Chlorine: Description

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278 / Chitosan / Monographs FCC 9
chromatogram between Va and Vb (NLT 40). [NOTE—If

Chlorine
.
n is sufficiently large, the use of area segments Ai or

peak heights Hi will yield equivalent results.] First Published: Prior to FCC 6
Calculate the molecular weight distribution or
polydispersity for the sample taken:
Cl2 Formula wt 70.91
Result = Mw/Mn INS: 925 CAS: [7782-50-5]
UNII: 4R7X1O2820 [chlorine]
Acceptance criteria: The values of apparent weight-
average molecular weight and polydispersity are DESCRIPTION
85%–115% of the respective values claimed by the Chlorine occurs as a green-yellow gas, normally packaged as
Monographs
manufacturer. a liquid under pressure in containers approved by the U.S.

• LOSS ON DRYING, Appendix IIC: 1 g is dried at 100°–105° Department of Transportation. At 60° F, it has a vapor
for 7 h. pressure of 70.91 psig. Its vapor density is about 2.5 times
Acceptance criteria: NMT 5.0% that of air. About 0.8 lb (0.362 kg) is soluble in 100 lb
• DEGREE OF DEACETYLATION BY 1H NMR SPECTROSCOPY, (45.4 kg) of water at 60° F under atmospheric pressure.
Nuclear Magnetic Resonance, Appendix IIC Function: Antimicrobial agent; bleaching agent; oxidizing
Solvent: Deuterated formic acid agent
NMR reference: Tetramethylsilane. [NOTE—If Packaging and Storage: Store in suitable pressure
tetramethylsilane is not used as the NMR reference, a containers, observing applicable U.S. Department of
suitable signal of the solvent itself can be used as a Transportation regulations pertaining to shipping
reference.] containers.
Sample: 5–10 mg [CAUTION—Chlorine gas is a respiratory irritant. Large
Sample solution: In a 20-mL scintillation vial with a amounts cause coughing, labored breathing, and irritation
screw cap, dissolve the Sample in a sufficient quantity of of the eyes. In extreme cases, the difficulty in breathing
deuterated formic acid containing 0.5%–1.0% of may cause death due to suffocation. Liquid Chlorine
tetramethylsilane to obtain 1 mL of solution. Tightly causes skin and eye burns on contact. (Safety precautions
close the vial, and dissolve the Sample using a magnetic to be observed in handling the material are specified in
stirrer. It may take up to 48 h to completely dissolve the Chlorine Manual, available from The Chlorine Institute,
the Sample, during which time any clumps formed in Inc., 1300 Wilson Blvd, Arlington, VA 22209,
the vial should be broken up with a spatula. Use the www.chlorineinstitute.org.)]
clear, viscous solution obtained.
Analysis: Transfer 0.5–1.0 mL of the Sample solution to a
IDENTIFICATION
• CHLORIDE, Appendix IIIA
standard 5-mm NMR sample tube. Collect 1H NMR
Sample preparation: Cautiously pass a few mL of
data for the Sample solution, scanning the region from
sample gas through 10 mL of 1 N sodium hydroxide
0–7 ppm. Record the spectrum and data obtained.
that has previously been chilled in an ice bath.
Identify the tetramethylsilane singlet at 0 ppm, as well
Acceptance criteria: The Sample preparation gives
as the signals due to the methyl groups of the acetyl
positive tests for Chloride, and it darkens starch iodide
units at about 2 ppm.
paper.
Calculate the percentage of deacetylation degree in the
Sample taken: ASSAY
• PROCEDURE
Result = [1 − (7 × A2)/(3 × A1)] × 100
Analysis: Determine by ASTM Method E 412-93, “Assay
A2 = average area of the signals due to the of Liquid Chlorine, Zinc Amalgam Method.”
methyl groups of the acetyl units Acceptance criteria: NLT 99.5%, by volume
A1 = average area of the composite band from
about 6–3 ppm (representing the seven
IMPURITIES
Inorganic Impurities
protons with oxygen neighbors in the
• LEAD, Lead Limit Test, Appendix IIIB
sugar ring)
Sample stock solution: Dissolve the residue obtained
Acceptance criteria: The degree of deacetylation is
under the test for Residue (below) in 2.5 mL of freshly
70.0%–95.0%.
prepared aqua regia, and dilute with water to a
OTHER REQUIREMENTS volume, in mL, equivalent to the weight, in g, of the
• LABELING: Label to indicate the apparent weight-average initial sample. One mL of the final dilution is equivalent
molecular weight and polydispersity, as well as the to 1 g of sample.
source (crab or shrimp). Sample solution: Mix a 1.0-mL portion of the Sample
stock solution with 5 mL of water and 11 mL of 2.7 N
hydrochloric acid.
Control: 10 µg Pb (10 mL of Diluted Standard Lead
Solution)
Acceptance criteria: NMT 10 mg/kg
• MERCURY, Mercury Limit Test, Method I, Appendix IIIB
FCC 9 Monographs / Choline Bitartrate / 279
Sample stock solution: Dissolve the residue obtained Acceptance criteria: The spectrum of the sample
under the test for Residue (below) in 2.5 mL of freshly exhibits maxima at the same wavelengths as those in
prepared aqua regia, and dilute with water to a the spectrum of the Reference standard.
volume, in mL, equivalent to the weight, in g, of the • B. PROCEDURE
initial sample. One mL of the final dilution is equivalent Sample solution: 0.2 mg/mL in 50% acetic acid
to 1 g of sample. Analysis: To 1 mL of the Sample solution add 1 mL of a
Sample solution: Transfer 2.0 mL of the Sample stock 1:100 furfural solution. Cool in an ice bath for 5 min,
solution to a 50-mL beaker and add 10 mL of water, 1 add 15 mL of 1:2 sulfuric acid, mix, and warm in a
mL of 1:5 sulfuric acid, and 1 mL of a 40 mg/mL water bath at 70° for 10 min. Immediately cool in an
potassium permanganate solution. Cover the beaker ice bath, and stir for 2 min.
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with a watch glass, boil for a few seconds, and cool. Acceptance criteria: A blue color appears.
ASSAY
SPECIFIC TESTS • PROCEDURE
• MOISTURE Sample: 400 mg
Analysis: Determine by ASTM Method E 410-92, Analysis: Transfer the Sample into a 250-mL Erlenmeyer
“Moisture and Residue in Liquid Chlorine.” flask, add 20 mL of water and 40 mL of alcohol, cover
[NOTE—Retain the residue obtained for use in tests for with a watch glass, heat gently on a steam bath until
Lead and Mercury (above).] dissolved, and cool. Add 5 drops of phenolphthalein TS
Acceptance criteria: NMT 0.015%, by weight and, using a 10-mL microburet, titrate with 0.1 N
• RESIDUE sodium hydroxide to the first pink color that persists for
Analysis: Determine by ASTM Method E 410-92, 15 s. Perform a blank determination (see General
“Moisture and Residue in Liquid Chlorine.” Provisions), and make any necessary correction. Each mL
[NOTE—Retain the residue obtained for use in tests for of 0.1 N sodium hydroxide is equivalent to 40.86 mg
Lead and Mercury (above).] of C24H40O5.
Acceptance criteria: NMT 0.015%, by weight, of Acceptance criteria: NLT 98.0% of C24H40O5, calculated
nonvolatile matter on the dried basis
IMPURITIES
• LEAD, Lead Limit Test, Flame Atomic Absorption
Cholic Acid
.
Spectrophotometric Method, Appendix IIIB

First Published: Prior to FCC 6 Sample: 10 g
Last Revision: Third Supplement, FCC 7 Acceptance criteria: NMT 4 mg/kg
Cholalic Acid SPECIFIC TESTS

3,7,12-Trihydroxycholanic Acid • LOSS ON DRYING, Appendix IIC: 140° under a vacuum of
NMT 5 mm Hg, for 4 h
Acceptance criteria: NMT 0.5%
• MELTING RANGE OR TEMPERATURE DETERMINATION, Appendix
IIB
Acceptance criteria: Between 197° and 202°
• OPTICAL (SPECIFIC) ROTATION, Appendix IIB
Sample solution: 20 mg/mL in alcohol
C24H40O5 Formula wt 408.58 Acceptance criteria: [α]D25 NLT +37°, calculated on the
INS: 1000 CAS: [81-25-4] dried basis
UNII: G1JO7801AE [cholic acid] • RESIDUE ON IGNITION (SULFATED ASH), Appendix IIC
Sample: 2 g
DESCRIPTION Acceptance criteria: NMT 0.1%
Cholic Acid occurs as colorless plates or as a white,
crystalline powder. One g dissolves in about 30 mL of
alcohol or acetone and in about 7 mL of glacial acetic acid.
Choline Bitartrate
.
It is very slightly soluble in water.

Function: Emulsifier First Published: Prior to FCC 6
Packaging and Storage: Store in tight containers. Last Revision: FCC 7
IDENTIFICATION
• A. INFRARED ABSORPTION, Spectrophotometric Identification (2-Hydroxyethyl)trimethylammonium-L-(+)-tartrate Salt
Tests, Appendix IIIC
Reference standard: USP Cholic Acid RS
Sample and standard preparation: K
280 / Choline Bitartrate / Monographs FCC 9
C9H19NO7 Formula wt 253.25 SPECIFIC TESTS

INS: 1001(v) CAS: [87-67-2] • OPTICAL (SPECIFIC) ROTATION, Appendix IIB
UNII: 6K2W7T9V6Y [choline bitartrate] Sample solution: 400 mg/mL
Acceptance criteria: [α]D25 between 17.5° and 18.5°
DESCRIPTION • RESIDUE ON IGNITION (SULFATED ASH), Appendix IIC
Choline Bitartrate occurs as a white, hygroscopic, crystalline Sample: 2 g
powder. It is freely soluble in water, slightly soluble in Acceptance criteria: NMT 0.1%
alcohol, and insoluble in ether and in chloroform. • WATER, Water Determination, Appendix IIB
Function: Nutrient Sample solution: 2 g of sample in 50 mL of methanol.
Packaging and Storage: Store in tight containers. [NOTE—Alternatively, the Water Determination can be
Monographs
made by drying the sample in a vacuum desiccator

IDENTIFICATION
over phosphorus pentoxide for 4 h.]
• A. INFRARED ABSORPTION,Spectrophotometric Identification
Reference standard: USP Choline Bitartrate RS
Sample and standard preparation: K
Acceptance criteria: The spectrum of the sample
Choline Chloride
.
exhibits maxima at the same wavelengths as those in

the spectrum of the Reference standard. First Published: Prior to FCC 6
• B. PROCEDURE Last Revision: FCC 7
Sample: 500 mg
Analysis: Dissolve the Sample in 2 mL of iodine TS. A (2-Hydroxyethyl)trimethylammonium Chloride
red-brown precipitate forms immediately. Add 5 mL of
1 N sodium hydroxide. The precipitate dissolves, and
the solution becomes clear yellow. Heat the solution.
Acceptance criteria: A pale yellow precipitate forms
following the heating step. C5H14ClNO Formula wt 139.65
• C. PROCEDURE INS: 1001(iii) CAS: [67-48-1]
Sample solution: 10 mg/mL UNII: 45I14D8O27 [choline chloride]
Analysis: Add 1 mL of the Sample solution and 2 mL of a
20 mg/mL solution of potassium ferrocyanide to 2 mL DESCRIPTION
of cobaltous chloride TS. Choline Chloride occurs as colorless or white crystals or as a
Acceptance criteria: An emerald green color develops crystalline powder. It is hygroscopic, and is very soluble in
immediately. water and in alcohol.
Function: Nutrient
ASSAY Packaging and Storage: Store in tight containers.
• PROCEDURE
Sample: 500 mg IDENTIFICATION
Analysis: Transfer the Sample into a 250-mL Erlenmeyer • A. CHLORIDE, Appendix IIIA
flask. Add 50 mL of glacial acetic acid and warm on a Sample solution: 50 mg/mL
steam bath until dissolution is complete. Cool, add 2 Acceptance criteria: Passes tests
drops of crystal violet TS, and titrate with 0.1 N • B. INFRARED ABSORPTION, Spectrophotometric Identification
perchloric acid in glacial acetic acid to a green Tests, Appendix IIIC
endpoint. [CAUTION—Handle perchloric acid in an Reference standard: USP Choline Chloride RS
appropriate fume hood.] Perform a blank determination Sample and standard preparation: K
(see General Provisions), and make any necessary Acceptance criteria: The spectrum of the sample
correction. Each mL of 0.1 N perchloric acid is exhibits maxima at the same wavelengths as those in
equivalent to 25.36 mg of C9H19NO7. the spectrum of the Reference standard.
Acceptance criteria: NLT 98.0% of C9H19NO7, • C. PROCEDURE
calculated on the anhydrous basis Sample: 500 mg
Analysis: Dissolve the Sample in 2 mL of iodine TS. A
IMPURITIES red-brown precipitate forms immediately. Add 5 mL of
Inorganic Impurities 1 N sodium hydroxide. The precipitate dissolves, and
• LEAD, Lead Limit Test, Flame Atomic Absorption the solution becomes clear yellow. Heat the solution.
Spectrophotometric Method, Appendix IIIB Acceptance criteria: A pale yellow precipitate forms
Sample: 5 g following the heating step.
Acceptance criteria: NMT 2 mg/kg • D. PROCEDURE
Organic Impurities Sample solution: 10 mg/mL
• 1,4-DIOXANE, Appendix IIIB Analysis: Add 1 mL of the Sample solution and 2 mL of a
Acceptance criteria: Passes test 20 mg/mL solution of potassium ferrocyanide to 2 mL
of cobaltous chloride TS.
FCC 9 Monographs / Chromic Chloride / 281
Acceptance criteria: An emerald green color develops [CAUTION—Avoid contact with skin and eyes. Avoid
immediately. formation of dust and aerosols. Handle under appropriate
exhaust ventilation.]
ASSAY
• PROCEDURE IDENTIFICATION
Sample: 300 mg • A. PROCEDURE
Analysis: Transfer the Sample into a 250-mL Erlenmeyer Sample solution: Dissolve 1 g of the sample in water to
flask. Add 50 mL of glacial acetic acid and warm on a a final concentration of 4 mg/mL.
steam bath until dissolution is complete. Cool, add 10 Analysis: In a test tube, add 1 mL of 5 N sodium
mL of mercuric acetate and 2 drops of crystal violet TS, hydroxide and 10 drops of 30% hydrogen peroxide to
Monographs
and titrate with 0.1 N perchloric acid in glacial acetic 5 mL of the Sample solution, and heat gently for about
acid to a green endpoint. [CAUTION—Handle perchloric 2 min.
acid in an appropriate fume hood.] Perform a blank Acceptance criteria: A yellow color develops.
determination (see General Provisions), and make any • B. CHLORIDE, Appendix IIIA
necessary correction. Each mL of 0.1 N perchloric acid Sample solution: 4 mg/mL
is equivalent to 13.96 mg of C5H14ClNO. Acceptance criteria: Passes test
Acceptance criteria: 98.0%–100.5% of C5H14ClNO,
calculated on the anhydrous basis ASSAY
• PROCEDURE
IMPURITIES Analysis: In a glass-stoppered, 500-mL conical flask,
Inorganic Impurities dissolve 0.4 g of the sample in 100 mL of water. Then,
• LEAD, Lead Limit Test, Flame Atomic Absorption add 5 mL of 5 N sodium hydroxide, and mix. Pipet
Spectrophotometric Method, Appendix IIIB slowly 4 mL of 30% hydrogen peroxide into the flask,
Sample: 5 g and boil the solution for 5 min. Cool the solution
Acceptance criteria: NMT 2 mg/kg slightly, and add 5 mL of nickel sulfate solution (50 mg
• WATER, Water Determination, Appendix IIB /mL). Boil the solution until no more oxygen is evolved,
[NOTE—Alternatively, the Water Determination can be cool, and add 2 N sulfuric acid dropwise until the color
made by drying the sample in a vacuum desiccator of the solution changes from yellow to orange. Add to
over phosphorus pentoxide for 4 h.] the flask a freshly prepared solution of 4 g of potassium
Acceptance criteria: NMT 0.5% iodide and 2 g of sodium bicarbonate in 100 mL of
Organic Impurities water, then add 6 mL of hydrochloric acid. Immediately
• 1,4-DIOXANE, Appendix IIIB insert the stopper in the flask, and allow to stand in the
Acceptance criteria: Passes test dark for 10 min. Rinse the stopper and the sides of the
flask with a few mL of water, and titrate the liberated
SPECIFIC TESTS iodine with 0.1 N sodium thiosulfate VS to an orange
• RESIDUE ON IGNITION (SULFATED ASH), Appendix IIC color. Add 3 mL of starch TS, and continue the titration
Sample: 4 g to a blue-green endpoint. Each mL of 0.1 N sodium
Acceptance criteria: NMT 0.05% thiosulfate is equivalent to 8.882 mg of CrCl3 · 6H2O.
Acceptance criteria: 98%–101.0%
IMPURITIES
Chromic Chloride
.
• ARSENIC, Elemental Impurities by ICP, Appendix IIIC
First Published: First Supplement, FCC 8
• CADMIUM, Elemental Impurities by ICP, Appendix IIIC
Chromium chloride (III) Acceptance criteria: NMT 1 mg/kg
Chromium trichloride hexahydrate • LEAD, Elemental Impurities by ICP, Appendix IIIC
Chromium chloride hexahydrate Acceptance criteria: NMT 1 mg/kg
Chromium chloride hexahydrate (III) • MERCURY, Elemental Impurities by ICP, Appendix IIIC
CrCl3 · 6H2O Formula wt 266.45 Acceptance criteria: NMT 1 mg/kg
CAS: [10060-12-5]
UNII: KB1PCR9DMW [chromic chloride] SPECIFIC TESTS
• WATER-INSOLUBLE MATTER
DESCRIPTION Analysis: Transfer 10 g of the sample to a 250-mL
Chromic chloride hexahydrate occurs as very dark green to beaker, add 100 mL of water, cover the beaker, and
violet crystals or crystalline powder. It is hygroscopic, and heat to boiling. Digest the hot solution on a steam bath
freely soluble in water. It is soluble in ethanol and insoluble for 30 min, and filter through a tared Gooch crucible.
in ether and acetone. Rinse the beaker with hot water, passing the rinsings
Function: Nutrient through the crucible, and wash the crucible with hot
Packaging and Storage: Store in tightly sealed containers water until the last washing is colorless. Dry the crucible
in a cool, dry place, away from moisture. at 105°.
282 / Chromic Chloride / Monographs FCC 9
Acceptance criteria: The weight of the residue does not Analysis: To 5 mL of the Sample solution, add 1 mL of
exceed 1 mg (NMT 0.01%). 5 N sodium hydroxide and 10 drops of 30% hydrogen
• SUBSTANCES NOT PRECIPITATED BY AMMONIUM HYDROXIDE peroxide, and heat gently for 2 min.
Analysis: Dissolve 2.0 g of the sample in 80 mL of Acceptance criteria: A yellow color develops.
water, heat the solution to boiling, and add a slight
excess of ammonium hydroxide to ensure the formation ASSAY
of a precipitate. Continue heating to remove the excess • PROCEDURE
ammonia, cool, dilute with water to 100.0 mL, and Standard stock solution: Prepare a solution containing
mix. Pass through a retentive filter, and transfer 50.0 100 µg/mL of chromium as follows. Transfer 0.283 g of
mL of the clear filtrate to an evaporating dish that potassium dichromate, previously dried at 120° for 4 h,
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previously has been ignited and tared. Add 0.5 mL of to a 1000-mL volumetric flask, and dilute with water to
sulfuric acid to the filtrate, evaporate on a steam bath volume. Store the solution in a polyethylene bottle.
to dryness, heat gently to remove the excess acid, and [CAUTION—Care should be taken when handling
ignite gently. potassium dichromate as it is a known human
Acceptance criteria: The weight of the residue does not carcinogen and mutagen. Use proper personal
exceed 2 mg (NMT 0.20% as sulfate). protective devices as described in the material’s MSDS.]
Standard solutions: Individually prepare solutions
containing 1.0 µg/mL, 2.0 µg/mL, 3.0 µg/mL, and 4.0
µg/mL of chromium as follows. Separately transfer 1.0
mL and 2.0 mL of the Standard stock solution to
Chromium Picolinate
.
separate 100-mL volumetric flasks, and transfer 1.5 mL

