregon Health and
Science University
DNA binding of radiolabeled benzene National Center for

Research Resources
Monks TJ
University of Texas at
Austin
Ross D
Texas A&M University

System
Smith MT
University of California
Berkeley
Source: FEDRIP 2005
Carcinogen inactivation of tumor

suppressor genes in p53 mice
Sponsor
Sciences
Mechanisms of leukemogenesis in
genetically-altered mice
Sciences
Biomarkers of exposure and effect in National Institute of
neurotoxicants including neurotic
benzene derivatives
Sciences
Benzene metabolites and
hematotoxicity
Sciences
Protective effect of NQ01 against
benzene toxicity
Sciences
Biomarkers of benzene exposure and National Institute of
genotoxicity
Sciences
BENZENE
238
3. HEALTH EFFECTS
BENZENE
239
4. CHEMICAL AND PHYSICAL INFORMATION

4.1
CHEMICAL IDENTITY
Information regarding the chemical identity of benzene is located in Table 4-1. Although the term benzol
is found in older literature for the commercial product, benzene is the name presently approved by the
International Union of Pure and Applied Chemistry (IUPAC), the Chemical Manufacturers Association
(CMA), and the American Society for Testing and Materials (ASTM) for the pure product.
4.2
PHYSICAL AND CHEMICAL PROPERTIES
Information regarding the physical and chemical properties of benzene is located in Table 4-2. The major
impurities found in commercial products are toluene, xylene, phenol, thiophene, carbon disulfide,
acetylnitrile, and pyridine (NIOSH 1974). Commercial refined benzene-535 is free of hydrogen sulfide
and sulfur dioxide, but contains a maximum of 1 ppm thiophene and a maximum of 0.15% nonaromatics.
Refined nitration-grade benzene is free of hydrogen sulfide and sulfur dioxide. Benzene is also
commercially available as thiophene-free, 99 mole%, 99.94 mole%, and nanograde quality (HSDB 2007).
BENZENE
240
Table 4-1. Chemical Identity of Benzene

Characteristic
Information
Reference
Chemical name
Synonym(s)
Benzene
Annulene, benzeen (Dutch), benzen
(Polish), benzol, benzole; benzolo (Italian),
coal naphtha, cyclohexatriene, fenzen
(Czech), phene, phenyl hydride,
pyrobenzol, pyrobenzole
Polystream
C6H6
HSDB 2007
HSDB 2007
71-43-2
CY-1400000
NA
No Data
UN1114; IMO3.2
35
C55276
1066
HSDB 2007
HSDB 2007
Registered trade name(s)

Chemical formula
Chemical structure
Identification numbers:
CAS registry
NIOSH RTECS
EPA hazardous waste
OHM/TADS
DOT/UN/NA/IMCO shipping
HSDB
NCI
Merck
IARC 1982
Budavari et al. 2001
HSDB 2007
HSDB 2007
HSDB 2007
CAS = Chemical Abstracts Service; DOT/UN/NA/IMO=Department of Transportation/United Nations/North America/

Intergovernmental Maritime Dangerous Goods Code; EPA = Environmental Protection Agency; HSDB=Hazardous
Substances Data Bank; NCI = National Cancer Institute; NIOSH= National Institute for Occupational Safety and
Health; OHM/TADS = Oil and Hazardous Materials/Technical Assistance Data System; RTECS = Registry of Toxic
Effects of Chemical Substances
BENZENE
241
Table 4-2. Physical and Chemical Properties of Benzene

Property
Information
Reference
Molecular weight
Color
Physical state
Melting point
Boiling point
Density at 15 C, g/cm3
Odor
Odor threshold:
Water
Aira
78.11
Clear, colorless liquid
colorless to light yellow liquid
5.5 C
80.1 C
0.8787
Aromatic

HSDB 2007
NFPA 1994
2.0 mg/L
Detection range: 34119 ppm
(geometric mean: 61 ppm)
Recognition: 97 ppm
0.54.5 mg/L
HSDB 2007
AIHA 1989
w/w: 0.188%
Alcohol, chloroform, ether, carbon
disulfide, acetone, oils, carbon,
tetrachloride, glacial acetic acid

Taste threshold:
Solubility:
Water at 25 C
Organic solvents
Partition coefficients:
Log Kowb
Log Kocc
Vapor pressure at 20 C
Henry's law constant at 25 C
Autoignition temperature
Flashpoint
NFPA hazard classification:
Health
Flammability
Reactivity
Flammability limits in air
Conversion factors
Explosive limits
HSDB 2007
HSDB 2007; Karickhoff 1981;

Kenaga 1980
2.13
1.81.9
75 mm Hg
5.5x10-3 atm-m3/mol
498 C
-11 C (closed cup)
2.2
3.3
0.0
1.2% (lower limit; 7.8% (upper limit)
1 ppm=3.26 mg/m3 at 20 C and
1 atm pressure; 1 mg/m3=0.31 ppm
1.4% (lower limit); 8% (upper limit)
NFPA 1994
Mackay and Leinonen 1975
NFPA 1994
HSDB 2007
NFPA 1994
HSDB 2007
HSDB 2007
Odor threshold values considered by AIHA (1989) to be acceptable based on review of peer-reviewed reports of
odor thresholds for benzene (range 0.78100 ppm).
b
Kow = octanol-water partitioning coefficient
c
Koc = soil adsorption coefficient
NFPA = National Fire Protection Association
BENZENE
242
BENZENE
243
5. PRODUCTION, IMPORT/EXPORT, USE, AND DISPOSAL

5.1
PRODUCTION
In 1825, Faraday isolated benzene from a liquid condensed by compressing oil gas. Benzene was first
synthesized by Mitscherlich in 1833 by distilling benzoic acid with lime. Benzene was first commercially
recovered from light oil derived from coal tar in 1849 and from petroleum in 1941 (IARC 1982a).
Several years after the end of World War II, the rapidly expanding chemical industry created an increased
demand for benzene that the coal carbonization industry could not fulfill. To meet this demand, benzene
was produced by the petroleum and petrochemical industries by recovery from reformat and liquid
byproducts of the ethylene manufacturing process (Fruscella 1992).
Currently, benzene is commercially recovered from both coal and petroleum sources. More than 98% of
the benzene produced in the United States is derived from the petrochemical and petroleum refining
industries (OSHA 1987). These sources include refinery streams (catalytic reformats), pyrolysis gasoline,
and toluene hydrodealkylation. Catalytic reformat is the major source of benzene (Greek 1990). Between
1978 and 1981, catalytic reformats accounted for approximately 4450% of the total U.S. benzene
production (Fishbein 1988). During catalytic reforming, cycloparaffins (also known by the obsolescent
term "naphthenes") such as cyclohexane, methyl cyclohexane, and dimethylcyclohexane are converted to
benzene by isomerization, dehydrogenation, and dealkylation, and paraffins in naphtha (such as hexane)
are converted to benzene by cyclodehydrogenation. The process conditions and the catalyst determine
which reaction will predominate. The benzene is recovered by solvent extraction (e.g., with sulpholane or
tetraethylene glycol). Pyrolysis gasoline is a liquid byproduct produced by the steam cracking of lower
paraffins (gas oil) or heavier hydrocarbons (heavy naphtha). Pyrolysis gasoline contains unsaturated
aliphatic hydrocarbons (such as ethylene and propylene) and aromatics. Several integrated pyrolysis
gasoline treatment processes are available including partial hydrogenation and extractive distillation;
hydrogenation, hydrodesulfurization, and solvent extraction; or partial hydrogenation, desulfurization,
hydrocracking, hydrodealkylation, and distillation for the optimization of benzene yield and the recovery
of benzene (Fruscella 1992; IARC 1982a). In the toluene hydrodealkylation process, toluene or
toluene/xylene mixtures are reacted with hydrogen at temperatures ranging from 500 to 595 C with usual
pressures of 46 mPa (4060 atm), and demethylated to produce benzene and methane. Another process
whereby toluene is converted to benzene and xylenes by transalkylation or disproportionation is also used
for the production of benzene (Fruscella 1992). Small quantities of benzene are also produced from
destructive distillation of coal used for coke manufacture. Benzene is derived from the light oil fraction
produced during the coking process (Greek 1990; Fruscella 1992). New coking, gasification, and
BENZENE
244
liquefaction processes for coal are all potential sources of benzene (IARC 1982a). Of the total U.S.
production capacity of 3.109 billion gallons in 2004, catalytic reformats constituted 45%, toluene and
xylene 30%, pyrolysis gasoline 23%, and coke oven <2% (SRI 2004).
Benzene ranks in the top 20 most abundantly produced chemicals in the United States (C&EN 1994;
Kirschner 1995; Reisch 1994). Production data from 1984 to 1994 indicate that the production of
benzene increased by about 4% annually (C&EN 1995).
According to the Toxics Release Inventory (TRI), 2,528 facilities in the United States produced or
processed benzene in 2004 (TRI04 2006). Table 5-1 lists the facilities in each state that manufacture or
process benzene, the intended use, and the range of maximum amounts of benzene that are stored on site.
The TRI data listed in Table 5-1 should be used with caution since only certain types of facilities were
required to report. Therefore, this is not an exhaustive list.
SRI (2004) lists the companies producing benzene in the United States in 2004. This list is summarized
in Table 5-2. Facilities listed as being responsible for producing the top 50% of regional capacity in the
United States include: Exxon Mobil Chemical Company, 518 million gallons (16%); Equistar Chemicals
LP, 330 million gallons (11%), Dow Chemical Company, 300 million gallons (10%); Flint Hills
Resources LP, 250 million gallons (8%), and BP Oil Company 230 million gallons (7%) (SRI 2004).
5.2
IMPORT/EXPORT
Benzene is imported and exported to the United States as both the pure chemical and as a mixture of
mineral fuels. The import of pure benzene into the United States is dependent on domestic production
and demand. Imports of benzene (from mineral fuels and pure benzene) into the United States were
approximately 5,252 million L (10,176 million pounds) in 2004, 6,024 million L (11,672 million pounds)
in 2003, and 5,912 million L (11,455 million pounds) in 2002 (USITC 2005). The largest exporters of
benzene to the United States during 2002-2004 were Canada, Iraq, Israel, Kuwait, Venezuela, and Saudi
Arabia (USITC 2005).
As in the case of import, the export of benzene from the United States to other countries is dependent on
domestic and world production and demand. The 20022004 data indicate an increase in export volumes
for benzene during this period. Exports of benzene (both pure benzene and benzene derived from mineral
fuels) to other countries were approximately 11 million L (21 million pounds) in 2002, 150 million L
BENZENE
245
Table 5-1. Facilities that Produce, Process, or Use Benzene
Statea
Minimum
Number of amount on site
facilities
in poundsb
Maximum
amount on site
in poundsb
Activities and usesc
AK
AL
AR
AS
AZ
CA
CO
CT
DE
FL
GA
GU
HI
IA
ID
IL
IN
KS
KY
LA
MA
MD
ME
MI
MN
MO
MP
MS
MT
NC
ND
NE
NH
NJ
NM
NV
NY
OH
26
85
24
1
30
172
23
8
13
36
33
6
15
21
3
102
80
59
66
212
15
20
9
108
36
30
7
43
27
34
9
11
9
61
32
5
48
101
99,999,999
499,999,999
9,999,999
999,999
9,999,999
99,999,999
9,999,999
9,999,999
49,999,999
499,999,999
49,999,999
9,999,999
49,999,999
10,000,000,000
9,999,999
99,999,999
99,999,999
499,999,999
99,999,999
10,000,000,000
99,999,999
49,999,999
9,999,999
499,999,999
99,999,999
9,999,999
999,999
499,999,999
499,999,999
49,999,999
49,999,999
9,999,999
999,999
99,999,999
9,999,999
99,999
49,999,999
99,999,999
1, 2, 3, 4, 5, 7, 8, 9, 11, 12
1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14
1, 2, 3, 4, 6, 8, 9, 11, 12, 13
9
1, 2, 4, 5, 7, 9, 10, 11, 12, 13, 14
1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 13, 14
1, 2, 3, 4, 5, 6, 7, 8, 9, 12, 13
1, 4, 6, 7, 9, 10, 12
1, 2, 3, 4, 5, 6, 7, 8, 9, 11, 12, 13
1, 2, 3, 4, 5, 7, 8, 9, 10, 12, 13, 14
1, 2, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14
2, 3, 4, 7, 9, 12
1, 2, 3, 4, 5, 6, 7, 9, 12, 13, 14
1, 2, 3, 4, 5, 7, 9, 10, 11, 12, 13, 14
7, 9, 12
1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14
1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14
1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 13, 14
1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14
1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14
2, 4, 7, 8, 9
1, 2, 3, 4, 5, 6, 7, 8, 9, 11, 12
2, 3, 4, 7, 9, 12, 13
1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14
1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14
1, 5, 7, 8, 9, 11, 12
2, 3, 4, 7, 9
1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 13, 14
1, 2, 3, 4, 5, 6, 7, 8, 9, 12, 13, 14
1, 2, 3, 4, 5, 6, 7, 9, 10, 11, 12, 13
1, 2, 3, 4, 5, 6, 9, 12, 13
1, 7, 10, 12, 13
1, 2, 7, 9, 12, 13
1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 13
1, 2, 3, 4, 5, 6, 7, 8, 9, 12, 13, 14
1, 2, 4, 8, 9, 12, 13
1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 13
1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14
0
0
100
100,000
100
0
0
1,000
1,000
0
0
100
100
0
10,000
0
0
0
0
0
100
100
100
0
100
0
0
0
10,000
0
0
0
0
0
100
100
0
0
BENZENE
246
Table 5-1. Facilities that Produce, Process, or Use Benzene
Statea
Minimum
Number of amount on site
facilities
in poundsb
Maximum
amount on site
in poundsb
Activities and usesc
OK
OR
PA
PR
RI
SC
SD
TN
TX
UT
VA
VI
WA
WI
WV
WY
48
12
100
52
9
26
8
53
361
42
36
8
57
27
32
37
499,999,999
9,999,999
99,999,999
10,000,000,000
9,999,999
9,999,999
999,999
9,999,999
10,000,000,000
49,999,999
49,999,999
499,999,999
499,999,999
9,999,999
99,999,999
9,999,999
1, 2, 3, 4, 5, 6, 7, 8, 9, 11, 12, 13, 14

1, 2, 3, 4, 5, 7, 9, 12, 13
1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14
1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 13
1, 2, 4, 7, 8, 9
1, 2, 3, 4, 5, 6, 8, 9, 10, 11, 12, 13
2, 3, 4, 7, 8, 11, 12, 13, 14
1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14
1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14
1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 13
1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 13
1, 2, 3, 4, 6, 7, 8, 9
1, 2, 3, 4, 5, 6, 7, 8, 9, 12, 13, 14
1, 4, 5, 7, 8, 9, 11, 12, 13, 14
1, 2, 4, 5, 6, 7, 8, 9, 12, 13
1, 2, 3, 4, 5, 6, 7, 9, 12, 13, 14
0
1,000
0
0
10,000
0
100
0
0
1,000
0
10,000
0
0
0
0
Post office state abbreviations used

Amounts on site reported by facilities in each state
c
Activities/Uses:
1. Produce
6. Impurity
2. Import
7. Reactant
3. Onsite use/processing
8. Formulation Component
4. Sale/Distribution
9. Article Component
5. Byproduct
10. Repackaging
b
Source: TRI04 2006 (Data are from 2004)
11.
12.
13.
14.
Chemical Processing Aid

Manufacturing Aid
Ancillary/Other Uses
Process Impurity
BENZENE
247
Table 5-2. Current U.S. Manufacturers of Benzenea

Company
ATOFINA Petrochemicals, Inc.
BASF FINA Petrochemicals, Inc.
BP America, Inc.
Ceveron Phillips Chemical Company, LP
Olefins and Polyolefins Business Unit
CITGO Petroleum Corporation
Conoco Phillips Refining Marketing and

Transportation Division
Dow Chemical USA
Equistar Chemicals, LP
ExxonMobil Chemical Company
Flint Hills Resources, LP

Frontier El Dorado Refining Company
HOVENSA, LLC
Huntsman, LLC
Lyondell-Citgo Refining, LP
Marathon Ashland Petroleum, LLC
Motiva Enterprises, LLC
NOVA Chemicals Corp.
The Premcor Refining Group Inc.
Shell Chemical Company
Sunoco, Inc.
Valero Energy Corporation

Eagle-Pitcher Industries, Inc.
Eagle-Pitcher Technologies, LLC
Chem Syn Laboratiories
(14C-benzene)
a
Location (annual capacity millions of gallons)
Port Arthur, Texas (87)
Texas City, Texas (230)
Pascagoula, Mississippi (150)

Corpus Christi, Texas (78)
Lake Charles, Louisiana (55)
Lemont, Illinois (19)
Alliance Louisiana (65)
Sweeny, Texas (39)
Wood River Illinois (60)
Freeport, Texas (80)
Plaquemine, Louisiana (317)
Alvin, Texas (137)
Channelveiw, Texas (90)
Baton Rouge Louisiana (100)
Baytown, Texas (180)
Beaumont, Texas (182)
Chalmette, Louisiana (56)
El Dorado, Kansas (15)
St. Croix, Virgin Islands (60)
Houston, Texas (50)
Catlettsburg, Kentucky (58)
Texas City, Texas (3)
Delaware City, Delaware (15)
Bayport, Texas (15)
Lima, Ohio (145)
Deer Park, Texas (180)
Marcus Hook, Pennsylvania (60)
Philadelphia, Pennsylvania (24)
Toledo Ohio (20)
Westville, New Jersey (18)

Three Rivers, Texas (20)
Lenexa, Kansas
Derived from Stanford Research Institute (SRI) 2004, receipt where otherwise noted. SRI reports production of
chemicals produced in commercial quantities (defined as exceeding 5,000 pounds or $10,000 in value annually) by
the companies listed.
BENZENE
248
(290 million pounds) in 2003, and 75 million L (145 million pounds) in 2004. These numbers are up
from 15 million L (29 million pounds) in 2001 and 3.3 million L (6.4 million pounds) in 1993 (USITC
2005). The largest importers of benzene from the United States are Canada, the Netherlands, Taiwan,
Spain, and Korea (USITC 2005).
5.3
USE
Benzene has been used extensively as a solvent in the chemical and drug industries, as a starting material
and intermediate in the synthesis of numerous chemicals, and as a gasoline additive (NTP 1994).
Benzene recovered from petroleum and coal sources is used primarily as an intermediate in the
manufacture of other chemicals and end products. The major uses of benzene are in the production of
ethylbenzene, cumene, and cyclohexane. Ethylbenzene (55% of benzene production volume) is an
intermediate in the synthesis of styrene, which is used to make plastics and elastomers. Cumene (24%) is
used to produce phenol and acetone. Phenols are used in the manufacture of phenolic resins and nylon
intermediates; acetone is used as a solvent and in the manufacture of pharmaceuticals. Cyclohexane
(12%) is used to make nylon resins. Other industrial chemicals manufactured from benzene include
nitrobenzene (5%), which is used in the production of aniline, urethanes, linear alkylbenzene sulfonates,
chlorobenzene, and maleic anhydride (Eveleth 1990; Greek 1990; HSDB 2007). Benzene is also a
component of gasoline since it occurs naturally in crude oil and since it is a byproduct of oil refining
processes (Brief et al. 1980; Holmberg and Lundberg 1985). Benzene is especially important for
unleaded gasoline because of its anti-knock characteristics. For this reason, the concentration of
aromatics, such as benzene, in unleaded fuels has increased (Brief et al. 1980). The percentage by
volume of benzene in unleaded gasoline is approximately 12% (NESCAUM 1989).
The EPA has listed benzene as a hazardous air pollutant and a hazardous waste (EPA 1977, 1981). In
addition, there is sufficient evidence to support classifying benzene as a human carcinogen (Group A)
(IRIS 2007). One result of EPA's action is that the widespread use of benzene as a solvent has decreased
in recent years. Many products that used benzene as solvents in the past have replaced it with other
organic solvents; however, benzene may still occur as a trace impurity in these products. Less than 2% of
the amount produced is used as a solvent in products such as trade and industrial paints, rubber cements,
adhesives, paint removers, artificial leather, and rubber goods. Benzene has also been used in the shoe
manufacturing and rotogravure printing industries (EPA 1978; OSHA 1977). In the past, certain
consumer products (such as some paint strippers, carburetor cleaners, denatured alcohol, and rubber
BENZENE
249
cement used in tire patch kits and arts and crafts supplies) contained small amounts of benzene (Young et
al. 1978). Other consumer products that contained benzene were certain types of carpet glue, textured
carpet liquid detergent, and furniture wax (Wallace et al. 1987).
The Consumer Products Safety Commission (CPSC) withdrew an earlier proposal to ban consumer
products, except gasoline and laboratory reagents, that contained benzene as an intentional ingredient or
as a contaminant at >0.1% by volume. The withdrawal of the rulemaking was based on CPSC findings
that benzene was no longer used as an intentional ingredient and that the contaminant levels remaining in
certain consumer products were unlikely to result in significant exposures (NTP 1994). Products
containing >5% benzene, and paint solvents and thinners containing <10% of petroleum distillates such as
benzene, are required to meet established labeling requirements. In a guidance document targeting school
science laboratories, the CPSC recommended that benzene not be used or stored in schools. The
document identified benzene as a carcinogen and ascertained that the hazards posed by its use in high
school laboratories may be greater than its potential usefulness.
The U.S. Food and Drug Administration (FDA) regulates benzene as an indirect food additive under the
Food, Drug, and Cosmetics Act (FDCA). Under the FDCA, benzene is restricted to use only as a
component of adhesives used on articles intended for packaging, transport, or holding foods (FDA 1977).
5.4
DISPOSAL
Benzene-containing wastes, such as commercial chemical products, manufacturing chemical

intermediates, and spent solvents, are subject to federal and/or state hazardous waste regulations (HSDB
2007). Regulations governing the treatment and disposal of benzene-containing wastes are presented in
Chapter 8. Waste byproducts from benzene production processes include acid and alkali sludges, liquidsolid slurries, and solids (EPA 1982b; Saxton and Narkus-Kramer 1975). In the past, landfilling and
lagooning have been the major methods of disposal of benzene-containing industrial wastes (EPA 1982b).
In addition to biodegradation, a portion of the benzene is expected to be lost due to volatilization.
Unfortunately benzene, along with other hazardous contaminants, also leaches into groundwater from the
lagooned wastes. Currently, the recommended method of disposal is to incinerate solvent mixtures and
sludges at a temperature that ensures complete combustion. The recommended methods for combustion
are liquid injection incineration at a temperature range of 6501,600 C and a residence time of 0.1
2 seconds; rotary kiln incineration at a temperature range of 8201,600 C and residence times of seconds
for liquids and gases, and hours for solids; and fluidized bed incineration at a temperature range of 450
BENZENE
250
980 C and residence times of seconds for liquids and gases and longer for solids (HSDB 2007). Since
benzene burns with a very smoky flame, dilution with alcohol or acetone is suggested to minimize smoke.
Small quantities of benzene waste can be destroyed by chemical reaction. For example, treating benzene
with dichromate in strong sulfuric acid for 12 days is sufficient for total destruction (HSDB 2007).
Underground injection also appears to be an important disposal method in some states. Approximately
436,000 pounds of benzene (6% of the total environmental release) was disposed of by underground
injection. This disposal via underground injection in 2006 was higher than the amount (356,000 pounds)
released in 1992 (TRI90 1992), but lower than the release in 1990 (654,000 pounds) (TRI90 1992) and
2002 (692,000 pounds) (TRI02 2005). In addition, 24,000 pounds of benzene (0.3% of the total
environmental release) was disposed of via land disposal (TRI04 2006). The amount discharged to soil in
2002 was less than one quarter of the amount (724,000 pounds) discharged in 1990 (TRI90 1992) and less
than one third of the amount (340,000 pounds) discharged in 1992 (TRI92 1994).
Several methods exist for the treatment of waste water that contains benzene: biological treatment
(aeration or activated sludge process), solvent extraction, air and/or steam stripping, and activated carbon
process (EPA 1994a; IRPTC 1985). Full-scale chemical treatability studies have demonstrated 95100%
reductions in benzene concentrations for industrial waste waters receiving biological treatments (HSDB
2007). A combination of steam stripping and air stripping, and a vapor extraction system that removes
the separated benzene vapor may be suitable for the treatment of contaminated groundwater and soil (Naft
1992). An in situ bioremediation process has been used to decontaminate a site by delivering a controlled
amount of nitrate (to accelerate biodegradation of benzene) to the site under hydraulic control (Kennedy
and Hutchins 1992).
BENZENE
251
6. POTENTIAL FOR HUMAN EXPOSURE

6.1
OVERVIEW
Benzene has been identified in at least 1,000 of the 1,684 hazardous waste sites that have been proposed
for inclusion on the EPA National Priorities List (NPL) (HazDat 2006). However, the number of sites
evaluated for benzene is not known. The frequency of these sites can be seen in Figure 6-1. Of these
sites, 995 are located within the United States, 3 are located in the Commonwealth of Puerto Rico (not
shown), and 2 are located in the Virgin Islands (not shown).
Benzene is released to the environment by both natural and industrial sources, although the anthropogenic
emissions are undoubtedly the most important. Emissions of benzene to the atmosphere result from
gasoline vapors, auto exhaust, and chemical production and user facilities. EPA's estimate of nationwide
benzene atmospheric emissions from various sources was 34,000 metric tons/year (EPA 1989).
According to the Toxics Release Inventory, releases to the air from manufacturing and processing
facilities were about 6,7 million pounds (3,055 metric tons) in 2004 (TRI04 2006). Releases to air
accounted for about 93% of the total industry-related releases to the environment (TRI04 2006). Benzene
is released to water and soil from industrial discharges, landfill leachate, and gasoline leaks from
underground storage tanks.
Chemical degradation reactions, primarily reaction with hydroxyl radicals, limit the atmospheric
residence time of benzene to only a few days, and possibly to only a few hours. Benzene released to soil
or waterways is subject to volatilization, photooxidation, and biodegradation. Biodegradation, principally
under aerobic conditions, is an important environmental fate process for water- and soil-associated
benzene.
Benzene is ubiquitous in the atmosphere. It has been identified in air samples of both rural and urban
environments and in indoor air. Although a large volume of benzene is released to the environment,
environmental levels are low because of efficient removal and degradation processes. Benzene partitions
mainly into air (99.9%) and inhalation is the dominant pathway of human exposure accounting for >99%
of the total daily intake of benzene (Hattemer-Frey et al. 1990; MacLeod and MacKay 1999). The
general population is exposed to benzene primarily by tobacco smoke (both active and passive smoking)
and by inhaling contaminated air (particularly in areas with heavy motor vehicle traffic and around filling
stations). Air around manufacturing plants that produce or use benzene and air around landfills and
hazardous waste sites that contain benzene are additional sources of exposure. Exposure to benzene can
252
BENZENE
Figure 6-1. Frequency of NPLSites with Benzene Contamination
. ''
Frequency of
NPL Sites
Derived from HazDat 2006
tm]1-10
~11-19
~21-28
l!l31-45
IIllJI]61-74
~96
BENZENE
253
also result from ingestion of contaminated food or water. Use of contaminated tap water for cooking,
showering, etc., can also be a source of inhalation exposure since benzene can volatilize from water.
Compared to inhalation, dermal exposure accounts for a minor portion of the total exposure of the general
population. The magnitude of exposure is greatest for those individuals occupationally exposed to
benzene; however, a far greater number of individuals are exposed as a result of benzene released from
smoking tobacco products, from gasoline filling stations, and from auto exhaust. Smoking was found to
be the largest anthropogenic source of direct human exposure to benzene (Duarte-Davidson et al. 2001;
Hattemer-Frey et al. 1990).
6.2
RELEASES TO THE ENVIRONMENT
The Toxics Release Inventory (TRI) data should be used with caution because only certain types of
facilities are required to report (EPA 2005g). This is not an exhaustive list. Manufacturing and
processing facilities are required to report information to the TRI only if they employ 10 or more full-time
employees; if their facility is included in Standard Industrial Classification (SIC) Codes 10 (except 1011,
1081, and 1094), 12 (except 1241), 2039, 4911 (limited to facilities that combust coal and/or oil for the
purpose of generating electricity for distribution in commerce), 4931 (limited to facilities that combust
coal and/or oil for the purpose of generating electricity for distribution in commerce), 4939 (limited to
facilities that combust coal and/or oil for the purpose of generating electricity for distribution in
commerce), 4953 (limited to facilities regulated under RCRA Subtitle C, 42 U.S.C. section 6921 et seq.),
5169, 5171, and 7389 (limited S.C. section 6921 et seq.), 5169, 5171, and 7389 (limited to facilities
primarily engaged in solvents recovery services on a contract or fee basis); and if their facility produces,
imports, or processes 25,000 pounds of any TRI chemical or otherwise uses >10,000 pounds of a TRI
chemical in a calendar year (EPA 2005g).
6.2.1
Air
Estimated releases of 6.7 million pounds (~ 3,055 metric tons) of benzene to the atmosphere from
968 domestic manufacturing and processing facilities in 2004, accounted for about 93% of the estimated
total environmental releases from facilities required to report to the TRI (TRI04 2006). These releases are
summarized in Table 6-1.
Benzene is released into the atmosphere from both natural and industrial sources. Natural sources include
crude oil seeps, forest fires, and plant volatiles (Brief et al. 1980; Graedel 1978). Major anthropogenic
sources of benzene include environmental tobacco smoke, automobile exhaust, automobile refueling
BENZENE
254
Table 6-1. Releases to the Environment from Facilities that Produce, Process, or
Use Benzenea
Reported amounts released in pounds per yearb

Total release
Statec RFd
AK
AL
AR
AZ
CA
CO
CT
DE
FL
GA
GU
HI
IA
ID
IL
IN
KS
KY
LA
MA
MD
ME
MI
MN
MO
MP
MS
MT
NC
ND
NE
NJ
NM
NV
NY
OH
OK
7
15
8
13
65
11
9
5
26
15
3
9
14
2
35
30
19
22
62
10
11
5
41
16
17
2
13
5
12
2
9
20
9
3
44
52
11
Aire
19,568
290,660
33,137
4,910
57,611
19,630
2,552
3,425
396,187
36,893
7,106
11,171
42,671
1,732
171,104
211,476
69,823
77,345
605,128
5,556
12,644
3,027
345,255
16,614
98,498
710
59,076
17,242
17,290
6,400
1,333
51,054
38,730
1,005
60,682
183,972
114,701
Waterf
UIg
19
0
290
0
66
0
0
0
129
228
0
0
2
0
6,006
0
41
0
128
0
1
0
30
0
1
0
No data
0
102
0
805 14,001
164
231
799
0
1,075 122,723
55
0
8
0
23
0
8
0
2,724
0
0
0
0
0
25
0
5
0
0
0
0
0
No data
0
443
0
No data
7
No data
0
56
0
67
5
14
0
Landh
375
280
0
9
1,292
10
0
26
0
9
0
12
0
0
2,152
4,654
259
766
1,688
26
184
0
127
131
0
0
14
15
5
1
0
150
2
0
10
730
278
Otheri
0
2
0
0
598
36
2
0
157
0
401
0
0
0
104
29
0
20
968
537
1
467
2,295
1,561
0
0
0
0
48
0
6
169
0
0
1,335
82
71
On-sitej
19,962
290,955
33,203
4,915
57,970
19,630
2,554
9,431
396,228
37,026
7,107
11,201
42,672
1,732
172,227
227,079
70,226
78,904
729,109
5,611
12,821
3,049
345,363
19,360
98,498
710
59,102
17,248
17,295
6,400
1,333
51,510
38,739
1,005
60,738
184,049
114,984
Off-sitek
0
277
0
4
1,888
46
2
26
157
4
401
12
0
0
1,235
3,886
251
26
2,473
563
15
467
2,322
1,670
0
0
13
14
48
1
6
306
0
0
1,345
807
80
On- and offsite

19,962
291,232
33,203
4,919
59,858
19,676
2,556
9,457
396,385
37,030
7,508
11,213
42,672
1,732
173,462
230,965
70,477
78,930
731,582
6,173
12,836
3,516
347,686
21,030
98,498
710
59,115
17,262
17,343
6,401
1,339
51,816
38,739
1,005
62,084
184,856
115,064
BENZENE
255
Table 6-1. Releases to the Environment from Facilities that Produce, Process, or
Use Benzenea
Reported amounts released in pounds per yearb

Total release
Statec RFd
OR
PA
PR
RI
SC
SD
TN
TX
UT
VA
VI
WA
WI
WV
WY
Total
6
39
10
2
6
11
17
150
11
17
3
18
14
5
7
968
Aire
4,651
234,647
49,879
1,841
64,627
272
73,926
2,681,863
31,314
183,748
20,576
50,667
141,183
76,556
24,986
6,736,655
Waterf
UIg
8
0
419
0
7
0
4
0
6
0
1
0
63
0
621 298,595
750
0
787
0
0
0
14
0
0
0
284
0
0
0
16,051 435,790
Landh
Otheri
41
0
345 11,389
0
0
0
820
250 1,023
0
0
280
0
8,326 1,713
809
233
26
0
9
0
95
48
251
63
393
0
1
0
24,033 24,179
On-sitej
4,659
235,066
49,886
1,845
64,883
273
73,989
2,935,539
32,069
184,535
20,576
50,770
141,183
76,840
24,986
7,147,049
Off-sitek
41
11,733
0
820
1,023
0
280
55,579
1,038
26
9
54
314
393
1
89,659
On- and offsite

4,700
246,800
49,886
2,665
65,906
273
74,269
2,991,118
33,107
184,561
20,585
50,824
141,496
77,233
24,987
7,236,707
The TRI data should be used with caution since only certain types of facilities are required to report. This is not an
exhaustive list. Data are rounded to nearest whole number.
b
Data in TRI are maximum amounts released by each facility.
c
Post office state abbreviations are used.
d
Number of reporting facilities.
e
The sum of fugitive and point source releases are included in releases to air by a given facility.
f
Surface water discharges, waste water treatment-(metals only), and publicly owned treatment works (POTWs)
(metal and metal compounds).
g
Class I wells, Class II-V wells, and underground injection.
h
Resource Conservation and Recovery Act (RCRA) subtitle C landfills; other on-site landfills, land treatment, surface
impoundments, other land disposal, other landfills.
i
Storage only, solidification/stabilization (metals only), other off-site management, transfers to waste broker for
disposal, unknown
j
The sum of all releases of the chemical to air, land, water, and underground injection wells.
k
Total amount of chemical transferred off-site, including to POTWs.
RF = reporting facilities; UI = underground injection

Source: TRI04 2006 (Data are from 2004)
BENZENE
256
operations, and industrial emissions. Using source exposure modeling, it was estimated that benzene
emissions were highest from coke oven blast furnaces (Edgerton and Shah 1992). Other sources that
contributed to emissions of benzene include automobiles, petrochemical industries, waste water treatment
plants, and petroleum industries (Edgerton and Shah 1992).
Industrial and automotive sources of benzene are well monitored. EPA (1989) estimates of nationwide
benzene atmospheric emissions, in metric tons/year (kkg/year), from various industrial sources include:
(1) coke byproduct recovery plants (17,000 kkg/year), (2) benzene waste operations (5,300 kkg/year),
(3) gasoline marketing systems, including bulk gasoline terminals and plants, service stations, and
delivery tank trucks (4,800 kkg/year), (4) transfer operations at chemical production facilities, bulk
terminals, and coke byproduct recovery plants (4,600 kkg/year), (5) benzene storage vessels (620
1,290 kkg/year), (6) industrial solvent use (450 kkg/year), (7) chemical manufacturing process vents
(340 kkg/year), and (8) ethylbenzene/styrene process vents (135 kkg/year). Benzene is present in
passenger car tailpipe emissions at compositions ranging from 2.9 to 15% of the total tailpipe
hydrocarbon composition (Black et al. 1980). The 2002 benzene industrial emission inventory for
California totaled 266 metric tons/year (CARB 2005); these numbers did not include motor vehicle
exhaust,which accounted for 71% of emissions in 1984 (Cal EPA 1987). The contribution of mobile
source hazardous air pollutant emissions has been compared to that of stationary sources in the SeattleTacoma area. Mobile sources were estimated to contribute approximately 83% of the benzene from
stationary areas and mobile sources combined, with major stationary point sources excluded (EPA
1994d). Benzene is also released by off-gassing from particle board (Glass et al. 1986), vaporization
from oil spills, and emissions from landfills (Bennett 1987; Wood and Porter 1987). While all of these
sources release more benzene into the environment, a large percentage of the benzene inhaled by humans
comes from cigarette smoke. Exhaled breath of smokers contains benzene (Wallace 1989a, 1989b;
Wallace and Pellizzari 1986; Wester et al. 1986).
According to the Texas Natural Resource Conservation Commission (TNRCC), of the 93 districts in
Texas reporting benzene emissions from industrial sources, in 1980, 1985, and 1988, 10 districts reported
0 tons/year, 79 districts reported 0.1100 tons/year, 4 districts reported 100400 tons/year, 2 districts
reported 4001,000 tons/year, and 1 district had emissions projected at above 1,000 tons/year (Pendleton
1995).
The natural sources most monitored for benzene released to air are fires (Austin et al. 2001; Lowry et al.
1985). A study of nine municipal fires in Canada found a mean concentration of 3.45 ppm of benzene
BENZENE
257
and had very high relative concentration of other volatile organic compounds (VOCs) monitored (Austin
et al. 2001).
There is a potential for atmospheric release of benzene from hazardous waste sites. Benzene has been
detected in air in 200 of the 1,684 current and former NPL sites where it has been detected in some
medium (HazDat 2006).
6.2.2
Water
Estimated releases of 16,051 pounds (~7 metric tons) of benzene to surface water from 968 domestic
manufacturing and processing facilities in 2004, accounted for about 0.2% of the estimated total
environmental releases from facilities required to report to the TRI (TRI04 2006). These releases are
summarized in Table 6-1.
Benzene is released to water from the discharges of both treated and untreated industrial waste water,
gasoline leaks from underground storage tanks, accidental spills during marine transportation of chemical
products, and leachate from landfills and other contaminated soils, (CDC 1994; Crawford et al. 1995;
EPA 1979; NESCAUM 1989; Staples et al. 1985). A fire in a tire dump site in western Frederick County,
Virginia, produced a free-flowing oily tar containing benzene among other chemicals. The seepage from
this site contaminated nearby surface water (EPA 1993). Between 1986 and 1991, 3,000 gallons of
benzene were accidentally released into Newark Bay and its major tributaries. Another 3,000 gallons
were released in 1991 (Crawford et al. 1995).
There is a potential for release of benzene to water from hazardous waste sites. Benzene has been
detected in groundwater samples collected at 832 of the 1,684 current and former NPL sites and in surface
water samples collected at 208 of the 1,684 sites (HazDat 2006).
6.2.3
Soil
Estimated releases of 24,033 pounds (~11 metric tons) of benzene to soils from 968 domestic
manufacturing and processing facilities in 2004, accounted for about 0.3% of the estimated total
environmental releases from facilities required to report to the TRI (TRI04 2006). An additional 435,000
pounds (~197 metric tons), constituting about 6% of the total environmental emissions, were released via
underground injection (TRI04 2006). These releases are summarized in Table 6-1.
BENZENE
258
Benzene is released to soils through industrial discharges, land disposal of benzene-containing wastes,
and gasoline leaks from underground storage tanks. In northern Virginia, approximately 200,000 gallons
of liquid hydrocarbons were released from a fuel-storage terminal into the underlying soil (Mushrush et
al. 1994).
There is a potential for release of benzene to soil from hazardous waste sites. Benzene has been detected
in soil samples collected at 436 of the 1,684 sites, and in sediment samples collected at 145 of the
1,684 sites where benzene has been detected in some medium (HazDat 2006).
6.3
6.3.1
ENVIRONMENTAL FATE
Transport and Partitioning
The high volatility of benzene is the controlling physical property in the environmental transport and
partitioning of this chemical. Benzene is considered to be highly volatile with a vapor pressure of
95.2 mm Hg at 25 C. Benzene is moderately soluble in water, with a solubility of 1,780 mg/L at 25 C,
and the Henry's law constant for benzene (5.5x10-3 atm-m3/mole at 25 C) indicates that benzene
partitions readily to the atmosphere from surface water (Mackay and Leinonen 1975). Since benzene is
soluble in water, some minor removal from the atmosphere via wet deposition may occur. A substantial
portion of any benzene in rainwater that is deposited to soil or water will be returned to the atmosphere
via volatilization.
Benzene released to soil surfaces partitions to the atmosphere through volatilization, to surface water
through runoff, and to groundwater as a result of leaching. The soil organic carbon sorption coefficient
(Koc) for benzene has been measured with a range of 6083 (Karickhoff 1981; Kenaga 1980), indicating
that benzene is highly mobile in soil and readily leaches into groundwater. Other parameters that
influence leaching potential include the soil type (e.g., sand versus clay), amount of rainfall, depth of the
groundwater, and extent of degradation. In a study of the sorptive characteristics of benzene to
groundwater aquifer solids, benzene showed a tendency to adsorb to aquifer solids. Greater soil
adsorption was observed with increasing organic matter content (Uchrin and Mangels 1987). An
investigation of the mechanisms governing the rates of adsorption and desorption of benzene by dry soil
grains revealed that periods of hours are required to achieve equilibrium and that adsorption is much
faster than desorption (Lin et al. 1994). The rate of volatilization and leaching are the principal factors
that determine overall persistence of benzene in sandy soils (Tucker et al. 1986).
BENZENE
259
Studies suggest that benzene does not bioaccumulate in marine organisms. The bioconcentration/
bioaccumulation potential of benzene in aquatic organisms of the open coastal ocean was investigated by
sampling final effluent from the Los Angeles County waste water treatment plant quarterly from
November 1980 to August 1981 (Gossett et al. 1983). Benzene has a relatively low octanol/water
partition coefficient (log Kow=2.13 or 2.15) (Gossett et al. 1983; HSDB 2007). In the alga, Chlorella, a
bioaccumulation factor of 30 was determined experimentally (Geyer et al. 1984). An experimental
bioconcentration factor (BCF) of 4.27 was measured in goldfish reared in water containing 1 ppm of
benzene (Ogata et al. 1984). Based on these measured values, bioconcentration/bioaccumulation of
benzene in the aquatic food chains does not appear to be important. These results are consistent with the
fact that benzene has a relatively low octanol/water partition coefficient (Gossett et al. 1983; HSDB
2007), suggesting relatively low bioaccumulation. There is no evidence in the literature of
biomagnification of benzene in aquatic food chain.
Evidence exists for the uptake of benzene by cress and barley plants from soil (Scheunert et al. 1985;
Topp et al. 1989). BCFs for barley plants after 12, 33, 71, and 125 days were 17, 2.3, 2.9, and 4.6,
respectively. BCFs for cress plants after 12, 33, and 79 days were 10, 2.3, and 1.9, respectively. The
relative decrease in the BCFs with time was attributed to growth dilution (Topp et al. 1989). Since
benzene exists primarily in the vapor phase, air-to-leaf transfer is considered to be the major pathway of
vegetative contamination (Hattemer-Frey et al. 1990). Based on an equation to estimate vegetative
contamination, the total concentration of benzene on exposed food crops consumed by humans and used
as forage by animals was estimated to be 587 ng/kg, 81% of which was from air-to-leaf transfer and 19%
was from root uptake (Hattemer-Frey et al. 1990).
Benzene also accumulates in leaves and fruits of plants. After 40 days, plants grown in benzene-rich
environments showed bioaccumulation in the leaves and fruit that were greater than the air portioning
coefficient of benzene in the atmosphere. Blackberries exposed to 0.313 ppm and apples exposed to
2.75 ppm contained about 1,000 and 36 ng/g of benzene, respectively (Collins et al. 2000).
6.3.2
Transformation and Degradation
Benzene undergoes a number of different transformation and degradation reactions in the environment as
discussed in the following sections. The resulting environmental transformation products within different
media are shown in Figure 6-2.
BENZENE
260
Figure 6-2. Environmental Transformation Products of Benzene in Various Media

Air
NO2
OH
O
NO2
photooxidation BenzeneOH
OH
adduct
Phenol
C
O
Intermediate
ring-cleavage
Nitrobenzene
C
H
Bandow et al.
1985
Formaldehyde
Glyoxal
photooxidation
+
O
Maleic anhydride
NO2
NO
hr
OH
OH
OH
NO2
Water
p-Nitrophenol
o-Nitrophenol
2,4-Dinitrophenol
NO2
Nojima et al.
1975
2,6-Dinitrophenol
OH
OH
OH
Biodegradation
O2N
NO2
NO2
Nitrobenzene
OH
NO2
Harayama and Timmis

1992
or
OH
Phenol
Catechol
OH
Hydroquinone
Soil
microbial
O2
OH
Benzene-OH
adduct
OH
Intermediate
Catechol
Substrate
for ring
fusion
Hopper 1978
BENZENE
261
6.3.2.1 Air
Benzene in the atmosphere exists predominantly in the vapor phase (Eisenreich et al. 1981). The most
significant degradation process for benzene is its reaction with atmospheric hydroxyl radicals. The rate
constant for the vapor phase reaction of benzene with photochemically produced hydroxyl radicals has
been determined to be 1.3x10-12 cm3/molecule-second, which corresponds to a residence time of 8 days at
an atmospheric hydroxyl radical concentration of 1.1x106 molecules/cm3 (Gaffney and Levine 1979;
Lyman 1982). With a hydroxyl radical concentration of 1x108 molecules/cm3, corresponding to a
polluted atmosphere, the estimated residence time is shortened to 2.1 hours (Lyman 1982). Benzene may
also react with other oxidants in the atmosphere such as nitrate radicals and ozone; however, the rate of
degradation is considered insignificant compared to the rate of reaction with hydroxyl radicals.
Residence times of 472 years for rural atmospheres and 152 years for urban atmospheres were calculated
for the reaction of benzene with ozone (O3) using a rate constant for O3 of 7x10-23 cm3/molecule-second
(Pate et al. 1976) and atmospheric concentrations for O3 of 9.6x1011 molecules/cm3 (rural) and
3x1012 molecules/cm3 (urban) (Lyman 1982).
The reaction of benzene and nitric oxide in a smog chamber was investigated to determine the role of
benzene in photochemical smog formation (Levy 1973). The results showed that benzene exhibited low
photochemical smog reactivity in the four categories tested: rate of photooxidation of nitric oxide,
maximum oxidant produced, eye-irritation response time, and formaldehyde formation. The authors
concluded that benzene probably does not play a significant role in photochemical smog formation (Levy
1973). In the presence of active species such as nitrogen oxides and sulfur dioxide, the rate of
photodegradation of benzene in the gas phase was greater than that in air alone. Its half-life in the
presence of such active species (100 ppm benzene in the presence of 10110 ppm NOx or 10
100 ppm SO2) was 46 hours with 50% mineralization to carbon dioxide in approximately 2 days (Korte
and Klein 1982). Some of the products of the reaction of benzene with nitrogen monoxide gas (e.g.,
nitrobenzene, o- and p-nitrophenol, and 2,4- and 2,6-dinitrophenol) may have potentially adverse effects
on human health (Nojima et al. 1975); however, these species also have relatively short atmospheric
lifetimes, which should limit the potential exposure to these compounds. Photooxidation of benzene in a
nitrogen monoxide/nitrogen dioxide-air system formed formaldehyde, formic acid, maleic anhydride,
phenol, nitrobenzene, and glyoxal (ethane-1,2-dione) (Bandow et al. 1985).
BENZENE
262
Direct photolysis of benzene in the atmosphere is not likely because the upper atmosphere effectively
filters out wavelengths of light <290 nm, and benzene does not absorb wavelengths of light >260 nm
(Bryce-Smith and Gilbert 1976).
6.3.2.2 Water
Benzene is subject to indirect photolysis in sunlit surface water, but does not undergo direct photolysis.
For direct photolysis to occur, a substance must absorb photons of light >290 nm. During indirect
photolysis, light energy is absorbed by other constituents (photosensitizers) of the media (water, soil) and
the excited species can then transfer energy to benzene (indirectly promoting it to an excited electronic
state), or lead to the formation of reactive species, such as singlet oxygen or hydroxy radicals, which react
with benzene. Humic and fulvic acids are well-known photosensitizing agents and are practically
ubiquitous in natural waters. A half-life of 16.9 days was reported for photolysis of benzene dissolved in
oxygen-saturated deionized water and exposed to sunlight (Hustert et al. 1981).
Benzene is readily degraded in water under aerobic conditions. Results of a biochemical oxygen demand
(BOD) test determined that benzene was completely biodegradable after the second week of static
incubation at 25 C at benzene concentrations of 5 and 10 mg/L using domestic waste water as the
microbial inoculum (Tabak et al. 1981). A study of the degradation of benzene by the microbial
population of industrial waste water at 23 C using a shaker flask system showed that after 6 hours, only
8% (4 mg/L) of the initial 50 mg/L dose of benzene remained (Davis et al. 1981). Water from a
petroleum production site was successfully biotreated for complete removal of benzene using a
flocculated culture of Thiobacillus denitrificans strain F and mixed heterotrophs (Rajganesh et al. 1995).
Microbial degradation of benzene in aquatic environments is influenced by many factors including
microbial population, dissolved oxygen, nutrients, other sources of carbon, inhibitors, temperature, pH,
and initial concentration of benzene. Vaishnav and Babeu (1987) reported biodegradation half-lives for
benzene in surface water (river water) and groundwater of 16 and 28 days, respectively. Benzene was
found to be resistant to biodegradation in surface water taken from a harbor and supplemented with either
nutrients (nitrogen and phosphorus) or acclimated microbes, however, biodegradation did occur, with a
half-life of 8 days, in surface water enriched with both nutrients and microbes (Vaishnav and Babeu
1987). At very high levels, as may be the case of a petroleum spill, benzene (and other compounds
contained in petroleum) is toxic to microorganisms and the rate of degradation is slow compared to low
initial starting concentrations. In another study, Davis et al. (1994) observed rapid aerobic biodegradation
BENZENE
263
of benzene in aquifer groundwater samples and measured times for 50% disappearance ranging from
4 days for an initial benzene concentration of 1 mg/kg to 14 days for an initial benzene concentration of
10 mg/kg. Under acidic conditions (pH 5.3, 20 C), benzene was completely microbially degraded in
16 days in groundwater taken from a shallow well (Delfino and Miles 1985).
The aerobic biodegradation of benzene is also influenced by the presence of other aromatic hydrocarbons.
A bacterial culture grown with aromatic hydrocarbons plus nitrogen-, sulfur-, and oxygen-containing
aromatic compounds was much less efficient in degrading benzene than the culture grown with aromatic
hydrocarbons alone. Pyrrole strongly inhibited benzene degradation. Benzene degradation was high
when toluene and xylene were present (Arvin et al. 1989).
Laboratory studies on microbial degradation of benzene with mixed cultures of microorganisms in
gasoline-contaminated groundwater revealed that both oxygen and nitrogen concentrations are major
controlling factors in the biodegradation of benzene. Nitrogen enhanced the biodegradation rate of
benzene 4.5-fold, over innoculum-enriched water alone. More than 95% of the benzene in groundwater
was removed through microbial action within 73.5 hours (Karlson and Frankenberger 1989).
Benzene biodegradation under anaerobic conditions does not readily occur. When dissolved oxygen is
depleted, an alternative electron acceptor such as nitrate, carbonate, or iron(III) must be available, and
microbes capable of using the alternative electron acceptor to degrade the benzene must be present
(McAllister and Chiang 1994). Using aquifer material obtained from a landfill from Norman, Oklahoma,
no significant benzene biodegradation was reported during the first 20 weeks of incubation under
anaerobic conditions at 17 C; however, after 40 weeks of incubation, benzene concentrations were
reduced by 72 and 99% of the benzene was degraded after 120 weeks (Wilson et al. 1986). No
degradation of benzene was observed in 96 days under anaerobic conditions (20 C) using raw water
intake from a water treatment plant (Delfino and Miles 1985).
Use of water as an oxygen source in the anaerobic degradation of benzene has been demonstrated.
Experiments indicated that incorporation of 18O from 18O-labeled water is the initial step in the anaerobic
oxidation of benzene by acclimated methanogenic cultures. Phenol was the first major product (Vogel
and Grbi-Gali 1986).
BENZENE
264
6.3.2.3 Sediment and Soil

Benzene is biodegraded in soil under aerobic conditions. Microbial metabolism of benzene proceeds
through the formation of cis-dihydrodiols and, with further metabolism, to catechols, which are the
substrates for ring fission (Gibson 1980; Hopper 1978). Pseudomonas putida oxidized benzene through
cis-1,2-dihydroxy-1,2-dihydrobenzene (Gibson 1977; Hopper 1978). After acclimation, Norcardia
species and Pseudomonas species, effectively degraded benzene to carbon dioxide within 7 days (45
90%). A strain of Rhodococcus isolated from contaminated river sediment mineralized 71% of benzene
at an initial concentration of 0.7 mg/L in 14 days (Malachowsky et al. 1994). The soil bacterium
Nitrosomonas europaea catabolized benzene to phenol and hydroquinone (Keener and Arp 1994).
Another new mixotrophic bacteria, a strain of Pseudomonas sp. isolated from contaminated soil, grew
under both anaerobic and aerobic conditions and used benzene for its growth (Morikawa and Imanaka
1993). The biodegradation of 2 mg of radiolabeled benzene in 100 g of soil with a mixed microbial
population transformed 47% of the added radioactivity to carbon dioxide after 10 weeks (Haider et al.
1981). The authors concluded that specific organisms that mineralize benzene were present in the soil in
only small numbers.
ULTRA, a fate and transport model used to predict the environmental fate of benzene following leakage
of gasoline from an underground storage tank into shallow sandy soil, indicated that only about 1% of the
benzene in the gasoline would be degraded over a 17-month period, and 3% would remain in the soil
(Tucker et al. 1986). According to this model, most of the benzene that would be leaked in soil would
either volatilize (67%) or move into groundwater (29%).
Salanitro (1993) summarized the aerobic degradation rates for BTEX in laboratory subsoil-groundwater
slurries and aquifers. The data indicate that decay rates for benzene are highest (1952% per day) for
benzene concentrations <1 ppm when initial dissolved oxygen levels are about 8 ppm. Rates are
significantly reduced (01.1% per day) when benzene levels are 12 ppm, and no degradation was
observed when benzene levels were >2 ppm. This is particularly relevant in the case of petroleum spills
as high concentrations of petroleum compounds are toxic to organisms and decrease the rate of
biodegradation.
Benzene has been shown to be anaerobically transformed by mixed methanogenic cultures derived from
ferulic acid-degrading sewage sludge enrichments. In most of the experiments, benzene was the only
semicontinuously supplied energy sources in the defined mineral medium (Grbi-Gali and Vogel 1987).
BENZENE
265
After an initial acclimation time of 11 days, at least 50% of the substrate was converted to CO2 and
methane. The intermediates were consistent with benzene degradation via initial oxidation by ring
hydroxylation.
It has been demonstrated that when mixtures of benzene, toluene, xylenes, and ethylbenzene are present
in an anaerobic environment, there is a sequential utilization of the substrate hydrocarbons, with toluene
usually being the first to be degraded, followed by the isomers of xylene in varying order. Benzene and
ethylbenzene tend to be degraded last, if they are degraded at all (Edwards and Grbi-Gali 1992).
Similar studies using only benzene at initial concentrations ranging from 40 to 200 M showed
degradation rates ranging from 0.36 to 3.7 M/day depending upon substrate concentration and the
presence of other carbon sources (Edwards and Grbi-Gali 1992).
6.3.2.4 Other Media
Twenty-day-old spinach leaves placed in a hermetic chamber containing vapors of 14C-labeled benzene
were shown to assimilate benzene, which was subsequently metabolized to various nonvolatile organic
acids (Ugrekhelidze et al. 1997).
6.4
LEVELS MONITORED OR ESTIMATED IN THE ENVIRONMENT
Reliable evaluation of the potential for human exposure to benzene depends in part on the reliability of
supporting analytical data from environmental samples and biological specimens. Concentrations of
benzene in unpolluted atmospheres and in pristine surface waters are often so low as to be near the limits
of current analytical methods. In reviewing data on benzene levels monitored or estimated in the
environment, it should also be noted that the amount of chemical identified analytically is not necessarily
equivalent to the amount that is bioavailable. The analytical methods available for monitoring benzene in
a variety of environmental media are detailed in Chapter 7.
6.4.1
Air
Benzene is ubiquitous in the atmosphere. It has been identified in outdoor air samples of both rural and
urban environments and in indoor air. Table 6-2 lists benzene levels in outdoor air from various cities in
the United States.
BENZENE
266
Table 6-2. Benzene Levels in Air Samples

Location
Concentration (ppb)
Comments
References
Outdoor air
San Francisco, California
0.85.2 (range); 2.61.3a
Wester et al. 1986
Stinson Beach, California
0.380.39a
Houston, Texas
1.218.7 (range); 37.5b
Houston, Texas
0.556.3; 1.73b
St. Louis, Missouri
0.6312.1 (range); 1.85b
Denver, Colorado
3.06.6 (range); 4.1b
Data from six different urban

locations in 1984; n=25
Data from remote coastal
area in 1984; n=21
EPA survey, urban area;
n=14; summer 1986
Semirural area; n=22;
19901991
n=18; summer 1985
n=13; summer 1986
n=14; summer 1985
n=12; summer 1986
n=14; summer 1986
Urban area; n=22; 1990
1991
Urban area; spring 1984
Philadelphia, Pennsylvania 0.323.0 (range); 1b

0.885.3 (range); 1.75b
New York (Manhattan),

New York
Chicago, Illinois
0.635.05 (range); 3.45b
Boston, Massachusetts
0.693.1; 1.06b
New York (Staten Island),

New York
73 km northwest of Denver,
Colorado
Elizabeth and Bayonne,
New Jersey
Personal air
New Jersey
New Jersey
0.134 (range); 4.46.6a
Los Angles, California
0.020.85 (range)
Night 2.7d, 28.5 (max);
day, 3.0d, 13.8 (max)
Night 9.7d, 159.6 (max);
day, 8.5d, 84.5 (max)
Winter 5.2,
summer 2.7,
fall 4.8
Winter 4.5, spring 2.2,
fall 2.8
Wester et al. 1986

EPA 1987ac
Kelly et al. 1993
EPA 1987ac
EPA 1987ac
EPA 1987ac
EPA 1987ac
EPA 1987ac
Kelly et al. 1993
Singh et al. 1985
Rural area; May 1981

Roberts et al. 1985
December 1982
Night, n=8186; day, n=86 Wallace et al. 1985
90; fall 1981
Night, n=346348; day,
n=339341; fall 1981
Wallace et al. 1985