First Published: Third Supplement, FCC 8 and 2.0 mL of the Standard stock solution to separate
50-mL volumetric flasks. Add 1.0 mL of nitric acid to
Chromium(III) Picolinate each flask, and dilute the contents of each flask with
Chromium Tripicolinate water to volume.
Chromium(III) Trispicolinate Sample solution: Transfer 200 mg of the sample,
Picolinic Acid, Chromium(III) Salt previously dried, to a 100-mL beaker, and add 25 mL
2-Pyroxidinecarboxylic Acid, Chromium Salt of water. Slowly add 10 mL of nitric acid to the beaker,
and boil for 10 min with constant swirling. Cool the
solution, quantitatively transfer it to a 500-mL
volumetric flask, and dilute with water to volume. Filter
a portion of the solution, and transfer 5.0 mL of the
filtrate to a 100-mL volumetric flask. Add 1 mL of nitric
acid to the flask, and dilute with water to volume.
Analysis: Using a suitable atomic absorption
spectrophotometer equipped with a chromium hollow-
cathode lamp and an air–acetylene flame set to the
C18H12N3O6Cr Formula wt 418.31 chromium emission line of 357.9 nm, determine the
CAS: [14639-25-9]
absorbances of the Standard solutions and the Sample
UNII: S71T8B8Z6P [chromium picolinate] solution using 10% nitric acid as the blank. Plot the
absorbances of the Standard solutions versus the
DESCRIPTION
chromium concentration, in µg/mL, and draw the
Chromium Picolinate occurs as a reddish, free-flowing
straight line best fitting the plotted points. From the
powder. It is manufactured using picolinic acid and a
graph so obtained, determine the chromium
chromium salt such as potassium chromium sulfate.
concentration, in µg/mL, of the Sample solution.
Chromium Picolinate is slightly soluble in water (at neutral
Calculate the percentage of chromium picolinate
pH) and slightly soluble in chloroform.
(C18H12N3O6Cr) in the portion of the sample taken:
Function: Source of chromium
Packaging and Storage: Store in tight containers. Result = (CCr/CU) × (Mr/ACr) × 100
IDENTIFICATION CCr = concentration of chromium in the Sample
• A. INFRARED ABSORPTION, Spectrophotometric Identification solution, as obtained from the standard
Tests, Appendix IIIC curve (µg/mL)
Reference standard: USP Chromium Picolinate RS CU = concentration of the Sample solution (µg/mL)
Sample and standard preparation: M Mr = molecular weight of chromium picolinate,
Acceptance criteria: The spectrum of the sample 418.31
exhibits maxima at the same wavelengths as those in ACr = atomic weight of chromium, 51.996
the spectrum of the Reference standard. Acceptance criteria: 98.0%–102.0% on the dried basis
• B. PROCEDURE
Sample solution: 4 mg/mL
FCC 9 Monographs / Cinnamaldehyde / 283
IMPURITIES
• CHLORIDE, Chloride and Sulfate Limit Tests, Chloride Limit
Test, Appendix IIIB
Sample solution: Dissolve 30 mg of the sample in
C9H8O Formula wt 132.16
30–40 mL of water, and heat to 70°. Cool overnight,
FEMA: 2286
and filter to remove precipitate.
Control: 0.25 mL of 0.002 hydrochloric acid (18 µg of UNII: SR60A3XG0F [cinnamaldehyde]
chloride)
DESCRIPTION
Analysis: Proceed as directed, substituting the Control in
Cinnamaldehyde occurs as a yellow, strongly refractive
Monographs
place of the Standard Chloride Solution.
liquid. It may contain a suitable antioxidant.
Acceptance criteria: Any turbidity produced by the
Odor: Cinnamon, burning aromatic taste
Sample solution does not exceed that in the Control
Solubility: Miscible in alcohol, chloroform, ether, fixed and
(NMT 0.06%).
volatile oils; 1 g dissolves in 700 mL of water.
• LEAD, Elemental Impurities by ICP, Method I, Appendix IIIC
Boiling Point: ∼248°
Solubility in Alcohol, Appendix VI: One mL dissolves in 5
• SULFATE, Chloride and Sulfate Limit Tests, Sulfate Limit Test,
mL of 60% alcohol.
Appendix IIIB
Function: Flavoring agent
Sample solution: Dissolve 100 mg of the sample in
30–40 mL of water, and heat to 90°. Cool overnight, IDENTIFICATION
and filter to remove precipitate. • INFRARED SPECTRA, Spectrophotometric Identification Tests,
Control: 0.2 mL of 0.02 N sulfuric acid (200 µg of Appendix IIIC
sulfate) Acceptance criteria: The spectrum of the sample
Analysis: Proceed as directed, substituting the Control in exhibits relative maxima at the same wavelengths as
place of the Standard Sulfate Solution. those of the spectrum below.
Acceptance criteria: Any turbidity produced by the
Sample solution does not exceed that in the Control ASSAY
(NMT 0.2%). • PROCEDURE: Proceed as directed under M-1b, Appendix
XI.
SPECIFIC TESTS Acceptance criteria: NLT 98.0% of C9H8O
• LOSS ON DRYING, Appendix IIC: 105° for 4 h
Acceptance criteria: NMT 4.0% SPECIFIC TESTS
• ACID VALUE, FLAVOR CHEMICALS (OTHER THAN ESSENTIAL
OILS), M-15, Appendix XI
Acceptance criteria: NMT 10.0
Cinnamaldehyde • REFRACTIVE INDEX, Appendix II: At 20°
.
Acceptance criteria: Between 1.619 and 1.623

First Published: Prior to FCC 6
• SPECIFIC GRAVITY: Determine at 25° by any reliable
Last Revision: First Supplement, FCC 6
method (see General Provisions).
Cinnamal
Cinnamic Aldehyde
284 / Cinnamaldehyde / Monographs FCC 9
OTHER REQUIREMENTS
• CHLORINATED COMPOUNDS, Appendix VI
Acceptance criteria: Passes test
Monographs
Cinnamaldehyde
Standard and sample preparation: M

Cinnamic Acid
.

First Published: Prior to FCC 6 exhibits maxima at the same wavelengths as those in
Last Revision: First Supplement, FCC 8 the spectrum of the Reference standard.
ASSAY
3-Phenylpropenoic Acid • PROCEDURE: Proceed as directed under M-3b, Appendix
XI.
Acceptance criteria: NLT 99.0% on the dried basis
OTHER REQUIREMENTS
C9H8O2 Formula wt 148.16 IIB
FEMA: 2288 Acceptance criteria: NLT 130°
UNII: U14A832J8D [cinnamic acid] • RESIDUE ON IGNITION (SULFATED ASH), Appendix IIC
Sample: 2 g
DESCRIPTION Acceptance criteria: NMT 0.05%
Cinnamic Acid occurs as white crystalline scales.
Odor: Honey-floral
Solubility: Soluble in acetic acid, acetone, benzene, most
Cinnamon Bark Oil, Ceylon Type
.
fixed oils; 1 g dissolves in 2000 mL of water.

Boiling Point: ∼300° First Published: Prior to FCC 6
Solubility in Alcohol, Appendix VI: One g dissolves in 7
mL of 95% alcohol.
FEMA: 2290
Function: Flavoring agent CAS: [8015-91-6]
IDENTIFICATION UNII: XE54U569EC [cinnamon bark oil]
• INFRARED ABSORPTION, Spectrophotometric Identification
DESCRIPTION
Cinnamon Bark Oil, Ceylon Type occurs as a yellow liquid
Reference standard: USP Cinnamic Acid RS
with an odor of cinnamon and a spicy burning taste. It is
FCC 9 Monographs / Cinnamon Leaf Oil / 285
the volatile oil obtained by steam distillation from the dried Analysis: Use 66.10 as the equivalence factor (e) in the
inner bark of the clipped cinnamon shrub Cinnamomum calculation.
zeylanicum Nees (Fam. Lauraceae). It is soluble in most Acceptance criteria: NLT 55.0% and NMT 78.0% of
fixed oils and in propylene glycol. It is insoluble in glycerin aldehydes, calculated as cinnamic aldehyde (C9H8O)
and in mineral oil.
Function: Flavoring agent SPECIFIC TESTS
Packaging and Storage: Store in a cool place protected • ANGULAR ROTATION, Optical (Specific) Rotation, Appendix
from light in full, tight containers that are made from glass IIB: Use a 100-mm tube.
or aluminum or that are lined with tin. Acceptance criteria: Between −2° and 0°
• REFRACTIVE INDEX, Appendix IIB
Monographs
IDENTIFICATION [NOTE—Use an Abbé or other refractometer of equal or
• INFRARED SPECTRA, Spectrophotometric Identification Tests, greater accuracy.]
Appendix IIIC Acceptance criteria: Between 1.573 and 1.591 at 20°
Acceptance criteria: The spectrum of the sample • SOLUBILITY IN ALCOHOL, Appendix VI
exhibits relative maxima at the same wavelengths as Acceptance criteria: One mL of sample dissolves in 3
those of the spectrum below. mL of 70% alcohol.
• SPECIFIC GRAVITY: Determine by any reliable method (see
ASSAY General Provisions).
• ALDEHYDES, Appendix VI Acceptance criteria: Between 1.010 and 1.030
Sample: 2.5 g
Cinnamon Bark Oil, Ceylon Type
and twigs of the true cinnamon shrub, Cinnamomum

Cinnamon Leaf Oil
.
zeylanicum Nees (Fam. Lauraceae). The commercial oils,

First Published: Prior to FCC 6 according to the geographical origin, are designated as
Cinnamon Leaf Oil, Ceylon, or Cinnamon Leaf Oil,
Seychelles, and the two types differ in physical and
FEMA: 2292
CAS: [8015-91-6] chemical properties. Cinnamon Leaf Oil is soluble in most
UNII: S92U8SQ71V [cinnamon leaf oil] fixed oils and in propylene glycol. It is soluble, with
cloudiness, in mineral oil, but is insoluble in glycerin.
DESCRIPTION Function: Flavoring agent
Cinnamon Leaf Oil occurs as a light to dark brown liquid Packaging and Storage: Store in a cool place protected
with a spicy cinnamon–clove odor and taste. It is the from light in full, tight containers that are made from glass
volatile oil obtained by steam distillation from the leaves or aluminum or that are lined with tin.
286 / Cinnamon Leaf Oil / Monographs FCC 9
IDENTIFICATION • REFRACTIVE INDEX, Appendix IIB

• INFRARED SPECTRA, Spectrophotometric Identification Tests, [NOTE—Use an Abbé or other refractometer of equal or
Appendix IIIC greater accuracy.]
Acceptance criteria: The spectrum of the sample Acceptance criteria
exhibits relative maxima at the same wavelengths as Ceylon type: Between 1.529 and 1.537
those of the spectrum below. Seychelles type: Between 1.533 and 1.540 at 20°
• SOLUBILITY IN ALCOHOL, Appendix VI
ASSAY Acceptance criteria
• PHENOLS, Appendix VI Ceylon type: One mL of sample dissolves in 1.5 mL of
Sample: Pretreat the oil by shaking a suitable quantity 70% alcohol. The solution may cloud upon further
Monographs
with about 2% powdered tartaric acid, and filter. dilution.

Acceptance criteria Seychelles type: One mL of sample dissolves in 1 mL
Ceylon type: NLT 80.0% and NMT 88.0%, by volume, of 70% alcohol. The solution may cloud upon further
of phenols dilution.
Seychelles type: NLT 87.0% and NMT 96.0%, by • SPECIFIC GRAVITY: Determine by any reliable method (see
volume, of phenols General Provisions).
Acceptance criteria
SPECIFIC TESTS Ceylon type: Between 1.030 and 1.050
• ANGULAR ROTATION, Optical (Specific) Rotation, Appendix
Seychelles type: Between 1.040 and 1.060
IIB: Use a 100-mm tube.
Acceptance criteria: OTHER REQUIREMENTS
Ceylon type: Between −2° and +1° • LABELING: Indicate whether the oil is the Ceylon or
Seychelles type: Between −2° and 0° Seychelles type.
Cinnamon Leaf Oil

FCC 9 Monographs / Cinnamyl Acetate / 287

Cinnamyl Acetate
.
IDENTIFICATION
• INFRARED SPECTRA, Spectrophotometric Identification Tests,
Appendix IIIC
exhibits relative maxima at the same wavelengths as
those of the spectrum below.
ASSAY
• PROCEDURE: Proceed as directed under M-1b, Appendix
Monographs
C11H12O2 Formula wt 176.22
FEMA: 2293 XI.
UNII: LFJ36XSV8K [cinnamyl acetate] Acceptance criteria: NLT 98.0% of C11H12O2
DESCRIPTION SPECIFIC TESTS

Cinnamyl Acetate occurs as a colorless to slightly yellow • ACID VALUE, FLAVOR CHEMICALS (OTHER THAN ESSENTIAL
liquid. OILS), M-15, Appendix XI
Odor: Sweet, balsamic, floral Acceptance criteria: NMT 1.0
Solubility: Miscible in alcohol, chloroform, ether, most • REFRACTIVE INDEX, Appendix II: At 20°
fixed oils; insoluble or practically insoluble in glycerin, Acceptance criteria: Between 1.539 and 1.543
water • SPECIFIC GRAVITY: Determine at 25° by any reliable
Boiling Point: ∼264° method (see General Provisions).
Solubility in Alcohol, Appendix VI: One mL dissolves in 5 Acceptance criteria: Between 1.050 and 1.054
mL of 70% alcohol.
Cinnamyl Acetate
288 / Cinnamyl Alcohol / Monographs FCC 9

Cinnamyl Alcohol
.
IDENTIFICATION
Appendix IIIC
Cinnamic Alcohol Acceptance criteria: The spectrum of the sample
ASSAY
Monographs

FEMA: 2294 XI.
UNII: SS8YOP444F [cinnamyl alcohol] Acceptance criteria: NLT 98.0% of C9H10O
DESCRIPTION OTHER REQUIREMENTS

Cinnamyl Alcohol occurs as a white to slightly yellow • ALDEHYDES, M-1b, Appendix XI
crystalline solid. Acceptance criteria: NMT 1.5%
Odor: Balsamic • CHLORINATED COMPOUNDS, Appendix VI
Solubility: Soluble in most fixed oils, propylene glycol; Acceptance criteria: Passes test
insoluble or practically insoluble in glycerin • SOLIDIFICATION POINT, Appendix IIB
Boiling Point: ∼258° Acceptance criteria: NLT 31°
Solubility in Alcohol, Appendix VI: One g dissolves in 1
mL of 70% alcohol, and remains in solution on dilution to
10 mL.
Cinnamyl Alcohol
FCC 9 Monographs / Cinnamyl Butyrate / 289
IDENTIFICATION
Cinnamyl Butyrate
.

First Published: Prior to FCC 6 Appendix IIIC
ASSAY
XI.
Monographs
C13H16O2 Formula wt 204.27 Acceptance criteria: NLT 96.0% of C13H16O2
FEMA: 2296
UNII: TKZ9V37P1G [cinnamyl butyrate] SPECIFIC TESTS
DESCRIPTION OILS), M-15, Appendix XI
Cinnamyl Butyrate occurs as a colorless to pale yellow Acceptance criteria: NMT 1.0
liquid. • REFRACTIVE INDEX, Appendix II: At 20°
Odor: Fruity, balsamic Acceptance criteria: Between 1.525 and 1.530
Boiling Point: ∼300° • SPECIFIC GRAVITY: Determine at 25° by any reliable
Solubility in Alcohol, Appendix VI: One mL dissolves in 1 method (see General Provisions).
mL of 95% alcohol. Acceptance criteria: Between 1.010 and 1.015
Cinnamyl Butyrate
290 / Cinnamyl Cinnamate / Monographs FCC 9

Cinnamyl Cinnamate
.
IDENTIFICATION
Appendix IIIC
ASSAY
Monographs

FEMA: 2298 XI.
UNII: F1438569N2 [cinnamyl cinnamate] Acceptance criteria: NLT 95.0% of C18H16O2

Cinnamyl Cinnamate occurs as a mixture of (Z) and (E) • ACID VALUE, FLAVOR CHEMICALS (OTHER THAN ESSENTIAL
isomers; low-melting solid. OILS), M-15, Appendix XI
Boiling Point: ∼370° Acceptance criteria: NMT 2.0
mL of 95% alcohol.
Cinnamyl Cinnamate
Solubility: Miscible in alcohol, chloroform, ether, most

Cinnamyl Formate
.
fixed oils; insoluble or practically insoluble in water

First Published: Prior to FCC 6 Boiling Point: ∼250°
mL of 80% alcohol to give a clear solution.
IDENTIFICATION
Appendix IIIC
C10H10O2 Formula wt 162.19 Acceptance criteria: The spectrum of the sample
FEMA: 2299 exhibits relative maxima at the same wavelengths as
UNII: 896AGS89RD [cinnamyl formate] those of the spectrum below.
DESCRIPTION ASSAY
Cinnamyl Formate occurs as a colorless to slightly yellow • PROCEDURE: Proceed as directed under M-1a, Appendix
liquid. XI.
Odor: Green, herbaceous, balsamic Acceptance criteria: NLT 92.0% of C10H10O2
FCC 9 Monographs / Cinnamyl Isobutyrate / 291
SPECIFIC TESTS Acceptance criteria: Between 1.077 and 1.082

OILS), M-15, Appendix XI OTHER REQUIREMENTS
Acceptance criteria: NMT 3.0 • CINNAMYL ALCOHOL, M-1a, Appendix XI
• REFRACTIVE INDEX, Appendix II: At 20° Acceptance criteria: NMT 8.0%
Monographs
Cinnamyl Formate
IDENTIFICATION
Cinnamyl Isobutyrate
.