Rappaport and
Kupper 2004
Rappaport and
Kupper 2004
Averagestandard deviation
Median
c
Concentrations reported as ppb carbon; values presented in table represent conversion equivalents as ppb
benzene.
d
Weighted average
b
EPA = Environmental Protection Agency; max = maximum; N = number of samples
BENZENE
267
Volatile organic compounds, including benzene, were measured at 11 monitoring sites in Anchorage,
Alaska, in a year-long study ending in April 1994 (Taylor and Morris 1995). Average annual benzene
concentrations ranged from a minimum of 1.15 parts per billion by volume (ppbv) in a low density
residential area to 5.44 ppbv near a major midtown intersection. In a neighborhood where residents were
complaining of petroleum odors, the highest benzene concentrations (annual average 3.74 ppbv) were
measured on a cliff over the petroleum tank farm, within 50 meters of a petroleum storage tank.
Ambient air samples from 44 sites in 39 U.S. urban areas were collected from 6 to 9 a.m. during June
through September of 1984, 1985, and 1986. Benzene was present in every sample. The median benzene
site concentrations ranged from 0.8 to 6 ppb, with the overall median being 2.1 ppb (estimated detection
limit ~0.04 ppb carbon; conversion equivalent 0.007 ppb benzene). The data indicated that mobile
sources were the major source of benzene in the vast majority of samples (EPA 1987a). Benzene
concentrations were in the range of >1<5 ppbv for all of the 13 sites tested in a 1996 study. The sites
included: Baton Rouge, Louisiana; Brownsville, Texas; Brattleboro, Vermont; Burlington, Vermont;
Camden, New Jersey; El Paso, Texas; Garyville, Louisiana; Galveston, Texas; Hanville, Louisiana; Port
Neches, Texas; Rutland, Vermont; Underhill, Vermont; and Winooski, Vermont (Mohamed et al. 2002).
The following daily median benzene air concentrations were reported in the Volatile Organic Compound
National Ambient Database (19751985): remote (0.16 ppb), rural (0.47 ppb), suburban (1.8 ppb), urban
(1.8 ppb), indoor air (1.8 ppb), and workplace air (2.1 ppb). The outdoor air data represent 300 cities in
42 states while the indoor air data represent 30 cities in 16 states (Shah and Singh 1988). In a NHEXAS
study of six states in the Great Lakes region, benzene was found in 99.7% of the 386 personal air samples
taken with an average concentration of 2.35 ppb. Benzene was also found in 100% of both indoor and
outdoor samples with concentrations averaging 2.25 and 1.13 ppb, respectively (Clayton et al. 1999).
An EPA study of the concentration of various pollutants in 1990 showed an average concentration of
benzene of 0.948 ppb. The source of benzene included manufacturing facilities and manufacturers that
emit benzene to the environment (17%), mobile (60%), and background (23%) sources (Morello-Frosch
et al. 2000).
In California, motor vehicle exhaust accounted for over 70% of the nonsmoking populations exposure to
ambient benzene (Cal EPA 1987). The 1984 population-weighted average benzene concentration in
California was estimated to be 3.3 ppb (Cal EPA 1987). Benzene emissions in a Los Angeles roadway
tunnel were measured at a concentration of 382 mg/L (118,420 ppm) (Fraser et al. 1998).
BENZENE
268
Industrial sites that produce benzene are monitored often. Analysis of ambient air samples collected in
industrial areas showed benzene levels ranging from 0.13 to 5 ppb in Iberville Parish, Louisiana, an area
that included many organic chemical and petroleum producer, user, and storage facilities located along
the Mississippi River (Pellizzari 1982). Indoor and outdoor air measurements were made in August 1987
in the Kanawha Valley region of West Virginia, which is the center of a heavily industrialized area known
for its chemical manufacturing. The mean, maximum, and median indoor concentrations of benzene were
2.1, 14.9, and 0.64 ppb, respectively. The median outdoor ambient air concentration of benzene was
0.78 ppb (Cohen et al. 1989).
An analysis of gas from 20 Class II (municipal) landfills revealed a maximum concentration of 32 ppm
for benzene (Wood and Porter 1987). Benzene was measured in the vicinity of the BKK landfill, a
hazardous waste landfill in California, at a maximum concentration of 1.2 ppb (Bennett 1987). Maximum
estimated levels of benzene in air near uncontrolled (Superfund) hazardous waste sites were 59.5 ppb at
the Kin-Buc Landfill (Edison, New Jersey) and 162.8 ppb in Love Canal basements (Niagara Falls, New
York) (Bennett 1987; Pellizzari 1982).
According to the National Cancer Institute, the average concentration of benzene in 1998 was 0.58 ppb in
metropolitan areas (NCI 2003). A study of New York City air found mean values of 0.80, 1.87, and
1.47 ppb for outdoor (home), indoor (home), and personal air, respectively (Kinney et al. 2002).
According to the New York Department of Health, the air from about 50% of oil fuel heated homes
between the years 1997 and 2003 contained benzene concentrations 0.69 ppb inside the home, and
0.47 ppb in the area outside their homes. New York City was not included in this study (NYSDOH
2005). Concentrations of VOCs were measured in 12 northern California office buildings with three
different types of ventilation. Benzene concentrations ranged from <0.1 to 2.7 ppb, with a geometric
mean of 0.98 ppb (Daisey et al. 1994).
Population-weighted personal exposures to benzene exceeded the outdoor air concentrations in data from
EPA's Total Exposure Assessment Methodology (TEAM) study. The overall mean personal exposure
was about 4.7 ppb, compared to an overall mean outdoor concentration of only 1.9 ppb (Wallace 1989a).
The study also reported that the median level of benzene in 185 homes without smokers was 2.2 ppb, and
the median level of benzene in 343 homes with one or more smokers was 3.3 ppb (Wallace 1989a). This
finding points to the possible significance of passive smoking as a source of benzene exposure. Indoor air
samples taken from a smoke-filled bar contained 8.0811.3 ppb of benzene (Brunnemann et al. 1989). A
BENZENE
269
study conducted by R.J. Reynolds Tobacco Company in smoking and nonsmoking homes revealed that
benzene levels were elevated in smoking homes. In 24 nonsmoking homes, the mean benzene
concentration was 1.21 ppb with a maximum of 5.93 ppb. In 25 smoking homes, the mean benzene
concentration was 1.73 ppb, with a maximum of 8.43 ppb. However, benzene was not significantly
correlated or associated with 3-ethenylpyridine, a proposed vapor phase environmental tobacco smoke
marker (Heavner et al. 1995).
A British study measured benzene concentrations from both rural and urban areas in 1995. The highest
concentrations of benzene were in Southampton urban center, London roadside, and Liverpool, which
contained measured benzene levels of 2.53, 1.69,, and 1.60 ppb, respectively. The lowest measured
benzene concentrations were in rural Hartwell and urban Edinborough, which contained 0.4 and 0.69 ppb
of benzene, respectively (Duarte-Davidson 2001).
6.4.2
Water
Data from EPA's STOrage and RETrieval (STORET) database (20032005) showed that benzene was
positively detected in 38% of the surface water samples collected at 571 observation stations ranging
from concentrations too low to quantify to 100 g/L (100 ppb) in one Utah test site. The sampling sites in
the STORET database include both ambient and pipe sites. Ambient sites include streams, lakes, ponds,
wells, reservoirs, canals, estuaries, and oceans and are intended to be indicative of general U.S. waterway
conditions. Pipe sites refer to municipal or industrial influents or effluents (EPA 2005f).
Benzene was found in 904 of the 8,200 samples tested by the U.S. Geological Survey across the United
States and Canada (USGS 2005). The EPA found benzene in 13,919 and 1,119 groundwater and surface
water samples, respectively (EPA 2001).
Benzene is found in both polluted and unpolluted waters and rainwater. Measured benzene levels in open
ocean from the Gulf of Mexico in 1977 in relatively unpolluted and polluted waters were 515 ng/kg (5
15 parts per trillion [ppt]), and 540 ng/kg (540 ppt), respectively (Sauer 1981). Benzene levels
measured in coastal surface waters of the Gulf of Mexico were 6 ng/kg (6 ppt) in relatively unpolluted
waters and 50175 ng/kg (50175 ppt) in polluted coastal waters (Sauer 1981). Benzene has been
detected in rainwater in the United Kingdom at a concentration of 87.2 ppb (Colenutt and Thorburn
1980).
BENZENE
270
Benzene levels in water in the vicinity of five industrial facilities using or producing benzene ranged from
<1 ppb to a high of 179 ppb found in plant effluent. In general, benzene in plant effluents quickly
dispersed in rivers or streams to levels of 12 ppb or less (EPA 1979). The maximum benzene levels
observed in monitoring wells in plumes from fuel spills at gasoline service stations ranged from 1,200 to
19,000 ppb (Salanitro 1993). A monitoring well in the vicinity of a bulk storage facility had a maximum
benzene level of 45,000 ppb (Salanitro 1993). In northern Virginia, approximately 200,000 gallons of
liquid hydrocarbons were released from a fuel-storage terminal into the underlying soil. A dichloromethane extract of groundwater from a monitoring well in the same area gave a benzene concentration of
52.1 ppm (Mushrush et al. 1994). Benzene has been detected at concentrations ranging from 16 to
110 ppb in landfill leachate from a landfill that accepted both municipal and industrial wastes (Cline and
Viste 1985).
Composite data from the Comprehensive Emergency Response, Compensation, and Liability Act
(CERCLA) monitoring program indicate that benzene was detected at a frequency of 11.2% in
groundwater in the vicinity of 178 inactive hazardous waste disposal sites (Plumb 1987). Data from a
1980 national survey by the Council on Environmental Quality on groundwater and surface water
contamination showed benzene concentrations in contaminated drinking water wells in New York, New
Jersey, and Connecticut ranged from 30 to 330 ppb, with the highest concentration in drinking water from
surface water sources reported to be 4.4 ppb (Burmaster 1982). Benzene has also been identified but not
quantified as one of the major organic constituents in commercially bottled artisan water in the United
States (Dowty et al. 1975). At detection limits ranging from 0.2 to 1.0 ppb, benzene was detected at a
concentration of 2 ppb in only one sample of 182 samples of bottled drinking water (Page et al. 1993). In
six states of the Great Lakes region, unspecified amounts of benzene were found in 5.7% of the
247 drinking water sites tested (Clayton et al. 1999).
Because the New Jersey Department of Environmental Protection identified serious water issues in a
930 km2 area within metropolitan Philadelphia, tests have been done to find other water sources such as
the Kirkwood-Cohansey aquifer system. A study of 57 water samples taken from the shallow ground
water supply at Kirkwood Cohansey aquifer in New Jersey showed that the majority of samples (50)
contained between 0.2 and 1 ppbv of benzene, three samples contained <0.1 ppbv, and four samples
contained >1 ppbv (Baehr et al. 1999).
BENZENE
271
6.4.3
Sediment and Soil
A Canadian study estimated that benzene is absorbed by the soil at a rate of 100 tons/year (MacLeod and
MacKay 1999). Data from EPA's STORET database (20032005) showed that benzene had been
positively detected in sediment samples taken at 9% of 355 observation stations with a median level of
<5 ppb (EPA 2005f). The concentration of benzene in soil near factories where benzene was produced or
used ranged from 2 to 191 ppb (IARC 1982).
Benzene levels ranging from <2 to 191 ppb were recorded in the vicinity of five industrial facilities using
or producing benzene (EPA 1979). Sediment levels ranging from 8 to 21 ppb were detected in Lake
Pontchartrain in Louisiana (Ferrario et al. 1985). In northern Virginia, approximately 200,000 gallons of
liquid hydrocarbons were released from a fuel-storage terminal into the underlying soil. Soil about
1,000 feet from that storage terminal contained benzene gas at a concentration of 1,500 ppm at a depth of
10 feet (Mushrush et al. 1994).
6.4.4
Other Environmental Media
Benzene has been detected in a variety of food. Benzene has been reported to occur in fruits, fish,
vegetables, nuts, dairy products, beverages, and eggs (EPA 1982a). Although benzene has been detected
in dairy products, there is no evidence of the presence of benzene in either cow's milk or human breast
milk (Hattemer-Frey et al. 1990). Eggs had the highest concentrations (2,100 ppb [uncooked] and 500
1,900 ppb [hard-boiled]), followed by haddock (100200 ppb), Jamaican rum (120 ppb), irradiated beef
(19 ppb), heat-treated canned beef (2 ppb), and butter (0.5 ppb). Lamb, mutton, veal, and chicken all had
<10 ppb benzene (when the meats were cooked) (EPA 1980b, 1982a). A survey of more than 50 foods
collected and analyzed from 1991 to early 1992 (McNeal et al. 1993) revealed that foods (including eggs)
without added benzoates contained benzene at levels 2 ng/g. The level of benzene in foods containing
added benzoates in addition to ascorbates (e.g., imitation strawberry preserves, taco sauce, and duck
sauce) ranged from <1 to 38 ng/g. In many foods, the presence of benzene is likely to be due to uptake
from the air (Grob et al. 1990). This conclusion was supported by the fact that the uptake decreased with
a decrease in exposed surface area of foods and contact time with air (Grob et al. 1990).
The U.S. Food and Drug Administration sponsored a 5-year study to determine the amount of volatile
organics in food from 1996 to 2000. The results are shown in Table 6-3 (Fleming-Jones and Smith 2003).
Benzene was found in cheddar cheese, cream cheese, margarine, butter, sour cream, ground beef,
bologna, hamburger, cheeseburger, pork, beef frankfurters, tuna canned in oil, chicken nuggets, chocolate
BENZENE
272
Table 6-3. Benzene in Fooda
Food
Cheddar cheese
Mixed nuts
Ground beef
Pork bacon
Banana, raw
Frankfurters, beef
Cream cheese
Chocolate cake icing
Tuna canned in oil
Fruit flavored cereal
Eggs scrambled
Peanut butter
Avocado, raw
Popcorn, popped in oil
Blueberry muffin
Strawberries, raw
Cola carbonated
Orange, raw
Coleslaw with dressing
Sweet roll danish
Potato chips
Fruit flavored sherbet
Quarter pound hamburger cooked
Margarine
Sandwich cookies
Butter
Chocolate chip cookies
Sour cream
Apple pie fresh/frozen
Chicken nuggets fast food
Graham crackers
French fries fast food
Cheeseburger quarter pound
Cheese pizza
Bologna
Cheese and pepperoni pizza
Olive/safflower oil
Sugar cookies
Cake doughnuts with icing
Popscicle
a
Number of cases found

(n=14)
2
3
12
6
13
4
3
2
7
5
4
5
10
3
3
1
3
2
14
1
2
3
11
1
3
6
2
2
4
4
2
3
8
2
4
4
6
3
2
4
Derived from Fleming-Jones and Smith 2003
Concentration minimummaximum in ppb

2047
16
9190
217
11132
211
117
223
413
221
240
225
330
422
38
1
1138
1115
11102
33
27
361
447
7
139
4-22
18
315
211
25
19
256
544
12
244
830
146
830
3
110
BENZENE
273
cake icing, sandwich cookie, chocolate chip cookies, graham crackers, sugar cookies, cake doughnuts
with icing, french fries, apple pie, cola carbonated beverages, sweet roll Danish, potato chips, cheese
pizza, cheese and pepperoni pizza, mixed nuts, fruit-flavored cereal, fruit flavored sorbet, popsicles,
olive/safflower oil, scrambled eggs, peanut butter, popcorn popped in oil, blueberry muffins, coleslaw
with dressing, raw banana, avocado, oranges, and strawberries. American cheese was the only food tested
that did not contain benzene. Foods with the greatest maximum concentration of benzene included
ground beef (maximum 190 ppb), raw bananas (maximum 132 ppb), carbonated cola (maximum
138 ppb), and coleslaw with dressing (maximum 102 ppb).
As part of a program to identify possible exposures that may be important in the high incidence of lung
cancer among women in Shanghai, China, Pellizzari et al. (1995) qualitatively identified the volatile
components emitted during heating of cooking oils to 265 C. This study found that, on a relative basis,
the intensity of the benzene peak in the total ion current chromatogram of vapors from Chinese rapeseed
oil (commonly used in wok cooking) was 14-, 6.6-, and 1.7-fold greater than in vapors from peanut,
soybean, and other canola (rapeseed) oils, respectively.
Cigarette smoke remains an important source of human exposure to benzene. The amount of benzene
measured in mainstream smoke ranged from 5.9 to 73 g/cigarette (Brunnemann et al. 1990). Larger
amounts of benzene were found in sidestream smoke, ranging from 345 to 653 g/cigarette (Brunnemann
et al. 1990). Benzene has been found in vapor from cigarette smoke in concentrations ranging from 3.2 to
61.2 g/cigarette depending on the brand of cigarette. The amount of tar in the cigarette was not directly
related to the benzene concentration (Darrall et al. 1998).
6.5
GENERAL POPULATION AND OCCUPATIONAL EXPOSURE
Exposure to low levels of environmental benzene is unavoidable due to the ubiquitous presence of
benzene in the environment from a variety of anthropogenic sources. Benzene can be detected in blood
samples. In a study designed to establish reference blood concentrations of benzene and other selected
volatile organic compounds in the general population of the United States, Ashley et al. (1994) measured
blood benzene levels in 883 people (not occupationally exposed) in the United States who had
participated in the Third National Health and Nutrition Examination Survey (NHANES III). Within this
group of nonoccupationally exposed subjects, the mean and median blood benzene levels were 0.13 and
0.061 ppb, respectively. Seven nonsmoking subjects from this study group were assessed for blood
benzene levels just prior to entering examination vans with limited potential for benzene exposure and at
BENZENE
274
the end of a 3-hour period in this environment. Mean blood benzene levels prior to van entry and after
3 hours in the van measured 0.046 ppb (range from not detected to 0.061 ppb) and 0.033 ppb (range from
not detected to 0.055 ppb), respectively. Lemire et al. (2004) reported a median blood benzene level of
0.047 ppb in a subgroup of 546 nonsmoking and nonoccupationally exposed subjects from the NHANES
III study group. Elevated blood benzene levels may be expected among subjects with potential for
elevated exposure to benzene, such as smokers, commuters, gas station attendants, and people who use
products that emit benzene.
The Total Exposure Assessment Monitoring (TEAM) studies, carried out by the EPA between 1980 and
1990, suggested that for many chemicals, including benzene, the most important sources of pollution are
small and close to the person, and that exposures are not clearly correlated with emissions. For example,
the TEAM study findings indicated that nearly 85% of atmospheric benzene in outdoor air is produced by
cars burning petroleum products and the remaining 15% is produced by industry. Despite the fact that
petroleum products contribute to the majority of benzene in the atmosphere, half of the total national
personal exposure to benzene comes from cigarette smoke (Wallace 1995). In fact, breath measurements
of benzene provided by the TEAM study between 1979 and 1988 identified smoking as the single most
important source of benzene exposure for about 40 million U.S. smokers (Wallace 1989b). Even passive
exposure to cigarette smoke is responsible for more benzene exposure (about 5% of the total) than the
emissions from the entire industrial capacity of the United States (about 3% of the total) (Wallace 1995).
A breakdown of the emissions and exposure sources for benzene that was derived from the Los Angeles
TEAM study data (Wallace et al. 1991) is given in Figure 6-3. The reason that a relatively small source
of emissions can have such a large effect on exposure is the efficiency of delivery. Wallace (1995)
reports that one cigarette delivers an average of 55 g of benzene with nearly 100% efficiency to the
smoker. Benzene from industrial sources is dissipated into the atmosphere.
Smokers (n=200) in the TEAM study had a mean breath concentration of 15 g/m3 (4.7 ppb), almost
10 times the level of 1.52 g/m3 (0.470.63 ppb) observed in more than 300 nonsmokers (Wallace
1989b). Smokers also had about 610 times as much benzene in their blood as non-smokers (Wallace
1995). In another study, benzene concentrations were compared in the breath of smokers and nonsmokers
and in ambient air in both an urban area of San Francisco and in a more remote area of Stinson Beach,
California (Wester et al. 1986). In the urban area, benzene in smokers' breath (6.83.0 ppb) was greater
than in nonsmokers' breath (2.50.8 ppb) and smokers' ambient air (3.30.8 ppb). In the remote area, the
same pattern was observed. This suggests that smoking represented an additional source of benzene
above that of outdoor ambient air (Wester et al. 1986). In 10 of 11 homes inhabited by tobacco smokers,
BENZENE
275
Figure 6-3. Benzene Emissions and Exposures
Benzene Emissions
Industry, 14%
Personal and
Hom e, 3%
Cigarettes,
0.1%
Cars, 82%
Benzene Exposures
Cigare tte s ,
40%
Indus try 3%,

ETS, 5%
Car Exhaus t,
18%
Pe rs onal
Activitie s ,
18%
Hom e
Source s , 16%
Note: A comparison of benzene emission sources (home sources include paints and petroleum products; personal
activities include driving and use of consumer products that contain benzene). Data taken from Wallace et al. (1991).
BENZENE
276
mean indoor and personal benzene concentrations were 25 times higher than outdoor levels (Thomas et
al. 1993).
Assuming that the average sales-weighted tar and nicotine cigarette yields 57 g benzene in mainstream
smoke, Wallace (1989a) estimated that the average smoker (32 cigarettes per day) takes in about 1.8 mg
benzene per day from smoking. This is nearly 10 times the average daily intake of nonsmokers (Wallace
1989a). On the assumption that intake of benzene from each cigarette is 30 g, Fishbein (1992) has
calculated that a smoker who consumes two packs of cigarettes per day will have an additional daily
intake of 1,200 g.
A British study of rural and urban environments suggests that benzene exposure is greatly affected by
proximity to smokers (Duarte-Davidson et al. 2001). Air concentrations of benzene at an urban center in
South Hampton averaged about 2.5 ppb, while in a rural location in Hartwell, the average amount of
benzene in the air was 0.41 ppb. Air at a smoky pub was found to contain 22 ppb of benzene. Comparing
the daily doses of rural nonsmokers, urban nonsmokers, urban passive smokers, and urban smokers, very
little difference between the rural nonsmokers 24 ppb daily dose and the urban nonsmokers 30 ppb daily
dose was found. Passive urban smokers, on average, have a daily benzene exposure dose of 38 ppb of
benzene while smokers have a daily exposure dose of benzene of 163 ppb. On average, nonsmokers in
urban and rural environments have estimated benzene intakes of 1.15 and 1.5 g/kg body weight/day.
Women tend to intake more of benzene per kg body weight than men. Passive smokers estimated daily
intake averages are 2.10 and 1.74 g/kg body weight/day for women and men, respectively. Urban
women and men smokers estimated intakes are estimated at 9.00 and 7.46 g/kg body weight/day,
respectively; this is equivalent to an atmospheric concentration of 8.2 ppb (Duarte-Davidson et al. 2001).
In 1990, a study in Germany analyzed factors that predicted peoples exposures to VOCs and found that
while smoking was the most significant determinant of benzene exposure, automobile-related activities,
such as refueling and driving, were also significant (Hoffmann et al. 2000). Virtually all (99.9%) of the
benzene released into the environment finally distributes itself into the air. The general population may
be exposed to benzene through inhalation of contaminated air, particularly in areas of heavy motor
vehicle traffic and around gas stations. Compared to inhalation, dermal exposure probably constitutes a
minor portion of benzene exposure for the general population. Personal sources account for 18% of the
total exposure of the general population to benzene. The main personal sources (other than smoking
BENZENE
277
cigarettes) are driving or riding in automobiles and using products that emit benzene (paints, adhesives,
marking pens, rubber products, and tapes) (Wallace 1989a).
Benzene constitutes 12% of most blends of gasoline and is released as a vapor from vehicular emissions.
Since benzene is a constituent of auto exhaust and fuel evaporation, people who spend more time in cars
or in areas of heavy traffic have increased personal exposure to benzene. Assuming an average benzene
concentration of 40 g/m3 (12.5 ppb) for a moving automobile and an exposure duration of 1 hour/day,
the calculated intake for driving or riding in an automobile is 40 g/day (Wallace 1989a). In an
investigation of exposure to methyl tertiary-butyl ether (MTBE) in oxygenated gasoline in Stamford,
Connecticut, venous blood samples were collected from 14 commuters and from 30 other persons who
worked in the vicinity of traffic or automobiles. In addition to MTBE, the samples were analyzed for five
chemicals, including benzene. Levels of benzene in the blood of 11 nonsmoking men and women
commuters ranged from 0.10 to 0.20 g/L (median=0.12 g/L). Blood benzene levels of 0.29, 0.14, and
0.58 g/L were measured in one female and two male smoking commuters, respectively. In
12 nonsmoking male car repair workers, blood benzene levels ranged from 0.11 to 0.98 g/L
(median=0.19 g/L); in 8 smoking male car repair workers, levels ranged from 0.17 to 0.67 g/L
(median=0.42 g/L). Three nonsmoking male gasoline attendants had blood benzene levels ranging from
0.32 to 0.47 g/L (median=0.36 g/L) (White et al. 1993).
Pumping gasoline can also be a significant source of exposure to benzene. A study conducted between
July 1998 and March 1999 that comprised of 39 customers of gasoline self service stations from North
Carolina, measured the benzene level in the air around the station as well as the levels of benzene in
customers breath prior to and immediately after fueling (Egeghy et al. 2000). Benzene levels in the air
around the station ranged from <0.02 to 11.16 ppm, with a mean (1 standard deviation) of
0.91 (1.8) ppm. The range of benzene levels in the breath of customers prior to fueling was <0.001
0.022 ppm with a mean (1 standard deviation) of 0.0027 (0.0034) ppm while the range of benzene
levels in the breath of customers after re-fueling was <0.0010.434 ppm with a mean (1 standard
deviation) of 0.05 (0.081) ppm (Egeghy et al. 2000). Another study reported a benzene concentration of
1 ppm at the breathing-level of a person pumping gas (EPA 1986). Using this concentration and an
estimated 70 minutes/year of time spent pumping gasoline, a benzene intake of 10 g/day has been
calculated (Wallace 1989a). In a group of 26 subjects who were not occupationally exposed to benzene,
but were exposed to benzene during refueling in Fairbanks, Alaska, median blood benzene levels prior to
and immediately following refueling were 0.19 ppb (range 0.080.65 ppb) and 0.54 ppb (range 0.13
1.70 ppb), respectively (Backer et al. 1997).
BENZENE
278
Gasoline vapors vented into the home from attached garages can also increase indoor air exposure to
benzene (Wallace 1989a, 1989b). Depending on airflow from garage to living areas, mean indoor
benzene concentrations in houses with a garage were 25 times higher than outdoor levels in most homes
(Thomas et al. 1993). Benzene levels in four garages during different times in a day ranged from 0.94 to
61.3 ppb. The higher concentrations of benzene in these garages were not only from vehicular activity,
but also in varying proportions from stored gasoline, paints, and benzene-containing consumer products
(Thomas et al. 1993). Inhalation exposure to off-gassing from benzene-containing products and to
evaporative emissions from automobiles in attached garages has been estimated to be 150 g/day
(Wallace 1989a).
A study of human exposure to benzene in the California South Coast Air Basin showed that benzene
exposure in that area has decreased greatly since 1989. Adult smokers saw a 28% decrease in benzene
exposure from 1989 to 1997, while adult nonsmokers and adolescents saw a decrease in benzene exposure
of 67 and 55%, respectively. Where people were being exposed to benzene also changed during that
time. Ambient air was the source of 49% of the benzene that nonsmokers were exposed to in 1997, which
is less than the 59% from 1987. For smokers in the region, 85% of their benzene exposure was from
smoking and 4% was from the ambient air in 1997. This shows an increase in benzene exposure from
smoking since 1987 (78%) and a decrease in exposure from the ambient air (12%) (Fruin et al. 2001).
The main sources credited with this decrease in exposure are reformulated gasoline and stricter air
emission laws as well as smoking control measures.
Another source of exposure to benzene for the general population is the use of domestic wood stoves. It
has been estimated that approximately 10% of the space heating in urban areas of the northern United
States is from wood burning, with up to 50% in smaller, rural towns (Larson and Koenig 1994). Benzene
has been found to be a major component of the emissions from wood burning, especially from efficient
flame combustion, and constituted roughly 1020% by weight of total non-methane hydrocarbons
(Barrefors and Petersson 1995). It should be noted, however, that chimney emissions result in much
lower human exposure than equally large emissions at the ground level.
Other sources of inhalation exposure to benzene include air around hazardous waste sites, industrial
facilities, off-gassing from particle board, and off-gassing from contaminated water during showering and
cooking. Based on the TEAM study findings, it appears that the following are not important sources of
BENZENE
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exposure to benzene on a nationwide basis: chemical manufacturing facilities, petroleum refining

operations, oil storage tanks, drinking water, food, and beverages (Wallace 1989a).
Average water intake of benzene (assuming a typical drinking water concentration of 0.1 ppb and a
consumption of 2 L/day) is 0.2 g/day (HSDB 2007). According to another estimate, the daily intakes of
benzene for a nonsmoking individual (not exposed to secondary smoke) are 1550 g (Fishbein 1992).
While most human exposure to benzene is believed to be through inhalation, studies show that benzene
can permeate skin with a permeability factor of about 0.140.18 cm/hour at 25 C. The permeability
factor was not affected by moisturizer, baby oil, or insect repellent; however, it was affected by
temperature (50 C) and sunscreen with the permeability factors increasing to 0.26 and 0.24 cm/hour,
respectively (Nakai et al. 1997).
Individuals employed in industries that use or make benzene or products containing benzene may be
exposed to the highest concentrations of benzene. The National Occupational Exposure Survey (NOES),
conducted by NIOSH from 1981 to 1983, estimated that approximately 272,300 workers employed in
various professions were potentially exposed to benzene in the United States. Approximately half of
these workers were employed in general medical and surgical hospitals, and their occupations included
nurses and aides, physicians, technicians, technologists, therapists, dieticians, pharmacists, and janitors
(NIOSH 1990). The NOES database does not contain information on the frequency, concentration, or
duration of exposure; the survey provides only estimates of workers potentially exposed to chemicals in
the workplace. The current OSHA permissible limit for an 8-hour TWA exposure to benzene is 1 ppm
and a short-term exposure limit in any 15-minute period is 5 ppm (OSHA 2003). The NIOSH
recommended exposure limit is 0.1 ppm for an 8-hour TWA and 1 ppm for short-term exposure (NIOSH
1992b). In 1987, OSHA estimated that approximately 238,000 workers were exposed to benzene in seven
major industry sectors, including petrochemical plants, petroleum refineries, coke and coal chemicals, tire
manufacturers, bulk terminals, bulk plants, and transportation via tank trucks (Table 6-4) (OSHA 1987).
Approximately 10,000 workers were estimated to be exposed to TWA concentrations in excess of the
1 ppm standard. This estimate did not include firms covered by the exclusions, firms under jurisdiction of
other agencies, or firms involved in the use of products containing small quantities of benzene. The
uptake of benzene by workers in a municipal waste incinerator in Germany was assessed by measuring
benzene levels in blood (Angerer et al. 1991). No significant difference (p<0.05) in blood benzene levels
between workers and controls were detected (mean 0.22 g/L for nonsmoking workers versus 0.25 g/L
BENZENE
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Table 6-4. Percentage of Employees Exposed to Benzene by Exposure Level and
Industry Divisiona
Industry sector
Petrochemical plantsb
Petroleum refineriesc,d
Coke and coal chemicalse
Tire manufacturersc
Bulk terminalsc
Bulk plantsc
Transportation via tank truckc
Total
a
Percentage of observations in each exposure category

according to range of 8-hour TWA benzene
Total
concentrations (ppm)
number of
employees
0.00.1 0.110.5 0.511.0 1.15.0 5.110 10+
64.6
0.0
53.4
57.8
57.8
68.4
74.6
26.1
39.3
37.5
32.8
32.8
23.1
4.6
27.6
6.3
5.3
5.3
5.3
23.0
3.8
27.5
2.8
3.7
3.7
2.9
2.4
0.5
4.4
0.0
0.3
0.3
0.1
Derived from OSHA 1987
Percentages represent the portion of workers whose average exposures are in each category.
c
Percentages represent the portion of sampling results in each category.
d
Data do not reflect respirator use and sampling biases.
e
Percentages represent the portion of workers whose average exposures are in each category.
f
Excludes workers employed at the coke ovens.
TWA = time-weighted average
0.0
0.4
1.3
0.0
0.1
0.1
0.2
4,300
47,547
947f
65,000
27,095
45,323
47,600
237,812
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for nonsmoking controls). OSHA requires the use of engineering controls and/or respiratory protection in
situations where compliance with the TWA is not feasible (OSHA 1987).
Certain jobs, such as gasoline station workers, firefighters, and dry cleaners, are believed to put people at
a higher risk of benzene exposure. In an analysis of literature, it was estimated that workers in the area of
crude petroleum and natural gas are exposed to 0.04 ppm benzene, while workers in petroleum refining,
gas stations, and crude petroleum pipelines are exposed to 0.22, 0.12 and 0.25 ppm benzene, respectively.
This study also showed that fire fighters are exposed to an average of 0.38 ppm benzene (van
Wijngaarden and Stewart 2003). Workers from four different dry cleaning facilities in Korea had mean
benzene air concentrations ranging from 2.7 to 3.2 ppb. Their exposure to benzene was dependent upon
the type of solvent used for cleaning (Jo and Kim 2001). Benzene concentrations of 25.46 and
1,331.29 ppb were found near the kiln and at the rotary line, respectively, inside a hazardous waste
incinerator in Turkey (Bakoglu et al. 2004).
A study comparing workers who were exposed to benzene regularly at work to people who were not
exposed to benzene at work showed that while the general population in Italy had average blood benzene
concentration of 165 ng/L, the people who were exposed to high benzene levels at work had an average
benzene blood concentration of 186 ng/L. Immediately following their shift, the average benzene blood
level samples from of benzene-exposed workers was 420 ng/L. The average blood benzene levels for
smoking and nonsmoking occupationally exposed workers were 264 and 123 ng/L respectively
(Brugnone et al. 1998).
6.6
EXPOSURES OF CHILDREN
This section focuses on exposures from conception to maturity at 18 years in humans. Differences from
adults in susceptibility to hazardous substances are discussed in Section 3.7, Childrens Susceptibility.
Children are not small adults. A childs exposure may differ from an adults exposure in many ways.
Children drink more fluids, eat more food, breathe more air per kilogram of body weight, and have a
larger skin surface in proportion to their body volume. A childs diet often differs from that of adults.
The developing humans source of nutrition changes with age: from placental nourishment to breast milk
or formula to the diet of older children who eat more of certain types of foods than adults. A childs
behavior and lifestyle also influence exposure. Children crawl on the floor, put things in their mouths,
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sometimes eat inappropriate things (such as dirt or paint chips), and spend more time outdoors. Children
also are closer to the ground, and they do not use the judgment of adults to avoid hazards (NRC 1993).
Children can be subject to increased benzene exposure by inhalation of second-hand smoke. In a study of
nonsmoking rural families, urban families, and urban smoking families, infant exposure to benzene was
estimated at doses of 15.3, 19.7, and 25.9 g/day, respectively, with daily intakes of 1.68, 2.16, and
2.55 g/kg body weight/day, respectively. For children of the same classification, benzene exposure was
measured at doses of 29.3, 37.6, and 49.3 g/day, respectively, with daily intakes of 0.71, 0.91, and
1.20 g/kg bodyweight/day, respectively. For all infants and children, benzene exposure predominantly
comes from the indoors (Duarte-Davidson et al. 2001).
Depending on a childs living environment, they may have higher exposure to benzene than adults. In a
study of two lower-income areas of Minneapolis, children were found to have average personal benzene
exposures of 0.66 and 0.53 ppb in the winter and spring, respectively. The highest concentration of
benzene in their environment came from the home with winter and spring concentrations of 0.69 and
0.66 ppb respectively, while the outdoor and school benzene concentrations were 0.41 and 0.19 ppb,
respectively (Adgate et al. 2004). In Italy, concentrations of the benzene metabolite, trans,trans-muconic
acid (MA), was measured in the urine of children from both urban areas in Naples and rural areas in
Pollica. The mean urinary concentrations of MA detected for rural and urban children were 48.4 and
98.7 g/L (Amodio-Cocchieri et al. 2001). These studies also found no strong link between passive
smoking and MA levels. The only factor that affected levels of MA in urine samples was how close the
family lived to the road. A study in Rouen, France, compared benzene exposure and concentrations in
nonsmoking parents and their children. Despite the fact that the children were exposed to slightly less
benzene 3.47 ppb (11.1 g/m3) than their parents 4.51 ppb (14.4 g/m3), there was no significant
correlation between exposure means and urinary metabolite levels (Kouniali et al. 2003).
Benzene concentrations in women and their tissue as well as breast milk have been a major concern. In a
study of South Korean housewives living near service stations, indoor, outdoor and breath benzene
concentrations averaged 5.73, 3.63, and 3.29 ppm, respectively (Jo and Moon 1999). Benzene has been
found in mothers milk (Pellizzari 1982). Benzene was found at mean concentrations of 0.06 g/kg in
23 samples of breast milk taken from a childrens hospital in Rome (Fabietti et al. 2004). While this may
provide a mechanism by which infants are exposed to benzene, these concentrations are lower than in
other foods.
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6.7
POPULATIONS WITH POTENTIALLY HIGH EXPOSURES
Individuals who live near hazardous waste sites or near leaking underground fuel storage tanks might be
exposed to potentially high concentrations of benzene in their drinking water if they obtain tap water from
wells located near these sources. In a series of experiments conducted in a single-family residence from
June 11 to 13, 1991, exposure to benzene through contaminated residential water was monitored
(Lindstrom et al. 1994). The residential water was contaminated with benzene and other hydrocarbons in
1986. Periodic testing conducted from 1986 to 1991 showed benzene concentrations ranging from 33 to
673 g/L (ppb). The experiment involved an individual taking a 20-minute shower with the bathroom
door closed, followed by 5 minutes for drying and dressing; then the bathroom door was opened and this
individual was allowed to leave the house. Integrated 60- and 240-minute whole-air samples were
collected from the bathroom, an adjacent bedroom, living room, and in ambient air. Glass, gas-tight
syringe grab samples were simultaneously collected from the shower, bathroom, bedroom, and living
room at 0, 10, 18, 20, 25, 25.5, and 30 minutes. Two members of the monitoring team were measured for
6 hours using personal Tenax gas GC monitors. For the first 30 minutes of each experiment, one member
was based in the bathroom and the other in the living room. Benzene concentrations in the shower head
ranged from 185 to 367 g/L (ppb), while drain level samples ranged from below the detectable limit
(0.6 g/L or ppb) to 198 g/L (ppb). Analysis of the syringe samples suggested a pulse of benzene
moving from the shower stall to the rest of the house over approximately 60 minutes. Peak benzene
levels were measured in the shower stall at 1820 minutes (7581,670 g/m3), in the bathroom at 10
25 minutes (366498 g/m3), in the bedroom at 25.530 minutes (81146 g/m3), and in the living room
at 3670 minutes (4062 g/m3). The total benzene dose resulting from the shower was estimated to be
approximately 281 g, with 40% via inhalation and 60% via the dermal pathway (Lindstrom et al. 1994).
The major source of exposure to benzene is cigarette smoke. A smoker of 32 cigarettes per day (the U.S.
average per smoker) would have a benzene intake of approximately 1.8 mg/day (at least 10 times the
average nonsmoker's intake) (Wallace 1989a). Median benzene concentrations in 343 homes with
smokers averaged 3.3 ppb compared to 2.2 ppb in 185 homes without smokers. This represents a 50%
increase in benzene exposure for nonsmokers exposed to passive smoke compared to nonsmokers not
exposed to passive smoke (Wallace 1989a). In a study in Germany, the mean benzene concentrations for
frequent smokers and nonsmokers were 6.1 and 2.4 ppb, respectively (Hoffmann et al. 2000).
BENZENE
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6.8
ADEQUACY OF THE DATABASE
Section 104(i)(5) of CERCLA, as amended, directs the Administrator of ATSDR (in consultation with the
available, ATSDR, in conjunction with NTP, is required to assure the initiation of a program of research
designed to determine the health effects (and techniques for developing methods to determine such health
effects) of benzene.
6.8.1
Identification of Data Needs
Physical and Chemical Properties.
The physical and chemical properties of benzene are well
characterized and allow prediction of the transport and transformation of the compound in the
environment.
Production, Import/Export, Use, Release, and Disposal.
According to the Emergency
Planning and Community Right-to-Know Act of 1986, 42 U.S.C. Section 11023, industries are required
to submit substance release and off-site transfer information to the EPA. The TRI, which contains this
information for 2004, became available in May of 2006. This database is updated yearly and should
provide a list of industrial production facilities and emissions.
Benzene is one of the top 20 highest volume chemicals produced in the United States. In 1994, the U.S.
production volume of benzene was 14.7 billion pounds (C&EN 1995). The production volume during the
19841994 period has increased by 4% annually (C&EN 1995). The United States currently has a
benzene production capacity of 11.8 billion liters (SRI 2004). Imports of benzene into the United States
have generally ranged from 10,176 to 11,672 million pounds during 20022004 (USITC 2005). Exports
increased from 21 million pounds in 2002 to 290 million pounds in 2003, but decreased to 145 million
pounds in 2004 (USITC 2005). The major use of benzene is in the production of other chemicals
(primarily ethylbenzene, cumene, and cyclohexane), accounting for approximately 91% of benzene
BENZENE
285
production volume. Benzene is also used in chemical laboratories as a solvent and a reactant (OSHA
1977, 1987), and as an anti-knock agent in unleaded gasoline (Brief et al. 1980; EPA 1985a). The
widespread use of benzene as a solvent has decreased in recent years. Many products that used benzene
as a solvent in the past have replaced it with other organic solvents; however, benzene may still occur as a
trace impurity in these products. Less than 2% of the amount of benzene produced is used as a solvent in
such products as trade and industrial paints, rubber cements, adhesives, paint removers, artificial leather,
and rubber goods. Benzene has also been used in the shoe manufacturing and rotogravure printing
industries (EPA 1978; OSHA 1977). In the past, certain consumer products (such as some paint strippers,
carburetor cleaners, denatured alcohol, and rubber cement used in tire patch kits and arts and crafts
supplies) contained small amounts of benzene (Young et al. 1978). Other consumer products that
contained benzene were certain types of carpet glue, textured carpet liquid detergent, and furniture wax
(Wallace et al. 1987). The use of benzene in certain pesticides has been canceled. Benzene-containing
wastes, such as commercial chemical products, manufacturing chemical intermediates, and spent solvents,
are subject to federal and/or state hazardous waste regulations (HSDB 2007). Currently, the
recommended method of disposal is to incinerate solvent mixtures and sludges at a temperature that
ensures complete combustion. No additional information on the production, import/export, use, release,
or disposal of benzene is needed at this time.
Environmental Fate. Benzene released to the environment partitions mainly to the atmosphere
(Mackay and Leinonen 1975). However, the compound can also be found in surface water and
groundwater. Benzene is mobile in soil (Karickhoff 1981; Kenaga 1980); however, there is a need for
more information on the leachability potential of benzene to groundwater in different soil types. Benzene
is transformed in the atmosphere by photooxidation. Biodegradation, principally aerobic, is the most
important fate process of benzene in water (Delfino and Miles 1985; McAllister and Chiang 1994;
Salanitro 1993) and soil (Gibson 1980; Hopper 1978; Salanitro 1993). Benzene can persist in
groundwater. No additional information on the environmental fate of benzene is needed at this time.
Bioavailability from Environmental Media.
Benzene can be absorbed following oral exposure
(Thienes and Haley 1972), dermal exposure (Blank and McAuliffe 1985; Franz 1984; Laitinen et al.
1994; Lindstrom et al. 1994; Lodn 1986; Susten et al. 1985), and inhalation exposure (Ashley et al.
1994; Avis and Hutton 1993; Boogaard and van Sittert 1995; Brunnemann et al. 1989; Byrd et al. 1990;
Etzel and Ashley 1994; Fustinoni et al. 1995; Ghittori et al. 1995; Gordian and Guay 1995; Hajimiragha
et al. 1989; Hanzlick 1995; HazDat 2006; Karacic et al. 1995; Kok and Ong 1994; Lagorio et al. 1994a;
Laitinen et al. 1994; Lauwerys et al. 1994; Lindstrom et al. 1994; Mannino et al. 1995; Nomiyama and
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286
Nomiyama 1974a; Ong and Lee 1994; Ong et al. 1995; Pekari et al. 1992; Popp et al. 1994; Rauscher et
al. 1994; Rothman et al. 1995; Ruppert et al. 1995; Scherer et al. 1995; Shamy et al. 1994; Srbova et al.
1950). These routes of exposure may be of concern to humans because of the potential for benzene to
contaminate the air (Bennett 1987; Black et al. 1980; Brief et al. 1980; Cal EPA 1987; Edgerton and Shah
1992; EPA 1989, 1994d; Glass et al. 1986; Graedel 1978; Mayer et al. 1994; TRI02 2005; Wallace 1989a,
1989b; Wallace and Pellizzari 1986; Wester et al. 1986; Wood and Porter 1987), drinking water (CDC
1994; EPA 1979), and soil (HazDat 2006; Mushrush et al. 1994; TRI02 2005). Information on inhalation
exposure and on the absorption of benzene following ingestion of plants grown in contaminated
environments near hazardous waste sites would be helpful in determining bioavailability of the compound
in these media.
Food Chain Bioaccumulation.
Benzene has an estimated low-to-moderate bioconcentration
potential in aquatic organisms (Miller et al. 1985; Ogata et al. 1984) and some plants (Geyer et al. 1984).
Most of the benzene accumulation on vegetation results from air-to-leaf transfer. Root uptake is not
believed to be important (Hattemer-Frey et al. 1990). Biomagnification in aquatic food chains does not
appear to be important (Ogata et al. 1984). No further information appears to be needed.
Exposure Levels in Environmental Media.
Reliable monitoring data for the levels of benzene in
contaminated media at hazardous waste sites are needed so that the information obtained on levels of
benzene in the environment can be used in combination with the known body burden of benzene to assess
the potential risk of adverse health effects in populations living in the vicinity of hazardous waste sites.
Benzene is widely distributed in the environment and has been detected in air (Clayton et al. 1999; EPA
1987a; Mohamed et al. 2002; Morello-Frosch et al. 2000), water (EPA 2001, 2005f; Sauer 1981, USGS
2005), soil (EPA 1979; Ferrario et al. 1985; MacLeod and MacKay 1999; Staples et al. 1985), sediment,
and some foods (EPA 1980b, 1982a; Fleming-Jones and Smith 2003). The levels of benzene in air and
water are well documented, but there is a need for more current information. Benzene is not expected to
be a significant contaminant in aquatic foods (Geyer et al. 1984; Gossett et al. 1983; Miller et al. 1985;
Ogata et al. 1984); however, some contamination of food crops consumed by humans may occur,
primarily from air-to-leaf transfer (Hattemer-Frey et al. 1990). The total concentration of benzene on
exposed food crops consumed by humans was estimated to be 587 ng/kg (Hattemer-Frey et al. 1990).
Humans are at risk of exposure to benzene because of its widespread distribution in the environment,
particularly in the atmosphere. Releases to the air from gasoline, smoking, and automobile exhaust
constitute the major risk of potential exposure for the general population (Wallace 1995). Additional data
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characterizing the concentration of benzene in drinking water, air, and soil surrounding hazardous waste
sites would be helpful in assessing human exposure for populations living near these waste sites. In
addition, more current data on levels of benzene in foods would be helpful in estimating intake of benzene
from food.
Exposure Levels in Humans.
Benzene has been detected in human body fluids and tissues such as
blood, urine, and fat (Brugnone et al. 1989; Chao et al. 1993; Karacic et al. 1987). Most of the
monitoring data have come from occupational studies of specific worker populations exposed to benzene
(Inoue et al. 1989b; Karacic et al. 1987; OSHA 1987; van Wijngaarden and Stewart 2003). Biological
monitoring studies exist for the general population (Melikian et al. 1994). There is information for
background levels in breath of smokers and nonsmokers (Wallace 1989b), baseline blood levels (Karacic
et al. 1987), and levels of urinary metabolites in unexposed people (Inoue et al. 1989b). Information on
exposure levels for populations living in the vicinity of hazardous waste sites would be helpful in
estimating exposure in these groups.
This information is necessary for assessing the need to conduct health studies on these populations.
Exposures of Children.
Benzene levels have been monitored in children and the environments in
which they live. This information gives levels found for infants and children in rural and urban area as
well as the levels found for children in homes of parents who smoke (Duarte-Davidson et al. 2001).
There have been many studies relating oil and petroleum exposure to childhood leukemia and other
diseases; however, the majority of these studies have not recorded benzene levels. More information
about the exposures of children, particularly those subject to high exposures such as smoking, busy roads,
and gasoline stations, are needed.
Child health data needs relating to susceptibility are discussed in Section 3.12.2, Identification of Data
Needs: Childrens Susceptibility.
Exposure Registries. In 2001, 1,143 people were included in the benzene subset of the Volatile
Organics Compounds subregistry of the National Exposure Registry. These people were exposed to
benzene at a site in Texas. Demographic and health information was obtained on all the exposed persons;
the information will be updated longitudinally. For those who were identified as exposed and were
deceased, a death certificate will be obtained to ascertain cause of death. This activity was carried out by
the Exposure and Disease Registry Branch (EDRB), Division of Health Studies (DHS), ATSDR. The
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288
data became part of public-user data files maintained by ATSDR. The statistical analyses of the baseline
data was completed and published (Agency for Toxic Substances and Disease Registry 1995). The
information that was amassed in the National Exposure Registry will be used to facilitate the
epidemiological research needs to assess adverse health outcomes that may be related to the exposure to
this substance.
6.8.2
Ongoing Studies
The Federal Research in Progress (FEDRIP 2005) database provides additional information obtainable
from a few ongoing studies that may fill in some of the data needs identified in Section 6.8.1. These
studies are summarized in Table 6-5.
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Table 6-5. Ongoing Studies on the Potential for Human Exposure to Benzenea
Investigator
Affiliation
Cho, CY
CHA Corporation; Laramie, Study of the use of microwave

Wyoming
technology for superfund site
remediation
Understanding potential health
hazards from groundwater and
soil contamination of chemicals
commonly found at hazardous
waste sites
Study of critical steps in the
ring-opening metabolism of the
human carcinogen benzene to
muconaldehyde
University of El Paso Texas; Modeling of organic
El Paso, Texas
compounds in drinking water
Fischer, LJ
Greenberg, A
Gurian, PL
Nylander-French, LA
Rothman, N
Scow, KM
Stenzel, PS
Thrall, KD
Weisel, CP
University of North Carolina; Assessment of dermal

Chapel Hill, North Carolina exposure to benzene and
naphthalene
Study of occupational and
environmental exposures
University of California;
Davis, California
National Institute of Health
Molecular characterization of
aquifer microbial communities
Study of the link between
occupational exposure of
carcinogens and cancer
Oregon Health and Science Assessment of human volatile
University; Portland, Oregon organic compounds exposure
near Superfund sites
Study of the modulation of
benzene metabolism by
exposure to environmental
mixtures
Sponsor
NIEHS
USDA
USDA
General Medical
Sciences
NIEHS
Division of cancer
epidemiology and
genetics
NIEHS
Division of cancer
epidemiology and
genetics
NIEHS
USDA
FEDRIP 2005
NIEHS = National Institute of Environmental Health Sciences; USDA = U.S. Department of Agriculture
BENZENE
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7. ANALYTICAL METHODS
The purpose of this chapter is to describe the analytical methods that are available for detecting,
measuring, and/or monitoring benzene, its metabolites, and other biomarkers of exposure and effect to
benzene. The intent is not to provide an exhaustive list of analytical methods. Rather, the intention is to
identify well-established methods that are used as the standard methods of analysis. Many of the
analytical methods used for environmental samples are the methods approved by federal agencies and
organizations such as EPA and the National Institute for Occupational Safety and Health (NIOSH). Other
methods presented in this chapter are those that are approved by groups such as the Association of
Official Analytical Chemists (AOAC) and the American Public Health Association (APHA).
Additionally, analytical methods are included that modify previously used methods to obtain lower
detection limits and/or to improve accuracy and precision.
7.1
BIOLOGICAL MATERIALS
Analytical methods have been developed to measure benzene levels in exhaled breath, blood, and various
body tissues. The primary method of analyzing for benzene in exhaled breath, body fluids, and tissues is
gas chromatography (GC) coupled with either flame ionization detection (FID), photoionization detection
(PID), or mass spectrometry (MS). Rigorous sample collection and preparation methods must be
followed when analyzing for benzene to prevent contamination of the sample. A summary of commonly
used methods of measuring benzene in biological samples is presented in Table 7-1.
Breath samples are collected on a solid sorbent (EPA 1987b; Gruenke et al. 1986; Pellizzari et al. 1988;
Wallace et al. 1985), in canisters (Thomas et al. 1991), or in a breath sampling tube and analyzed directly
(Sherwood and Carter 1970). Samples collected on Tenax sorbent are subjected to a thermal
desorption/cryofocussing step prior to analysis by capillary GC/MS (EPA 1987b; Pellizzari et al. 1988;
Wallace et al. 1985). Techniques involving headspace analysis of benzene adsorbed on silica gel have
also been used (Gruenke et al. 1986). MS detection generally provides the most sensitivity, from the low
to sub-ppb. The selectivity of the methods is improved if capillary GC columns are used (Pellizzari et al.
1988). Extraction of benzene from blood is frequently accomplished by either purge-and-trap or
headspace analysis. In purge-and-trap analysis, an inert gas such as helium or nitrogen is passed through
the sample, and purged volatiles are trapped on an appropriate solid sorbent (Antoine et al. 1986; Ashley
et al. 1992, 1994; Michael et al. 1980). Recent improvements in the method have resulted in excellent
sensitivity (300 ppt) and acceptable precision and accuracy (Ashley et al. 1992, 1994). The purge-andtrap method has also been used to analyze breast milk for other volatile organic compounds and could be
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Table 7-1. Analytical Methods for Determining Benzene in Biological Samples

Sample
matrix
Preparation method
Analytical
method
Sample
Percent
detection limit recovery
HRGC/MS
(IARC
Method 5)
GC/FID
3 ppt
70130
estimated
Pellizzari et al.
1988
100 ppb
100
1.6 ppb
(5-L sample)
8690
Sherwood and
Carter 1970
Wallace et al.
1986, 1985
0.1 ppb
NR
Gruenke et al.
1986
30 ppt
112128
HRGC/PID
0.4 g/L
NR
Ashley et al.
1992; 1994
Pekari et al.
1989
GC/MS-SIM
2 g/L
NR
Gruenke et al.
1986
GC/MS
0.5 g/L
NR
Antoine et al.
1986
GC/FID
100 g/L
98100
Jirka and
Bourne 1982
ITMS
90 ppt
NR
HPLC
20 pmol/g
globin
NR
St-Germain et
al. 1995
Hanway et al.
2000
GC/PID
0.51 nmol/L
>90
GC/MS-SIM
NR
NR
Breath
Collection on Tenax GC;

thermal desorption
Breath
Collection in sampling tube;

direct injection of sample
Collection on Tenax GC;
HRGC/MS
thermal desorption to on
column cryogenic trap
Collection in bags, adsorption GC/MS-SIM
on silica gel; desorption to
headspace vial; analysis of
headspace gases
Purge and trap
HRGC/MS
Breath
Breath
Blood
Blood
Heparinization; transfer to
isotonic saline in headspace
vial; equilibration with heat
Blood
Collection and transfer to
headspace vial; analysis of
headspace gases
Blood
Purging with nitrogen;
collection on Tenax GC-silica
gel
Blood
Extraction with toluene + HCl;
centrifugation; analysis of
toluene layer
Blood
Extraction device coupled to
MS
Blood
Centrifugation; dialysis;
precipitation; acidic
hydrolysis; Sep Pak cartridge
column purification
Urine
Incubation; analysis of
headspace gases
Tissues
Homogenization with internal
(bone
standard; centrifugation;
marrow, fat) analysis of supernatant
Tissues
Homogenization in buffer;
(lung, liver) centrifugation; analysis of
supernatant
RID-preparative 20 pg/g
HPLC/UV
NR
Reference
Kok and Ong

1994
Rickert et al.
1979
Bechtold et al.
1988
FID = flame ionization detection; GC = gas chromatography; HCl = hydrochloric acid; HPLC = high-performance
liquid chromatography; HRGC = high resolution gas chromatography; IARC = International Agency for Research on
Cancer; ITMS = ion trap mass spectrometry; MS = mass spectrometry; NR = not reported; PID = photoionization
detection; RID = reverse isotope dilution; SIM = selected ion monitoring; UV = ultraviolet detection
BENZENE
293
used for analyzing benzene (Michael et al. 1980; Pellizzari 1982). For headspace analysis, the samples
are placed in a special vial, and the gas generated above the liquid sample under equilibrium conditions is
analyzed (Gruenke et al. 1986; Pekari et al. 1989). Sensitivity is in the sub- to low-ppb range. A third
method of sample preparation involves extraction of the blood sample with an organic solvent (Jirka and
Bourne 1982) and analysis of the organic fraction. These methods are generally less sensitive, with
reported detection limits usually in the low- to mid-ppb range. Selectivity is improved with use of high
resolution gas chromatography (HRGC). Accuracy and precision could not be adequately compared
given the limited data available.
Screening methods are available for analysis of benzene in feces and urine (Ghoos et al. 1994) and body
fluids (Schuberth 1994). Both employ analysis by capillary GC with an ion trap detector (ITD). Benzene
in urine has been determined by trapping benzene stripped from the urine on a Carbotrap tube, followed
by thermal desorption GC/FID. Care must be taken when preparing benzene metabolite samples from
urine and bodily fluids in order to protect against enzymatic and oxidative degradation. These samples
are often treated to denature enzymes and avoid oxidation by hydroquinone. The detection limit is
50 ng/L and the average recovery is approximately 82% (Ghittori et al. 1993). Benzene in urine has also
been determined using headspace analysis with capillary GC/PID. The detection limit is 40 ng/L (Kok
and Ong 1994).
Methods are also available for determining metabolites of benzene in urine. A summary of available
methods is shown in Table 7-2. Both GC/FID or GC/MS and high-performance liquid chromatography
(HPLC) with ultraviolet detection (UV) have been used to measure urinary metabolites.
The primary metabolite of benzene is phenol. Phenol is excreted as glucuronide and sulphate conjugates
in urine. Total phenolic metabolites (phenol, phenyl sulfate, and phenyl glucuronide) have been
determined by hydrolyzing urine samples either enzymatically or by acid, then extracting the phenol with
solvent. Phenol is then measured by GC or HPLC techniques. Enzymatic hydrolysis coupled with
GC/FID has been reported; the detection limit is 1 mg/L and recovery is excellent (9298%) (IARC
1988). Sulfate and glucuronide conjugates have been determined directly by HPLC/UV (Ogata and
Taguchi 1987). The normal baseline levels of urinary phenolic metabolites from humans are usually 2
18 mg/L (Ong and Lee 1994). The available methods are sensitive enough to measure these relatively
high amounts accurately.
BENZENE
294
Table 7-2. Analytical Methods for Determining Metabolites of Benzene in Urine

Analytical
method
Sample
Percent
detection limit recovery
Centrifugation
HPLC/UV
4 mg/L (PS);
5 mg/L (PG)
100.5 (PS); Ogata and

101.8 (PG) Taguchi 1987
Digestion (enzymatic and

with acid); extraction with
diethyl ether
GC/FID
(IARC
Method 6)
1 mg/L
9298
IARC 1988
NR
NR
NIOSH 1974
2 mg/L
NR
Roush and Ott

1977
50 ppb
97.7
Jen and Tsai

1994
0.1 mg/L
NR
Inoue et al.
1989
GC/MS
10 g/L
NR
Bechtold et al.
1991
GC/MS
0.01 mg/L
93106
Ruppert et al.
1995
HPLC
1 g/L
NR
HPLC/UV
5 ppb
NR
Einig and
Dehnen 1995
Weaver et al.
2000
Sample matrix Preparation method

Urine (phenol,
phenyl sulfate,
phenyl
glucuronide)
Urine (phenol,
phenyl sulfate,
phenyl
glucuronide)
Urine (phenols
and cresols)
Urine (phenols
and cresols)
Urine (phenol)
Urine
(trans,trans
muconic acid)
Urine (muconic
acid, phenol)
Hydrolysis with perchloric GC/FID

acid; extraction with
diisopropyl ether
Hydrolysis with perchloric GC/FID
acid; saturation with NaCl;
extraction with diisopropyl
ether
Enzymatic reaction
HPLC/fluoro
metric
detection
Mixing with methanol;
HPLC/UV
centrifugation
Mixing with formic acid;

extraction (twice) with ethyl
ether; evaporation of
combined extracts
Urine
Cleanup on anion
(trans,trans
exchange resin;
muconic acid)
derivitization
Urine (S-phenyl- Solid-phase extraction;
mercapturic acid hydrolysis; derivatization
Urine
Cleanup on anion
(trans,trans
exchange resin
muconic acid)
Reference
FID = flame ionization detection; GC = gas chromatography; HPLC = high performance liquid chromatography;
HRGC = high resolution gas chromatography; IARC = International Agency for Research on Cancer; MS = mass
spectrometry; NaCl = sodium chloride; NR = not reported; PG = phenyl glucuronide; PS = phenyl sulfate;
UV = ultraviolet detection
BENZENE
295
Analysis of urinary trans,trans-muconic acid seems to be a better indicator than phenol for assessing
exposure to low levels of benzene (Ducos et al. 1990). However, muconic acid is a minor metabolic route
and background levels of muconic acid in urine are much lower than levels of phenolic metabolites and
are frequently below the limit of detection of the method used to determine them (Inoue et al. 1989b).
The detection of low levels of trans,trans-muconic acid in urine was difficult by earlier methods because
of low recovery of trans,trans-muconic acid (37% with ether) by the commonly used solvent extraction
method (Gad-El-Karim et al. 1985). An improved method for the determination of urinary trans,transmuconic acid utilizes solid phase extraction with SAX sorbent in combination with the HPLC/UV for
quantitation. The detection limit is 0.060.1 mg/L, and recovery is very good (90%) (Boogaard and van
Sittert 1995; Ducos et al. 1990). Weaver et al. (2000) used an anion exchange column to extract the urine
sample, which was then analyzed using HPLC-UV; detection limits were around 5 ppb. The relative
standard deviation of the method was 5% in the concentration range 120 ng/L. trans,trans-muconic acid
has been determined directly by HPLC/UV with similar sensitivity (detection limit=0.1 mg/L) (Inoue et
al. 1989b). The detection limit and specificity for the determination of urinary trans,trans-muconic acid
may be improved by using HPLC with diode array detector, GC/FID of the methylated product, or
GC/MS of trimethylsilyated product (Bartczak et al. 1994). Both GC/FID and HPLC/diode array
detection are capable of detecting urinary trans,trans-muconic acid at concentrations above 40 g/L, but
GC/MS is capable of detecting the metabolite at concentrations below 40 g/L (Bartczak et al. 1994).
The metabolite, S-phenyl-mercaptic acid, may be an indicator of exposure to benzene. It can be detected
at low levels (1 g/L) in urine using solid phase extraction and determination by HPLC (Einig and
Dehnen 1995). After purification by reverse phase cartridge chromatography, S-phenyl cysteine was
detected with sensitivity of about 20 pmol/g globin using HPLC (Hanway et al. 2000).
7.2
ENVIRONMENTAL SAMPLES
Methods exist for determining benzene in air (ambient, occupational, and industrial), water, sediment,
soil, foods, cigarette smoke, gasoline, and jet fuel. Most involve separation by GC with detection by FID,
PID, or MS. HPLC/UV and spectrophotometry have also been used. Table 7-3 summarizes several of
the methods that have been used to analyze for benzene in environmental samples.
Numerous methods exist for detecting and measuring benzene in ambient air. Air samples for benzene
analysis may be preconcentrated by passing the sample through a trap containing a solid adsorbent (Bayer
et al. 1988; EPA 1979, 1980a; Fung and Wright 1986; Gruenke et al. 1986; Harkov et al. 1985; Reineke
BENZENE
296
Table 7-3. Analytical Methods for Determining Benzene in Environmental
Samples
Sample
matrix
Air
Air
Air
Preparation method
Analytical
method
Sample trapped on silica gel; GC/MS

thermal desorption
Cryogenically trap; thermal
GC/FID
desorption
Direct on-line analysis
GC/FID
Air (ambient) Direct injection of ambient air GC/PID
Sample
Percent
detection limit recovery Reference
0.1 ppb
88105
NR
85115
NR
NR
0.25 ppb
NR
Air (ambient) Direct analysis of ambient air Electrochemical NR
NR
Air (ambient) Sample collection in Tedlar

bag; Cryogenically trap;
thermally desorb
Air (ambient) Sample collection in stainless
steel canisters or sorbent
tubes; trap cryogenically;
thermal desorption
Air (ambient) Collect on charcoal (vapor
badge or tube); desorb with
carbon disulfide
Air (ambient) Cryogenically trap; thermal
desorption
Air (ambient) Collection in canisters;
preconcentration using
2-stage trap
Air (ambient) Direct analysis
Air (ambient) Sample collection onto oncolumn cryogenic sample loop
or into stainless steel
containers
Air (ambient) Sample trapped on Tenax
GC; thermal desorption to oncolumn cryogenic trap
Air (ambient) Sample collected on Tenax
GC; thermal desorion to oncolumn cryogenic trap
Air (ambient) Sample trapped on Tenax
Air (consumer Sample trapped on solid
products)
sorbent; thermal desorption
Gruenke et al.
1986
Singh et al.
1985
Bayer et al.
1988
Clark et al.
1984
Stetter et al.
1986
Kowalski et
al. 1985
GC/PID
0.5 ppb
NR
HRGC/PID or
HRGC/FID
5 ppt (PID)
24 ppt (FID)
97104 Reineke and