ASSAY
XI.
FEMA: 2297
UNII: C286CMK03D [cinnamyl isobutyrate] SPECIFIC TESTS
Cinnamyl Isobutyrate occurs as a colorless to pale yellow Acceptance criteria: NMT 3.0
liquid. • REFRACTIVE INDEX, Appendix II: At 20°
Odor: Sweet, balsamic, fruity Acceptance criteria: Between 1.523 and 1.528
Solubility in Alcohol, Appendix VI: One mL dissolves in method (see General Provisions).
one mL of 95% alcohol. Acceptance criteria: Between 1.006 and 1.009
292 / Cinnamyl Isobutyrate / Monographs FCC 9
Monographs
Cinnamyl Isobutyrate

Cinnamyl Isovalerate
.
IDENTIFICATION
Appendix IIIC
ASSAY
C14H18O2 Formula wt 218.30 • PROCEDURE: Proceed as directed under M-1b, Appendix
FEMA: 2302 XI.
UNII: 5JHK9Y2XRM [cinnamyl isovalerate] Acceptance criteria: NLT 95.0% of C14H18O2 (one major
isomer)
DESCRIPTION
Cinnamyl Isovalerate occurs as a colorless to slightly yellow SPECIFIC TESTS
liquid. • ACID VALUE, FLAVOR CHEMICALS (OTHER THAN ESSENTIAL
Odor: Spicy, floral, fruity OILS), M-15, Appendix XI
Solubility: Miscible in alcohol, chloroform, most fixed oils, Acceptance criteria: NMT 3.0
ether; insoluble or practically insoluble in glycerin, • REFRACTIVE INDEX, Appendix II: At 20°
propylene glycol, water Acceptance criteria: Between 1.518 and 1.524
FCC 9 Monographs / Cinnamyl Propionate / 293
Monographs
Cinnamyl Isovalerate
IDENTIFICATION
Cinnamyl Propionate
.

ASSAY
XI.
C12H14O2 Formula wt 190.24 Acceptance criteria: NLT 98.0% of C12H14O2 (one
FEMA: 2301 isomer)
UNII: OI92915815 [cinnamyl propionate]
SPECIFIC TESTS
DESCRIPTION • ACID VALUE, FLAVOR CHEMICALS (OTHER THAN ESSENTIAL
Cinnamyl Propionate occurs as a colorless to pale yellow OILS), M-15, Appendix XI
liquid. Acceptance criteria: NMT 3.0
Odor: Spicy, fruity, balsamic • REFRACTIVE INDEX, Appendix II: At 20°
Solubility: Miscible in alcohol, chloroform, ether, most Acceptance criteria: Between 1.532 and 1.537
fixed oils; insoluble or practically insoluble in glycerin, • SPECIFIC GRAVITY: Determine at 25° by any reliable
propylene glycol, water method (see General Provisions).
Boiling Point: ∼289° Acceptance criteria: Between 1.029 and 1.035
294 / Cinnamyl Propionate / Monographs FCC 9
Monographs
Cinnamyl Propionate

Citral
.

First Published: Prior to FCC 6 mL of 70% alcohol.
Last Revision: First Supplement, FCC 6 Function: Flavoring agent
IDENTIFICATION
Mixture of Geranial [(E)-3,7-dimethyl-2,6-octadien-1-al] and • INFRARED SPECTRA, Spectrophotometric Identification Tests,
Neral [the (Z) isomer] Appendix IIIC
ASSAY
XI.
Acceptance criteria: NLT 96.0% of C10H16O (sum of
neral and geranial)
FEMA: 2303 SPECIFIC TESTS
UNII: T7EU0O9VPP [citral] • REFRACTIVE INDEX, Appendix II: At 20°
DESCRIPTION • SPECIFIC GRAVITY: Determine at 25° by any reliable
Citral occurs as a pale yellow liquid. It may contain a method (see General Provisions).
suitable antioxidant. Acceptance criteria: Between 0.885 and 0.891
Odor: Strong, lemon
Solubility: Soluble in most fixed oils, mineral oil, propylene
glycol; insoluble or practically insoluble in glycerin
FCC 9 Monographs / Citric Acid / 295
Monographs
Citral

Citric Acid
.
ASSAY
• PROCEDURE
Sample: 3 g
Analysis: Dissolve the Sample in 40 mL of water, add
phenolphthalein TS and titrate with 1 N sodium
hydroxide. Each mL of 1 N sodium hydroxide is
equivalent to 64.04 mg of C6H8O7.
Acceptance criteria: NLT 99.5% and NMT 100.5% of
C6H8O7, calculated on the anhydrous basis
C6H8O7 Formula wt, anhydrous 192.13
C6H8O7 · H2O Formula wt, monohydrate 210.14 IMPURITIES
INS: 330 CAS: anhydrous [77-92-9] Inorganic Impurities
monohydrate [5949-29-1] • LEAD, Lead Limit Test, Atomic Absorption Spectrophotometric
UNII: 2968PHW8QP [citric acid monohydrate] Graphite Furnace Method, Method I, Appendix IIIB
UNII: XF417D3PSL [anhydrous citric acid] Acceptance criteria: NMT 0.5 mg/kg
Organic Impurities
DESCRIPTION • TRIDODECYLAMINE (FOR SOLVENT-EXTRACTED CITRIC ACID
Citric Acid occurs as colorless, translucent crystals or as a ONLY)
white, granular to fine, crystalline powder. It is anhydrous Buffered indicator solution: Prepare a mixture
or contains one molecule of water of hydration. The consisting of 700 mL of 0.1 M citric acid (anhydrous,
hydrous form is efflorescent in dry air. It is odorless and reagent grade), 200 mL of 0.2 M disodium phosphate,
has a strongly acid taste. One g is soluble in about 0.5 mL and 50 mL each of 0.2% bromophenol blue and of
of water, in about 2 mL of alcohol, and in about 30 mL of 0.2% bromocresol green in spectrograde methanol.
ether. Non-indicator buffer solution: Prepare a mixture
Function: Sequestrant; dispersing agent; acidifier; flavoring consisting of 700 mL of 0.1 M citric acid (anhydrous,
agent reagent grade), 200 mL of 0.2 M disodium phosphate,
Packaging and Storage: Store in tight containers. and 100 mL of spectrograde methanol.
Standard stock solution: 0.08 mg/mL tridodecylamine
IDENTIFICATION in isopropyl alcohol [NOTE—Discard after 3 weeks.]
• CITRATE, Appendix IIIA
296 / Citric Acid / Monographs FCC 9
Standard solution: 4 µg/mL tridodecylamine in Acceptance criteria

isopropyl alcohol: from Standard stock solution [NOTE— Absorbance: NMT 0.52
Prepare this solution fresh on the day of use.] Transmission: NLT 30%
Sample solution: Either 160 g of anhydrous sample in • RESIDUE ON IGNITION (SULFATED ASH), Appendix IIC
320 mL of water, or dissolve 174 g of monohydrate Sample: 4 g
sample in 306 mL of water. Acceptance criteria: NMT 0.05%
Analysis: Dissolve 160 g of anhydrous, reagent-grade • WATER, Water Determination, Appendix IIB
citric acid in 320 mL of water, and divide the solution Acceptance criteria
equally between two 250-mL separatory funnels, S1 Anhydrous: NMT 0.5%
and S2. Add 5 mL of Non-indicator buffer solution to S1. Monohydrate: NMT 8.8%
Monographs
Add 2.0 mL of Standard solution and 5 mL of Buffered

indicator solution to S2. Divide the Sample solution OTHER REQUIREMENTS
equally between two additional 250-mL separatory • LABELING: Indicate whether it is anhydrous or hydrous.
funnels, S3 and S4. Add 5 mL of Non-indicator buffer
solution to S3 and 5 mL of Buffered indicator solution to
S4 .
Citric and Fatty Acid Esters of Glycerol
.
Add 20 mL of a spectrograde chloroform and n-heptane

(1:1, v/v) to each of the four separatory funnels, shake First Published: Third Supplement, FCC 7
for 15 min on a mechanical shaker, and allow the
phases to separate for 45 min. Drain all except the last CITREM
few drops of the lower (aqueous) phases, and discard. Citric Acid Esters of Mono- and Diglycerides
Add 25 mL of 0.05 N sulfuric acid to the organic Citroglycerides
phases in each separatory funnel, hand-shake for 30 s,
INS: 472c
and allow the phases to separate for 30 min. Drain all
except the last few drops of the lower (organic) phases DESCRIPTION
through dry Whatman No. 40 (or equivalent) paper Citric and Fatty Acid Esters of Glycerol occurs as a yellowish
and collect the filtrates in separate, small, glass- to light brown liquid of variable viscosity or as a solid. It
stoppered containers. consists of mixed esters of citric acid and edible fatty acids
Using a suitable spectrophotometer, determine the with glycerol. It may contain minor amounts of free fatty
absorbance of each solution against chloroform and acids, free glycerol, free citric acid, and mono- and
heptane (1:1, v/v) in a 5-cm cell at 400 nm. diglycerides, and may be fully or partially neutralized with
Calculate the Sample solution net absorbance (AU) as the substances suitable for the purpose (as declared on the
absorbance of S4 minus the absorbance of S3. Calculate label). It is obtained by esterification of glycerol with citric
the Standard solution net absorbance (AS) as the acid and edible fatty acids or by reaction of a mixture of
absorbance of S2 minus the absorbance of S1. mono- and diglycerides of edible fatty acid with citric acid.
Acceptance criteria: AU is NMT AS (NMT 0.1 mg/kg) Citroglycerides can be differentiated from stearyl citrate by
the distinctive amount of stearyl alcohol in the latter.
SPECIFIC TESTS Because the mono- or diglycerides in citroglycerides may
• OXALATE
include either one or two fatty acids, and because there is
a variety of edible fatty acids with chain lengths ranging
Analysis: Neutralize 10 mL of the Sample solution with
most commonly from 12 to 18, there is no single
6 N ammonium hydroxide, add 5 drops of 2.7 N
molecular or structural formula. It forms a dispersion in hot
hydrochloric acid, cool, and add 2 mL of calcium
water; is soluble in oils and fats; insoluble in cold water
chloride TS.
and in cold ethanol.
Acceptance criteria: No turbidity forms.
Function: Stabilizer; emulsifier; dough conditioner;
• READILY CARBONIZABLE SUBSTANCES
antioxidant synergist
Sample: 1.00 ± 0.01 g, finely powdered
Packaging and Storage: Store in well-closed containers.
Analysis: Transfer the Sample into a 150-mm × 18-mm
(od) tube previously rinsed with 10 mL of 95% sulfuric ASSAY
acid at 90° or used exclusively for this test. Add • TOTAL GLYCERIN
10 ± 0.1 mL of 95% sulfuric acid, carefully agitate the Solution A: Mix exactly 99 mL (from a buret) of
tube until solution is complete, and immerse the tube chloroform and 25 mL of glacial acetic acid in a 1-L
in a water bath at 90° ± 1° for 1 h. Occasionally volumetric flask.
remove the tube from the water bath, and carefully Sample solution: Weigh accurately 2 g of sample into a
agitate it to ensure that the sample is dissolved and saponification flask, add 50 mL of ethanolic potassium
gaseous decomposition products are allowed to escape hydroxide solution (0.5 M), and gently boil for 30 min.
to the atmosphere. Cool the tube to ambient Quantitatively transfer the content of the saponification
temperature, carefully shake it to ensure that all gases flask to the 1-L volumetric flask with Solution A, using
are removed, and using an adequate three 25-mL portions of water. Add about 500 mL of
spectrophotometer, measure the absorbance and water, and shake vigorously for about 1 min. Dilute
transmission of the solution at 470 nm in a 1-cm cell. with water to volume, mix thoroughly, and set aside for
FCC 9 Monographs / Citric and Fatty Acid Esters of Glycerol / 297
separation of layers. The aqueous layer result is the Standard stock solution: 3 mg/mL of USP Citric Acid
Sample solution. RS solution
Analysis: Pipet 50 mL of acetic periodic acid TS into a Sample solution: Weigh accurately 1 g of sample into a
series of 400-mL beakers. Prepare two blanks by adding round-bottomed flask, add 25 mL of 0.5 M ethanolic
50 mL of water to each. Pipet 50 mL of the Sample potassium hydroxide, and reflux for 30 min. Acidify the
solution into one of the beakers containing 50 mL of mixture with hydrochloric acid, and evaporate in a
acetic periodic acid TS, shake gently to mix, cover with rotary evaporator or by another suitable method.
a watch glass, and allow to stand 30 min but NMT 90 Quantitatively transfer the contents of the flask to a
min. Add 20 mL of 15% ethanolic potassium iodide separator, using NMT 50 mL of water, and extract with
solution, shake gently to mix, and allow to stand at three 50-mL portions of heptane, discarding the
Monographs
least 1 min but NMT 5 min. Do not allow to stand in extracts. Transfer the aqueous layer to a 100-mL
bright or direct sunlight. Add 100 mL of water, and volumetric flask, neutralize, dilute with water to volume,
titrate with 0.1 N sodium thiosulfate VS. Use a variable and mix. Transfer 1 mL of this mixture and 1 mL of the
speed electric stirrer to keep the solution thoroughly Internal standard solution into a 10-mL round-bottom
mixed. Continue the titration until the brown iodine flask, and evaporate to dryness. Add to the flask 1.0 mL
color disappears from the aqueous layer. Add 2 mL of of pyridine, 0.2 mL of trimethyl-chlorosilane, 0.4 mL of
starch TS, and continue the titration until the blue hexamethyl-disilazane, and 0.1 mL of N-methyl-N-
iodine-starch color disappears from both the thin trimethylsilyl-tri-fluoroacetamide. Cap the flask tightly,
chloroform layer (separated during titration) and the and swirl carefully to dissolve completely. Heat the flask
aqueous layer. in an oven at 60° for 1 h.
Calculate the percentage of total glycerol in the sample Standard solution: Transfer 1 mL of the Standard stock
taken: solution and 1 mL of the Internal standard solution into a
10-mL round-bottom flask, and evaporate to dryness.
Result = [(B − S) × N × K]/W Add to the flask 1.0 mL of pyridine, 0.2 mL of
trimethyl-chlorosilane, 0.4 mL of hexamethyl-disilazane,
B = volume of titrant consumed by the blank
and 0.1 mL of N-methyl-N-trimethylsilyl-tri-
containing 50 mL of water (mL)
fluoroacetamide. Cap the flask tightly, and swirl
S = volume of titrant consumed by the Sample
carefully to dissolve completely. Heat the flask in an
solution (mL)
oven at 60° for 1 h.
N = exact normality of 0.1 N thiosulfate
Chromatographic system, Appendix IIA
K = molecular weight of glycerin divided by 40,
Mode: Gas chromatography
2.302
Detector: Flame ionization
W = weight of the original sample taken (g)
Column: 1.8-m × 2.0-mm (id) glass column packed
Acceptance criteria: 8%–33%
with 10% DC-200 on 80- to 100-mesh, chromosorb
• TOTAL FATTY ACID
Q, or equivalent
Analysis: Transfer 5 g of sample into a 250-mL round-
Temperature
bottomed flask, add 50 mL of 1 N ethanolic potassium
Oven: 165°
hydroxide, and reflux for 1 h on a water bath.
Injection block: 240°
Quantitatively transfer the contents of the saponification
Detector block: 240°
flask to a 1000-mL separating funnel, using three 25-mL
Carrier gas: Nitrogen
portions of water, and add 5 drops of methyl orange
Flow rate: 24 mL/min
TS. Cautiously add concentrated hydrochloric acid until
Injection volume: 5 µL
the solution color changes clearly to red, and shake well
Analysis: Separately inject derivatized Sample solution
to separate fatty acids. Extract the separated fatty acids
and Standard solution into the chromatograph. Measure
with three 100-mL portions of diethyl ether. Combine
each peak area by a suitable method, and calculate the
the extracts, and wash with 50-mL portions of 10%
percentage of citric acid in the sample taken:
sodium chloride solution until the washed sodium
[NOTE—The retention times for citric acid/tartaric acid
chloride solution becomes neutral. Dry the ether
and tartaric acid are about 2.3 min and 12 min,
solution with anhydrous sodium sulfate. Then evaporate
respectively.]
off ether on a steam bath, leave an additional 10 min
on the steam bath, and weigh the residue. Result = RS × 100 × RO × 100 × (WO/WS)
Acceptance criteria: 37%–81%
• TOTAL CITRIC ACID RS = peak area ratio of citric acid and tartaric acid
[NOTE—In this test, the sample is saponified with an from the Sample solution
alcoholic potassium hydroxide solution and the fatty RO = peak area ratio of tartaric acid and citric acid
acids are removed by extraction. The citric acid present from the Standard solution
is converted to trimethylsilyl derivatives and analyzed by WO = weight of USP Citric Acid RS used in the
gas liquid chromatography.] Standard solution (g)
Internal standard solution: 1 mg/mL of tartaric acid WS = sample weight (g)
solution Acceptance criteria: 13%–50%
298 / Citric and Fatty Acid Esters of Glycerol / Monographs FCC 9
IMPURITIES Odor: Intense lemon-citronella-rose

Inorganic Impurities Solubility: Soluble in alcohol, most fixed oils; slightly
• LEAD, Lead Limit Test, Flame Atomic Absorption soluble in propylene glycol; insoluble or practically
Spectrophotometric Method, Appendix IIIB insoluble in glycerin, water
Sample: 10 g Boiling Point: ∼206°
Acceptance criteria: NMT 2 mg/kg Solubility in Alcohol, Appendix VI: One mL dissolves in 5
mL of 70% alcohol, and remains clear on dilution.
SPECIFIC TESTS Function: Flavoring agent
• RESIDUE ON IGNITION (SULFATED ASH), Method 1, Appendix
IIC IDENTIFICATION
Monographs
Sample: 2 g • INFRARED SPECTRA, Spectrophotometric Identification Tests,

Acceptance criteria: NMT 0.5% for non-neutralized Appendix IIIC
products, NMT 10% for partially or wholly neutralized Acceptance criteria: The spectrum of the sample
products exhibits relative maxima at the same wavelengths as
• FREE GLYCERIN, Free Glycerin or Propylene Glycol, Appendix those of the spectrum below.
VII
Acceptance criteria: NMT 4% ASSAY
OTHER REQUIREMENTS XI.
• LABELING: Indicate substances used to neutralize material. Acceptance criteria: NLT 85.0% of aldehydes as
C10H18O (one isomer)
SPECIFIC TESTS
Citronellal
.