(PID) 96 Bchmann
104 (FID) 1985
GC/FID
0
.3 ppb
(estimated)
NR
Fung and
Wright 1986
GC/PID/FID
1 ppt
70130
HRGC/ITD
sub-ppb level
813%
bias
Nutmagul and
Cronn 1985
Kelly et al.
1993
ALMS
GC/PID/FID
250 ppb
1 ppt
NR
70130
EPA 1985d
Nutmagul and
Cronn 1985
HRGC
<0.1 ppb
69126
EPA 1979
GC/FID
0.03 ppb
56144
EPA 1980a
HRGC/FID;
conf. by
HRGC/MS
GC/MS
3 ppt
NR
Roberts et al.
1984
NR
NR
Bayer et al.
1988
BENZENE
297
Samples
Sample
matrix
Air (at waste
sites and
landfills)
Air
(occupational)
Preparation method
Sample trapped on Tenax
GC; thermal desorption
Sample trapped on charcoal;

desorption with carbon
disulfide
Air
Sample collection on silica
(occupational) gel; desorption with ethanol
Air
(occupational)
Air
(occupational)
Analytical
method
Sample
Percent
HRGC/FID/ECD; 0.05 ppb

conf. by
HRGC/MS
GC/FID (NIOSH 10100 ppb
Methods 1500
and 1501)
GC/FID
100 ppb
Sample collection in Tedlar

GC/PID (NIOSH
bag; direct injection
Method 3700)
Sample collection on charcoal GC/PID
disk in miniature passive
dosimeter; thermal desorption
Air
Sample collection on
HPLC/UV
(occupational; charcoal; desorption with
jet fuel fumes) methyl chloride-ethyl acetate
Soil air
Sample collection on activated GC/FID
charcoal; desorption with
carbon disulfide
Drinking
Purge and trap
GC/MS
water
Drinking
Purge and trap
HRGC/MS (EPA
water
Method 524.2)
Water
Purge and trap onto Tenax
HRGC/MS
Water
Purge and trap on Tenax GC; HRGC
thermal desorption to oncolumn cryogenic trap
Water
Purge and trap onto Tenax
GC/MS
GC; thermal desorption
Water
Solvent extraction with
GC/MS
dichloromethane;
concentration
Water
Purge and trap on adsorbent GC/PID (EPA
column; thermal desorption
Method 602)
Water
Purge and trap on activated
GC/FID; conf. by
carbon; desorption with
GC/MS
carbon disulfide
Water
Purge and trap on Tenax;
GC/FID
thermal desorption
NR
Harkov et al.
1985
NR
NIOSH 1984
90
Sherwood

and Carter
1970
NIOSH 1994
10 ppb
NR
60 ppb
85115
0.08 ppm
94112 Dibben et al.

1989
NR
97100
Colenutt
and
Davies 1980
0.2 g/L
NR
Brass et al.
1977
EPA 1992a
0.030.04 g/L 9799
Gonzalez and
Levine 1986
0.110 g/L
7478
Michael et al.
1988
<1 g/L
69126
EPA 1979

NR
85125
15 g/L
NR
Harland et al.
1985
Sporstl et al.
1985
0.2 g/L
106
NEMI

2005a
NR
9699
0.001 g/L
94111
Colenutt and
Thorburn
1980
Hammers and
Bosman 1986
BENZENE
298
Samples
Sample
matrix
Preparation method
Water
Permeation of benzene
through a silicone
polycarbonate membrane into
an inert gas stream
Waste water Purge and trap onto
adsorbent column; thermal
desorption
Waste water Addition of isotopically labeled
benzene analog; purge and
trap onto adsorbent column;
thermal desorption
Water,
industrial
thermal desorption
effluents
Landfill
Purge sample and trap on
leachate
Tenax-silica gel; thermally
desorb
Landfill
Extract sample with pentane
leachate
Analytical
method
Sample
Percent
GC/FID
7.2 g/L
NR
Blanchard

and Hardy
1986
GC/MS (EPA
Method 624)
4.4 g/L
113
NEMI 2005b
GC/IDMS (EPA 10 g/L

Method 1624)
65141
NEMI 2005c
GC/MS
<5 g/L
95106
Pereira
and
Hughes 1980
GC/FID/FID
1 g/L
NR
EPA
1984b
GC/MS
1,000
10,000 g/L
NR
Schultz and
Kjeldsen
1986
EPA 1994c
Solid wastes Purge and trap, direct

injection, vacuum distillation,
or headspace
Soil
Sample mixed with NaOH
solution; equilibration;
analysis of headspace gases
Soil
thermal desorption to oncolumn cryogenic trap
Soil
thermal desorption
Soil
Supercritical fluid extraction
PID (EPA
0.009 g/L
Method 8021B)
99
HRGC/FID;
conf. by
HRGC/MS
HRGC
0.02 ng/mL
7598
<0.1 ppb
69126 EPA 1979
GC/FID
1 ppt
52
HRGC/FID
low ppb
7781
Sediment and Purge and trap on Tenax GCbiota

silica gel; thermal desorption
Sediment
thermal desorption
Fruits and
Mix with water and methanol;
vegetables
filter; distill azeotrope
Shellfish
Tissue homogenized; purge
with inert gas and trap on
Tenax GC-silica gel; thermally
desorb
HRGC/MS
NR
NR
GC/MS
NR
64
GC/FID
NR
8496
GC/MS
NR
NR
Kiang and
Grob 1986
Hammers and
Bosman 1986
Burford et al.
1994
Ferrario et al.
1985
Harland et al.
1985
Kozioski 1985
Ferrario et al.
1985
BENZENE
299
Samples
Sample
matrix
Mainstream
cigarette
smoke
Cigarette
smoke
Gasoline
Gasoline
Preparation method
Collection on filters and
impingers; [2H6]-benzene
added to impinger
Mainstream smoke filtered
and analyzed directly; sidestream smoke and smokepolluted air filtered and
collected in cryogenic
methanol-filled impingers
Dilute sample with hexane
Analytical
method
Sample
Percent
HRGC/IDMSSIM
0.05 g/
cigarette
HRGC/MS-SIM 0.1 g/
cigarette
GC/FID
Dilute sample with methanol; HPLC/UV

elute benzene to analytical
column with 50% methanol;
back-flush guard column with
100% methanol
7585
Byrd et al.
(trapping 1990
efficiency)
NR
Brunnemann
et al. 1989,
1990
NR
NR
NR
NR
Poole et al.
1988
Ludwig and
Eksteen 1988
ECD = electron capture detection; EPA = Environmental Protection Agency; FID = flame ionization detection;
GC = gas chromatography; HPLC = high-performance liquid chromatography; HRGC = high resolution gas
chromatography; IARC = International Agency for Research on Cancer; IDMS = isotope dilution mass spectrometry;
ITD = ion trap mass spectrometry; LRS = laser Raman spectroscopy; MS = mass spectrometry; NaOH = sodium
hydroxide; NIOSH = National Institute of Occupational Safety and Health; NR = not reported; PID = photoionization
detection; SIM = selected ion monitoring; UV = ultraviolet detection
BENZENE
300
and Bchmann 1985; Roberts et al. 1984). Commonly used adsorbents are Tenax resins (e.g., Tenax TA,
Tenax GC), silica gel, activated carbon, and carbonaceous polymeric compounds. Benzene in ambient air
can be collected in stainless steel canisters (EPA 1988a; Kelly et al. 1993) or Tedlar bags (Kowalski et al.
1985) and can be analyzed with or without preconcentration. Preconcentration of benzene can be
accomplished by direct on-column cryogenic trapping (EPA 1985c; Kowalski et al. 1985; Nutmagul and
Cronn 1985; Reineke and Bchmann 1985; Singh et al. 1985), or samples may be analyzed directly
without preconcentration (Bayer et al. 1988; Clark et al. 1984).
The most common methods of analysis for benzene in air are GC/PID, GC/FID, and GC/MS. The limit
of detection for GC/FID and GC/PID ranges from low ppb to low ppt. GC/MS is generally considered to
be more reliable than GC/FID or GC/PID in identifying benzene in samples, particularly those containing
multiple components having similar GC elution characteristics. Benzene has been quantified in ambient
air samples at sub-ppb levels by GC/MS (Gruenke et al. 1986) and ion trap mass spectrometry (Kelly et
al. 1993). The ion trap detector has the advantage of remaining largely unaffected by water vapor in the
sample. A continuous monitoring instrument using Fourier transform infrared (FTIR) spectroscopy is
being developed for measuring the levels of benzene or other toxic chemicals in exhaust emissions from
hazardous waste incinerators (DOE 1992).
Several analytical methods are available for determining atmospheric levels of benzene in the workplace.
The OSHA recommended procedure involves the collection of the sample vapors on charcoal adsorption
tubes, and then desorption followed by GC/MS analysis (OSHA 1985). Samples desorbed from charcoal
are also analyzed by GC/FID (NIOSH 1984) or HPLC/UV (Dibben et al. 1989). Detection limits are in
the ppb range (Dibben et al. 1989; NIOSH 1984). Passive dosimeters are also utilized, with GC/PID
quantitation; detection limits are in the ppb range (Gonzalez and Levine 1986). Other acceptable methods
include portable direct reading instruments and real-time continuous monitoring systems; these methods
generally have a sensitivity in the ppm range.
The most frequently used analytical methods for water samples containing benzene are GC/MS, GC/FID,
and GC/PID (Blanchard and Hardy 1986; Colenutt and Thorburn 1980; DOI 1984; EPA 1984, 1992;
Hammers and Bosman 1986; Harland et al. 1985; Lysyj et al. 1980; Michael et al. 1988; Pereira and
Hughes 1980; Sporstol et al. 1985). Benzene is usually isolated from aqueous media by the purge-andtrap method (Brass et al. 1977; Colenutt and Thorburn 1980; DOI 1984; EPA 1979, 1984, 1992;
Hammers and Bosman 1986; Harland et al. 1985; Michael et al. 1988). An inert gas such as nitrogen is
used to purge the sample. The purged benzene is trapped on an adsorbent substance, such as Tenax GC
BENZENE
301
or activated charcoal, and thermally desorbed. Recovery, where reported, ranges from acceptable (70%)
(EPA 1979, 1984; Michael et al. 1988) to very good (90%) (Colenutt and Thorburn 1980; EPA 1984,
1992; Hammers and Bosman 1986). Detection limits in the sub-ppb to ppt range may be attained with
HRGC/MS techniques (EPA 1992a); Michael et al. 1988). Liquid-liquid extraction procedures (Harrison
et al. 1994; Schultz and Kjeldsen 1986; Sporstol et al. 1985) are less commonly used, having been
replaced by more sensitive purge-and-trap methods. Interference from contamination can occur with all
methods if extreme care is not used in the handling of samples and cleaning of all equipment.
Solid samples, such as soil, sediment, and foods, are most frequently prepared for analysis using the
purge-and-trap method (EPA 1979, 1994c; Ferrario et al. 1985; Hammers and Bosman 1986; Harland et
al. 1985), although supercritical fluid extraction has recently been utilized (Burford et al. 1994).
Detection and quantitation of benzene may be GC/FID, GC/PID, or GC/MS. Detection limits as low as
1 ppt have been reported, but recoveries and precision have frequently been low. Improvements in the
method, including analysis by HRGC/PID, have resulted in low detection limits (9 ppt) and excellent
recovery (99%) for benzene (EPA 1994c). Screening methods are available for benzene; some may be
used at field sites. Immunoassay may be used as a screening and semiquantitative tool (Van Emon and
Gerlach 1995).
Methods exist for detection of benzene in other environmental media such as cigarette smoke, gasoline,
and jet fuel and its fumes (Brunnemann et al. 1989; Byrd et al. 1990; Ludwig and Eksteen 1988; Poole et
al. 1988). HPLC/UV, GC/FID, and GC/MS separation and detection techniques have been used for these
analyses. Sensitivity and reliability of these methods cannot be compared because of the lack of data.
Few methods have been reported for measurement of benzene in foods; performance data are generally
lacking.
7.3
ADEQUACY OF THE DATABASE
Section 104(i)(5) of CERCLA, as amended, directs the Administrator of ATSDR (in consultation with the
available, ATSDR, in conjunction with NTP, is required to assure the initiation of a program of research
designed to determine the health effects (and techniques for developing methods to determine such health
effects) of benzene.
BENZENE
302
7.3.1
Identification of Data Needs
Methods for Determining Biomarkers of Exposure and Effect.

Exposure. Methods exist for measuring benzene in breath (Gruenke et al. 1986; Pellizzari et al. 1988;
Sherwood and Carter 1970; Wallace et al. 1986), blood (Antoine et al. 1986; Ashley et al. 1992, 1994;
Gruenke et al. 1986; Jirka and Bourne 1982; Pekari et al. 1989), and tissues (Bechtold et al. 1988; Rickert
et al. 1979). The methods for breath are sensitive and accurate for determining exposure levels of
benzene at which health effects have been observed to occur, as well as for background levels in the
general population. The methods are relatively precise and selective. Methods for determining benzene
in blood are sensitive; based on the limited recovery data available, they appear to be accurate. More
information on the performance obtained with different methods would be helpful. The application of
GC/MS techniques to the analysis of blood specimens has resulted in a rapid, cost-effective, clinical
screening test for common volatile organic compounds, including benzene (DeLeon and Antoine 1985).
This test, the VOST (Volatile Organics Screening Test), has demonstrated the presence (down to 0.1 ppb)
of a variety of toxic volatile organics in the blood of environmentally sensitive patients and has provided
preliminary baseline concentration levels for the test population (DeLeon and Antoine 1985). The data on
determination of benzene in urine and tissue samples are very limited. In general, the available methods
have limits of detection that are too high to be useful in other than acute exposure situations. Methods
that could be used to measure low levels in human tissues would be useful for determining the
relationship between chronic low-level exposure and the effects observed in specific tissues.
Methods are available for measuring phenolic benzene metabolites in urine (Bechtold et al. 1991; IARC
1988; Jen and Tsai 1994; Jongeneelen et al. 1987; NIOSH 1974; Ogata and Taguchi 1987). Available
methods for determining most benzene metabolites in urine are sufficiently sensitive and reliable to allow
measurement of background concentrations in nonoccupationally exposed individuals. However, the
phenolic metabolites are not unique to benzene. Improved methods to detect phenolic metabolites are not
needed. Sensitive assays have been developed for detection of urinary trans,trans-muconic acid
BENZENE
303
(detection limit 10 g/L) (Bechtold et al. 1991; Ruppert et al. 1995). Since urinary trans,trans-muconic
acid concentration can be correlated with benzene exposure, this may provide a useful biomarker of
exposure on an individual basis (Bechtold et al. 1991; EPA 1992b; Weaver et al. 2000). In addition,
information is needed to assess the effect of co-exposure to other chemicals (e.g., toluene) on urinary
muconic acid levels. Also needed are specific biomarkers of cumulative exposure to benzene, based on
albumin or hemoglobin adducts, and lymphocyte DNA adducts of N-7-phenylguanine. It would also be
useful to develop specific biomarkers of acute- and chronic-duration exposure to benzene based on
adducts of muconaldehyde. The levels of such biomarkers formed in vivo would be useful later for
correlation with toxic effects of acute- or chronic-duration exposure to benzene. S-phenyl-mercapturic
acid has also been useful in biological monitoring of benzene exposure in humans and animals. S-phenylmercapturic acid can be measured with a sensitivity of about 20 pmol/g globin using HPLC (Hanway et
al. 2000). S-phenyl-mercapturic acid levels can also be correlated to environmental benzene exposure
(Popp et al. 1994), which may indicate its utility as a biomarker.
Effect. Methods for determining benzene in breath, blood, and tissues and for determining its metabolites
in urine could also be used as biomarkers of effect. However, efforts to correlate these measures with
observed toxic effects of benzene exposure have been unsuccessful. Other biomarkers of effect (e.g.,
complete blood cell counts, red and white blood cell counts, chromosomal aberrations, sister chromatid
exchanges, and examination of bone marrow) have been suggested for benzene, but they are not specific
for benzene exposure. Further development of methods for determining reliable unique biomarkers of
effect for benzene would be useful.
Methods for Determining Parent Compounds and Degradation Products in Environmental
Media.
Methods for determining benzene in air (Clark et al. 1984; Gruenke et al. 1986) and water
(Brass et al. 1977; EPA 1979, 1984; Hammers and Bosman 1986; Pereira and Hughes 1980), the media of
most concern for human exposure, are sensitive enough to measure background levels in the environment
and levels at which health effects might occur. Their reliability is limited primarily by the ubiquitous
presence of benzene in the environment, which makes contamination a constant problem. The accuracy
and precision of some methods for water analyses (e.g., GC/MS) need to be improved to produce more
reliable results. Methods for soil and other solid media appear to have the same problems as those for air
and water. In addition, there is a lack of information on methods for determining benzene in media such
as shellfish, fish, foods, and plants. Although exposure to benzene via ingestion of food is believed to be
minimal, standardized methods for these media are needed to better assess the extent of benzene
contamination in the environment and the resulting risk of exposure.
BENZENE
304
7.3.2
Ongoing Studies
The information in Table 7-4 was found as a result of a search of the Federal Research in Progress
database (FEDRIP 2005).
The Environmental Health Laboratory Sciences Division of the National Center for Environmental
Health, Centers for Disease Control and Prevention, is developing methods for the analysis of benzene
and other volatile organic compounds in blood. These methods use purge and trap methodology, highresolution gas chromatography, and magnetic sector mass spectrometry, which give detection limits in the
low parts per trillion (ppt) range.
BENZENE
305
Table 7-4. Ongoing Studies on Benzene, Analytical Methodsa

Investigator
Affiliation
Fales, HM
Analysis of proteins, peptides,

and metabolites for carcinogen
exposure using mass
spectrometry
University of Washington
Study of the use of human
Seattle, Washington
dosimetry for assessment of
exposure to volatile compounds
Study of glutathione biosynthesis
as a biomarker of toxic exposure
Assessment Environmental
Health Breath using PF-GC
VOC Technologies Inc.;
Assessment Environmental
Portland, Oregon
Health Breath using PF-GC
University of North Carolina; Development and application of
Chapel Hill, North Carolina biomarkers in the study of
exposure to carcinogens
Oregon Health Science
Study of biomarkers of
University; Portland, Oregon neurotoxicant exposure and
neurodegeneration
VOC Technologies Inc.;
A PF-GC for Environmental
Portland, Oregon
Health Breath Assessment
University of California at
Study of biomarkers of benzene
Berkeley
exposure and genotoxicity
Oregon Health Sciences
Study of neurotoxic chemicals
University; Portland, Oregon and biomarkers at superfund
sites
Use of biomarkers to study
occupational exposure of
carcinogens in order to enhance
exposure assessment
University of California at
Study and development of
Berkeley
biomarkers for exposure using
accelerator mass spectrometry
Use of biomarkers to study
benzene exposure in inner city
residents
Use of molecular biomarkers of
occupational benzene exposure
Kalman, DA
Kavanaugh, TJ
OBrien, RJ
Pavel, K
Rappaport, SM
Sabri, MI
Smejtek, PK
Smith, MT
Spencer, PS
Stewart, P
Turtletaub, KW
Weaver, VM
Wiencke, JK
a
Sponsor
USDA
NIEHS
NSF
NIEHS
NIEHS
NIEHS
NIH
NIEHS
NIEHS
NIH
Division of cancer
epidemiology and
genetics
NIEHS
USDA
USDA
FEDRIP 2005
NIEHS = National Institute of Environmental Health Sciences; NIH = National Institutes of Health; NSF = National
Science Foundation; USDA = U.S. Department of Agriculture
BENZENE
306
BENZENE
307
8. REGULATIONS AND ADVISORIES
The international and national regulations and guidelines regarding benzene in air, water, and other media
are summarized in Table 8-1.
ATSDR has derived an acute-duration inhalation MRL of 0.009 ppm for benzene based on a LOAEL of
10.2 ppm for immunological effects in mice exposed for 6 hours/day for 6 consecutive days (Rozen et al.
1984). The LOAEL of 10.2 ppm was adjusted from intermittent to continuous exposure
(LOAELADJ=2.55 ppm) and converted to a human equivalent concentration (LOAELHEC=2.55 ppm) using
EPA (1994b) methodology for a category 3 gas; an uncertainty factor of 300 (10 for use of a LOAEL,
3 for extrapolation from animals to humans using dosimetric conversion, and 10 to protect sensitive
individuals) was applied.
ATSDR has derived an intermediate-duration inhalation MRL of 0.006 ppm for benzene based on a
LOAEL of 10 ppm for significantly delayed splenic lymphocyte reaction to foreign antigens evaluated in
in vitro mixed lymphocyte reaction following the exposure of male C57Bl/6 mice to benzene vapors for
6 hours/day, 5 days/week for 20 exposure days (Rosenthal and Snyder 1987). The concentration was
adjusted from intermittent to continuous exposure (LOAELADJ=1.8 ppm) and converted to a human
equivalent concentration (LOAELHEC=1.8 ppm) using EPA (1994b) methodology for a category 3 gas; an
uncertainty factor of 300 (10 for the use of LOAEL, 3 for extrapolation from animals to humans using
dosimetric conversion, and 10 for human variability) was applied.
ATSDR has derived a chronic-duration inhalation MRL of 0.003 ppm for benzene based on the results of
benchmark dose (BMD) modeling of B cell counts in workers of shoe manufacturing industries in
Tianjin, China (Lan et al. 2004a). The resulting BMCL0.25sd of 0.10 ppm was adjusted from intermittent
to continuous exposure (BMCL0.25sdADJ=0.03 ppm) using EPA (1994b) methodology; an uncertainty factor
of 10 (to protect sensitive individuals) was applied.
ATSDR has derived a chronic-duration oral MRL of 0.0005 mg/kg/day for benzene based on estimation
of equivalent chronic-duration oral dosing that would result in effects similar to those observed in the
occupationally-exposed workers assessed by Lan et al. (2004a, 2004b). The BMCL0.25sdADJ of 0.03 ppm
(0.096 mg/m3) was converted to an equivalent BMDL0.25sdADJ of 0.014 mg/kg/day for ingested benzene
using EPA (1988b) human reference values for inhalation rate (20 m3/day) and body weight (70 kg) and a
factor of 0.5 to adjust for differences in absorption of benzene following inhalation versus oral exposure;
BENZENE
308
Table 8-1. Regulations and Guidelines Applicable to Benzene

Agency
Description
INTERNATIONAL
Guidelines:
IARC
Carcinogenicity classification
WHO
Air quality guidelines

Drinking water quality guidelines
NATIONAL
Regulations and Guidelines:
a. Air
ACGIH
TLV (TWA)
STEL
EPA
Hazardous air pollutant
NAS/NRC
NIOSH
OSHA
AEGL-1d
10 minutes
30 minutes
60 minutes
4 hours
8 hours
AEGL-2d
10 minutes
30 minutes
60 minutes
4 hours
8 hours
AEGL-3d
10 minutes
30 minutes
60 minutes
4 hours
8 hours
REL (10-hour TWA)
STEL
IDLH
PEL (8-hour TWA) for general industry
PEL (8-hour TWA) for construction

industry
Information
Reference
Group 1a
IARC 1987, 2004,

2007
WHO 2000
WHO 2004
6x10-6 unit risk

0.01 mg/Lb
0.5 ppmc
2.5 ppmc
Yes
ACGIH 2006
EPA 2004b
42 USC 7412
EPA 2005a
130 ppm
73 ppm
52 ppm
18 ppm
9.0 ppm
2,000 ppme
1,100 ppm
800 ppm
400 ppm
200 ppm
9,700 ppmf
5,600 ppme
4,000 ppme
2,000 ppme
990 ppm
0.1 ppmg
1.0 ppmg
500 ppmg
1 ppm
1 ppm
NIOSH 2005
OSHA 2005b, 2005e

29 CFR 1910.1000
29 CFR 1910.1028
OSHA 2005d, 2005f
29 CFR 1926.55,
Appendix A
29 CFR 1926.1128
BENZENE
309

Agency
Description
Information
NATIONAL (cont.)
OSHA
PEL (8-hour TWA) for shipyard industry 1 ppm
b. Water
EPA
c. Food
FDA
d. Other
ACGIH
EPA
Reference
OSHA 2005a,2005b
29 CFR 1915.1000
29 CFR 1915.1028
Designated as hazardous substances Yes

in accordance with Section 311(b)(2)(A)
of the Clean Water Act
Drinking water standards and health
advisories
1-day health advisory for a 10-kg
0.2 mg/L
child
10-day health advisory for a 10-kg
0.2 mg/L
child
DWEL
0.1 mg/L
-4
0.1 mg/L
10 Cancer risk
National primary drinking water
standards
MCLG
Zero
MCL
0.005 mg/L
Reportable quantities of hazardous
10 pounds
substances designated pursuant to
Section 311 of the Clean Water Act
Water quality criteria for human health
consumption of:
Water + organism
2.2 g/Lh
Organism only
51 g/Lh
EPA 2005b
40 CFR 116.4
Bottled drinking water
0.005 mg/L
FDA 2004
21 CFR 165.110
Biological exposure indices (end of
shift)
S-phenylmercapturic acid in urine
t,t-Muconic acid in urine
Oral slope factor
A1i
ACGIH 2006
Inhalation unit risk

RfC
RfD
25 g/g creatinine
500 g/g creatinine
Group Aj
1.5x10-25.5x10-2
per (mg/kg)/day
2.2x10-67.8x10-6
per g/m3
0.03 mg/m3
4x10-3 mg/kg/day
EPA 2004a
EPA 2002a
EPA 2005c
40 CFR 117.3
EPA 2002b
IRIS 2007
BENZENE
310

Agency
Description
NATIONAL (cont.)
EPA
Superfund, emergency planning, and
community right-to-know
Designated CERCLA hazardous
substance
Reportable quantity
RCRA hazardous waste number
Effective date of toxic chemical
release reporting
NTP
Information
Reference
EPA 2005d
40 CFR 302.4
10 poundsk
U019
01/01/87
Known human
carcinogen
EPA 2005e
40 CFR 372.65
NTP 2005
Group 1: carcinogenic to humans

For substances that are considered to be carcinogenic, the guideline value is the concentration in drinking water
associated with an upper-bound excess lifetime cancer risk of 10-5 (one additional cancer per 100,000 of the
population ingesting drinking water containing the substance at the guideline value for 70 years). Concentrations
associated with upper-bound estimated excess lifetime cancer risks of 10-4 and 10-6 can be calculated by multiplying
and dividing, respectively, the guideline value by 10.
c
Skin notation: refers to the potential significant contribution to the overall exposure by the cutaneous route,
including mucous membranes and the eyes, either by contact with vapors or, of probable greater significance, by
direct skin contact with the substance.
d
AEGL-1 is the airborne concentration of a substance above which it is predicted that the general population,
including susceptible individuals, could experience notable discomfort, irritation, or certain asymptomatic nonsensory
effects. AEGL-2 is the airborne concentration of a substance above which it is predicted that the general population,
including susceptible individuals, could experience irreversible or other serious, long-lasting adverse health effects or
an impaired ability to escape. AEGL-3 is the airborne concentration of a substance above which it is predicted that
the general population, including susceptible individuals, could experience life-threatening health effects or death.
e
Values denoted as having safety considerations against the hazard of explosion, whereas the Lower Explosive Limit
(LEL) =14,000 ppm and each value should be 10% LEL.
f
Value denoted as having extreme safety considerations against the hazard of explosion must be taken into account,
whereas the LEL =14,000 ppm and each value should be 50% LEL.
g
NIOSH potential occupational carcinogen
h
This criterion is based on carcinogenicity of 10-6 risk.
i
A1: confirmed human carcinogen
j
Group A: known human carcinogen
k
Designated CERCLA hazardous substance pursuant to Section 311(b)(2) and 307(a) of the Clean Water Act,
Section 112 of the Clean Air Act, and Section 3001 of RCRA.
b
ACGIH = American Conference of Governmental Industrial Hygienists; AEGL = Acute Emergency Exposure
Guideline Levels; CERCLA = Comprehensive Environmetnal Response, Compensation, and Liability Act;
CFR = Code of Federal Regulations; DWEL = drinking water equivalent level; EPA = Environmental Protection
Agency; FDA = Food and Drug Administration; IARC = International Agency for Research on Cancer;
IDLH = immediately dangerous to life or health; IRIS = Integrated Risk Information System; MCL = maximum
contaminant level; MCLG = maximum contaminant level goal; NAS/NRC = National Academy of Sciences/National
Research Council; NIOSH = National Institute for Occupational Safety and Health; NTP = National Toxicology
Program; OSHA = Occupational Safety and Health Administration; PEL = permissible exposure limit;
RCRA = Resource Conservation and Recovery Act; REL = recommended exposure limit; RfC = inhalation reference
concentration; RfD = oral reference dose; STEL = short-term expsoure limit; TLV = threshold limit values;
TWA = time-weighted average; USC = United States Code; WHO = World Health Organization
BENZENE
311
an uncertainty factor of 30 (10 to protect sensitive individuals and 3 for uncertainty in route-to-route
extrapolation) was applied.
EPA (IRIS 2007) derived an inhalation reference concentration (RfC) for benzene of 0.03 mg/m3
(0.009 ppm) based on the results of BMD modeling of absolute lymphocyte (ALC) data from the
occupational epidemiologic study of Rothman et al. (1996a), in which workers were exposed to benzene
by inhalation. The resulting BMCL of 7.2 ppm for decreased lymphocyte count was converted to
23.0 mg/m3 and adjusted from intermittent to continuous exposure (BMCLADJ=8.2 mg/m3); a total
uncertainty factor of 300 (3 for effect-level extrapolation, 10 to protect sensitive individuals, 3 for
subchronic-to-chronic extrapolation, and 3 for database deficiencies) was applied.
EPA (IRIS 2007) derived an oral reference dose (RfD) for benzene of 0.004 mg/kg/day, based on the
results of BMD modeling of ALC data from the occupational epidemiologic study of Rothman et al.
(1996a), in which workers were exposed to benzene by inhalation. The resulting BMCL of 7.2 ppm for
decreased lymphocyte count was converted to 23.0 mg/m3 and adjusted from intermittent to continuous
exposure (BMCLADJ=8.2 mg/m3). Route-to-route extrapolation methodology was applied to convert from
inhalation to equivalent oral exposure, resulting in an equivalent oral dose rate of 1.2 mg/kg/day. This
value was divided by a total uncertainty factor of 300 (3 for effect-level extrapolation, 10 to protect
sensitive individuals, 3 for subchronic-to-chronic extrapolation, and 3 for database deficiencies).
The International Agency for Research on Cancer (IARC) classifies benzene as a Group 1 carcinogen
(carcinogenic to humans) (IARC 2004). EPA classified benzene in Category A (known human
carcinogen) based on convincing evidence in humans supported by evidence from animal studies. Under
EPAs most recent guidelines for carcinogen risk assessment, benzene is characterized as a known human
carcinogen for all routes of exposure based on convincing human evidence as well as supporting evidence
from animal studies (IRIS 2007). The National Toxicology Programs lists benzene as a "substance
known to be carcinogenic," that is, a substance for which the evidence from human studies indicates that
there is a causal relationship between exposure to the substance and human cancer (NTP 2005).
The EPA has a current maximum contaminant level (MCL) of 0.005 mg/L for benzene in drinking water
(EPA 2002a). The World Health Organization (WHO) has established a guideline value of 0.01 mg/L for
benzene in drinking water (WHO 2004).
BENZENE
312
Benzene is on the list of chemicals in "The Emergency Planning and Community Right-to-Know Act of
1986" (EPA 2005d). Section 313 of Title III of the Superfund Amendments and Reauthorization Act
(SARA) requires owners and operators of certain facilities that manufacture, import, process, or otherwise
use the chemicals on this list to report annually any release of those chemicals to any environmental
media over a specified threshold level (U.S. Congress 1986).
OSHA requires employers of workers who are occupationally exposed to benzene to institute engineering
controls and work practices to reduce and maintain employee exposure at or below permissible exposure
limits (PEL). If an employer can document that benzene is used in the workplace <30 days/year, the
employer can use any combination of engineering controls, work practice controls, or respirators to
reduce employee exposure to or below the PEL of 1 ppm (8-hour TWA). Respirators must be provided
and used during the time period necessary to install or implement feasible engineering and work practice
controls, or where controls are not yet sufficient. Respirators are also required when the employer
determines that compliance with the PEL is not feasible with engineering or work practice controls, such
as maintenance and repair activities, vessel cleaning, or other operations where exposures are intermittent
and limited in duration, and in emergencies (OSHA 2005c).
ACGIH limits exposure to benzene to 0.5 ppm (8-hour TWA) (ACGIH 2006). The National Institute for
Occupational Safety and Health (NIOSH 2005) has established a recommended exposure level (REL) of
0.1 ppm (15-minute ceiling limit).
BENZENE
313
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_______________________
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*
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microsomal system. Mechanistic studies and formation of a new metabolite. Biochem Pharmacol
50(10):1607-1617.
Zhang Z, Xiang Q, Glatt H, et al. 1995b. Studies of pathways of ring opening of benzene in a Fenton
system. Free Radic Biol Med 18(3):411-419.
Zhu H, Li Y, Trush MA. 1995. Differences in xenobiotic detoxifying activities between bone marrow
stromal cells from mice and rats: Implications for benzene-induced hematotoxicity. J Toxicol Environ
Health 46(2):183-201.
Ziegler EE, Edwards BB, Jensen RL, et al. 1978. Absorption and retention of lead by infants. Pediatr
Res 12:29-34.
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9. REFERENCES
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10. GLOSSARY
AbsorptionThe taking up of liquids by solids, or of gases by solids or liquids.
Acute ExposureExposure to a chemical for a duration of 14 days or less, as specified in the
Toxicological Profiles.
AdsorptionThe adhesion in an extremely thin layer of molecules (as of gases, solutes, or liquids) to the
surfaces of solid bodies or liquids with which they are in contact.
Adsorption Coefficient (Koc)The ratio of the amount of a chemical adsorbed per unit weight of
organic carbon in the soil or sediment to the concentration of the chemical in solution at equilibrium.
Adsorption Ratio (Kd)The amount of a chemical adsorbed by sediment or soil (i.e., the solid phase)
divided by the amount of chemical in the solution phase, which is in equilibrium with the solid phase, at a
fixed solid/solution ratio. It is generally expressed in micrograms of chemical sorbed per gram of soil or
sediment.
Benchmark Dose (BMD)Usually defined as the lower confidence limit on the dose that produces a
specified magnitude of changes in a specified adverse response. For example, a BMD10 would be the
dose at the 95% lower confidence limit on a 10% response, and the benchmark response (BMR) would be
10%. The BMD is determined by modeling the dose response curve in the region of the dose response
relationship where biologically observable data are feasible.
Benchmark Dose ModelA statistical dose-response model applied to either experimental toxicological
or epidemiological data to calculate a BMD.
Bioconcentration Factor (BCF)The quotient of the concentration of a chemical in aquatic organisms
at a specific time or during a discrete time period of exposure divided by the concentration in the
surrounding water at the same time or during the same period.
BiomarkersBroadly defined as indicators signaling events in biologic systems or samples. They have
been classified as markers of exposure, markers of effect, and markers of susceptibility.
Cancer Effect Level (CEL)The lowest dose of chemical in a study, or group of studies, that produces
significant increases in the incidence of cancer (or tumors) between the exposed population and its
appropriate control.
CarcinogenA chemical capable of inducing cancer.
Case-Control StudyA type of epidemiological study that examines the relationship between a
particular outcome (disease or condition) and a variety of potential causative agents (such as toxic
chemicals). In a case-controlled study, a group of people with a specified and well-defined outcome is
identified and compared to a similar group of people without outcome.
Case ReportDescribes a single individual with a particular disease or exposure. These may suggest
some potential topics for scientific research, but are not actual research studies.
Case SeriesDescribes the experience of a small number of individuals with the same disease or
exposure. These may suggest potential topics for scientific research, but are not actual research studies.
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10. GLOSSARY
Ceiling ValueA concentration of a substance that should not be exceeded, even instantaneously.
Chronic ExposureExposure to a chemical for 365 days or more, as specified in the Toxicological
Profiles.
Cohort StudyA type of epidemiological study of a specific group or groups of people who have had a
common insult (e.g., exposure to an agent suspected of causing disease or a common disease) and are
followed forward from exposure to outcome. At least one exposed group is compared to one unexposed
group.
Cross-sectional StudyA type of epidemiological study of a group or groups of people that examines
the relationship between exposure and outcome to a chemical or to chemicals at one point in time.
Data NeedsSubstance-specific informational needs that if met would reduce the uncertainties of human
health assessment.
Developmental ToxicityThe occurrence of adverse effects on the developing organism that may result
from exposure to a chemical prior to conception (either parent), during prenatal development, or
postnatally to the time of sexual maturation. Adverse developmental effects may be detected at any point
in the life span of the organism.
Dose-Response RelationshipThe quantitative relationship between the amount of exposure to a
toxicant and the incidence of the adverse effects.
Embryotoxicity and FetotoxicityAny toxic effect on the conceptus as a result of prenatal exposure to
a chemical; the distinguishing feature between the two terms is the stage of development during which the
insult occurs. The terms, as used here, include malformations and variations, altered growth, and in utero
death.
Environmental Protection Agency (EPA) Health AdvisoryAn estimate of acceptable drinking water
levels for a chemical substance based on health effects information. A health advisory is not a legally
enforceable federal standard, but serves as technical guidance to assist federal, state, and local officials.
EpidemiologyRefers to the investigation of factors that determine the frequency and distribution of
disease or other health-related conditions within a defined human population during a specified period.
GenotoxicityA specific adverse effect on the genome of living cells that, upon the duplication of
affected cells, can be expressed as a mutagenic, clastogenic, or carcinogenic event because of specific
alteration of the molecular structure of the genome.
Half-lifeA measure of rate for the time required to eliminate one half of a quantity of a chemical from
the body or environmental media.
Immediately Dangerous to Life or Health (IDLH)The maximum environmental concentration of a
contaminant from which one could escape within 30 minutes without any escape-impairing symptoms or
irreversible health effects.
Immunologic ToxicityThe occurrence of adverse effects on the immune system that may result from
exposure to environmental agents such as chemicals.
Immunological EffectsFunctional changes in the immune response.
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10. GLOSSARY
IncidenceThe ratio of individuals in a population who develop a specified condition to the total
number of individuals in that population who could have developed that condition in a specified time
period.
Intermediate ExposureExposure to a chemical for a duration of 15364 days, as specified in the
Toxicological Profiles.
In VitroIsolated from the living organism and artificially maintained, as in a test tube.
In VivoOccurring within the living organism.
Lethal Concentration(LO) (LCLO)The lowest concentration of a chemical in air that has been reported
to have caused death in humans or animals.
Lethal Concentration(50) (LC50)A calculated concentration of a chemical in air to which exposure for
a specific length of time is expected to cause death in 50% of a defined experimental animal population.
Lethal Dose(LO) (LDLo)The lowest dose of a chemical introduced by a route other than inhalation that
has been reported to have caused death in humans or animals.
Lethal Dose(50) (LD50)The dose of a chemical that has been calculated to cause death in 50% of a
defined experimental animal population.
Lethal Time(50) (LT50)A calculated period of time within which a specific concentration of a chemical
is expected to cause death in 50% of a defined experimental animal population.
Lowest-Observed-Adverse-Effect Level (LOAEL)The lowest exposure level of chemical in a study,
or group of studies, that produces statistically or biologically significant increases in frequency or severity
of adverse effects between the exposed population and its appropriate control.
Lymphoreticular EffectsRepresent morphological effects involving lymphatic tissues such as the
lymph nodes, spleen, and thymus.
MalformationsPermanent structural changes that may adversely affect survival, development, or
function.
Minimal Risk Level (MRL)An estimate of daily human exposure to a hazardous substance that is
likely to be without an appreciable risk of adverse noncancer health effects over a specified route and
duration of exposure.
Modifying Factor (MF)A value (greater than zero) that is applied to the derivation of a Minimal Risk
Level (MRL) to reflect additional concerns about the database that are not covered by the uncertainty
factors. The default value for a MF is 1.
MorbidityState of being diseased; morbidity rate is the incidence or prevalence of disease in a specific
population.
MortalityDeath; mortality rate is a measure of the number of deaths in a population during a specified
interval of time.
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10. GLOSSARY
MutagenA substance that causes mutations. A mutation is a change in the DNA sequence of a cells
DNA. Mutations can lead to birth defects, miscarriages, or cancer.
NecropsyThe gross examination of the organs and tissues of a dead body to determine the cause of
death or pathological conditions.
NeurotoxicityThe occurrence of adverse effects on the nervous system following exposure to a
chemical.
No-Observed-Adverse-Effect Level (NOAEL)The dose of a chemical at which there were no
statistically or biologically significant increases in frequency or severity of adverse effects seen between
the exposed population and its appropriate control. Effects may be produced at this dose, but they are not
considered to be adverse.
Octanol-Water Partition Coefficient (Kow)The equilibrium ratio of the concentrations of a chemical
in n-octanol and water, in dilute solution.
Odds Ratio (OR)A means of measuring the association between an exposure (such as toxic substances
and a disease or condition) that represents the best estimate of relative risk (risk as a ratio of the incidence
among subjects exposed to a particular risk factor divided by the incidence among subjects who were not
exposed to the risk factor). An OR of greater than 1 is considered to indicate greater risk of disease in the
exposed group compared to the unexposed group.
Organophosphate or Organophosphorus CompoundA phosphorus-containing organic compound
and especially a pesticide that acts by inhibiting cholinesterase.
Permissible Exposure Limit (PEL)An Occupational Safety and Health Administration (OSHA)
allowable exposure level in workplace air averaged over an 8-hour shift of a 40-hour workweek.
PesticideGeneral classification of chemicals specifically developed and produced for use in the control
of agricultural and public health pests.
PharmacokineticsThe dynamic behavior of a material in the body, used to predict the fate
(disposition) of an exogenous substance in an organism. Utilizing computational techniques, it provides
the means of studying the absorption, distribution, metabolism, and excretion of chemicals by the body.
Pharmacokinetic ModelA set of equations that can be used to describe the time course of a parent
chemical or metabolite in an animal system. There are two types of pharmacokinetic models: data-based
and physiologically-based. A data-based model divides the animal system into a series of compartments,
which, in general, do not represent real, identifiable anatomic regions of the body, whereas the
physiologically-based model compartments represent real anatomic regions of the body.
Physiologically Based Pharmacodynamic (PBPD) ModelA type of physiologically based doseresponse model that quantitatively describes the relationship between target tissue dose and toxic end
points. These models advance the importance of physiologically based models in that they clearly
describe the biological effect (response) produced by the system following exposure to an exogenous
substance.
Physiologically Based Pharmacokinetic (PBPK) ModelComprised of a series of compartments
representing organs or tissue groups with realistic weights and blood flows. These models require a
variety of physiological information: tissue volumes, blood flow rates to tissues, cardiac output, alveolar
BENZENE
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10. GLOSSARY
ventilation rates, and possibly membrane permeabilities. The models also utilize biochemical
information, such as air/blood partition coefficients, and metabolic parameters. PBPK models are also
called biologically based tissue dosimetry models.
PrevalenceThe number of cases of a disease or condition in a population at one point in time.
Prospective StudyA type of cohort study in which the pertinent observations are made on events
occurring after the start of the study. A group is followed over time.
q1*The upper-bound estimate of the low-dose slope of the dose-response curve as determined by the
multistage procedure. The q1* can be used to calculate an estimate of carcinogenic potency, the
incremental excess cancer risk per unit of exposure (usually g/L for water, mg/kg/day for food, and
g/m3 for air).
Recommended Exposure Limit (REL)A National Institute for Occupational Safety and Health
(NIOSH) time-weighted average (TWA) concentration for up to a 10-hour workday during a 40-hour
workweek.
Reference Concentration (RfC)An estimate (with uncertainty spanning perhaps an order of
magnitude) of a continuous inhalation exposure to the human population (including sensitive subgroups)
that is likely to be without an appreciable risk of deleterious noncancer health effects during a lifetime.
The inhalation reference concentration is for continuous inhalation exposures and is appropriately
expressed in units of mg/m3 or ppm.
Reference Dose (RfD)An estimate (with uncertainty spanning perhaps an order of magnitude) of the
daily exposure of the human population to a potential hazard that is likely to be without risk of deleterious
effects during a lifetime. The RfD is operationally derived from the no-observed-adverse-effect level
(NOAEL, from animal and human studies) by a consistent application of uncertainty factors that reflect
various types of data used to estimate RfDs and an additional modifying factor, which is based on a
professional judgment of the entire database on the chemical. The RfDs are not applicable to
nonthreshold effects such as cancer.
Reportable Quantity (RQ)The quantity of a hazardous substance that is considered reportable under
the Comprehensive Environmental Response, Compensation, and Liability Act (CERCLA). Reportable
quantities are (1) 1 pound or greater or (2) for selected substances, an amount established by regulation
either under CERCLA or under Section 311 of the Clean Water Act. Quantities are measured over a
24-hour period.
Reproductive ToxicityThe occurrence of adverse effects on the reproductive system that may result
from exposure to a chemical. The toxicity may be directed to the reproductive organs and/or the related
endocrine system. The manifestation of such toxicity may be noted as alterations in sexual behavior,
fertility, pregnancy outcomes, or modifications in other functions that are dependent on the integrity of
this system.
Retrospective StudyA type of cohort study based on a group of persons known to have been exposed
at some time in the past. Data are collected from routinely recorded events, up to the time the study is
undertaken. Retrospective studies are limited to causal factors that can be ascertained from existing
records and/or examining survivors of the cohort.
RiskThe possibility or chance that some adverse effect will result from a given exposure to a chemical.
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10. GLOSSARY
Risk FactorAn aspect of personal behavior or lifestyle, an environmental exposure, or an inborn or

inherited characteristic that is associated with an increased occurrence of disease or other health-related
event or condition.
Risk RatioThe ratio of the risk among persons with specific risk factors compared to the risk among
persons without risk factors. A risk ratio greater than 1 indicates greater risk of disease in the exposed
group compared to the unexposed group.
Short-Term Exposure Limit (STEL)The American Conference of Governmental Industrial
Hygienists (ACGIH) maximum concentration to which workers can be exposed for up to 15 minutes
continually. No more than four excursions are allowed per day, and there must be at least 60 minutes
between exposure periods. The daily Threshold Limit Value-Time Weighted Average (TLV-TWA) may
not be exceeded.
Standardized Mortality Ratio (SMR)A ratio of the observed number of deaths and the expected
number of deaths in a specific standard population.
Target Organ ToxicityThis term covers a broad range of adverse effects on target organs or
physiological systems (e.g., renal, cardiovascular) extending from those arising through a single limited
exposure to those assumed over a lifetime of exposure to a chemical.
TeratogenA chemical that causes structural defects that affect the development of an organism.
Threshold Limit Value (TLV)An American Conference of Governmental Industrial Hygienists
(ACGIH) concentration of a substance to which most workers can be exposed without adverse effect.
The TLV may be expressed as a Time Weighted Average (TWA), as a Short-Term Exposure Limit
(STEL), or as a ceiling limit (CL).
Time-Weighted Average (TWA)An allowable exposure concentration averaged over a normal 8-hour
workday or 40-hour workweek.
Toxic Dose(50) (TD50)A calculated dose of a chemical, introduced by a route other than inhalation,
which is expected to cause a specific toxic effect in 50% of a defined experimental animal population.
ToxicokineticThe absorption, distribution, and elimination of toxic compounds in the living organism.
Uncertainty Factor (UF)A factor used in operationally deriving the Minimal Risk Level (MRL) or
Reference Dose (RfD) or Reference Concentration (RfC) from experimental data. UFs are intended to
account for (1) the variation in sensitivity among the members of the human population, (2) the
uncertainty in extrapolating animal data to the case of human, (3) the uncertainty in extrapolating from
data obtained in a study that is of less than lifetime exposure, and (4) the uncertainty in using lowestobserved-adverse-effect level (LOAEL) data rather than no-observed-adverse-effect level (NOAEL) data.
A default for each individual UF is 10; if complete certainty in data exists, a value of 1 can be used;
however, a reduced UF of 3 may be used on a case-by-case basis, 3 being the approximate logarithmic
average of 10 and 1.
XenobioticAny chemical that is foreign to the biological system.
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APPENDIX A. ATSDR MINIMAL RISK LEVELS AND WORKSHEETS

The Comprehensive Environmental Response, Compensation, and Liability Act (CERCLA) [42 U.S.C.
9601 et seq.], as amended by the Superfund Amendments and Reauthorization Act (SARA) [Pub. L. 99
499], requires that the Agency for Toxic Substances and Disease Registry (ATSDR) develop jointly with
the U.S. Environmental Protection Agency (EPA), in order of priority, a list of hazardous substances most
commonly found at facilities on the CERCLA National Priorities List (NPL); prepare toxicological
profiles for each substance included on the priority list of hazardous substances; and assure the initiation
of a research program to fill identified data needs associated with the substances.
The toxicological profiles include an examination, summary, and interpretation of available toxicological
information and epidemiologic evaluations of a hazardous substance. During the development of
toxicological profiles, Minimal Risk Levels (MRLs) are derived when reliable and sufficient data exist to
identify the target organ(s) of effect or the most sensitive health effect(s) for a specific duration for a
given route of exposure. An MRL is an estimate of the daily human exposure to a hazardous substance
that is likely to be without appreciable risk of adverse noncancer health effects over a specified duration
of exposure. MRLs are based on noncancer health effects only and are not based on a consideration of
cancer effects. These substance-specific estimates, which are intended to serve as screening levels, are
used by ATSDR health assessors to identify contaminants and potential health effects that may be of
concern at hazardous waste sites. It is important to note that MRLs are not intended to define clean-up or
action levels.
MRLs are derived for hazardous substances using the no-observed-adverse-effect level/uncertainty factor
approach. They are below levels that might cause adverse health effects in the people most sensitive to
such chemical-induced effects. MRLs are derived for acute (114 days), intermediate (15364 days), and
chronic (365 days and longer) durations and for the oral and inhalation routes of exposure. Currently,
MRLs for the dermal route of exposure are not derived because ATSDR has not yet identified a method
suitable for this route of exposure. MRLs are generally based on the most sensitive chemical-induced end
point considered to be of relevance to humans. Serious health effects (such as irreparable damage to the
liver or kidneys, or birth defects) are not used as a basis for establishing MRLs. Exposure to a level
above the MRL does not mean that adverse health effects will occur.
MRLs are intended only to serve as a screening tool to help public health professionals decide where to
look more closely. They may also be viewed as a mechanism to identify those hazardous waste sites that
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APPENDIX A
are not expected to cause adverse health effects. Most MRLs contain a degree of uncertainty because of
the lack of precise toxicological information on the people who might be most sensitive (e.g., infants,
elderly, nutritionally or immunologically compromised) to the effects of hazardous substances. ATSDR
uses a conservative (i.e., protective) approach to address this uncertainty consistent with the public health
principle of prevention. Although human data are preferred, MRLs often must be based on animal studies
because relevant human studies are lacking. In the absence of evidence to the contrary, ATSDR assumes
that humans are more sensitive to the effects of hazardous substance than animals and that certain persons
may be particularly sensitive. Thus, the resulting MRL may be as much as 100-fold below levels that
have been shown to be nontoxic in laboratory animals.
Proposed MRLs undergo a rigorous review process: Health Effects/MRL Workgroup reviews within the
Division of Toxicology and Environmental Medicine, expert panel peer reviews, and agency-wide MRL
Workgroup reviews, with participation from other federal agencies and comments from the public. They
are subject to change as new information becomes available concomitant with updating the toxicological
profiles. Thus, MRLs in the most recent toxicological profiles supersede previously published levels.
For additional information regarding MRLs, please contact the Division of Toxicology and
Environmental Medicine, Agency for Toxic Substances and Disease Registry, 1600 Clifton Road NE,
Mailstop F-32, Atlanta, Georgia 30333.
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APPENDIX A
MINIMAL RISK LEVEL (MRL) WORKSHEET

Chemical Name:
CAS Numbers:
Date:
Profile Status:
Route:
Duration:
Graph Key:
Species:
Benzene
71-43-2
August 2007
Post Public, Final Draft
[x] Inhalation [ ] Oral
[x] Acute [ ] Intermediate [ ] Chronic
46
Mouse
Minimal Risk Level: 0.009 [ ] mg/kg/day [x] ppm

Reference: Rozen MG, Snyder CA, Albert RE. 1984. Depression in B- and T-lymphocyte mitogeninduced blastogenesis in mice exposed to low concentrations of benzene. Toxicol Lett 20:343-349.
Experimental design: Male C57BL/6J mice (78/group) were exposed to benzene (0, 10.2, 31, 100, or
301 ppm) in whole-body dynamic inhalation chambers for 6 hours/day for 6 consecutive days. Control
mice were exposed to filtered, conditioned air only. Erythrocyte counts were depressed in C57BL/6 mice
only at 100 and 301 ppm. The 10.2 ppm exposure level resulted in significant depression of femoral
lipopolysaccharide-induced B-colony-forming ability in the absence of a significant depression of total
numbers of B cells. At 31 ppm, splenic phytohemagglutinin-induced blastogenesis was significantly
depressed without a concomitant significant depression in numbers of T-lymphocytes. Peripheral
lymphocyte counts were depressed at all exposure levels. These results demonstrate that short-term
inhaled benzene even at low exposure concentrations can alter certain immune associated processes.
Effect noted in study and corresponding doses:
10.2 ppm =
No adverse effect on erythrocytes, depressed peripheral lymphocytes and mitogeninduced blastogenesis of femoral B-lymphocytes (less serious LOAEL).
31 ppm =
No adverse effect on erythrocytes, depression of mitogen-induced blastogenesis of

splenic T-cells.
100 ppm =
Depressed erythrocyte counts.
Dose and end point used for MRL derivation:

[ ] NOAEL [x] LOAEL
Uncertainty Factors used in MRL derivation: 300
[ ] 1 [ ] 3 [x] 10 (for use of a LOAEL)
[ ] 1 [x] 3 [ ] 10 (for extrapolation from animals to humans using dosimetric conversion)
[ ] 1 [ ] 3 [x] 10 (for human variability)
Was a conversion factor used from ppm in food or water to a mg/body weight dose? Not applicable.
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APPENDIX A
If an inhalation study in animals, list conversion factors used in determining human equivalent dose: The
concentration was adjusted for intermittent exposure by multiplying the LOAEL (10.2 ppm) by 6/24 to
correct for less than a full day of exposure. The resulting LOAELADJ is 2.55 ppm.
According to current EPA (1994b) methodology for calculating a human equivalent concentration (HEC)
for extrarespiratory effects of a category 3 gas (such as benzene):
LOAELHEC = LOAELADJ x ([Hb/g]A/[Hb/g]H)
where:
LOAELHEC
= The LOAEL dosimetrically adjusted to a human equivalent concentration
LOAELADJ
= The LOAEL adjusted from intermittent to continuous exposure
[Hb/g]A/[Hb/g]H = The ratio of the blood:gas partition coefficient of the chemical for the laboratory
animal species to the human value
default value of 1 is used for the ratio. According to Wiester et al. (2002), benzene blood:gas partition
coefficients for mice and humans are 17.44 and 8.12, respectively. Therefore the default value of 1 is
applied, in which case, the LOAELHEC is equivalent to the LOAELADJ = 2.55 ppm.
Was a conversion used from intermittent to continuous exposure? The concentration was adjusted for
intermittent exposure by multiplying the LOAEL (10.2 ppm) by 6/24 to correct for less than a full day of
exposure. The resulting LOAELADJ is 2.55 ppm.
Other additional studies or pertinent information that lend support to this MRL: Increased number of
micronucleated polychromatic erythrocytes (MN-PCEs), decreased numbers of granulopoietic stem cells
(Toft et al. 1982), lymphopenia (Cronkite et al. 1985), lymphocyte depression, and increased
susceptibility to bacterial infection (Rosenthal and Snyder 1985) are among the adverse hematological
and immunological effects observed in several other acute-duration inhalation studies. The study by
Rozen et al. (1984) shows benzene immunotoxicity (reduced mitogen-induced lymphocyte proliferation)
at a slightly lower exposure level than these other studies. C57BI/6J mice were exposed to 0, 10.2, 31,
100, and 301 ppm benzene for 6 days at 6 hours/day. Lymphocyte counts were depressed at all exposure
levels while erythrocyte counts were elevated at 10.2 ppm, equal to controls at 31 ppm, and depressed at
100 and 301 ppm. Femoral B-lymphocyte and splenic B-lymphocyte numbers were reduced at 100 ppm.
Levels of circulating lymphocytes and mitogen-induced blastogenesis of femoral B-lymphocytes were
depressed after exposure to 10.2 ppm benzene for 6 days. Mitogen-induced blastogeneses of splenic
T-lymphocytes were depressed after exposure to 31 ppm of benzene for 6 days. In another study, mice
exhibited a 50% decrease in the population of erythroid progenitor cells (CFU-E) after exposure to
10 ppm benzene for 5 days, 6 hours/day (Dempster and Snyder 1991). In a study by Wells and Nerland
(1991), groups of 45 male Swiss-Webster mice were exposed to 0, 3, 25, 55, 105, 199, 303, 527, 1,150,
or 2,290 ppm benzene for 6 hours/day for 5 days. The number of leukocytes in peripheral blood and
spleen weights were significantly decreased compared with untreated controls at all concentrations
25 ppm. Therefore, 3 ppm was the NOAEL and 25 ppm was the LOAEL for these effects. Other end
points were not monitored in this study. These data support the choice of Rozen et al. (1984) as a critical
study.
Agency Contacts (Chemical Managers): Sharon Wilbur, M.A., Sam Keith, M.S., C.H.P., Obaid Faroon,
Ph.D.
BENZENE
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APPENDIX A

Chemical Name:
CAS Numbers:
Date:
Profile Status:
Route:
Duration:
Graph Key:
Species:
Benzene
71-43-2
August 2007
[ ] Acute [x] Intermediate [ ] Chronic
126
Mouse
Minimal Risk Level: 0.006 [ ] mg/kg/day [x] ppm