First Published: Prior to FCC 6 Acceptance criteria: NMT 3.0
Last Revision: First Supplement, FCC 6 • REFRACTIVE INDEX, Appendix II: At 20°
3,7-Dimethyl-6-octen-1-al • SPECIFIC GRAVITY: Determine at 25° by any reliable
OTHER REQUIREMENTS
FEMA: 2307
Acceptance criteria: Between −1° and +11°
UNII: QB99VZZ7GZ [citronellal]
DESCRIPTION
Citronellal occurs as a colorless to slightly yellow liquid. It
may contain a suitable antioxidant.
FCC 9 Monographs / Citronellol / 299
Monographs
Citronellal

Citronellol
.

First Published: Prior to FCC 6 those of the spectrum below.
ASSAY
3,7-Dimethyl-6-octen-1-ol • PROCEDURE: Proceed as directed under Total Alcohols,
Appendix VI.
Sample: 1.2 g
Analysis: Use 78.13 as the equivalence factor (e).
Acceptance criteria: NLT 90.0% of total alcohols as
C10H20O Formula wt 156.27 C10H20O
FEMA: 2309
UNII: P01OUT964K [β-citronellol, (r)-] SPECIFIC TESTS
• REFRACTIVE INDEX, Appendix II: At 20°
DESCRIPTION Acceptance criteria: Between 1.454 and 1.462
Citronellol occurs as a colorless, oily liquid. • SPECIFIC GRAVITY: Determine at 25° by any reliable
Odor: Rosy method (see General Provisions).
Solubility: Soluble in most fixed oils, propylene glycol; Acceptance criteria: Between 0.850 and 0.860
slightly soluble in water; insoluble or practically insoluble in
glycerin OTHER REQUIREMENTS
Boiling Point: ∼225° • ALDEHYDES, M-2c, Appendix XI
Solubility in Alcohol, Appendix VI: One mL dissolves in 2 Sample: 5 g
mL of 70% alcohol and remains in solution on dilution to Analysis: Use 66.08 as the equivalence factor (e).
10 mL. Acceptance criteria: NMT 1.0% as citronellal
Function: Flavoring agent • ESTERS, Appendix VI
Sample: 5 g
IDENTIFICATION Analysis: Use 99.15 as the equivalence factor (e).
• INFRARED SPECTRA, Spectrophotometric Identification Tests, Acceptance criteria: NMT 1.0% as citronellyl acetate
Appendix IIIC
300 / Citronellol / Monographs FCC 9
Monographs
Citronellol

Citronellyl Acetate
.

ASSAY
3,7-Dimethyl-6-octen-1-yl Acetate • PROCEDURE: Proceed as directed under Esters, Appendix
VI.
Sample: 1.4 g
Acceptance criteria: NLT 92.0% of total esters as
C12H22O2 Formula wt 198.31 C12H22O2
FEMA: 2311
UNII: IZ420RT3OY [citronellyl acetate] SPECIFIC TESTS
Citronellyl Acetate occurs as a colorless liquid. Acceptance criteria: NMT 1.0
Odor: Fruity • REFRACTIVE INDEX, Appendix II: At 20°
Solubility: Soluble in alcohol, most fixed oils; insoluble or Acceptance criteria: Between 1.440 and 1.450
practically insoluble in glycerin, propylene glycol, water • SPECIFIC GRAVITY: Determine at 25° by any reliable
mL of 70% alcohol.
IDENTIFICATION
Appendix IIIC
FCC 9 Monographs / Citronellyl Butyrate / 301
Monographs
Citronellyl Acetate

Citronellyl Butyrate
.

ASSAY
3,7-Dimethyl-6-octen-1-yl Butyrate • PROCEDURE: Proceed as directed under Esters, Appendix
VI.
Sample: 1.5 g
C14H26O2 Formula wt 226.36 C14H26O2
FEMA: 2312
UNII: IY68QPD54V [citronellyl butyrate] SPECIFIC TESTS
Citronellyl Butyrate occurs as a colorless liquid. Acceptance criteria: NMT 1.0
Odor: Strong, fruity-rosy • REFRACTIVE INDEX, Appendix II: At 20°
Solubility: Miscible in alcohol, ether, most fixed oils, Acceptance criteria: Between 1.444 and 1.448
chloroform; insoluble or practically insoluble in water • SPECIFIC GRAVITY: Determine at 25° by any reliable
IDENTIFICATION
Appendix IIIC
302 / Citronellyl Butyrate / Monographs FCC 9
Monographs
Citronellyl Butyrate
IDENTIFICATION
Citronellyl Formate
.

3,7-Dimethyl-6-octen-1-yl Formate exhibits relative maxima at the same wavelengths as
ASSAY
• PROCEDURE: Proceed as directed under Esters, Appendix
VI.
C11H20O2 Formula wt 184.28 Sample: 1.0 g
FEMA: 2314 Analysis: Use 92.14 as the equivalence factor (e).
UNII: 7B1MY2BRDK [citronellyl formate] Acceptance criteria: NLT 86.0% of total esters as
C11H20O2
DESCRIPTION
Citronellyl Formate occurs as a colorless liquid. SPECIFIC TESTS
Odor: Strong, fruity, floral • ACID VALUE, FLAVOR CHEMICALS (OTHER THAN ESSENTIAL
Solubility: Soluble in alcohol, most fixed oils; slightly OILS), M-15, Appendix XI
soluble in propylene glycol; insoluble or practically Acceptance criteria: NMT 3.0
insoluble in glycerin, water • REFRACTIVE INDEX, Appendix II: At 20°
Solubility in Alcohol, Appendix VI: One mL dissolves in 3 • SPECIFIC GRAVITY: Determine at 25° by any reliable
mL of 80% alcohol, and remains in solution upon dilution method (see General Provisions).
to 10 mL. Acceptance criteria: Between 0.890 and 0.903
FCC 9 Monographs / Citronellyl Isobutyrate / 303
Monographs
Citronellyl Formate

Citronellyl Isobutyrate
.

ASSAY
3,7-Dimethyl-6-octen-1-yl Isobutyrate • PROCEDURE: Proceed as directed under Esters, Appendix
VI.
Sample: 1.5 g
C14H26O2
FEMA: 2313 SPECIFIC TESTS
UNII: 5RZR3JKW1P [citronellyl isobutyrate] • ACID VALUE, FLAVOR CHEMICALS (OTHER THAN ESSENTIAL
DESCRIPTION Acceptance criteria: NMT 1.0
Citronellyl Isobutyrate occurs as a colorless liquid. • REFRACTIVE INDEX, Appendix II: At 20°
Odor: Fruity-rosy Acceptance criteria: Between 1.440 and 1.448
Solubility: Miscible in alcohol, chloroform, ether, most • SPECIFIC GRAVITY: Determine at 25° by any reliable
fixed oils; insoluble or practically insoluble in water method (see General Provisions).
IDENTIFICATION
Appendix IIIC
304 / Citronellyl Isobutyrate / Monographs FCC 9
Monographs
Citronellyl Isobutyrate
IDENTIFICATION
Citronellyl Propionate
.

Citronellyl Propanoate exhibits relative maxima at the same wavelengths as
3,7-Dimethyl-6-octen-1-yl Propionate those of the spectrum below.
ASSAY
• PROCEDURE: Proceed as directed under Esters, Appendix
VI.
Sample: 1.2 g
C13H24O2 Formula wt 212.33 Analysis: Use 95.12 as the equivalence factor (e).
FEMA: 2316 Acceptance criteria: NLT 90.0% of total esters as
UNII: 87R1092U7J [citronellyl propionate] C13H24O2

Citronellyl Propionate occurs as a colorless liquid. • ACID VALUE, FLAVOR CHEMICALS (OTHER THAN ESSENTIAL
Odor: Fruity-rosy OILS), M-15, Appendix XI
Solubility: Miscible in alcohol, most fixed oils; insoluble or Acceptance criteria: NMT 1.0
practically insoluble in water • REFRACTIVE INDEX, Appendix II: At 20°
Solubility in Alcohol, Appendix VI: One mL dissolves in 4 • SPECIFIC GRAVITY: Determine at 25° by any reliable
mL of 80% alcohol to give a clear solution. method (see General Provisions).
Function: Flavoring agent Acceptance criteria: Between 0.877 and 0.886
FCC 9 Monographs / Clary Oil / 305
Monographs
Citronellyl Propionate
ASSAY
Clary Oil
.
• ESTERS, Ester Determination, Appendix VI

First Published: Prior to FCC 6 Sample: 2 g
Analysis: Use 98.15 as the equivalence factor (e) in the
Clary Sage Oil calculation.
CAS: [8016-63-5] Acceptance criteria: NLT 48.0% and NMT 75.0% of
UNII: 87L0D4U3M0 [clary sage oil] esters, calculated as linalyl acetate (C12H20O2)

Clary Oil occurs as a pale yellow to yellow liquid with a • ACID VALUE (ESSENTIAL OILS AND FLAVORS), Appendix VI
herbaceous odor and a winy bouquet. It is the oil obtained Acceptance criteria: NMT 2.5
by steam distillation from the flowering tops and leaves of • ANGULAR ROTATION, Optical (Specific) Rotation, Appendix
the clary sage plant, Salvia sclarea L. (Fam. Labiatae). It is IIB: Use a 100-mm tube.
soluble in most fixed oils, and in mineral oil up to 3 Acceptance criteria: Between −6° and −20°
volumes, but it becomes opalescent on further dilution. It • REFRACTIVE INDEX, Appendix IIB
is insoluble in glycerin and in propylene glycol. [NOTE—Use an Abbé or other refractometer of equal or
Function: Flavoring agent greater accuracy.]
Packaging and Storage: Store in a cool place protected Acceptance criteria: Between 1.458 and 1.473 at 20°
from light in full, tight containers that are made from steel • SOLUBILITY IN ALCOHOL, Appendix VI
or aluminum and that are suitably lined. Acceptance criteria: One mL of sample dissolves in 3
mL of 90% alcohol and becomes opalescent on further
IDENTIFICATION dilution.
• INFRARED SPECTRA, Spectrophotometric Identification Tests, • SPECIFIC GRAVITY: Determine by any reliable method (see
Appendix IIIC General Provisions).
Acceptance criteria: The spectrum of the sample Acceptance criteria: Between 0.886 and 0.929
306 / Clary Oil / Monographs FCC 9
Monographs
Clary Oil
Analysis: Modify the test by heating the cassia flask in a

Clove Leaf Oil
.
boiling water bath for 10 min after shaking the

First Published: Prior to FCC 6 pretreated Sample with 1 N potassium hydroxide.
Remove from the boiling water bath, cool, and proceed
CAS: [8015-97-2] as directed.
UNII: VCA5491KVF [clove leaf oil] Acceptance criteria: NLT 84.0% and NMT 88.0%, by
volume, of phenols
DESCRIPTION
Clove Leaf Oil occurs as a pale yellow liquid with a sharp,
SPECIFIC TESTS
spicy, peppery odor and taste. It is the volatile oil obtained
by steam distillation of the leaves of Eugenia caryophyllata
Acceptance criteria: Between −2° and 0°
Thunberg (Eugenia aromatica L. Baill.) (Fam. Myrtaceae). It
is soluble in propylene glycol and in most fixed oils with
[NOTE—Use an Abbé or other refractometer of equal or
slight opalescence, and it is relatively insoluble in glycerin
greater accuracy.]
and in mineral oil.
Acceptance criteria: Between 1.531 and 1.535 at 20°
Packaging and Storage: Store in a cool place protected
Acceptance criteria: One mL of sample dissolves in 2
from light in full, tight containers that are made from steel
mL of 70% alcohol. A slight opalescence may occur
or aluminum and that are suitably lined.
when additional solvent is added.
IDENTIFICATION • SPECIFIC GRAVITY: Determine by any reliable method (see
• INFRARED SPECTRA, Spectrophotometric Identification Tests, General Provisions).
Appendix IIIC Acceptance criteria: Between 1.036 and 1.046
ASSAY
• PHENOLS, Appendix VI
Sample: Pretreat a suitable quantity of sample by
shaking it with 2% powdered tartaric acid for about 2
min, and filtering.
FCC 9 Monographs / Clove Oil / 307
Monographs
Clove Leaf Oil
SPECIFIC TESTS
Clove Oil
.

First Published: Prior to FCC 6 IIB: Use a 100-mm tube.
Acceptance criteria: Between −1.5° and 0°
Clove Bud Oil • PHENOLS
CAS: [8000-34-8] Sample: 1 mL
UNII: 578389D6D0 [clove oil] Analysis: Shake the Sample with 20 mL of hot water.
The water shows no more than a scarcely perceptible
DESCRIPTION acid reaction with blue litmus paper. Cool the mixture,
Clove Oil occurs as a colorless or pale yellow liquid with a pass the water layer through a wetted filter, and treat
sharp, spicy odor and taste. It is the volatile oil obtained by the clear filtrate with 1 drop of ferric chloride TS.
steam distillation from the dried flower buds of Eugenia Acceptance criteria: The mixture has only a transient
caryophyllata Thunberg (Eugenia aromatica L. Baill.) (Fam. gray-green color, but not a blue or violet color.
Myrtaceae). It darkens and thickens upon aging or • REFRACTIVE INDEX, Appendix IIB
exposure to air. [NOTE—Use an Abbé or other refractometer of equal or
Function: Flavoring agent greater accuracy.]
Packaging and Storage: Store in aluminum or tin- or Acceptance criteria: Between 1.527 and 1.535 at 20°
epoxyphenolic-lined, tight, light-resistant containers, and • SOLUBILITY IN ALCOHOL, Appendix VI
avoid exposure to excessive heat. Acceptance criteria: One mL of sample dissolves in 2
mL of 70% alcohol.
Appendix IIIC Acceptance criteria: Between 1.038 and 1.060
ASSAY
Acceptance criteria: NLT 85.0%, by volume, of phenols
308 / Clove Oil / Monographs FCC 9
Monographs
Clove Oil
ASSAY
Clove Stem Oil
.
First Published: Prior to FCC 6 Sample: Pretreat a suitable quantity of sample by
shaking it with 2% powdered tartaric acid for about 2
CAS: [8015-98-3] min and filtering.
UNII: 9368YZM9M4 [clove stem oil] Analysis: Modify the test by heating the cassia flask in a
boiling water bath for 10 min after shaking the
DESCRIPTION pretreated Sample with 1 N potassium hydroxide.
Clove Stem Oil occurs as a yellow to light brown liquid with Remove from the boiling water bath, cool, and proceed
a sharp, spicy odor and taste. It is the volatile oil obtained as directed.
by steam distillation from the dried stems of the buds of Acceptance criteria: NLT 89.0% and NMT 95.0%, by
Eugenia caryophyllata Thunberg (Eugenia aromatica L. Baill.) volume, of phenols
(Fam. Myrtaceae). It is soluble in fixed oils and in
propylene glycol, but it is relatively insoluble in glycerin SPECIFIC TESTS
and in mineral oil. • ANGULAR ROTATION, Optical (Specific) Rotation, Appendix
Function: Flavoring agent IIB: Use a 100-mm tube.
Packaging and Storage: Store in a cool place protected • REFRACTIVE INDEX, Appendix IIB
from light in full, tight containers that are made from steel [NOTE—Use an Abbé or other refractometer of equal or
or aluminum and that are suitably lined. greater accuracy.]
Appendix IIIC
FCC 9 Monographs / Cocoa Butter Substitute / 309
Monographs
Clove Stem Oil
Weight %
Cocoa Butter Substitute
.
Fatty Acid (Range)