Reference: Rosenthal GJ, Snyder CA. 1987. Inhaled benzene reduces aspects of cell-mediated tumor
surveillance in mice. Toxicol Appl Pharmacol 88:35-43.
Experimental design: Male C57Bl/6 mice were exposed to 10, 30, or 100 ppm of benzene by inhalation
6 hours/day, 5 days/week for 20 exposure days. The number of lymphocytes and their functional
capacities were evaluated in spleens of exposed mice. Following the 20 days of exposure, functional
capacity of splenic lymphocytes was evaluated in two in vitro assays: mixed-lymphocyte culture (MLC)
and 51Cr-release cytotoxicity assay. Measured mean daily benzene concentrations in the 10, 30, and
100 ppm groups were 11.1 (1.5) ppm, 29.5 (4.4) ppm, and 99.7 (7.0) ppm, respectively. No changes
were observed in the relative proportions of splenic leukocytes, in the percentage of T-cell subsets or in
the ratio of T-helper and T-suppressor cells, even at the highest exposure level (100 ppm). Therefore, the
functional assays could be normalized for particular lymphocyte populations by using equal numbers of
splenic cells. MLC is an in vitro measure of alloreactivity (capacity to mount an immune response
against foreign antigens). The MLC activity of spleen lymphocytes from 10- and 100-ppm mice was
delayed on days 24 of culture (relative to air-exposed controls), indicating that benzene exposure causes
impaired in vitro alloreactivity (data for the 30-ppm mice were not included in the reported results). This
delayed alloreactivity was not due to spleen suppressor cells. The lymphocyte cytotoxic function
evaluated in the 51Cr-release assay was also altered; splenic lymphocytes from 100-ppm mice had a
significantly reduced lysing capacity. The results indicate that inhalation exposure of mice to benzene has
an immunodepressive effect on in vitro alloreactivity and cytotoxicity of splenic lymphocytes.
Effect noted in study and corresponding doses:
10 ppm =
Significantly delayed splenic lymphocyte reaction to foreign antigens evaluated in in

vitro mixed lymphocyte reaction (less serious LOAEL).
30 ppm =
Results not reported.
100 ppm =
Significantly delayed splenic lymphocyte reaction to foreign antigens evaluated in in

vitro mixed lymphocyte reaction.
Dose and end point used for MRL derivation:

[ ] NOAEL [x] LOAEL
BENZENE
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APPENDIX A

[ ] 1 [ ] 3 [x] 10 (for use of a LOAEL)
[ ] 1 [x] 3 [ ] 10 (for extrapolation from animals to humans using dosimetric conversion)
If an inhalation study in animals, list conversion factors used in determining human equivalent dose:
According to current EPA (1994b) methodology for calculating a human equivalent concentration (HEC)
for extrarespiratory effects of a category 3 gas (such as benzene):
LOAELHEC = LOAELADJ x ([Hb/g]A/[Hb/g]H)
where:
LOAELHEC
= The LOAEL dosimetrically adjusted to a human equivalent concentration
LOAELADJ
= The LOAEL adjusted from intermittent to continuous exposure
[Hb/g]A/[Hb/g]H = The ratio of the blood:gas partition coefficient of the chemical for the laboratory
animal species to the human value
default value of 1 is used for the ratio. According to Wiester et al. (2002), benzene blood:gas partition
coefficients for mice and humans are 17.44 and 8.12, respectively. Therefore, the default value of 1 is
applied, in which case, the LOAELHEC is equivalent to the LOAELADJ.
Was a conversion used from intermittent to continuous exposure? The concentration was adjusted for
intermittent exposure by multiplying the LOAEL (10 ppm) by 6 hours/24 hours to correct for less than a
full day of exposure and 5 days/7 days to correct for less than a full week of exposure. The resulting
LOAELADJ is 1.8 ppm.
Other additional studies or pertinent information that lend support to this MRL: Exposure of C57BL
mice to 10 ppm benzene for 6 hours/day, 5 days/week caused significant depressions in numbers of
lymphocytes (ca. 30% lower than controls) as early as exposure day 32; this effect was also noted at the
other scheduled periods of testing (exposure days 66 and 178) (Baarson et al. 1984). Splenic red blood
cells were significantly reduced (ca. 15% lower than controls) at exposure days 66 and 178. The failure
of the erythrons of benzene-exposed mice to support normal red cell mass was illustrated by the
significant reduction in peripheral red cell numbers in these animals at 66 and 178 days of benzene
exposure. Green et al. (1981a, 1981b) exposed male CD-1 mice to benzene vapors at concentrations of
0 or 9.6 ppm for 6 hours/day, 5 days/week for 50 days and assessed the effects of exposure on cellularity
in the spleen, bone marrow, and peripheral blood. Exposure-related effects included a 90% increase in
numbers of multipotential hematopoietic stem cells (CFU-S) (Green et al. 1981a), approximately 25%
increase in spleen weight and total splenic nucleated cellularity (Green et al. 1981b), and 80% increase in
nucleated RBCs (Green et al. 1981b). The results of Baarson et al. (1984) and Green et al. (1981a,
1981b) are limited for purposes of quantitative risk assessment because a single exposure level was
employed. However, they support the choice of Rosenthal and Snyder (1987) as the critical study, which
serves as the basis for the intermediate-duration inhalation MRL.
BENZENE
A-7
APPENDIX A
Ph.D.
BENZENE
A-8
APPENDIX A

Chemical Name:
CAS Numbers:
Date:
Profile Status:
Route:
Duration:
Graph Key:
Species:
Benzene
71-43-2
August 2007
[ ] Acute [ ] Intermediate [x ] Chronic
161
Human
Minimal Risk Level: 0.003 [ ] mg/kg/day [x] ppm [ ] mg/m3

Reference: Lan Q, Zhang L, Li G, et al. 2004a. Hematotoxicity in workers exposed to low levels of
benzene. Science 306:1774-1776.
Experimental design: A cross-sectional study was performed on 250 workers (approximately two-thirds
female) exposed to benzene at two shoe manufacturing facilities in Tianjin, China, and 140 age- and
gender-matched workers in clothing manufacturing facilities that did not use benzene. The benzeneexposed workers had been employed for an average of 6.12.9 years. Benzene exposure was monitored
by individual organic vapor monitors (full shift) 5 or more times during 16 months prior to phlebotomy.
Post-shift urine samples were collected from every worker. Urinary benzene concentrations were highly
correlated with mean individual air levels. Benzene was not found (detection limit 0.04 ppm) in
workplace and home air samples of control workers taken at three different time periods. Study subjects
were categorized into four groups (140 controls, 109 at <1 ppm, 110 at 1<10 ppm, and 31 at 10 ppm)
according to mean benzene exposure levels measured twice during the month prior to phlebotomy. Of the
250 exposed workers, 109 were exposed to <1 ppm benzene. Each of these individuals worked at the
larger of the two facilities included in the study. Exposure concentrations were generally higher at the
smaller facility due to a less adequate ventilation system. Complete blood count (CBC) and differential
were analyzed mechanically. Coefficients of variation for all cell counts were <10%.
Mean 1-month benzene exposure levels in the four groups (controls, <1 ppm, 1<10 ppm, and 10 ppm)
were <0.04, 0.570.24, 2.852.11, and 28.7320.74 ppm, respectively (see Table A-1). An evaluation of
potential confounding factors showed that age, gender, cigarette smoking, alcohol consumption, recent
infection, and body mass index were associated with at least one hematological end point. The values in
Table A-1 represent values that were adjusted to account for these variables. All types of white blood
cells (WBCs) and platelets were significantly decreased in the lowest exposure group (<1 ppm), ranging
in magnitude from approximately 8 to 15% lower than controls. Although similar statistical analyses for
the mid- and high-exposure groups were not included in the study report, decreases in all types of WBCs
and platelets were noted at these exposure levels as well; the decreases in the highest exposure group
ranged in magnitude from 15 to 36%. Lymphocyte subset analysis revealed significantly decreased
CD4+-T cells, CD4+/CD8+ ratio, and B cells. Hemoglobin concentrations were significantly decreased
only within the highest (10 ppm) exposure group. Tests for a linear trend using benzene air level as a
continuous variable were significant for platelets and all WBC measures except monocytes and
CD8+-T cells. Upon restricting the linear trend analyses to workers exposed to <10 ppm benzene,
excluding controls, inverse associations remained for total WBCs, granulocytes, lymphocytes, B cells,
and platelets. In order to evaluate the effect of past benzene exposures on the hematological effects
observed in this study, the authors compared findings for a group of workers who had been exposed to
<1 ppm benzene over the previous year (n=60) and a subset who also had <40 ppm-years lifetime
cumulative benzene exposure (n=50). The authors stated that the same cell types were significantly
reduced in these groups, but did not provide further information of the magnitude (i.e., percent change) of
BENZENE
A-9
APPENDIX A
the hematological effects observed. These data suggest that the 1-month benzene exposure results could
be used as an indicator of longer term low-level benzene hematotoxicity. To demonstrate that the
observed effects were attributable to benzene, significantly decreased levels of WBCs, granulocytes,
lymphocytes, and B cells were noted in a subgroup (n=30; mean 1-month exposure level of
0.290.15 ppm) of the <1 ppm group for which exposure to other solvents was negligible.
Table A-1. Significantly Reduced Blood Values in Workers Exposed to Benzene

in Tianjin, China (Adapted from Lan et al. 2004a)
End point
Mean exposure level in ppma (number of subjects)

<0.04 (140)
0.570.24 (109) 2.852.11 (110) 28.7320.74 (31)
WBCsb
Granulocytesb
Monocytesb
Lymphocytesb
CD4+-T cellsb
CD4+/CD8+ ratio
B cellsb
Plateletsc
6,4801,710
4,1101,410
24192
2,130577
742262
1.460.58
21894
23059.7
5,5401,220d
3,360948d
21797d
1,960541d
635187d
1.260.41d
18695d
21448.8d
5,6601,500
3,4801,170
22493
1,960533
623177
1.220.45
17075
20053.4
4,770892
2,790750
17974
1,800392
576188
1.090.35
140101
17244.8
Arithmetic mean of an average of two measurements per subject collected during the month prior to phlebotomy
Mean cell numbers per microliter bloodstandard deviation
c
Mean number of platelets (x103)
d
Statistically significantly lower than controls (p<0.05) by linear regression on ln of each end point
b
Effect noted in study and corresponding doses: As shown in Table A-1, exposure-response relationships
were noted for several blood factors. Benzene-induced decreased B cell count was selected as the critical
effect for benchmark dose (BMD) modeling because it represented the highest magnitude of effect (i.e., B
cell count in the highest exposure group was approximately 36% lower than that of controls). A BMD
modeling approach was selected to identify the point of departure because the critical study (Lan et al.
2004a) identified a LOAEL in the absence of a NOAEL.
Dose and end point used for MRL derivation: 0.10 ppm (BMCL0.25sd) for decreased B cell count.
All continuous variable models in the EPA Benchmark Dose Software (Version 1.3.2) were fit to the
B cell count data shown in Table A-1. Visual inspection of the plots of observed versus expected values
for B cell counts indicated that the Hill model provided the only adequate fit of the data set (see
Figure A-1). A benchmark response (BMR) of 0.25 sd below the control mean B cell count was selected
because it resulted in a BMC0.25sd of 0.42 ppm and its lower 95% confidence limit (BMCL0.25sd) of
0.10 ppm (Figure A-1), which are below the mean exposure level of the lowest exposure group
(0.57 ppm) for which a statistically significant decrease in mean B cell count (186 versus 218 in controls,
see Table A-1) was observed. Although Lan et al. (2004a, 2004b) noted significantly decreased levels of
WBCs, granulocytes, lymphocytes, and B cells in a subgroup (n=30; mean 1-month exposure level of
0.290.15 ppm) of the 0.57 ppm exposure group, this subgroup could not be included in the BMD
analysis because the study authors did not include the means and standard deviations for the decreased
blood factors, nor did they provide quantitative information regarding the remaining 70 subjects in the
0.57 ppm exposure group (n=109). Assuming that the 0.29 ppm exposure level may represent a
minimally adverse exposure level, it seems reasonable to accept the BMCL0.25sd of 0.10 ppm as the point
of departure for deriving a chronic-duration inhalation MRL for benzene.
BENZENE
A-10
APPENDIX A
Figure A-1. Observed and Predicted B Cell Counts in Human Subjects
Occupationally Exposed to Benzene. BMD=BMC0.25sd=0.42 ppm;
BMDL=BMCL0.25sd=0.10 ppm
Hill Model with 0.95 Confidence Level

240
Hill
Mean Response
220
200
180
160
140
120
100
BMDL BMD
0
08:53 04/11 2005
10
15
dose
20
25
30
The computer output for fitting of the Hill model to B cell counts in human subjects occupationally
exposed to benzene (Lan et al. 2004a) follows.
====================================================================
Hill Model. $Revision: 2.1 $ $Date: 2000/10/11 21:21:23 $
Input Data File: C:\ATSDR\BENZENE\BMD FILES\BENZENELANBCELLS.(d)
Gnuplot Plotting File: C:\ATSDR\BENZENE\BMD
FILES\BENZENELANBCELLS.plt
Mon Nov 20 09:27:13 2006
====================================================================
BMDS MODEL RUN
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
The form of the response function is:
Y[dose] = intercept + v*dose^n/(k^n + dose^n)
Dependent variable = MEAN
Independent variable = ppm
rho is set to 0
Power parameter restricted to be greater than 1
A constant variance model is fit
Total number of dose groups = 4
Total number of records with missing values = 0
Maximum number of iterations = 250
Relative Function Convergence has been set to: 1e-008
Parameter Convergence has been set to: 1e-008
BENZENE
A-11
APPENDIX A
Default Initial Parameter Values

alpha =
8224.64
rho =
0
Specified
intercept =
218
v =
-78
n =
0.572459
k =
1.5675
Asymptotic Correlation Matrix of Parameter Estimates
( *** The model parameter(s) -n have been estimated at a
boundary point, or have been specified by the user, and do not appear in the
correlation matrix )
alpha
rho
intercept
alpha
rho
intercept
Parameter Estimates
Variable
alpha
rho
intercept
v
n
k
Std. Err.
NA
Estimate
8027.09
0
217.113
-69.0144
1
0.878186
NA - Indicates that this parameter has hit a bound

implied by some inequality constraint and thus
has no standard error.
Table of Data and Estimated Values of Interest

Dose
------
N
---
Obs Mean
--------
0
0.57
2.85
28.73
140
109
110
31
218
186
170
140
Obs Std Dev

-----------
Est Mean
--------
Est Std Dev

-----------
Chi^2 Res.
----------
217
190
164
150
89.6
89.6
89.6
89.6
0.0099
-0.0441
0.063
-0.113
94
95
75
101
Model Descriptions for likelihoods calculated
Model A1:
Yij = Mu(i) + e(ij)
BENZENE
A-12
APPENDIX A
Var{e(ij)} = Sigma^2
Model A2:
Model
R:
Yij = Mu(i) + e(ij)
Var{e(ij)} = Sigma(i)^2
Yi = Mu + e(i)
Var{e(i)} = Sigma^2
Degrees of freedom for Test A1 vs fitted <= 0

Likelihoods of Interest
Model
A1
A2
fitted
R
Test 1:
Test 2:
Test 3:
Log(likelihood)
-1947.632025
-1943.411648
-1948.162584
-1962.157799
DF
5
8
4
2
AIC
3905.264050
3902.823297
3904.325168
3928.315597
Does response and/or variances differ among dose levels

(A2 vs. R)
Are Variances Homogeneous (A1 vs A2)
Does the Model for the Mean Fit (A1 vs. fitted)
Tests of Interest
Test
-2*log(Likelihood Ratio)
Test 1
Test 2
Test 3
37.4923
8.44075
1.06112
Test df
6
3
0
p-value
<.0001
0.03773
NA
The p-value for Test 1 is less than .05. There appears to be a

difference between response and/or variances among the dose levels.
It seems appropriate to model the data
The p-value for Test 2 is less than .05.
non-homogeneous variance model
Consider running a
NA - Degrees of freedom for Test 3 are less than or equal to 0.

Square test for fit is not valid
Benchmark Dose Computation
Specified effect =
Risk Type
The Chi-
0.25
Estimated standard deviations from the control mean
Confidence level =
0.95
BMC =
0.42196
BMCL =
0.104163
Although Test 3 (mean fit) produced an invalid Chi-Square test (degrees of freedom 0), visual inspection
of the observed vs expected B cell counts from the Hill model output (Figure A-1) resulted in the
determination that the predicted B cell counts adequately reflect the observed values and that the
BENZENE
A-13
APPENDIX A
associated BMCL0.25sd of 0.104163 provides an appropriate point of departure for deriving a chronicduration inhalation MRL for benzene.
[ ] NOAEL [ ] LOAEL
[ ] 1 [ ] 3 [ ] 10 (for use of a LOAEL)
[ ] 1 [ ] 3 [ ] 10 (for extrapolation from animals to humans)
If an inhalation study in animals, list conversion factors used in determining human equivalent dose: Not
applicable.
Was a conversion used from intermittent to continuous exposure? The BMCL0.25sd of 0.10 ppm was
adjusted from the 8-hour TWA to a continuous exposure concentration (BMCL0.25sdADJ) as follows:
BMCL0.25sdADJ = BMCL0.25sd x (8 hours/24 hours) x (6 days/7 days)
Therefore:
BMCL0.25sdADJ = 0.10 ppm x (8 hours/24 hours) x (6 days/7 days)
BMCL0.25sdADJ = 0.03 ppm
Other additional studies or pertinent information that lend support to this MRL: Lan et al. (2004a, 2004b)
was selected as the critical study for derivation of a chronic-duration inhalation MRL because it (1) was
well designed, (2) provided adequate exposure-response information, (3) employed individual exposure
monitoring data collected for up to 16 months prior to blood testing, (4) demonstrated effects that did not
appear to be significantly influenced by previous high-level exposures, and (5) included larger numbers of
subjects than previous studies (Qu et al. 2002, 2003; Rothman et al. 1996a, 1996b; Ward et al. 1996). In
addition, Lan et al. (2004a, 2004b) measured lymphocyte subsets and colony formation from
hematopoietic progenitor cells as measures of toxicity.
Previously conducted epidemiology studies provide support to the findings of Lan et al. (2004a). Qu et
al. (2002, 2003) compared hematologic values among 105 healthy workers (51 men, 54 women) in
industries with a history of benzene usage (Tianjin, China) and 26 age- and gender-matched workers in
industries that did not use benzene. Benzene-exposed workers were chosen based on at least 3 years of
exposure history. The mean duration of occupational exposure to benzene was 9.7 years (SD=6.2 years).
At the time of the study, benzene exposure was monitored by individual organic vapor monitors at
1-week intervals for 4 weeks prior to collection of blood samples for analysis. Measured benzene levels
were averaged for each individual to produce a 4-week mean exposure level. Exposure-response
relationships were assessed according to ranges of benzene levels (unexposed, >05, >515, >1530, and
>30 ppm). Benzene hematotoxicity was assessed by mechanical counts of total WBCs, red blood cells
(RBCs), and platelets. The WBC differential was hand-counted on a total of 900 cells. Calculations of
the numbers of various WBC types were based on total WBCs and differential counts. The mean 4-week
benzene level in the control group was 0.0040.003 ppm. Among all the benzene-exposed workers, the
mean 4-week benzene exposure level was 5.27.3 ppm. Within the >05, >515, >1530, and >30 ppm
exposure categories, mean 4-week benzene levels were 2.261.35, 8.672.44, 19.93.1, and
BENZENE
A-14
APPENDIX A
51.843.3 ppm, respectively. A significant exposure-related reduction in the numbers of neutrophils

(ranging in magnitude from 12% in the 2.26 ppm exposure group to 31% in the 51.8 ppm exposure
group) was observed in all four groups of benzene-exposed workers, relative to controls. Significantly
reduced numbers of RBCs (approximately 1116% lower than controls) were also noted in all benzeneexposed groups. Significantly reduced total WBCs were seen in the highest (>30 ppm) exposure group.
The study authors identified a subgroup (within the >05 ppm exposure group) of 16 women with no
measured exposure levels exceeding 0.5 ppm (4-week mean benzene exposure level of 0.140.04 ppm)
and reported significantly reduced total WBCs, neutrophils, and RBCs in this subgroup as well.
However, these results are based on a small number of workers within the larger group and the reduced
counts of total WBCs, neutrophils, and RBCs within this subgroup are much greater in magnitude than
those reported for the main (>05 ppm) exposure group, rendering the results in this subgroup of
questionable value for purposes of risk assessment. Qu et al. (2002, 2003) clearly identified a LOAEL of
2.26 ppm for significantly reduced total WBCs, neutrophils, and RBCs, and provided indication of
benzene-induced changes in some hematological values at exposure levels lower than the current industry
8-hour TWA of 1 ppm.
Rothman et al. (1996a, 1996b) performed a cross-sectional study in 1992 on 44 healthy workers
(23 males, 21 females) in Chinese (Shanghai) industries with a history of benzene usage and 44 age- and
gender-matched workers in industries that did not use benzene. The mean duration of occupational
exposure to benzene was 6.3 years (SD=4.4 years). At the time of the study, benzene exposure was
monitored by individual organic passive dosimetry badges on 5 separate days during 1 to 2 weeks prior to
the collection of blood and urine samples for analysis. Benzene hematotoxicity was assessed by
mechanical counts of total WBCs, absolute lymphocytes (ALC), RBCs, and platelets, as well as
hemoglobin value and mean corpuscular volume (MCV). The WBC differential was also hand-counted
on 100 cells. Abnormal counts were reviewed by hand. Mean (geometric mean of the five exposure
samples) 8-hour TWAs for the benzene-exposed workers ranged from 1 to 238 ppm (median 8-hour
TWA of 31 ppm). Benzene-exposed workers exhibited statistically significantly reduced numbers of total
WBCs, ALC, RBCs, and platelets (approximately 12, 21, 6, and 23% lower, respectively) and
significantly increased MCV (approximately 3% higher), relative to unexposed workers. The results were
comparable in both men and women. Among the benzene-exposed workers whose mean exposure levels
were >31 ppm (median 8-hour TWA of 91.9 ppm; n=22), all measured blood parameters were
significantly different from controls; only ALC, RBCs, and platelets were significantly lower in benzene
workers with mean exposures of <31 ppm (median 8-hour TWA of 13.6; n=22), compared with controls.
In a subgroup of benzene-exposed workers whose measured benzene exposure levels did not exceed
31 ppm on any of the five sampling days (median 8hour TWA of 7.6 ppm; n=11), significantly reduced
ALC (approximately 16% lower than controls) was noted.
In a nested case-control study of a cohort of workers in the Pliofilm production departments of a rubber
products manufacturer in Ohio (Ward et al. 1996), incident cases were defined as the first occurrence of a
low WBC or RBC count, and matched controls were chosen from those tested within approximately
6 months of the cases blood test date. Hematologic screening data were available for 657 of
1,037 individuals employed at the plant from 1939 through 1976. A total of 21,710 blood test records
were identified; the number of blood tests per individual ranged from 1 to 354, but the majority of
subjects had five or fewer blood tests. All blood tests were taken from 1940 through 1975, the majority
of which were routine hematological screening tests. Benzene exposures were estimated using a job
exposure matrix developed by Rinsky et al. (1987). The effects of benzene exposure in the 30, 90, and
180 days prior to the blood test date, as well as cumulative exposure up until the blood test date, were
examined using conditional logistic regression. A total of 78 cases and 5,637 controls were included in
the WBC analysis and 105 cases and 8,489 controls in the RBC analysis, all of whom had worked only
within the rubber hydrochloride departments during the 180 days prior to the selected blood sample date.
The maximum daily benzene exposure estimate was 34 ppm. A strong exposure-response relationship
BENZENE
A-15
APPENDIX A
was noted for WBCs, and all of the exposure metrics selected showed a significant relationship with low
blood count. A weak positive exposure-response relationship was observed for RBCs, which was
significant for the dose metric of cumulative exposure up until the blood test date. The study authors
noted that there was no evidence for a threshold for hematologic effects and suggested that exposure to
benzene levels <5 ppm may result in hematologic suppression.
Ph.D.
BENZENE
A-16
APPENDIX A

Chemical Name:
CAS Numbers:
Date:
Profile Status:
Route:
Duration:
Graph Key:
Species:
Benzene
71-43-2
August 2007
[ ] Inhalation [x] Oral
[ ] Acute [ ] Intermediate [x] Chronic
45
Human
Minimal Risk Level: 0.0005 [x] mg/kg/day [ ] ppm

Reference: Lan Q, Zhang L, Li G, et al. 2004a. Hematotoxicity in workers exposed to low levels of
benzene. Science 306:1774-1776.
Experimental design: The chronic-duration oral MRL for benzene is based on route-to-route
extrapolation of the results of benchmark dose analysis of a hematological endpoint (B cell count)
assessed in 250 workers (approximately two-thirds female) exposed to benzene at two shoe
manufacturing facilities in Tianjin, China, and 140 age- and gender-matched workers in clothing
manufacturing facilities that did not use benzene. See the MRL worksheet for the chronic-duration
inhalation MRL for details of study design.
Effect noted in study and corresponding doses: As described in the MRL worksheet for the chronicduration inhalation MRL (see also Table A-1), exposure-response relationships were noted for several
blood factors. Benzene-induced decreased B cell count was selected as the critical effect for benchmark
dose (BMD) modeling because it represented the highest magnitude of effect (i.e., B cell count in the
highest exposure group was approximately 36% lower than that of controls). A BMD modeling approach
was selected to identify the point of departure because the critical study (Lan et al. 2004a) identified a
LOAEL in the absence of a NOAEL.
Dose and end point used for MRL derivation: BMCL0.25sdADJ of 0.014 mg/kg/day for decreased B cell
count, resulting from route-to-route extrapolation of the BMCL0.25sdADJ of 0.03 ppm described in the MRL
worksheet for the chronic-duration inhalation MRL.
Results of toxicokinetic studies of inhaled benzene in humans (Nomiyama and Nomiyama 1974a; Pekari
et al. 1992; Srbova et al. 1950) and inhaled and orally-administered benzene in rats and mice (Sabourin et
al. 1987) indicate that absorption of benzene at relatively low levels of exposure is approximately 50% of
an inhaled dose and essentially 100% of an oral dose. Based on these assumptions, inhalation data can be
used to estimate equivalent oral doses that would be expected to similarly affect the critical targets of
benzene toxicity. Therefore, the point of departure for the chronic-duration inhalation MRL for benzene,
namely the BMCL0.25sdADJ of 0.03 ppm for decreased B cell counts in benzene-exposed workers (Lan et al.
2004a, 2004b), serves as the point of departure for deriving the chronic-duration oral MRL as well.
The point of departure (in ppm) was converted to mg/m3 using the molecular weight of 78.11 for benzene
and assuming 25 C and 760 mm Hg:
BMCL0.25sdADJ of 0.03 ppm x 78.11/24.45 = 0.096 mg/m3
The BMCL0.25sdADJ of 0.096 mg/m3 for inhaled benzene was converted to an equivalent BMDL0.25sdADJ for
ingested benzene using EPA (1988b) human reference values for inhalation rate (20 m3/day) and body
BENZENE
A-17
APPENDIX A
weight (70 kg) and a factor of 0.5 to adjust for differences in absorption of benzene following inhalation
versus oral exposure (50 versus 100%, respectively) as follows:
BMDL0.25sdADJ = BMCL0.25sdADJ of 0.096 mg/m3 x 20 m3/day x 0.5 70 kg = 0.014 mg/kg/day
[ ] NOAEL [ ] LOAEL
[]1
[]1
[]1
[]1
[ ] 3 [ ] 10 (for use of a LOAEL)
[ ] 3 [ ] 10 (for extrapolation from animals to humans)
[ ] 3 [x] 10 (for human variability)
[x] 3 [ ] 10 (for uncertainty in route-to-route extrapolation)
If an inhalation study in animals, list conversion factors used in determining human equivalent dose: Not
applicable.
Was a conversion used from intermittent to continuous exposure? The BMCL0.25sd of 0.10 ppm was
adjusted from the 8-hour TWA to a continuous exposure concentration (BMCL0.25sdADJ) as follows:
BMCL0.25sdADJ = BMCL0.25sd x (8 hours/24 hours) x (6 days/7 days)
Therefore:
BMCL0.25sdADJ = 0.10 ppm x (8 hours/24 hours) x (6 days/7 days)
BMCL0.25sdADJ = 0.03 ppm
Other additional studies or pertinent information that lend support to this MRL: Results of toxicokinetic
studies of inhaled benzene in humans (Nomiyama and Nomiyama 1974a; Pekari et al. 1992; Srbova et al.
1950) and inhaled and orally-administered benzene in rats and mice (Sabourin et al. 1987) indicate that
absorption of benzene at relatively low levels of exposure is approximately 50% of an inhaled dose and
essentially 100% of an oral dose. Based on these assumptions, inhalation data can be used to estimate
equivalent oral doses that would be expected to similarly affect the critical targets of benzene toxicity.
See the chronic-duration inhalation MRL worksheet for additional information that supports the selection
of the principal study and critical effect for deriving the chronic-duration inhalation MRL.
Ph.D.
BENZENE
A-18
APPENDIX A
BENZENE
B-1
APPENDIX B. USER'S GUIDE