18:2 0.5–1.5
≥20 0.3–0.7
DESCRIPTION IMPURITIES
Cocoa Butter Substitute occurs as a white, waxy solid that is
predominantly a mixture of triglycerides derived primarily
• LEAD, Lead Limit Test, Atomic Absorption Spectrophotometric
from palm, safflower, sunflower, or coconut oils. The
Graphite Furnace Method, Method II, Appendix IIIB
resulting products may be used directly or with cocoa
Sample: 5 g
butter in all proportions for the preparation of coatings. In
Acceptance criteria: NMT 0.1 mg/kg
contrast to many edible oils and hard butters, Cocoa Butter
• RESIDUAL CATALYST (AS FLUORIDE), Fluoride Limit Test,
Substitute has an abrupt melting range, changing from a
Method I, Appendix IIIB
rather firm, plastic solid below 32° to a liquid at about
Sample: Transfer 30 g of sample into a 250-mL
33.8° to 35.5°.
distillation flask having a side arm and a trap. Connect
Function: Coating agent; texturizer
the flask with a condenser, and fit it with a
thermometer and a capillary tube. Both of these should
IDENTIFICATION reach nearly to the bottom of the flask so that they
• FATTY ACID COMPOSITION, Appendix VII extend into the liquid during the distillation. Add 0.2 g
Acceptance criteria: Cocoa Butter Substitute exhibits of silver sulfate, three boiling beads, and 25 mL of 1:1
the following composition profile of fatty acids: sulfuric acid:water to the flask. Connect a dropping
funnel or a steam generator to the capillary tube. Distill
Weight % until the temperature reaches 135°. Then, through the
Fatty Acid (Range) capillary, add water from the funnel or introduce
≤12 0.0 steam, as necessary, to maintain the temperature as
12:0 0.0
close as possible to 135° until 250 mL of distillate has
been collected in a beaker. Cool the distillate. Add 3
14:0 0.0
mL of 30% hydrogen peroxide to remove any sulfites,
16:0 21–24 let it stand for 5 min, and evaporate the distillate in a
16:1 0.0 dish containing 15 mL of saturated calcium hydroxide
18:0 40–44 suspension. Ash the residue at 600° for 4 h. Use the
18:1 31–35 ashed residue so obtained as the sample in the Analysis
below.
310 / Cocoa Butter Substitute / Monographs FCC 9
Analysis: Proceed as directed beginning with from the Sample preparation at the same time as that
“… and 30 mL of water in a 125-mL distillation flask of the Standard aliquots, read directly from the
having a side arm and trap…” Standard curve the concentration, C, of hexane, in mg/
Acceptance criteria: The total volume of sodium kg, of the Sample preparation. Calculate the quantity of
fluoride TS required for the solutions from both hexane, in mg/kg, in the sample taken by the formula:
Distillate A and Distillate B should not exceed 0.75 mL
(NMT 0.5 mg/kg). Result = 25C/W
Organic Impurities
• HEXANE
W = weight of the sample introduced into the
Standard aliquots: Using a micropipet, transfer and
gas chromatograph (mg)
Monographs
dissolve 34 µL of hexane in 45 g of cold-pressed

cottonseed oil that has not been extracted with
hexane. As directed under Standard curve (below), SPECIFIC TESTS
analyze aliquots of 0.1, 0.25, 0.5, and 5.0 mg; the • COLOR (FATS AND RELATED SUBSTANCES), Appendix VII
aliquots correspond to 2, 5, 10, and 100 mg/kg, Acceptance criteria: NMT 2.5 red
respectively, of residual hexane in a 25-mg sample. • FREE FATTY ACIDS (AS OLEIC ACID), Appendix VII
Sample preparation: Pack the lower half of 8.5-cm × Sample: Use the diglyceride fraction obtained in the test
9.5-mm (od) borosilicate glass tubing (inlet liner) with for Total Glycerides (below).
glass wool that has been heated at 200° for 16 h to Analysis: Determine as directed, except add 2 mL of
expel volatiles. Transfer 25 mg of sample, accurately phenolphthalein TS, and titrate with the appropriate
weighed, into the glass tubing, and cover it with a normality of sodium hydroxide. Use the following
small plug of treated glass wool. equivalence factor (e) in the formula given in the
Standard curve: Chromatograph the Standard aliquots procedure:
as directed for the Sample preparation under Analysis Free fatty acids as oleic acid, e = 28.2
(below), using the same borosilicate glass tubing setup Acceptance criteria: NMT 1.0%
as used for the Sample preparation. Measure the peak • IODINE VALUE, Appendix VII
areas for each of the Standard aliquots. Plot a standard Acceptance criteria: Between 30 and 33
curve using the concentration, in mg/kg, of each of • PEROXIDE VALUE, Method II, Appendix VII
the Standard aliquots versus its corresponding peak Acceptance criteria: NMT 10 mEq/kg
area, and draw the best straight line. To ensure that • TOTAL GLYCERIDES, Total Monoglycerides, Appendix VII
the relative standard deviation does not exceed 2.0%, Analysis: Proceed as directed, using toluene instead of
chromatograph a sufficient number of replicates of benzene and saving all three elution fractions to
each of the Standard aliquots, and record the areas as determine the percentages of Monoglycerides,
directed under Analysis (below). Diglycerides, and Triglycerides. The diglyceride fraction
Chromatographic system, Appendix IIA also contains free fatty acids, the percentage of which is
Mode: Gas chromatography determined under Free Fatty Acids (above). Calculate the
Detector: Independent dual flame-ionization percentage of Total Glycerides (G), which is the sum of
Column: 0.6-m × 6.35-mm (od) stainless-steel U-tube, the percentages of Monoglycerides, Diglycerides, and
or equivalent, packed with Porapak P, or equivalent Triglycerides, by the following formulas:
Inlet temperature: 70°
Detectors temperature: 110° M = WM100/WU
Column temperature: Hold at 70° for 5 min followed
by a linear temperature gradient at 5°/min to 180°, D = (WD100/WU) − F
and finally hold at 180° or until the column is clean
Carrier gas: Helium
Carrier gas flow rate: 60 mL/min T = WT100/WU
Fuel gas: Hydrogen
Fuel gas flow rate: 52 mL/min (for each flame) G=M+D+T
Scavenger gas: Air
Scavenger gas flow rate: 500 mL/min (for each
flame) M =percentage of monoglycerides
Analysis: Insert the Sample preparation into the inlet WM =weight of monoglycerides (g)
liner of the gas chromatograph, immediately sealing WU =weight of the sample (g) taken
the base of the inlet and the lower lip of the glass D =percentage of diglycerides
tubing with a silicone O-ring (Applied Science WD =weight of diglycerides (g)
Laboratories, Inc., or equivalent) previously heated at F =percentage of free fatty acids (determined
200° for 2 h to remove volatile impurities. Immediately above in the test for Free Fatty Acids)
close the inlet liner with the septum and septum liner. T = percentage of triglycerides
Allow the carrier gas to flow through the Sample WT = weight of triglycerides (g)
preparation, chromatograph, and record the Acceptance criteria
chromatograms. Using the peak area of hexane eluting Total glycerides: NLT 98.0% of total
FCC 9 Monographs / Cognac Oil, Green / 311
Monoglycerides: NMT 1.0% Analysis: Proceed as directed under Procedure, except

Diglycerides: NMT 7.0% use 1 µg of As (1.0 mL of Standard Arsenic Solution),
Triglycerides: NLT 90.0% instead of 3 µg As.
• UNSAPONIFIABLE MATTER, Appendix VII Acceptance criteria: The absorbance caused by any red
Acceptance criteria: NMT 1.0% color from the Sample solution does not exceed that
• WATER, Water Determination, Appendix IIB produced by the 1.0 mL of Standard Arsenic Solution.
Analysis: Proceed as directed except use 50 mL of a 1:1 (NMT 0.5 mg/kg)
solution of chloroform in methanol to dissolve the • LEAD, Lead Limit Test, Atomic Absorption Spectrophotometric
sample instead of using 35 to 40 mL of methanol. Graphite Furnace Method, Method II, Appendix IIIB
Acceptance criteria: NMT 0.1% Acceptance criteria: NMT 0.1 mg/kg
Monographs
SPECIFIC TESTS
• COLOR (FATS AND RELATED SUBSTANCES), Appendix VII
Acceptance criteria: NMT 20 yellow/2.0 red
Coconut Oil (Unhydrogenated)
.
• FREE FATTY ACIDS, Appendix VII

First Published: Prior to FCC 6 Analysis: Determine as directed using the following
equivalence factor (e) in the formula given in the
CAS: [8001-31-8] procedure:
UNII: Q9L0O73W7L [coconut oil] Free fatty acids as oleic acid, e = 28.2
Free fatty acids as lauric acid, e = 20.0
DESCRIPTION Acceptance criteria:
Coconut Oil (Unhydrogenated) occurs as a viscous, white to Oleic acid: NMT 0.1%
light yellow-tan liquid. It is obtained from the kernel of the Lauric acid: NMT 0.07%
fruit of the coconut palm Cocos nucifera (Fam. Palmae). • IODINE VALUE, Appendix VII
The crude oil obtained by mechanically pressing dried Acceptance criteria: Between 6 and 11
coconut meat (copra) is refined, bleached, and deodorized • MELTING RANGE (FATS AND RELATED SUBSTANCES), Appendix
to substantially remove free fatty acids, phospholipids, VII
color, odor and flavor components, and other non-oil Acceptance criteria: Between 23.5° and 27°
materials. Compared with many natural fats, Coconut Oil • PEROXIDE VALUE, Method II, Appendix VII
(Unhydrogenated) has an abrupt melting range, changing Acceptance criteria: NMT 10 mEq/kg
from a rather firm, plastic solid at about 21° or below or to • UNSAPONIFIABLE MATTER, Appendix VII
a liquid at about 27°. Acceptance criteria: NMT 1.5%
Function: Coating agent; emulsifying agent; texturizer • WATER, Water Determination, Appendix IIB
Packaging and Storage: Store in well-closed containers. Analysis: Proceed as directed except use 50 mL of
IDENTIFICATION chloroform to dissolve the sample instead of using 35
• FATTY ACID COMPOSITION, Appendix VII to 40 mL of methanol.
Acceptance criteria: A sample exhibits the following Acceptance criteria: NMT 0.1%
typical composition profile of fatty acids:
Fatty Acid Weight % (Range)

Cognac Oil, Green
.
6:0 0–0.8
8:0 5–9 First Published: Prior to FCC 6
10:0 4–8
12:0 44–52
Wine Yeast Oil
CAS: [8016-21-5]
14:0 15–21
UNII: 930MLC8XGG [grape seed oil]
16:0 8–11
16:1 0–1 DESCRIPTION
18:0 1–4
Cognac Oil, Green occurs as a green to blue-green liquid
with the characteristic aroma of cognac. It is the volatile oil
18:1 5–8
obtained by steam distillation from wine lees. It is soluble
18:2 0–2.5 in most fixed oils and in mineral oil. It is very slightly
20:0 0–0.4 soluble in propylene glycol, and it is insoluble in glycerin.
Packaging and Storage: Store in full, tight containers in
IMPURITIES a cool place protected from light.
• ARSENIC, Arsenic Limit Test, Appendix IIIB IDENTIFICATION
Sample solution: Prepare using 2 g of sample • INFRARED SPECTRA, Spectrophotometric Identification Tests,
Appendix IIIC
312 / Cognac Oil, Green / Monographs FCC 9
Acceptance criteria: The spectrum of the sample Acceptance criteria: Between 200 and 245
exhibits relative maxima at the same wavelengths as • REFRACTIVE INDEX, Appendix IIB
those of the spectrum below. [NOTE—Use an Abbé or other refractometer of equal or
greater accuracy.]
SPECIFIC TESTS Acceptance criteria: Between 1.427 and 1.430 at 20°
• ACID VALUE (ESSENTIAL OILS AND FLAVORS), Appendix VI • SOLUBILITY IN ALCOHOL, Appendix VI
Acceptance criteria: Between 32 and 70 Acceptance criteria: One mL of sample dissolves in 2
• ANGULAR ROTATION, Optical (Specific) Rotation, Appendix mL of 80% alcohol.
IIB: Use a 100-mm tube. • SPECIFIC GRAVITY: Determine by any reliable method (see
Acceptance criteria: Between −1° and +2° General Provisions).
Monographs
• ESTER VALUE, Esters, Appendix VI Acceptance criteria: Between 0.864 and 0.870
Sample: 1 g
Cognac Oil, Green
IDENTIFICATION
Copaiba Oil
.

CAS: [8013-97-6] exhibits relative maxima at the same wavelengths as
UNII: 64VX45Y68N [copaiba oil] those of the spectrum below.

Copaiba Oil occurs as a colorless to slightly yellow liquid • ANGULAR ROTATION, Optical (Specific) Rotation, Appendix
with the characteristic odor of copaiba balsam and an IIB: Use a 100-mm tube.
aromatic, slightly bitter and pungent taste. It is the volatile Acceptance criteria: Between −7° and −33°
oil obtained by steam distillation of copaiba balsam, an • GURJUN OIL
exudate from the trunk of various South American species Analysis: Introduce 5 or 6 drops of sample into 10 mL
of Copaifera L. (Fam. Leguminosae). It is soluble in alcohol, of glacial acetic acid to which 5 drops of nitric acid has
in most fixed oils, and in mineral oil. It is insoluble in been added.
glycerin and practically insoluble in propylene glycol. Acceptance criteria: No purple color appears within 2
Function: Flavoring agent min, indicating the absence of gurjun oil.
Packaging and Storage: Store in a cool place protected • REFRACTIVE INDEX, Appendix IIB
from light in full, tight containers that are made from steel [NOTE—Use an Abbé or other refractometer of equal or
or aluminum and that are suitably lined. greater accuracy.]
FCC 9 Monographs / Copovidone / 313
• SPECIFIC GRAVITY: Determine by any reliable method (see Acceptance criteria: Between 0.880 and 0.907
General Provisions).
Monographs
Copaiba Oil
Packaging and Storage: Store in tight containers.

Copovidone
.
IDENTIFICATION
First Published: First Supplement, FCC 6
• A. PROCEDURE
PVP/VA Copolymer Analysis: To 5 mL of the Sample solution, add a few
Poly(1-vinyl-2-pyrrolidone)/(vinyl acetate) Copolymer drops of iodine TS.
1-Vinyl-2-pyrrolidone polymer with vinyl acetate Acceptance criteria: A deep red color is produced.
Acetic acid ethenyl ester polymer with 1-ethenyl-2- • B. INFRARED ABSORPTION, INFRARED SPECTRA,
pyrrolidone Spectrophotometric Identification Tests, Appendix IIIC
Copolyvidone Reference standard: USP Copovidone RS
Sample and Standard preparation: K
the spectrum of the Reference standard.
ASSAY
• NITROGEN DETERMINATION, Method II, Appendix IIIC
(C6H9NO)n/(C4H6O2)m CAS: [25086-89-9] Sample: 100 mg
UNII: D9C330MD8B [copovidone] Bromocresol green–methyl red solution: Dissolve 0.15
g of bromocresol green and 0.1 g of methyl red in 180
DESCRIPTION mL of alcohol, and dilute with water to 200 mL.
Copovidone occurs as a white to yellowish-white powder or
Analysis: Modify the procedure as follows: in the wet-
as flakes. It is a copolymer of 1-vinyl-2-pyrrolidone and
digestion step, use 5 g of a 33:1:1 mixture of potassium
vinyl acetate in the mass proportion of 3:2. It is freely
sulfate–cupric sulfate–titanium dioxide instead of the
soluble in water, in alcohol, and in methylene chloride, but
10:1 potassium sulfate–cupric sulfate mixture; and omit
practically insoluble in ether. It is hygroscopic. The pH of a
the use of hydrogen peroxide. Heat until the solution
1:10 aqueous solution is between 3 and 7.
has a clear, yellow-green color and the sides of the flask
Function: Coating for fresh and fresh-cut fruits and
are free from carbonaceous material. Then heat for an
vegetables, film-forming agent, binder, and crystallization
additional 45 min and continue as directed, beginning
inhibitor
with “Cautiously add 20 mL of water, cool, then…”.
314 / Copovidone / Monographs FCC 9
Use Bromocresol green–methyl red solution instead of volumetric flask, dilute with Pyrophosphate buffer to
methyl red–methylene blue TS. Titrate the distillate with volume, and mix.
0.05 N sulfuric acid until the color of the solution Sample solution: Transfer 1 g of sample into a 100-mL
changes from green through pale grayish-blue to pale volumetric flask, dissolve it in 50 mL of Pyrophosphate
grayish red-purple. Perform a blank determination, and buffer, dilute to volume with Pyrophosphate buffer, and
make any necessary correction. Each mL of 0.05 N mix. Stopper the flask loosely, heat at 60° for 1 h, and
sulfuric acid is equivalent to 0.7004 mg of nitrogen. cool to room temperature.
Acceptance criteria: NLT 7.0% and NMT 8.0%, Blank: Water
calculated on the dried basis Analysis: Pipet 0.5 mL each of the Standard solution,
• COPOLYMERIZED VINYL ACETATE the Sample solution, and Blank into separate 1-cm cells.
Monographs
Sample: 2 g Add 2.5 mL of Pyrophosphate buffer and 0.2 mL of NAD

Analysis: Transfer the Sample into a 250-mL borosilicate solution to each cell. Cover the cells to exclude oxygen.
glass flask, add 25 mL of 0.5 N alcoholic potassium Mix them by inversion and allow them to stand for 2
hydroxide and a few glass beads, and heat under reflux to 3 min at 22° ± 2°. Using a suitable
for 30 min. Add 1 mL of phenolphthalein TS, and spectrophotometer, measure the absorbances of the
titrate the excess 0.5 N alcoholic potassium hydroxide solutions at 340 nm. Add 0.05 mL of Aldehyde
immediately (while still hot) with 0.5 N hydrochloric dehydrogenase solution to each cell. Stopper the cells
acid. Perform a blank determination and make any tightly and mix by inversion. Allow them to stand for 5
necessary correction. Calculate the percentage of min at 22 ± 2°. Measure the absorbances of the
copolymerized vinyl acetate in the sample taken by the solutions as before. Calculate the percentage of
formula: Aldehydes (as acetaldehyde) in the sample taken by the
formula:
Result = 0.1 × (Mr1/Mr2) × (Mr2 × NA) × (VB − VU)/W
Result = 10(C/W){[(AU2 − AU1) − (AB2 − AB1)]/[(AS2 − AS1) −
(AB2 − AB1)]}
Mr1 = molecular weight of vinyl acetate, 86.09
Mr2 = molecular weight of potassium hydroxide,
56.11 C = concentration (mg/mL) of acetaldehyde in
NA = actual normality of the alcoholic potassium the Standard solution calculated from the
hydroxide weight of the acetaldehyde–ammonia
VB = volume (mL) of 0.5 N hydrochloric acid trimer trihydrate with the factor of 0.72
consumed for the blank determination W = weight (g) of sample taken to prepare the
VU = volume (mL) of 0.5 N hydrochloric acid Sample solution
consumed for the Sample determination AU1 = absorbance of the solution obtained from
W = weight (g) of Sample taken the Sample solution, before the Aldehyde
Acceptance criteria: NLT 35.3% and NMT 42.0%, dehydrogenase solution was added
calculated on the dried basis AS1 = absorbance of the solution obtained from
the Standard solution before the Aldehyde
IMPURITIES dehydrogenase solution was added
Inorganic Impurities AB1 = absorbance of the solution obtained from
• LEAD, Lead Limit Test, Flame Atomic Absorption the Blank, before the Aldehyde
Spectrometric Method, Appendix IIIB dehydrogenase solution was added
Sample: 10 g AU2 = absorbance of the solution obtained from
Acceptance criteria: NMT 2 mg/kg the Sample solution, after the Aldehyde
Organic Impurities dehydrogenase solution was added
• ALDEHYDES AS2 = absorbance of the solution obtained from
Pyrophosphate buffer, 0.05 M: Transfer 8.7 g of the Standard solution after the Aldehyde
potassium pyrophosphate into a 500-mL volumetric dehydrogenase solution was added
flask, and dissolve in 400 mL of water. Adjust, if AB2 = absorbance of the solution obtained from
necessary, to a pH of 9.0 with 1 N potassium the Blank, after the Aldehyde dehydrogenase
hydroxide, dilute to volume, and mix. solution was added
Aldehyde dehydrogenase solution: Transfer a quantity Acceptance criteria: NMT 0.05% (as acetaldehyde)
of lyophilized aldehyde dehydrogenase (Sigma A550, • HYDRAZINE, Thin-Layer Chromatography, Appendix IIA
or equivalent) equivalent to 70 units into a glass vial, Salicylaldazine standard solution: 9 µg/mL of
dissolve it in 10.0 mL of water, and mix. [NOTE—This salicylaldazine in toluene
solution is stable for 8 h at 4°.] Sample: An amount of sample equivalent to 2.5 g on
NAD solution: 4.0 mg/mL of nicotinamide adenine the dried basis
dinucleotide in Pyrophosphate buffer Sample solution: Transfer the Sample into a 50-mL
Standard solution: Dissolve 0.140 g of acetaldehyde centrifuge tube, add 25 mL of water, and mix to
ammonia trimer trihydrate in 200.0 mL of water, and dissolve. Add 500 µL of a 50 mg/mL solution of
mix. Pipet 1.0 mL of this solution into a 100-mL salicylaldehyde in methanol, swirl, and heat in a water
bath at 60° for 15 min. Allow the solution to cool, add
FCC 9 Monographs / Copovidone / 315
2.0 mL of toluene, insert a stopper in the tube, shake System suitability

the tube vigorously for 2 min, and centrifuge. The clear Sample: Standard solution
upper layer is the Sample solution. Resolution: NLT 2.0 between 2-pyrrolidone, vinyl
Adsorbent: 0.25-mm layer of dimethylsilanized acetate, and 1-vinyl-2-pyrrolidone peaks
chromatographic silica gel mixture with fluorescent Relative standard deviation: NMT 2.0% for each
indicator analyte for replicate injections
Developing solvent system: Methanol and water Analysis: Separately inject the Standard solution and the
(2:1) Sample solution into the chromatograph, record the
Application volume: 10 µL chromatograms, and measure the responses for the 2-
Detection/Visualization: UV 365 nm pyrrolidone, vinyl acetate, and 1-vinyl-2-pyrrolidone
Monographs
Analysis: Spot the Sample solution and the peak areas. [NOTE—The order of elution is 2-
Salicylaldazine standard solution onto the plate. pyrrolidone, vinyl acetate, and 1-vinyl-2-pyrrolidone.]
Following development, locate the spots on the plate [NOTE—After each injection of the Sample solution, wash
by examination under UV light. Salicylaldazine appears the polymeric material from the guard column by
as a fluorescent spot having an RF value of about 0.3. passing the mobile phase (100% Solution A) through
Acceptance criteria: The fluorescence of any the column backwards for about 30 min at the same
salicylaldazine spot from the Sample solution is not more flow rate.] Calculate the content of the three monomers
intense than that produced by the spot obtained from in the sample taken using the following equations:
the Salicylaldazine standard solution (NMT 1 mg/kg).
• MONOMERS (1-VINYL-2-PYRROLIDONE, VINYL ACETATE, AND 1-Vinyl-2-pyrrolidone (mg/kg) = (rUA/rSA) ×
2-PYRROLIDONE) (CSA/CU) × F1
Solution A: Acetonitrile, methanol, and water
(1:1:18) Vinyl acetate (mg/kg) = (rUB/rSB) × (CSB/CU) × F1
Solution B: Acetonitrile, methanol, and water
(9:1:10)
Standard stock solution: 0.5 mg/mL 1-vinyl-2- 2-Pyrrolidone (%) = (rUC/rSC) × (CSC/CU) ×
pyrrolidone, 0.5 mg/mL vinyl acetate, and 3 mg/mL of F2 × 100%
2-pyrrolidone in methanol
Standard solution: 0.25 µg/mL 1-vinyl-2-pyrrolidone, rUA = peak area responses from the Sample
0.25 µg/mL vinyl acetate, and 1.5 µg/mL of 2- rUB solution for 1-vinyl-2-pyrrolidone, vinyl
pyrrolidone prepared by diluting the Standard stock rUC acetate, and 2-pyrrolidone, respectively
solution with Solution A rSA = peak area responses from the Standard
Sample solution: Transfer 250 mg of sample into a 10- rSB solution for 1-vinyl-2-pyrrolidone, vinyl
mL volumetric flask, add 1 mL of methanol, mix rSC acetate, and 2-pyrrolidone, respectively
ultrasonically to dissolve, dilute with water to volume, CSA = concentrations of 1-vinyl-2-pyrrolidone,
and mix. If necessary, filter this solution to remove CSB vinyl acetate, and 2-pyrrolidone,
undissolved particles. CSC respectively, in the Standard solution (µg/
Chromatographic system, Appendix IIA mL)
Mode: High-performance liquid chromatography CU = Sample solution concentration (mg/mL)
Detectors: UV 205 nm and 235 nm F1 = correction factor to convert units from
Column: 4-mm × 250-mm stainless steel, packed with mg/g to mg/kg, 100
octadecylsilane silica gel (5 µm particle diameter)1, F2 = correction factor to convert units from, 1/
with 4-mm × 30-mm guard column with the same 1000
packing2 µg/mg to µg/µg
Column temperature: 30° Acceptance criteria
Flow rate: About 1.0 mL/min 1-Vinyl-2-pyrrolidone: NMT 10 mg/kg
Injection volume: About 10 µL Vinyl acetate: NMT 10 mg/kg
Mobile phase: See gradient table (below). 2-Pyrrolidone: NMT 0.5%
• PEROXIDES
Time Solution A Solution B
(min) (%) (%)
Sample stock solution: 40 mg/mL
Titanium trichloride solution: 150 mg/mL titanium
0 100 0
trichloride in 10% hydrochloric acid
2 100 0 Titanium trichloride–sulfuric acid solution: Mix
26 80 20 carefully 20 mL of Titanium trichloride solution in 13 mL
27 0 100 of sulfuric acid. Add sufficient 30% hydrogen peroxide
36 0 100 to produce a yellow color. Heat until white fumes are
evolved, allow to cool, and dilute with water. Repeat
38 100 0
the evaporation and addition of water until a colorless
solution is obtained. Dilute with water to 100 mL.
1 Aquasil C18 (Thermo-Hypersil), or equivalent. Sample solution: Transfer 25.0 mL of the Sample stock
2 Nucleosil 120-5 C18 (Machery-Nagel), or equivalent. solution to a 50-mL beaker, add 2 mL of Titanium
316 / Copovidone / Monographs FCC 9
trichloride–sulfuric acid solution, and mix. Allow to stand z = relative viscosity

for 30 min at room temperature. Acceptance criteria: NLT 90.0% and NMT 110.0% of
Blank solution: Transfer 25.0 mL of Sample stock solution the K-Value stated on the label
to a 50-mL beaker, add 2 mL of 13% sulfuric acid, and • LOSS ON DRYING, Appendix IIC : 105° for 3h
mix. Acceptance criteria: NMT 5.0%
Analysis: Measure the absorbances of the Sample • RESIDUE ON IGNITION (SULFATED ASH), Appendix IIC
solution and Blank solution in a 1-cm cell at the Sample: 2 g
wavelength of maximum absorbance (about 405 nm), Analysis: Proceed as directed, but igniting at
using a suitable spectrophotometer. 600° ± 50° for 30 min.
Acceptance criteria: The blank-corrected absorbance is Acceptance criteria: NMT 0.1%
Monographs
NMT 0.35 (corresponding to NMT 0.04%, expressed as

hydrogen peroxide). OTHER REQUIREMENTS
• LABELING: Indicate the nominal K-Value of the product.
SPECIFIC TESTS
• CLARITY AND COLOR
Copper Gluconate
.
Acceptance criteria: The Sample solution is clear or

slightly opalescent and colorless to pale yellow or pale
red.
• K-VALUE
[NOTE—The molecular weight of the sample is
characterized by its viscosity in aqueous solution,
relative to that of water, expressed as a K-Value.]
Sample: An amount of sample equivalent to 1.0 g on
the dried basis
Sample solution: Transfer the Sample into a 100-mL C12H22CuO14 Formula wt 453.84
CAS: [527-09-3]
volumetric flask, dissolve it in about 50 mL of water,
dilute with water to volume, mix thoroughly, and allow UNII: RV823G6G67 [copper gluconate]
it to stand for 1 h. Filter the solution. Pipet 15 mL of
DESCRIPTION
filtrate into a clean, dry Ubbelholde-type viscometer,
Copper Gluconate occurs as a fine, light blue powder. It is
and place the viscometer in a water bath maintained at
very soluble in water, and is very slightly soluble in alcohol.
25° ± 0.2°.
Function: Nutrient
Analysis: After allowing the viscometer and the Sample
solution to warm in the water bath for 10 min, draw
the solution by means of very gentle suction up IDENTIFICATION
through the capillary until the meniscus is above the • COPPER, Appendix IIIA
upper etched mark. Release suction, and after the Sample solution: 50 mg/mL
meniscus reaches the upper etched mark, begin timing Acceptance criteria: Passes tests
the flow through the capillary. Record the exact time • THIN-LAYER CHROMATOGRAPHY, Appendix IIA
when the meniscus reaches the lower etched mark, and Sample solution: 10 mg/mL in water
calculate the flow time to the nearest 0.01 s. Repeat Standard solution: 10 mg/mL of USP Potassium
this operation until at least three readings are obtained. Gluconate RS in water
The readings must agree within 0.1 s; if not, repeat the [NOTE—Sample solution and Standard solution (above), if
determination with additional 15-mL portions of the necessary, can be heated in a water bath at 60° to aid
Sample solution after recleaning the viscometer with dissolution.]
sulfuric acid–dichromate cleaning solution or with a Adsorbent: 0.25-mm layer of chromatographic silica gel
suitable laboratory cleaning compound that will remove Application volume: 5 µL
oils, greases, waxes, and other impurities. Calculate the Developing solvent system: Alcohol, water, ammonium
average flow time and then obtain the flow time in a hydroxide, and ethyl acetate (5:3:1:1)
similar manner for 15 mL of water. Calculate the Spray reagent: 25 mg/mL of ammonium molybdate
relative viscosity, z, of the Sample by dividing the and 10 mg/mL of ceric sulfate in 2 N sulfuric acid
average flow time of the Sample solution by that of the Analysis: Following development, dry the plate at 110°
water sample, and then calculate the K-Value by the for 20 min, and allow to cool. Spray the cooled plate
formula: with the Spray reagent. After spraying, heat the plate at
110° for about 10 min.
Acceptance criteria: The principal spot obtained from
the Sample solution corresponds in color, size, and RF
value to that of the Standard solution.
c = weight (g) of the sample, calculated on the

dried basis, in each 100.0 g of solution
FCC 9 Monographs / Copper Sulfate / 317
ASSAY DESCRIPTION
• PROCEDURE Copper Sulfate occurs as blue crystals, crystalline granules,
Sample: 1.5 g or powder. It effloresces slowly in dry air and is freely
Analysis: Dissolve the Sample in 100 mL of water in a soluble in water, soluble in glycerin, and slightly soluble in
250-mL Erlenmeyer flask, add 2 mL of glacial acetic alcohol.
acid and 5 g of potassium iodide, mix well, and titrate Function: Nutrient
with 0.1 N sodium thiosulfate to a light yellow color. Packaging and Storage: Store in tight containers.
Add 2 g of ammonium thiocyanate, mix, then add 3
mL of starch TS and continue titrating to a milk-white IDENTIFICATION
endpoint. Each mL of 0.1 N sodium thiosulfate is • A. COPPER, Appendix IIIA
Monographs
equivalent to 45.38 mg of C12H22CuO14. Sample solution: 50 mg/mL
Acceptance criteria: NLT 98.0% and NMT 102.0% Acceptance criteria: Passes tests
C12H22CuO14 • B. SULFATE, Appendix IIIA
IMPURITIES Acceptance criteria: Passes tests
• LEAD, Lead Limit Test, Appendix IIIB ASSAY
Sample solution: 1 g of sample in 25 mL of water • PROCEDURE
Control: 5 µg of Pb (5 mL of Diluted Standard Lead Sample: 1 g
Solution) Analysis: Dissolve the Sample in 50 mL of water. Add 4
Acceptance criteria: NMT 5 mg/kg mL of glacial acetic acid and 3 g of potassium iodide,
mix well, and titrate with 0.1 N sodium thiosulfate to a
SPECIFIC TESTS light yellow color. Add 2 g of ammonium thiocyanate,
• REDUCING SUBSTANCES mix, and then add 3 mL of starch TS, and continue
Sample: 1 g titrating to a milky white endpoint. Perform a blank
Analysis: Dissolve the Sample in 10 mL of water in a titration (see General Provisions) and make any
250-mL Erlenmeyer flask. Add 25 mL of alkaline cupric necessary correction. Each mL of 0.1 N sodium
citrate TS and cover the flask with a small beaker. Boil thiosulfate used is equivalent to 24.97 mg of CuSO4 ·
gently for exactly 5 min, and cool rapidly to room 5H2O.
temperature. Add 25 mL of a 1:10 solution of acetic Acceptance criteria: NLT 98.0% and NMT 102.0% of
acid, 10.0 mL of 0.1 N iodine, 10.0 mL of 2.7 N CuSO4 · 5H2O
hydrochloric acid, and 3 mL of starch TS and titrate
with 0.1 N sodium thiosulfate to the disappearance of IMPURITIES
the blue color. Calculate the weight, in mg, of reducing Inorganic Impurities
substances (as D-glucose) by the formula: • IRON
Sample: Residue from Substances Not Precipitated by
Result = (V1N1 − V2N2) × FE Hydrogen Sulfide (below)
Control: 0.033 mg iron
Analysis: Add 2 mL of hydrochloric acid and 0.1 mL of
V1 = volume of the iodine solution (mL) nitric acid to the Sample, cover with a watch glass, and
N1 = normality of the iodine solution digest on a steam bath for 20 min. Remove the watch
V2 = volume of the sodium thiosulfate solution glass, and evaporate to dryness. Dissolve the residue in
(mL) 1 mL of hydrochloric acid, and dilute to 60 mL with
N2 = normality of the sodium thiosulfate solution water. Dilute 5 mL of this solution to 40 mL with
FE = equivalence factor for D-glucose, 27 water, add 2 mL of hydrochloric acid, and dilute to 50
Acceptance criteria: NMT 1.0% mL with water. Add 40 mg of ammonium
peroxydisulfate crystals and 10 mL of ammonium
thiocyanate TS, and mix thoroughly. Repeat the
preceding using the Control in place of the Sample.
Copper Sulfate
.
Acceptance criteria: Any red color produced within 1 h

First Published: Prior to FCC 6 by the Sample does not exceed that produced by the
Control. (NMT 0.01%)
Cupric Sulfate • LEAD, Lead Limit Test, APDC Extraction Method, Appendix
IIIB
CuSO4 Formula wt, anhydrous 159.6 Acceptance criteria: NMT 4 mg/kg
CuSO4 · 5H2O Formula wt, pentahydrate 249.68
INS: 519 CAS: anhydrous [7758-98-7] SPECIFIC TESTS
pentahydrate [7758-99-8] • SUBSTANCES NOT PRECIPITATED BY HYDROGEN SULFIDE
UNII: KUW2Q3U1VV [cupric sulfate anhydrous] Sample: 5 g
UNII: LRX7AJ16DT [cupric sulfate] Analysis: Dissolve the Sample in 200 mL of 1:100
sulfuric acid, heat to 70°, and pass hydrogen sulfide
through the solution until the copper is completely
318 / Copper Sulfate / Monographs FCC 9
precipitated. Dilute to 250 mL, mix thoroughly, allow IDENTIFICATION

the precipitate to settle, and filter. Evaporate 200 mL of • INFRARED SPECTRA, Spectrophotometric Identification Tests,
the filtrate to dryness in a tared dish, ignite at 800° ± Appendix IIIC
25° for 15 min, cool, and weigh. [NOTE—Retain the Acceptance criteria: The spectrum of the sample
resulting residue for the Iron test (above).] exhibits relative maxima at the same wavelengths as
Acceptance criteria: NMT 0.3% those of the spectrum below.
SPECIFIC TESTS
Coriander Oil IIB: Use a 100-mm tube.
.
Monographs
Acceptance criteria: Between +8° and +15°

First Published: Prior to FCC 6 • REFRACTIVE INDEX, Appendix IIB
CAS: [8008-52-4] greater accuracy.]
UNII: 7626GC95E5 [coriander oil] Acceptance criteria: Between 1.462 and 1.472 at 20°
DESCRIPTION Acceptance criteria: One mL of sample dissolves in 3
Coriander Oil occurs as a colorless or pale yellow liquid with
mL of 70% alcohol.
the characteristic odor and taste of coriander. It is the
volatile oil obtained by steam distillation from the dried
ripe fruit of Coriandrum sativum L. (Fam. Umbelliferae).
Packaging and Storage: Store in full, tight containers
protected from light. Avoid exposure to excessive heat.
Coriander Oil
FCC 9 Monographs / Costus Root Oil / 319
Analysis: Proceed as directed except use 50 mL of

Corn Oil (Unhydrogenated)
.
chloroform to dissolve the sample instead of using 35

First Published: Prior to FCC 6 to 40 mL of methanol.
CAS: [8001-30-7]
SPECIFIC TESTS
UNII: 8470G57WFM [corn oil] • COLOR (FATS AND RELATED SUBSTANCES), Appendix VII
Acceptance criteria: NMT 5.0 red
DESCRIPTION
• FREE FATTY ACIDS (AS OLEIC ACID), Appendix VII
Corn Oil (Unhydrogenated) occurs as an amber-colored oil.
Analysis: Use 28.2 for the equivalence factor (e) in the
It is obtained from the corn plant Zea mays (Fam.
Monographs
formula given in the procedure.
Gramineae), usually by solvent extraction of the corn
germ. It is refined, bleached, and deodorized to
• IODINE VALUE, Appendix VII
substantially remove free fatty acids, phospholipids, color,
Acceptance criteria: Between 120 and 130
odor and flavor components, and other non-oil materials.
• LINOLENIC ACID, Fatty Acid Composition, Appendix VII
It is a liquid at 21° to 27°, but traces of wax, unless they
are removed by winterization, may cause the oil to cloud
• PEROXIDE VALUE, Method II, Appendix VII
when cooled to low temperature. It is free from visible
Acceptance criteria: NMT 10 mEq/kg
foreign material (other than wax) at 21° to 27°.
• UNSAPONIFIABLE MATTER, Appendix VII
Function: Coating agent; emulsifying agent; texturizer
IDENTIFICATION
• FATTY ACID COMPOSITION, Appendix VII
Costus Root Oil
.
Acceptance criteria: Corn Oil exhibits the following

typical composition profile of fatty acids: First Published: Prior to FCC 6
Fatty Acid Weight % (Range)