Chapter 1
Public Health Statement
This chapter of the profile is a health effects summary written in non-technical language. Its intended
audience is the general public, especially people living in the vicinity of a hazardous waste site or
chemical release. If the Public Health Statement were removed from the rest of the document, it would
still communicate to the lay public essential information about the chemical.
The major headings in the Public Health Statement are useful to find specific topics of concern. The
topics are written in a question and answer format. The answer to each question includes a sentence that
will direct the reader to chapters in the profile that will provide more information on the given topic.
Chapter 2
Relevance to Public Health
This chapter provides a health effects summary based on evaluations of existing toxicologic,
epidemiologic, and toxicokinetic information. This summary is designed to present interpretive, weightof-evidence discussions for human health end points by addressing the following questions:
1. What effects are known to occur in humans?
2. What effects observed in animals are likely to be of concern to humans?
3. What exposure conditions are likely to be of concern to humans, especially around hazardous
waste sites?
The chapter covers end points in the same order that they appear within the Discussion of Health Effects
by Route of Exposure section, by route (inhalation, oral, and dermal) and within route by effect. Human
data are presented first, then animal data. Both are organized by duration (acute, intermediate, chronic).
In vitro data and data from parenteral routes (intramuscular, intravenous, subcutaneous, etc.) are also
considered in this chapter.
The carcinogenic potential of the profiled substance is qualitatively evaluated, when appropriate, using
existing toxicokinetic, genotoxic, and carcinogenic data. ATSDR does not currently assess cancer
potency or perform cancer risk assessments. Minimal Risk Levels (MRLs) for noncancer end points (if
derived) and the end points from which they were derived are indicated and discussed.
Limitations to existing scientific literature that prevent a satisfactory evaluation of the relevance to public
health are identified in the Chapter 3 Data Needs section.
Interpretation of Minimal Risk Levels
Where sufficient toxicologic information is available, ATSDR has derived MRLs for inhalation and oral
routes of entry at each duration of exposure (acute, intermediate, and chronic). These MRLs are not
meant to support regulatory action, but to acquaint health professionals with exposure levels at which
adverse health effects are not expected to occur in humans.
BENZENE
B-2
APPENDIX B
MRLs should help physicians and public health officials determine the safety of a community living near
a chemical emission, given the concentration of a contaminant in air or the estimated daily dose in water.
MRLs are based largely on toxicological studies in animals and on reports of human occupational
exposure.
MRL users should be familiar with the toxicologic information on which the number is based. Chapter 2,
"Relevance to Public Health," contains basic information known about the substance. Other sections such
as Chapter 3 Section 3.9, "Interactions with Other Substances, and Section 3.10, "Populations that are
Unusually Susceptible" provide important supplemental information.
MRL users should also understand the MRL derivation methodology. MRLs are derived using a
modified version of the risk assessment methodology that the Environmental Protection Agency (EPA)
provides (Barnes and Dourson 1988) to determine reference doses (RfDs) for lifetime exposure.
To derive an MRL, ATSDR generally selects the most sensitive end point which, in its best judgement,
represents the most sensitive human health effect for a given exposure route and duration. ATSDR
cannot make this judgement or derive an MRL unless information (quantitative or qualitative) is available
for all potential systemic, neurological, and developmental effects. If this information and reliable
quantitative data on the chosen end point are available, ATSDR derives an MRL using the most sensitive
species (when information from multiple species is available) with the highest no-observed-adverse-effect
level (NOAEL) that does not exceed any adverse effect levels. When a NOAEL is not available, a
lowest-observed-adverse-effect level (LOAEL) can be used to derive an MRL, and an uncertainty factor
(UF) of 10 must be employed. Additional uncertainty factors of 10 must be used both for human
variability to protect sensitive subpopulations (people who are most susceptible to the health effects
caused by the substance) and for interspecies variability (extrapolation from animals to humans). In
deriving an MRL, these individual uncertainty factors are multiplied together. The product is then
divided into the inhalation concentration or oral dosage selected from the study. Uncertainty factors used
in developing a substance-specific MRL are provided in the footnotes of the levels of significant exposure
(LSE) tables.
Chapter 3
Health Effects
Tables and Figures for Levels of Significant Exposure (LSE)
Tables and figures are used to summarize health effects and illustrate graphically levels of exposure
associated with those effects. These levels cover health effects observed at increasing dose
concentrations and durations, differences in response by species, MRLs to humans for noncancer end
points, and EPA's estimated range associated with an upper- bound individual lifetime cancer risk of 1 in
10,000 to 1 in 10,000,000. Use the LSE tables and figures for a quick review of the health effects and to
locate data for a specific exposure scenario. The LSE tables and figures should always be used in
conjunction with the text. All entries in these tables and figures represent studies that provide reliable,
quantitative estimates of NOAELs, LOAELs, or Cancer Effect Levels (CELs).
The legends presented below demonstrate the application of these tables and figures. Representative
examples of LSE Table 3-1 and Figure 3-1 are shown. The numbers in the left column of the legends
correspond to the numbers in the example table and figure.
BENZENE
B-3
APPENDIX B
LEGEND
See Sample LSE Table 3-1 (page B-6)
(1)
Route of Exposure. One of the first considerations when reviewing the toxicity of a substance
using these tables and figures should be the relevant and appropriate route of exposure. Typically
when sufficient data exist, three LSE tables and two LSE figures are presented in the document.
The three LSE tables present data on the three principal routes of exposure, i.e., inhalation, oral,
and dermal (LSE Tables 3-1, 3-2, and 3-3, respectively). LSE figures are limited to the inhalation
(LSE Figure 3-1) and oral (LSE Figure 3-2) routes. Not all substances will have data on each
route of exposure and will not, therefore, have all five of the tables and figures.
(2)
Exposure Period. Three exposure periodsacute (less than 15 days), intermediate (15
364 days), and chronic (365 days or more)are presented within each relevant route of exposure.
In this example, an inhalation study of intermediate exposure duration is reported. For quick
reference to health effects occurring from a known length of exposure, locate the applicable
exposure period within the LSE table and figure.
(3)
Health Effect. The major categories of health effects included in LSE tables and figures are
death, systemic, immunological, neurological, developmental, reproductive, and cancer.
NOAELs and LOAELs can be reported in the tables and figures for all effects but cancer.
Systemic effects are further defined in the "System" column of the LSE table (see key number
18).
(4)
Key to Figure. Each key number in the LSE table links study information to one or more data
points using the same key number in the corresponding LSE figure. In this example, the study
represented by key number 18 has been used to derive a NOAEL and a Less Serious LOAEL
(also see the two "18r" data points in sample Figure 3-1).
(5)
Species. The test species, whether animal or human, are identified in this column. Chapter 2,
"Relevance to Public Health," covers the relevance of animal data to human toxicity and
Section 3.4, "Toxicokinetics," contains any available information on comparative toxicokinetics.
Although NOAELs and LOAELs are species specific, the levels are extrapolated to equivalent
human doses to derive an MRL.
(6)
Exposure Frequency/Duration. The duration of the study and the weekly and daily exposure
regimens are provided in this column. This permits comparison of NOAELs and LOAELs from
different studies. In this case (key number 18), rats were exposed to Chemical x via inhalation
for 6 hours/day, 5 days/week, for 13 weeks. For a more complete review of the dosing regimen,
refer to the appropriate sections of the text or the original reference paper (i.e., Nitschke et al.
1981).
(7)
System. This column further defines the systemic effects. These systems include respiratory,
cardiovascular, gastrointestinal, hematological, musculoskeletal, hepatic, renal, and
dermal/ocular. "Other" refers to any systemic effect (e.g., a decrease in body weight) not covered
in these systems. In the example of key number 18, one systemic effect (respiratory) was
investigated.
(8)
NOAEL. A NOAEL is the highest exposure level at which no harmful effects were seen in the
organ system studied. Key number 18 reports a NOAEL of 3 ppm for the respiratory system,
which was used to derive an intermediate exposure, inhalation MRL of 0.005 ppm (see
footnote "b").
BENZENE
B-4
APPENDIX B
(9)
LOAEL. A LOAEL is the lowest dose used in the study that caused a harmful health effect.
LOAELs have been classified into "Less Serious" and "Serious" effects. These distinctions help
readers identify the levels of exposure at which adverse health effects first appear and the
gradation of effects with increasing dose. A brief description of the specific end point used to
quantify the adverse effect accompanies the LOAEL. The respiratory effect reported in key
number 18 (hyperplasia) is a Less Serious LOAEL of 10 ppm. MRLs are not derived from
Serious LOAELs.
(10)
Reference. The complete reference citation is given in Chapter 9 of the profile.
(11)
CEL. A CEL is the lowest exposure level associated with the onset of carcinogenesis in
experimental or epidemiologic studies. CELs are always considered serious effects. The LSE
tables and figures do not contain NOAELs for cancer, but the text may report doses not causing
measurable cancer increases.
(12)
Footnotes. Explanations of abbreviations or reference notes for data in the LSE tables are found
in the footnotes. Footnote "b" indicates that the NOAEL of 3 ppm in key number 18 was used to
derive an MRL of 0.005 ppm.
LEGEND
See Sample Figure 3-1 (page B-7)
LSE figures graphically illustrate the data presented in the corresponding LSE tables. Figures help the
reader quickly compare health effects according to exposure concentrations for particular exposure
periods.
(13)
Exposure Period. The same exposure periods appear as in the LSE table. In this example, health
effects observed within the acute and intermediate exposure periods are illustrated.
(14)
Health Effect. These are the categories of health effects for which reliable quantitative data
exists. The same health effects appear in the LSE table.
(15)
Levels of Exposure. Concentrations or doses for each health effect in the LSE tables are
graphically displayed in the LSE figures. Exposure concentration or dose is measured on the log
scale "y" axis. Inhalation exposure is reported in mg/m3 or ppm and oral exposure is reported in
mg/kg/day.
(16)
N
OAEL. In this example, the open circle designated 18r identifies a NOAEL critical end point in
the rat upon which an intermediate inhalation exposure MRL is based. The key number 18
corresponds to the entry in the LSE table. The dashed descending arrow indicates the
extrapolation from the exposure level of 3 ppm (see entry 18 in the table) to the MRL of
0.005 ppm (see footnote "b" in the LSE table).
(17)
CEL. Key number 38m is one of three studies for which CELs were derived. The diamond
symbol refers to a CEL for the test species-mouse. The number 38 corresponds to the entry in the
LSE table.
BENZENE
B-5
APPENDIX B
(18)
Estimated Upper-Bound Human Cancer Risk Levels. This is the range associated with the upperbound for lifetime cancer risk of 1 in 10,000 to 1 in 10,000,000. These risk levels are derived
from the EPA's Human Health Assessment Group's upper-bound estimates of the slope of the
cancer dose response curve at low dose levels (q1*).
(19)
Key to LSE Figure. The Key explains the abbreviations and symbols used in the figure.
BENZENE
SAMPLE
1
Table 3-1. Levels of Significant Exposure to [Chemical x] Inhalation
Key to
figurea
2
Exposure
NOAEL
frequency/
Species duration
System (ppm)
Serious (ppm)
Reference
Systemic
18
Rat
13 wk
5 d/wk
6 hr/d
Resp
3b
10
10 (hyperplasia)
Nitschke et al. 1981
APPENDIX B
Less serious
(ppm)
5
LOAEL (effect)
CHRONIC EXPOSURE
11
Cancer
12
38
Rat
18 mo
5 d/wk
7 hr/d
20
(CEL, multiple
organs)
Wong et al. 1982
39
Rat
89104 wk
5 d/wk
6 hr/d
10
(CEL, lung tumors,

nasal tumors)
NTP 1982
40
Mouse
79103 wk
5 d/wk
6 hr/d
10
(CEL, lung tumors,

hemangiosarcomas)
NTP 1982
The number corresponds to entries in Figure 3-1.

Used to derive an intermediate inhalation Minimal Risk Level (MRL) of 5x10-3 ppm; dose adjusted for intermittent exposure and divided
by an uncertainty factor of 100 (10 for extrapolation from animal to humans, 10 for human variability).
b
B-6
BENZENE
SAMPLE
Figure 3-1. Levels of Significant Exposure to [Chemical X] - Inhalation

Acute ( :5;14 days)

Systemic
Systemic
E)-------+
0 -.ppm - . - -Q:v=e~.;s_ _
1 0000
<l 17 h
1000100
()15h
()14r
30r
10 .
16
037 h
e 32r
31r
() 35h
()33h
38m
+--- - - - -
~9m + 40 m
@]
17
10.1
APPENDIX B
11r
12 r
0.01
4-[!!J
0 .001
Estimated
Upper-Bound
Human Cancer
Risk Levels
0 .0001
0 .000010 .0000010 .0000001
ooses represent the lowest dose tested per study that produced a tumorigenic
response and do not imply the existence of a threshold for the cancer end point.
k-Monkey
g-Guinea P ig
r-Rat
h-Rabbit
m.Mouse
Cancer Effect
Leve~Ani mals
. LO AEL, More Serious- Animals

()LOA EL , Less Serious- A nimals
0-JOA EL - Anima ls
M inimal Risk Level

tor effects
.:. o ther than
Cancer
....-----~
B-7
BENZENE
B-8
APPENDIX B
BENZENE
C-1
APPENDIX C. ACRONYMS, ABBREVIATIONS, AND SYMBOLS
ACGIH
ACOEM
ADI
ADME
AED
AFID
AFOSH
ALT
AML
AOAC
AOEC
AP
APHA
AST
atm
ATSDR
AWQC
BAT
BCF
BEI
BMD
BMR
BSC
C
CAA
CAG
CAS
CDC
CEL
CELDS
CERCLA
CFR
Ci
CI
CL
CLP
cm
CML
CPSC
CWA
DHEW
DHHS
DNA
DOD
DOE
DOL
DOT
DOT/UN/
NA/IMCO
American Conference of Governmental Industrial Hygienists

American College of Occupational and Environmental Medicine
acceptable daily intake
absorption, distribution, metabolism, and excretion
atomic emission detection
alkali flame ionization detector
Air Force Office of Safety and Health
alanine aminotransferase
acute myeloid leukemia
Association of Official Analytical Chemists
Association of Occupational and Environmental Clinics
alkaline phosphatase
American Public Health Association
aspartate aminotransferase
atmosphere
Ambient Water Quality Criteria
best available technology
bioconcentration factor
Biological Exposure Index
benchmark dose
benchmark response
Board of Scientific Counselors
centigrade
Clean Air Act
Cancer Assessment Group of the U.S. Environmental Protection Agency
Chemical Abstract Services
Centers for Disease Control and Prevention
cancer effect level
Computer-Environmental Legislative Data System
Comprehensive Environmental Response, Compensation, and Liability Act
Code of Federal Regulations
curie
confidence interval
ceiling limit value
Contract Laboratory Program
centimeter
chronic myeloid leukemia
Consumer Products Safety Commission
Clean Water Act
Department of Health, Education, and Welfare
Department of Health and Human Services
deoxyribonucleic acid
Department of Defense
Department of Energy
Department of Labor
Department of Transportation
Department of Transportation/United Nations/
North America/Intergovernmental Maritime Dangerous Goods Code
BENZENE
C-2
APPENDIX C
DWEL
ECD
ECG/EKG
EEG
EEGL
EPA
F
F1
FAO
FDA
FEMA
FIFRA
FPD
fpm
FR
FSH
g
GC
gd
GLC
GPC
HPLC
HRGC
HSDB
IARC
IDLH
ILO
IRIS
Kd
kg
kkg
Koc
Kow
L
LC
LC50
LCLo
LD50
LDLo
LDH
LH
LOAEL
LSE
LT50
m
MA
MAL
mCi
MCL
MCLG
MF
drinking water exposure level

electron capture detection
electrocardiogram
electroencephalogram
Emergency Exposure Guidance Level
Environmental Protection Agency
Fahrenheit
first-filial generation
Food and Agricultural Organization of the United Nations
Food and Drug Administration
Federal Emergency Management Agency
Federal Insecticide, Fungicide, and Rodenticide Act
flame photometric detection
feet per minute
Federal Register
follicle stimulating hormone
gram
gas chromatography
gestational day
gas liquid chromatography
gel permeation chromatography
high-performance liquid chromatography
high resolution gas chromatography
Hazardous Substance Data Bank
International Agency for Research on Cancer
immediately dangerous to life and health
International Labor Organization
Integrated Risk Information System
adsorption ratio
kilogram
metric ton
organic carbon partition coefficient
octanol-water partition coefficient
liter
liquid chromatography
lethal concentration, 50% kill
lethal concentration, low
lethal dose, 50% kill
lethal dose, low
lactic dehydrogenase
luteinizing hormone
lowest-observed-adverse-effect level
Levels of Significant Exposure
lethal time, 50% kill
meter
trans,trans-muconic acid
maximum allowable level
millicurie
maximum contaminant level
maximum contaminant level goal
modifying factor
BENZENE
C-3
APPENDIX C
MFO
mg
mL
mm
mmHg
mmol
mppcf
MRL
MS
NAAQS
NAS
NATICH
NATO
NCE
NCEH
NCI
ND
NFPA
ng
NHANES
NIEHS
NIOSH
NIOSHTIC
NLM
nm
nmol
NOAEL
NOES
NOHS
NPD
NPDES
NPL
NR
NRC
NS
NSPS
NTIS
NTP
ODW
OERR
OHM/TADS
OPP
OPPT
OPPTS
OR
OSHA
OSW
OTS
OW
OWRS
PAH
mixed function oxidase

milligram
milliliter
millimeter
millimeters of mercury
millimole
millions of particles per cubic foot
Minimal Risk Level
mass spectrometry
National Ambient Air Quality Standard
National Academy of Science
National Air Toxics Information Clearinghouse
North Atlantic Treaty Organization
normochromatic erythrocytes
National Center for Environmental Health
National Cancer Institute
not detected
National Fire Protection Association
nanogram
National Health and Nutrition Examination Survey
National Institute of Environmental Health Sciences
National Institute for Occupational Safety and Health
NIOSH's Computerized Information Retrieval System
National Library of Medicine
nanometer
nanomole
no-observed-adverse-effect level
National Occupational Exposure Survey
National Occupational Hazard Survey
nitrogen phosphorus detection
National Pollutant Discharge Elimination System
National Priorities List
not reported
National Research Council
not specified
New Source Performance Standards
National Technical Information Service
National Toxicology Program
Office of Drinking Water, EPA
Office of Emergency and Remedial Response, EPA
Oil and Hazardous Materials/Technical Assistance Data System
Office of Pesticide Programs, EPA
Office of Pollution Prevention and Toxics, EPA
Office of Prevention, Pesticides and Toxic Substances, EPA
odds ratio
Occupational Safety and Health Administration
Office of Solid Waste, EPA
Office of Toxic Substances
Office of Water
Office of Water Regulations and Standards, EPA
polycyclic aromatic hydrocarbon
BENZENE
C-4
APPENDIX C
PBPD
PBPK
PCE
PEL
pg
PHS
PID
pmol
PMR
ppb
ppm
ppt
PSNS
RBC
REL
RfC
RfD
RNA
RQ
RTECS
SARA
SCE
SGOT
SGPT
SIC
SIM
SMCL
SMR
SNARL
SPEGL
STEL
STORET
TD50
TLV
TOC
TPQ
TRI
TSCA
TWA
UF
U.S.
USDA
USGS
VOC
WBC
WHO
physiologically based pharmacodynamic

physiologically based pharmacokinetic
polychromatic erythrocytes
permissible exposure limit
picogram
Public Health Service
photo ionization detector
picomole
proportionate mortality ratio
parts per billion
parts per million
parts per trillion
pretreatment standards for new sources
red blood cell
recommended exposure level/limit
reference concentration
reference dose
ribonucleic acid
reportable quantity
Registry of Toxic Effects of Chemical Substances
Superfund Amendments and Reauthorization Act
sister chromatid exchange
serum glutamic oxaloacetic transaminase
serum glutamic pyruvic transaminase
standard industrial classification
selected ion monitoring
secondary maximum contaminant level
standardized mortality ratio
suggested no adverse response level
Short-Term Public Emergency Guidance Level
short term exposure limit
Storage and Retrieval
toxic dose, 50% specific toxic effect
threshold limit value
total organic carbon
threshold planning quantity
Toxics Release Inventory
Toxic Substances Control Act
time-weighted average
uncertainty factor
United States
United States Department of Agriculture
United States Geological Survey
volatile organic compound
white blood cell
World Health Organization
BENZENE
C-5
APPENDIX C
>
=
<
m
g
q1*
+
(+)
()
greater than
greater than or equal to
equal to
less than
less than or equal to
percent
alpha
beta
gamma
delta
micrometer
microgram
cancer slope factor
negative
positive
weakly positive result
weakly negative result
BENZENE
C-6
APPENDIX C
BENZENE
D-1
APPENDIX D. INDEX
absorbed dose.................................................................................................... 155, 160, 162, 176, 179, 209
acetylcholine ............................................................................................................................................. 134
acetylcholinesterase .............................................................................................................................. 20, 91
adipose tissue ............................................................................................................................................ 165
adrenal gland............................................................................................................................................. 129
adrenals ....................................................................................................................................................... 73
adsorbed ............................................................................................................................ 159, 165, 179, 291
adsorption.................................................................................................................. 162, 241, 258, 292, 300
aerobic....................................................................................................................... 251, 262, 263, 264, 285
ambient air .......................................................................... 20, 157, 161, 268, 274, 278, 283, 295, 296, 300
anaerobic ................................................................................................................................... 263, 264, 265
anemia ..........................................5, 12, 13, 17, 19, 73, 74, 77, 78, 79, 86, 90, 127, 213, 215, 221, 224, 229
bioaccumulation........................................................................................................................................ 259
bioavailability ................................................................................................................... 159, 162, 165, 286
bioconcentration factor ............................................................................................................................. 259
biodegradation................................................................................................... 249, 250, 251, 262, 263, 264
biokinetics ......................................................................................................................................... 182, 192
biomarker .................12, 74, 85, 149, 170, 175, 189, 208, 209, 210, 211, 212, 231, 232, 233, 291, 303, 305
blood cell count................................................................................................................... 85, 141, 212, 303
body weight effects ............................................................................................... 32, 83, 105, 130, 131, 138
bone marrow aplasia ................................................................................................................................. 213
breast milk..........................................176, 177, 179, 182, 185, 186, 192, 193, 207, 235, 271, 281, 282, 291
cancer ........................................................................... 4, 6, 9, 16, 30, 31, 98, 101, 103, 137, 140, 166, 207,
208, 219, 221, 223, 224, 230, 273, 289, 305, 310, 311
carcinogen ..................................................6, 15, 16, 103, 136, 137, 223, 224, 248, 249, 289, 305, 310, 311
carcinogenic ...................................................................................... 6, 16, 21, 29, 30, 86, 98, 103, 310, 311
carcinogenicity.................................................12, 15, 16, 103, 136, 137, 141, 224, 225, 229, 233, 235, 310
carcinoma.................................................................................................................................... 15, 103, 136
cardiovascular ....................................................................................................................... 13, 73, 105, 138
cardiovascular effects.................................................................................................................... 13, 73, 105
chromosomal aberrations .................................................................................... 94, 141, 150, 212, 226, 303
clearance ................................................................................................................................................... 212
death.....5, 13, 14, 27, 29, 31, 73, 78, 82, 89, 98, 99, 104, 105, 128, 131, 137, 140, 163, 219, 221, 287, 310
deoxyribonucleic acid (see DNA)..................................................................................................... 145, 148
dermal effects........................................................................................................................ 32, 83, 130, 138
developmental effects ................................................................................... 94, 97, 135, 136, 140, 227, 235
DNA (see deoxyribonucleic acid).....142, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 168, 174,
201, 202, 203, 204, 209, 210, 212, 224, 225, 230, 232, 237, 303
dopamine................................................................................................................................................... 134
endocrine..................................................................................................................... 32, 105, 138, 205, 206
endocrine effects ....................................................................................................................................... 129
erythema.................................................................................................................................................... 138
fetus............................................................................................... 6, 7, 14, 93, 143, 151, 206, 208, 226, 227
gastrointestinal effects ........................................................................................................................ 73, 127
general population..................................3, 5, 11, 79, 208, 251, 253, 273, 276, 278, 281, 286, 287, 302, 310
genotoxic....................................................................................... 12, 29, 141, 152, 154, 204, 212, 219, 221
genotoxicity............................................................... 150, 151, 152, 168, 202, 215, 218, 224, 225, 237, 305
germinal epithelium .................................................................................................................................... 94
groundwater .......................................................229, 249, 250, 257, 258, 262, 263, 264, 269, 270, 285, 289
BENZENE
D-2
APPENDIX D
growth retardation....................................................................................................................................... 95
half-life.............................................................................................................. 159, 175, 176, 208, 261, 262
hematological effects ...........16, 17, 18, 25, 73, 75, 76, 77, 79, 127, 138, 212, 213, 221, 222, 223, 226, 230
hematopoietic....................................................................... 24, 73, 80, 85, 96, 97, 133, 136, 140, 155, 172,
200, 201, 208, 218, 221, 222, 227, 231, 233
hepatic effects ............................................................................................................................... 13, 82, 128
hydrolysis.......................................................................................................................... 194, 292, 293, 294
hydroxyl radical ................................................................................................................ 171, 233, 251, 261
immune system ........................................................................................... 6, 12, 20, 88, 221, 222, 227, 231
immunological ...................................................................... 18, 19, 22, 27, 29, 89, 131, 140, 222, 227, 307
immunological effects......................................................................... 18, 19, 22, 27, 89, 131, 222, 227, 307
Kow ............................................................................................................................................................ 259
LD50................................................................................................................................................... 104, 105
leukemia......................................................6, 9, 12, 14, 16, 17, 74, 78, 79, 97, 98, 100, 103, 104, 127, 137,
141, 147, 154, 207, 211, 223, 224, 230, 231, 233, 235, 287
leukopenia ................................................................. 12, 27, 73, 77, 78, 79, 85, 86, 127, 132, 211, 221, 233
lymphatic .......................................................................................................................................... 100, 140
lymphoreticular ......................................................................................................................................... 140
menstrual................................................................................................................................... 6, 92, 93, 225
micronuclei ....................................................................................................... 145, 149, 151, 152, 153, 224
milk ............................................................................................................................. 91, 192, 216, 271, 282
musculoskeletal effects ................................................................................................................. 81, 82, 128
neonatal ....................................................................................................................................................... 20
neoplastic .................................................................................................................... 82, 128, 129, 135, 226
neurobehavioral......................................................................................................................................... 206
neurological effects ................................................................. 13, 19, 20, 24, 89, 90, 92, 134, 135, 140, 228
neurotransmitter ........................................................................................................................................ 228
neutropenia.................................................................................................................................................. 82
non-Hodgkins lymphoma ............................................................................................................ 15, 97, 223
norepinephrine .......................................................................................................................................... 134
octanol-water partition coefficient ............................................................................................................ 199
ocular effects......................................................................................................................... 13, 83, 130, 138
odds ratio................................................................................................................................................... 102
pancytopenia ................................................................................................... 12, 17, 73, 77, 78, 79, 94, 226
partition coefficients ..................................................................................................... 22, 23, 182, 192, 199
pharmacodynamic ............................................................................................................................. 162, 181
pharmacokinetic................................................................................................ 181, 182, 183, 196, 207, 235
photolysis .................................................................................................................................................. 262
placenta ....................................................................................................................... 94, 163, 164, 207, 235
rate constant ...................................................................................................................................... 187, 261
renal effects................................................................................................................................... 13, 82, 129
reproductive effects....................................................................................................... 92, 94, 135, 140, 225
respiratory effects................................................................................................................................ 32, 105
retention .................................................................................................................................................... 204
solubility ........................................................................................................................... 159, 161, 200, 258
spermatogonia ........................................................................................................................................... 145
spermatozoa ........................................................................................................................................ 94, 225
systemic effects..................................................................................... 12, 32, 105, 131, 138, 219, 221, 222
T4 ................................................................................................................................................................ 88
thrombocytopenia ................................................................................................................... 12, 73, 77, 221
thyroid............................................................................................................................................... 129, 166
BENZENE
D-3
APPENDIX D
toxicokinetic........................................................................................................................................ 29, 154
tremors ........................................................................................................................ 5, 13, 20, 91, 134, 228
tumors ............................................................................................... 15, 16, 18, 89, 103, 141, 165, 228, 231
vapor phase ............................................................................................................................... 259, 261, 269
vapor pressure ........................................................................................................................................... 258
volatility .................................................................................................................................................... 258
volatilization ............................................................................................................................. 249, 251, 258
BENZENE
D-4

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TOXICOLOGICAL PROFILE FOR

U.S. DEPARTMENT OF HEALTH AND HUMAN SERVICES

Public Health Service

Agency for Toxic Substances and Disease Registry

Division of Toxicology and Environmental Medicine/Applied Toxicology Branch

1600 Clifton Road NE

Atlanta, Georgia 30333

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ATSDR Information Center

Managing Hazardous Materials Incidents is a three-volume set of recommendations for on-scene

Sam Keith, M.S., C.H.P.

Obaid Faroon, Ph.D.

ATSDR, Division of Toxicology and Environmental Medicine, Atlanta, GA

David Wohlers, Ph.D.

Julie Stickney, Ph.D.

Sari Paikoff, Ph.D.

Gary Diamond, Ph.D

Antonio Quiones-Rivera, Ph.D.

Syracuse Research Corporation, North Syracuse, NY

THE PROFILE HAS UNDERGONE THE FOLLOWING ATSDR INTERNAL REVIEWS:

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UPDATE STATEMENT .............................................................................................................................iii

QUICK REFERENCE FOR HEALTH CARE PROVIDERS....................................................................vii

PEER REVIEW ...........................................................................................................................................xi

LIST OF FIGURES ..................................................................................................................................xvii

1. PUBLIC HEALTH STATEMENT.......................................................................................................... 1

PROTECT HUMAN HEALTH?................................................................................................. 8

1.10 WHERE CAN I GET MORE INFORMATION? ....................................................................... 9

2. RELEVANCE TO PUBLIC HEALTH ................................................................................................. 11

8. REGULATIONS AND ADVISORIES ............................................................................................... 307

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3-1. Levels of Significant Exposure to Benzene Inhalation.................................................................... 66

3-2. Levels of Significant Exposure to Benzene Oral........................................................................... 122

3-3. Metabolic Pathways for Benzene ..................................................................................................... 167

3-4. Conceptual Representation of a Physiologically Based Pharmacokinetic (PBPK) Model for a

Hypothetical Chemical Substance.................................................................................................... 183

3-6. Existing Information on Health Effects of Benzene......................................................................... 220

6-1. Frequency of NPL Sites with Benzene Contamination .................................................................... 252

6-2. Environmental Transformation Products of Benzene in Various Media.......................................... 260

6-3. Benzene Emissions and Exposures................................................................................................... 275

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3-1. Levels of Significant Exposure to Benzene Inhalation.................................................................... 33

3-2. Levels of Significant Exposure to Benzene Oral........................................................................... 106

3-3. Levels of Significant Exposure to Benzene Dermal ...................................................................... 139

3-4. Genotoxicity of Benzene In Vivo...................................................................................................... 142

3-5. Genotoxicity of Benzene In Vitro..................................................................................................... 146

3-7. Ongoing Studies on the Health Effects of Benzene.......................................................................... 237

4-1. Chemical Identity of Benzene .......................................................................................................... 240

4-2. Physical and Chemical Properties of Benzene ................................................................................. 241

5-1. Facilities that Produce, Process, or Use Benzene ............................................................................. 245

5-2. Current U.S. Manufacturers of Benzene........................................................................................... 247

6-2. Benzene Levels in Air Samples........................................................................................................ 266

6-3. Benzene in Food ............................................................................................................................... 272

7-1. Analytical Methods for Determining Benzene in Biological Samples............................................. 292

7-2. Analytical Methods for Determining Metabolites of Benzene in Urine........................................... 294

7-3. Analytical Methods for Determining Benzene in Environmental Samples...................................... 296