CAS: [8023-88-9]
<14 <0.1 UNII: 2WF6750061 [costus root oil]
14:0 <1.0
16:0 8.0–19 DESCRIPTION
Costus Root Oil occurs as a light yellow to brown, viscous
16:1 <0.5
liquid with a peculiar, persistent odor reminiscent of violet,
18:0 0.5–4.0
orris, and vetivert. It is the volatile oil obtained by steam
18:1 19–50 distillation from the dried, triturated roots of the
18:2 38–65 herbaceous perennial plant Saussurea lappa Clarke (Fam.
18:3 <2.0 Compositae) or by a solvent extraction procedure followed
20:0 <1.0
by vacuum distillation of the resinoid extract. It is soluble
in most fixed oils and in mineral oil. It is insoluble in
20:1 <0.5
glycerin and in propylene glycol.
22:0 <0.3 Function: Flavoring agent
22:1 <0.1 Packaging and Storage: Store in a cool place protected
24:0 <0.4 from light in full, tight containers that are made from steel
or aluminum and that are suitably lined.
IMPURITIES IDENTIFICATION
Inorganic Impurities • INFRARED SPECTRA, Spectrophotometric Identification Tests,
• ARSENIC, Arsenic Limit Test, Appendix IIIB Appendix IIIC
Sample solution: Use 4 g of sample and prepare as Acceptance criteria: The spectrum of the sample
directed for organic compounds. exhibits relative maxima at the same wavelengths as
Analysis: Proceed as directed under Procedure, except those of the spectrum below.
use 2 µg of As (2.0 mL of Standard Arsenic Solution)
instead of 3 µg As. SPECIFIC TESTS
Acceptance criteria: The absorbance caused by any red • ACID VALUE (ESSENTIAL OILS AND FLAVORS), Appendix VI
color from the Sample solution does not exceed that Acceptance criteria: NMT 42
produced by the 2.0 mL of Standard Arsenic Solution. • ANGULAR ROTATION, Optical (Specific) Rotation, Appendix
(NMT 0.5 mg/kg) IIB: Use a 100-mm tube.
• LEAD, Lead Limit Test, Atomic Absorption Spectrophotometric Acceptance criteria: Between +10° and +36°
Graphite Furnace Method, Method II, Appendix IIIB • ESTERS, Ester Value, Appendix VI
Acceptance criteria: NMT 0.1 mg/kg Sample: 1 g
• WATER, Water Determination, Appendix IIB Acceptance criteria: Between 90 and 150
320 / Costus Root Oil / Monographs FCC 9
• REFRACTIVE INDEX, Appendix IIB upon further dilution, and paraffin crystals may
[NOTE—Use an Abbé or other refractometer of equal or occasionally separate.
greater accuracy.] • SPECIFIC GRAVITY: Determine by any reliable method (see
Acceptance criteria: Between 1.512 and 1.523 at 20° General Provisions).
• SOLUBILITY IN ALCOHOL, Appendix VI Acceptance criteria: Between 0.995 and 1.039
Acceptance criteria: One mL of sample dissolves in 0.5
mL of 90% alcohol, but the solution becomes cloudy
Monographs
Costus Root Oil
IDENTIFICATION
Cottonseed Oil (Unhydrogenated)
.
• FATTY ACID COMPOSITION, Appendix VII

First Published: Prior to FCC 6 Acceptance criteria: A sample exhibits the following
composition profile of fatty acids:
CAS: [8001-29-4]
Weight %
UNII: H3E878020N [cottonseed oil]
Fatty Acid (Range)
DESCRIPTION <14 <0.1
Cottonseed Oil (Unhydrogenated) occurs as a dark red- 14:0 0.5-2.0
brown oil. It is obtained from the seed of the cotton plant 16:0 17-29
Gossypium hirsutum (American) or Gossypium barbadense 16:1 <1.5
(Egyptian) by mechanical expression or solvent extraction.
18:0 1.0-4.0
It is refined, bleached, and deodorized to substantially
remove free fatty acids, phospholipids, color, odor and 18:1 13-44
flavor components, and miscellaneous other non-oil 18:2 40-63
materials. It is liquid at 21° to 27°, clouds at 21°, and 18:3 0.1-2.1
partially solidifies at storage temperatures below 10° to 20:0 <0.5
16°. It is free from visible foreign material at 23° to 27°.
20:1 <0.5
Function: Cooking or salad oil; component of margarine
22:0 <0.5
or shortening; tenderizer; carrier; stabilizer; thickener;
coating agent; texturizer 22:1 <0.5
Packaging and Storage: Store in well-closed containers. 24:0 <0.5
FCC 9 Monographs / p-Cresyl Acetate / 321
IMPURITIES C9H10O2 Formula wt 150.18

Inorganic Impurities FEMA: 3073
• LEAD, Lead Limit Test, Atomic Absorption Spectrophotometric UNII: 42I5PWW20C [p-cresyl acetate]
Acceptance criteria: NMT 0.1 mg/kg DESCRIPTION
• WATER, Water Determination, Appendix IIB p-Cresyl Acetate occurs as a colorless liquid.
Analysis: Proceed as directed, except use 50 mL of Odor: Strong, floral
chloroform to dissolve the sample instead of 35 to 40 Solubility: Soluble in most fixed oils, propylene glycol;
mL of methanol. insoluble or practically insoluble in glycerin
Acceptance criteria: NMT 0.1% Boiling Point: ∼212°
Monographs
Organic Impurities Solubility in Alcohol, Appendix VI: One mL dissolves in 2
• FREE FATTY ACIDS (AS OLEIC ACID), Free Fatty Acids, mL of 70% alcohol.
Appendix VII Function: Flavoring agent
Analysis: Use 28.2 for the equivalence factor (e) in the
formula given in the procedure.
IDENTIFICATION
Appendix IIIC
• LINOLENIC ACID, Fatty Acid Composition, Appendix VII
SPECIFIC TESTS those of the spectrum below.
• COLOR (FATS AND RELATED SUBSTANCES), Appendix VII
Acceptance criteria: NMT 70 yellow/4.5 red
ASSAY
XI.
Acceptance criteria: NLT 98.0% of C9H10O2 (one
• PEROXIDE VALUE
isomer)
Sample: 10 g
Analysis: To the Sample, add 30 mL of a glacial acetic SPECIFIC TESTS
acid and chloroform mixture (3:2) and mix. Add 1 mL • ACID VALUE, FLAVOR CHEMICALS (OTHER THAN ESSENTIAL
of a saturated solution of potassium iodide and mix for OILS), M-15, Appendix XI
1 min. Add 100 mL of water, begin titrating [NOTE—Use phenol red TS as the indicator.]
immediately with 0.05 N sodium thiosulfate, adding Acceptance criteria: NMT 1.0
starch TS as the endpoint is approached, and continue • REFRACTIVE INDEX, Appendix II: At 20°
the titration until the blue starch color has just Acceptance criteria: Between 1.499 and 1.502
disappeared. Perform a blank determination (see • SPECIFIC GRAVITY: Determine at 25° by any reliable
General Provisions), and make any necessary correction. method (see General Provisions).
Calculate the peroxide value, as mEq/kg of sample, by Acceptance criteria: Between 1.044 and 1.050
the formula:
OTHER REQUIREMENTS
Result = S × N × 1000/W • FREE CRESOL, M-17, Cresyl Acetate (Test for Free Cresol),
Appendix XI
S = net volume of sodium thiosulfate solution
required for the sample (mL)
N = exact normality of the sodium thiosulfate
solution
W = weight of sample taken (g)
Acceptance criteria: NMT 10 mEq/kg
p-Cresyl Acetate
.
p-Methylphenyl Acetate
p-Tolyl Acetate
322 / p-Cresyl Acetate / Monographs FCC 9
Monographs
p-Cresyl Acetate

Crospovidone
.

First Published: Prior to FCC 6 the spectrum of the Reference standard.
Last Revision: First Supplement, FCC 7 • B. PROCEDURE
Sample: 1 g
Analysis: Add 0.1 mL of iodine TS to a suspension of
Polyvinylpolypyrrolidone
the Sample in 10 mL of water, and shake the mixture
PVPP
for 30 s. [NOTE—The reagent is discolored, a distinction
1-Vinyl-2-pyrrolidone Crosslinked Insoluble Polymer
from povidone, which produces a red color.] Add 1 mL
of starch TS, and shake the mixture.
Acceptance criteria: No blue color appears.
ASSAY
• NITROGEN DETERMINATION, Method II, Appendix IIIC
INS: 1202 Sample: 100 mg
UNII: 68401960MK [crospovidone] Analysis: Determine as directed, except in the wet-
digestion step, repeat the addition of hydrogen
DESCRIPTION peroxide (usually three to six times) until a clear, light
Crospovidone occurs as a white to off-white, hygroscopic, green solution is obtained, then heat for an additional 4
free-flowing powder. It is a crosslinked homopolymer of h, and continue as directed, beginning with “Cautiously
purified vinylpyrrolidone, produced catalytically. It is add 20 mL of water”.
insoluble in water and in other common solvents. Acceptance criteria: NLT 11.0% and NMT 12.8% of
Function: Clarifying agent; stabilizer nitrogen (N)
Packaging and Storage: Store in tight containers.
IMPURITIES
IDENTIFICATION Inorganic Impurities
• A. INFRARED ABSORPTION, Spectrophotometric Identification • LEAD, Lead Limit Test, Flame Atomic Absorption
Tests, Appendix IIIC Spectrophotometric Method, Appendix IIIB
Reference standard: USP Crospovidone RS Sample: 10 g
Standard and sample preparation: K; previously dried Acceptance criteria: NMT 2 mg/kg
in vacuum at 105° for 1 h
FCC 9 Monographs / Cubeb Oil / 323
SPECIFIC TESTS Analysis: Separately inject the Standard solution and the
• ACID-ALCOHOL SOLUBLE SUBSTANCES Sample solution into the chromatograph, record the
Sample: 1 g chromatograms, and measure the responses for the
Solubility solution: 15 g of glacial acetic acid in 50 mL vinylpyrrolidone peak area. [NOTE—If necessary, after
of ethanol and sufficient water to make 500 mL of each injection of the Sample solution, wash the
solution polymeric material from the guard column by passing
Analysis: Place the Sample into a flask containing 500 the Mobile phase through the column backwards for
mL of the Solubility solution. Allow the contents of the about 30 min at the same flow rate.]
flask to rest for 24 h. Filter on a filter screen with a Calculate the concentration (mg/mL) of
porosity of 2.5 µm, then on a filter screen with a vinylpyrrolidinone in the sample taken:
Monographs
porosity of 0.8 µm. Concentrate the filtrate over a
water bath. Finish evaporation over the water bath in a Result = 1000 × (C/W) × (rU/rS)
70-mm diameter tared silica capsule.
Acceptance criteria: The dry residue remaining after C = concentration of vinylpyrrolidinone in the
evaporation must be less than 10 mg, taking into Standard solution (mg/mL)
account any residue left by the evaporation of 500 mL W = weight of sample taken to prepare the
of the acetic acid–ethanol mixture (NMT 1.0%). Sample solution (mg)
• PH, pH Determination, Appendix IIB rU = peak area response for vinylpyrrolidinone
Sample suspension: 1 g in 100 mL of water obtained from the Sample solution
Acceptance criteria: Between 5.0 and 11.0 rS = peak area response for vinylpyrrolidinone
• RESIDUE ON IGNITION (SULFATED ASH), Appendix IIC obtained from the Standard solution
Sample: 2 g Acceptance criteria: NMT 0.001%
Acceptance criteria: NMT 0.4% • WATER, Water Determination, Appendix IIB
• UNSATURATION (AS VINYLPYRROLIDONE) Acceptance criteria: NMT 6.0%
Mobile phase: Acetonitrile and water (10:90) • WATER SOLUBLE SUBSTANCES
System suitability solution: Transfer 10 mg of Sample: 10 g
vinylpyrrolidone and 500 mg of vinyl acetate to a 100- Analysis: Place the Sample into a 200-mL flask
mL volumetric flask, and dissolve in and dilute with containing 100 mL of water. Shake the flask, and allow
methanol to volume. Transfer 1.0 mL of this solution to the contents to rest for 24 h. Pass through a membrane
a 100-mL volumetric flask, dilute with Mobile phase to filter having a 0.45-µm porosity, protected against
volume, and mix. clogging by superimposing a membrane filter having a
Standard solution: Transfer 50 mg of vinylpyrrolidone 3-µm porosity. Evaporate the filtrate over a water bath
to a 100-mL volumetric flask, dilute with methanol to until dry.
volume, and mix. Transfer 1.0 mL of this solution to a Acceptance criteria: The residue left by evaporating the
100-mL volumetric flask, dilute with methanol to filtrate is less than 150 mg (NMT 1.5%).
volume, and mix. Transfer 5.0 mL of this solution to a
100-mL volumetric flask, dilute with Mobile phase to
volume, and mix.
Sample solution: Suspend 1.250 g in 50.0 mL of
Cubeb Oil
.
methanol, and shake for 60 min. Leave the bulk to

settle, and pass through a 0.2-µm filter. First Published: Prior to FCC 6
Mode: High-performance liquid chromatography CAS: [8007-87-2]
Detector: UV 235 nm UNII: HDE20IN4M5 [cubeb oil]
Column: Stainless steel column about 4-mm × 250-
mm, packed with octadecylsilanized silica gel (5 µm in DESCRIPTION
particle diameter), with a guard column about 4-mm Cubeb Oil occurs as a colorless or light green to blue-green
× 25-mm with the same packing liquid with a spicy odor and a slightly acrid taste. It is the
Column temperature: 40° volatile oil obtained by steam distillation from the mature,
Flow rate: Adjust so that the retention time of unripe, sun-dried fruit of the perennial vine Piper cubeba L.
vinylpyrrolidone is about 10 min. (Fam. Piperaceae). It is soluble in most fixed oils and in
Injection volume: About 50 µL mineral oil, but it is insoluble in glycerin and propylene
System suitability glycol.
Samples: System suitability solution and Standard Function: Flavoring agent
solution Packaging and Storage: Store in a cool place protected
Suitability requirement 1: The resolution, R, between from light in full, tight containers that are made from steel
vinylpyrrolidone and vinyl acetate for the System or aluminum and that are suitably lined.
suitability solution is NLT 2.0.
IDENTIFICATION
Suitability requirement 2: The relative standard
deviation for replicate injections of the Standard
Appendix IIIC
solution is NMT 2.0%.
324 / Cubeb Oil / Monographs FCC 9
Acceptance criteria: The spectrum of the sample Acceptance criteria: Between 1.492 and 1.502 at 20°
exhibits relative maxima at the same wavelengths as • SAPONIFICATION VALUE, Appendix VI
those of the spectrum below. Sample: 5 g
Acceptance criteria: NMT 8
SPECIFIC TESTS • SOLUBILITY IN ALCOHOL, Appendix VI
• ACID VALUE (ESSENTIAL OILS AND FLAVORS), Appendix VI Acceptance criteria: One mL of sample dissolves in 10
Acceptance criteria: NMT 2.0 mL of 90% alcohol.
• ANGULAR ROTATION, Optical (Specific) Rotation, Appendix • SPECIFIC GRAVITY: Determine by any reliable method (see
IIB: Use a 100-mm tube. General Provisions).
Acceptance criteria: Between −12° and −43° Acceptance criteria: Between 0.898 and 0.928
Monographs

greater accuracy.]
Cubeb Oil
IDENTIFICATION
Cumin Oil
.

CAS: [8014-13-9] exhibits relative maxima at the same wavelengths as
UNII: N356X94O43 [cumin oil] those of the spectrum below.
DESCRIPTION ASSAY
Cumin Oil occurs as a light yellow to brown liquid with a • ALDEHYDES, Appendix VI
strong and somewhat disagreeable odor. It is the volatile Sample: 1 g
oil obtained by steam distillation from the plant Cuminum Analysis: Proceed as directed, but allow the mixture to
cyminum L. (Fam. Umbelliferae). It is relatively soluble in stand for 30 min at room temperature before titrating.
most fixed oils and in mineral oil. It is very soluble in Use 74.10 as the equivalence factor (e) in the
glycerin and in propylene glycol. calculation.
Function: Flavoring agent Acceptance criteria: NLT 45.0% and NMT 54.0% of
Packaging and Storage: Store in a cool place protected aldehydes, calculated as cuminaldehyde (C10H12O)
from light in full, tight containers that are made from steel SPECIFIC TESTS
or aluminum and that are suitably lined. • ANGULAR ROTATION, Optical (Specific) Rotation, Appendix
FCC 9 Monographs / Cuminic Aldehyde / 325
Acceptance criteria: Between +3° and +8° • SOLUBILITY IN ALCOHOL, Appendix VI

• REFRACTIVE INDEX, Appendix IIB Acceptance criteria: One mL of sample dissolves in 8
[NOTE—Use an Abbé or other refractometer of equal or mL of 80% alcohol. The solution can become hazy on
greater accuracy.] the addition of more alcohol.
Acceptance criteria: Between 1.500 and 1.506 at 20° • SPECIFIC GRAVITY: Determine by any reliable method (see
Monographs
Cumin Oil

Cuminic Aldehyde
.

First Published: Prior to FCC 6 mL of 70% alcohol.
Last Revision: First Supplement, FCC 6 Function: Flavoring agent
IDENTIFICATION
Cuminal • INFRARED SPECTRA, Spectrophotometric Identification Tests,
Cuminaldehyde Appendix IIIC
p-Cuminic Aldehyde Acceptance criteria: The spectrum of the sample
p-Isopropylbenzaldehyde exhibits relative maxima at the same wavelengths as
ASSAY
• PROCEDURE: Proceed as directed under M-2a, Appendix
XI.
Acceptance criteria: NLT 95.0% of C10H12O
FEMA: 2341
SPECIFIC TESTS
UNII: O0893NC35F [cuminaldehyde]
DESCRIPTION Acceptance criteria: NMT 5.0
Cuminic Aldehyde occurs as a colorless to pale yellow liquid. • REFRACTIVE INDEX, Appendix II: At 20°
It may contain a suitable antioxidant. Acceptance criteria: Between 1.528 and 1.534
Odor: Strong, pungent, cumin oil • SPECIFIC GRAVITY: Determine at 25° by any reliable
Solubility: Soluble in alcohol, ether; insoluble or practically method (see General Provisions).
insoluble in water Acceptance criteria: Between 0.975 and 0.980
326 / Cuminic Aldehyde / Monographs FCC 9
OTHER REQUIREMENTS
• CHLORINATED COMPOUNDS, Appendix VI
Monographs
Cuminic Aldehyde
IDENTIFICATION
Curdlan
.
• A. PROCEDURE
First Published: Prior to FCC 6 Sample mixture: 2% aqueous suspension of sample
Analysis: Add 5 mL of sulfuric acid TS to 10 mL of
Beta-1,3-glucan Sample mixture, heat in a boiling water bath for 30 min,
and cool. Neutralize the mixture with barium
carbonate, and centrifuge it at 900 g for 10 min. Add 1
mL of the supernatant to 5 mL of hot alkaline cupric
tartrate TS.
Acceptance criteria: A copious red precipitate of
cuprous oxide forms.
• B. PROCEDURE
(C6H10O5)n CAS: [54724-00-4] Sample mixture: 2% aqueous suspension of sample
UNII: 6930DL209R [curdlan] Analysis: Heat the Sample mixture in a boiling water
bath for 10 min, then cool.
DESCRIPTION Acceptance criteria: A firm gel forms.
Curdlan occurs as a white to nearly white powder. It is a • C. PROCEDURE
high-molecular-weight polymer of glucose (β-1,3-glucan) Sample: 0.2 g
produced by pure-culture fermentation of a carbohydrate Analysis: Suspend the Sample in 5 mL of water, add 1
by a nonpathogenic and nontoxigenic strain of mL of 3 N sodium hydroxide, and shake.
Agrobacterium biobar 1 (formerly Alcaligenes faecalis var. Acceptance criteria: The sample dissolves.
myxogenes) or Agrobacterium radiobacter. Curdlan consists
of β-(1,3)-linked glucose residues and has the unusual ASSAY
property of forming an elastic gel when its aqueous • ANHYDROUS GLUCOSE CONTENT
suspension is heated to a temperature above 54°. It is Sample solution: Transfer 100 mg of sample into a 100-
insoluble in water, but is soluble in alkaline solutions. mL volumetric flask and dissolve in and dilute to
Function: Firming agent; gelling agent; stabilizer; thickener volume with 0.1 N sodium hydroxide. Transfer 5 mL of
Packaging and Storage: Store in airtight containers. this solution into a 100-mL volumetric flask, dilute to
volume with water, and mix. To 1 mL of this solution,
add 1 mL of a 50 mg/mL solution of reagent-grade
FCC 9 Monographs / Cyclamen Aldehyde / 327
phenol and 5 mL of sulfuric acid TS. Shake the flask 0.196 = area (cm2) of the plunger
vigorously, and cool it in ice-cold water. Acceptance criteria: NLT 600 g/cm2
Standard solution: Prepare as directed for the Sample • LOSS ON DRYING, Appendix IIC: 60° for 5 h in a vacuum
solution, using 100 mg of reagent-grade glucose in Acceptance criteria: NMT 10%
place of the sample. • MICROBIAL LIMITS
Blank solution: Prepare as directed for the Sample [NOTE—Current methods for the following tests may be
solution, using 0.1 mL of water in place of the sample. found in the Food and Drug Administration’s
Analysis: Use a suitable spectrophotometer set to 490 Bacteriological Analytical Manual online at www.fda.gov
nm with 1-cm sample cells and set to zero with the /Food/default.htm.]
Blank solution. Determine the absorbance of the Sample Acceptance criteria
Monographs
solution and the Standard solution. Calculate the percent Aerobic plate count: NMT 1000 CFU/g
curdlan (%C) in the sample taken using the following e. coli: Negative in 1 g
equation: • NITROGEN, Nitrogen Determination, Method II, Appendix
IIIC
%C = (A/AR) × (0.9 × WR/W) × 100% Sample: 1 g
• PH, pH Determination, Appendix IIB
A = absorbance of the Sample solution
Sample suspension: 1% aqueous suspension
AR = absorbance of the Standard solution
0.9 = molecular weight of anhydrous glucose
• RESIDUE ON IGNITION (SULFATED ASH), Appendix IIC
divided by the molecular weight of glucose
Sample: 1 g
WR = weight (mg) of the reagent-grade glucose
Acceptance criteria: NMT 6%
used to make the Standard solution
W = weight (mg) of the sample used to make
the Sample solution
Acceptance criteria: NLT 80% (calculated as anhydrous
Cyclamen Aldehyde
.
glucose)
IMPURITIES Last Revision: First Supplement, FCC 6
2-Methyl-3-(p-isopropylphenyl)propionaldehyde
SPECIFIC TESTS
• GEL STRENGTH
2% Sample suspension: Place 200 mg of sample into
the tube of a Potter-Elvehjem homogenizer, add 10 mL
of water, and homogenize at about 1500 g for 5 min. C13H18O Formula wt 190.29
Analysis: Transfer the 2% Sample suspension into a 16- FEMA: 2743
mm × 150-mm test tube, de-aerate in vacuum for 3 UNII: 4U37UX0E1E [cyclamen aldehyde]
min, and heat in a boiling water bath for 10 min to
form a gel. Cool in running water, let it stand for 30 DESCRIPTION
min, and remove the gel from the test tube. Accurately Cyclamen Aldehyde occurs as a colorless to pale yellow
cut the gel at distances of 20 mm and 30 mm from the liquid. It may contain a suitable antioxidant.
bottom to obtain a section 10 mm long. Determine the Odor: Strong, floral
gel strength using a Rheo Meter Model CR-200D (Sun Solubility: Soluble in most fixed oils; insoluble or
Scientific Co., Ltd., Japan; Load cell: 1000 g; set to a practically insoluble in glycerin, propylene glycol
measurement mode 4) or an equivalent instrument Boiling Point: ∼270°
capable of uniaxial compression and having a load cell Solubility in Alcohol, Appendix VI: One mL dissolves in 3
sensitivity of 500 to 1000 g. Use a cylindrical stainless mL of 80% alcohol.
steel plunger with a 0.5-cm diameter. Lower the Function: Flavoring agent
plunger into the gel at 250 mm/min. The resulting
force-time curve is recorded and used for gel strength
IDENTIFICATION
calculation. Calculate gel strength by the following
Appendix IIIC
equation:
gel strength (g force/cm2) = f/0.196 cm2 exhibits relative maxima at the same wavelengths as
f = force on the force-time curve that shows a ASSAY

sharp yielding downward trend associated • PROCEDURE: Proceed as directed under M-1b, Appendix
with rupture of the gel XI.
328 / Cyclamen Aldehyde / Monographs FCC 9
Acceptance criteria Acceptance criteria: NMT 5.0

Sum of two isomers: NLT 90.0% of C13H18O • REFRACTIVE INDEX, Appendix II: At 20°
Major isomer: NLT 85.0% of C13H18O Acceptance criteria: Between 1.503 and 1.508
SPECIFIC TESTS method (see General Provisions).
• ACID VALUE, FLAVOR CHEMICALS (OTHER THAN ESSENTIAL Acceptance criteria: Between 0.946 and 0.952
Monographs
Cyclamen Aldehyde
Sample and standard preparation: M

Cyclamic Acid
.

First Published: FCC 8 exhibits maxima at the same wavelengths as those in
Last Revision: Second Supplement, FCC 8 the spectrum of the Reference standard.
• B. PROCEDURE
Cyclohexanesulfamic Acid
Analysis: Acidify the Sample solution with hydrochloric
Cyclohexylsulfamic Acid
acid. Add 1 mL of barium chloride TS to the acidified
solution, then filter if any turbidity or precipitate forms.
When a clear solution is obtained, add 1 mL of 10%
sodium nitrite solution.
Acceptance criteria: A white precipitate forms.
C6H13NO3S Formula wt 179.24
INS: 952 CAS: [100-88-9] ASSAY
UNII: HN3OFO5036 [cyclamic acid] • PROCEDURE
Phenolphthalein solution: Dissolve 0.2 g of
DESCRIPTION phenolphthalein in 60 mL of 90% ethanol, and dilute
Cyclamic Acid occurs as a practically colorless, white with water to 100 mL.
crystalline powder. It is soluble in water and in ethanol. Sample: 350 mg
Function: Sweetener Analysis: Transfer the Sample to a 250-mL flask, and
Packaging and Storage: Store in well-closed containers. dissolve it in 50 mL of water. Titrate the solution with
0.1 N sodium hydroxide, using Phenolphthalein solution
IDENTIFICATION as the indicator. Each mL of 0.1 N sodium hydroxide is
• A. INFRARED ABSORPTION, Spectrophotometric Identification equivalent to 17.92 mg of C6H13NO3S.
Tests, Appendix IIIC Acceptance criteria: 98.0%–102.0% of C6H13NO3S,
Reference standard: USP Cyclamic Acid RS calculated on the dried basis
FCC 9 Monographs / alpha-Cyclodextrin / 329
IMPURITIES measure the responses. [NOTE—The approximate

Inorganic Impurities retention times (relative to cyclohexanamine, which has
• LEAD, Lead Limit Test, Flame Atomic Absorption a retention time of about 2.3 min) for aniline,
Spectrophotometric Method, Appendix IIIB tetradecane, and N-cyclohexylcyclohexanamine are
Sample: 5 g about 1.4, 4.3, and 4.5 min, respectively.]
Acceptance criteria: NMT 1.0 mg/kg Acceptance criteria
Organic Impurities Cyclohexanamine: NMT 10.0 mg/kg
• CYCLOHEXANAMINE, ANILINE, AND N- Aniline: NMT 1.0 mg/kg
CYCLOHEXYLCYCLOHEXANAMINE N-Cyclohexylcyclohexanamine: NMT 1.0 mg/kg
Internal standard solution: 0.02 µL/mL of tetradecane
Monographs
in methylene chloride SPECIFIC TESTS
Solution A: Dissolve 10 mg of cyclohexanamine, 1 mg • LOSS ON DRYING, Appendix IIC: 105° for 1 h
of N-cyclohexylcyclohexanamine, and 1 mg of aniline Acceptance criteria: NMT 1%
in water, then dilute with the same solvent to 1000
mL. Dilute 10 mL of this solution with water to 100
mL.
alpha-Cyclodextrin
.
Solution B: 42% (w/v) sodium hydroxide solution

Standard solution: To 20 mL of Solution A add 0.5 mL First Published: Second Supplement, FCC 6
of Solution B, and extract with 30 mL of toluene. Shake
20 mL of the upper layer with 4 mL of a mixture of α-Schardinger dextrin
equal volumes of water and an acetic acid solution α-Dextrin
(12% w/v). Separate the lower layer, add 0.5 mL of Cyclohexaamylose
Solution B and 0.5 mL of the Internal standard solution, Cyclomaltohexose
and shake. Use the lower layer immediately after α-Cycloamylose
separation.
Sample solution: Dissolve 2 g of sample in 20 mL of
water, add 0.5 mL of Solution B, and shake with 30 mL
of toluene. Shake 20 mL of the upper layer with 4 mL
of a mixture of equal volumes of an acetic acid solution
(12% w/v) and water. Separate the lower layer, add
0.5 mL of Solution B and 0.5 mL of the Internal
standard solution, and shake. Use the lower layer
immediately after separation.
Detector: Flame ionization
Column: 25-cm × 0.32-mm (i.d.) fused-silica column (C6H10O5)6 Formula wt 972.85
INS: 457 CAS: [10016-20-3]
with poly(dimethyl)(diphenyl)siloxane containing
95% of methyl groups and 5% of phenyl groups1 as UNII: Z1LH97KTRM [alfadex]
stationary phase (film thickness 0.51 µm)
DESCRIPTION
Carrier gas: Helium
Alpha-Cyclodextrin occurs as a virtually odorless, white or
Flow rate: 1.8 mL/min
almost white crystalline solid. It is a non-reducing cyclic
Temperatures
saccharide consisting of six α-(1→4)-linked D-
Injection port: 250°
glucopyranosyl units produced by the action of
Detector: 270°
cyclodextrin glucosyltransferase (CGTase, EC 2.4.1.19) on
Column: See the temperature program in the table
hydrolyzed starch. Recovery and purification of alpha-
below.
cyclodextrin may be carried out using one of the following
procedures: precipitation of a complex of alpha-
Time Temperature
(min) (°) cyclodextrin with 1-decanol, dissolution in water at
elevated temperature and re-precipitation, steam-stripping
0–1 85
of the complexant, and crystallization of alpha-cyclodextrin
1–9 85–150
from the solution; or chromatography with ion-exchange
9–13 150 or gel filtration followed by crystallization of alpha-
cyclodextrin from the purified mother liquor; or membrane
Injection volume: 1.5 µL. Use a split vent at a flow separation methods such as ultra-filtration and reverse
rate of 20 mL/min. osmosis. It is freely soluble in water and very slightly
Analysis: Separately inject equal volumes of the soluble in ethanol.
Standard solution and Sample solution into the Function: Carrier; encapsulating agent; stabilizer
chromatograph, record the chromatograms, and Packaging and storage: Store in tight containers in a dry
1 DB-5 available from J&W Scientific, SE-52 available from Restek Corp., or place.
equivalent.
330 / alpha-Cyclodextrin / Monographs FCC 9
IDENTIFICATION heating in a water bath. Prepare a solution of 16.0 g

• PROCEDURE sodium hydroxide in 200 mL of water and a solution of
Acceptance criteria: The retention time of the major 300 g sodium potassium tartrate in 500 mL of water.
peak in the chromatogram of the Sample solution is the Transfer both solutions to the 1000-mL flask. Dilute
same as that of the Standard solution in the Assay. with water to volume, shake the flask, and let it stand
for 24 h. Filter (paper) the reagent solution prior to use
ASSAY if a precipitate appears.
• PROCEDURE Standard stock solution: 10 mg/mL dextrose (on the
Mobile phase: Acetonitrile and water (67:33, v/v) anhydrous basis)
Standard solution: 10 mg/mL USP alpha-Cyclodextrin Standard solutions: 0 to 1.0 mg/mL dextrose prepared
Monographs
RS [NOTE—Ultra-sonication for 10–15 min may be as follows: weigh 1.0 g of alpha-cyclodextrin standard2
necessary to aid in complete dissolution.] into each of eleven 10-mL volumetric flasks (numbered
Sample solution: 10 mg/mL filtered through a 0.45-µm 0 to 10). Add 0, 0.1, 0.2, ..., 1.0 mL of Standard
filter [NOTE—Ultra-sonication for 10–15 min may be solution to flasks nos. 0, 1, ... to 10, respectively. Dilute
necessary to aid in complete dissolution.] all flasks with water to volume.
Chromatographic system, Appendix IIA Sample solutions: 100 mg/mL [NOTE—Ultra-sonication
Mode: High-performance liquid chromatography for 10–15 min at 30° may be necessary to aid in
Detector: Refractive index complete dissolution.]
Column: 25-cm × 4-mm, packed with a Calibration curve: Assemble a set of eleven 10-mL
monomolecular layer of aminopropylsilane chemically volumetric flasks. Transfer 1 mL of each of the eleven
bonded to totally porous silica gel support (10 µm Standard solutions into the flasks, and add 1 mL of
particle diameter)1 Reagent solution to each flask. Heat each flask in the
Column temperature: 40° boiling water bath for 10 min. Cool rapidly to room
Flow rate: 2.0 mL/min temperature, and dilute all flasks with water to volume.
Injection volume: 10 µL For each solution, measure the absorbance against
Analysis: Separately inject equal volumes of the Standard water at 545 nm. Generate a standard curve by
solution and Sample solution into the chromatograph, plotting absorbance vs. the concentration, in mg/mL,
and measure the responses for the major peaks on the of dextrose in the Standard solutions.
resulting chromatograms. Calculate the percentage of Analysis: Prepare a set of six 10-mL volumetric flasks
alpha-cyclodextrin in the portion of the sample taken (labeled a through f), and add 1 mL of Reagent solution
by the equation: to each. Transfer 1 mL of the Sample solution to flasks
“a,” “b,” and “c”. Transfer 1 mL of the Standard
Result = (rU/rS) × (CS/CU) × 100%
solutions numbered 0, 3, and 6 to flasks “d,” “e,” and
“f”. Thoroughly mix the contents of each flask, and
rU = peak response for alpha-cyclodextrin from place in a boiling water bath for 10 min. Then, cool
the Sample solution the flasks to room temperature, fill to the mark with
rS = peak response for alpha-cyclodextrin from water, and measure absorbance of the solutions against
the Standard solution water at 545 nm. The result is only valid if the
CU = concentration of the sample in the Sample absorbances of the solutions in flasks “d,” “e,” and “f”
solution (mg/mL) do not deviate more than 5% from the corresponding
CS = concentration of alpha-cyclodextrin in the absorbances from the Calibration curve. Calculate the
Standard solution (mg/mL) percentage reducing substance (as dextrose) in the
Acceptance criteria: NLT 98%, calculated on the sample taken using the following equation:
anhydrous basis
Reducing substances = CRS/CU × 100%
IMPURITIES
CRS = average of the reducing substance
concentrations (as dextrose), in mg/mL,
Graphite Furnace Method, Method I, Appendix IIIB
from flasks “a,” “b,” and “c,” calculated
using the Calibration curve
Organic Impurities
CU = concentration of the sample in the Sample
• REDUCING SUBSTANCES (AS DEXTROSE) [NOTE—Dextrose
solution (mg/mL)
levels are usually lower when determined by the
Acceptance criteria: NMT 0.5% (as dextrose)
following procedure in the presence of alpha-
• RESIDUAL COMPLEXANT (1-DECANOL)
cyclodextrin, compared to levels determined in its
Tris buffer solution: Dissolve 606 mg of tris (hydroxy-
absence. An alpha-cyclodextrin reference standard is
methyl) aminomethane and 430 mg of calcium sulfate
therefore utilized in this procedure for the calibration.]
dihydrate in 500 mL of water. Adjust the pH to 6.5
Reagent solution: Weigh 10.0 g of 3,5-dinitrosalicylic
with phosphoric acid.
acid in a 1000-mL volumetric flask. Add 80 mL of
water, and dissolve the 3,5-dinitrosalicylic acid by
2 Available from Consortium für Elektrochemische Industrie GmbH (München,
1 Nucleosil 100 NH2 (Macherey-Nagel Co, Düren, Germany), or equivalent. Germany), or Wacker Biochem Group (Adrian, MI, USA).
FCC 9 Monographs / beta-Cyclodextrin / 331
Internal standard solution: Add 50 mg of 1-octanol to Acceptance criteria: NMT 20 mg/kg

250 mL of tetrahydrofuran.
Standard stock solution: 750 µg/mL 1-decanol in SPECIFIC TESTS
Internal standard solution • MELTING RANGE OR TEMPERATURE DETERMINATION,
Standard solution: 7.5 µg/mL 1-decanol in Internal Appendix IIB
standard solution: diluted from Standard stock solution Acceptance criteria: Decomposes above 278°
Sample solution: Dissolve 750 mg of sample and 50 • RESIDUE ON IGNITION (SULFATED ASH), Appendix IIC
mg of glucoamylase3 (EC 3.2.1.3) in 7 mL of Tris buffer Sample: 1 to 2 g
solution. Add 100 µL of Internal standard solution and Acceptance criteria: NMT 0.1%
50 µL of cyclodextrin glucosyltransferase preparation • OPTICAL (SPECIFIC) ROTATION, Appendix IIB
Monographs
(500 U/mL).4 Close tightly, mix, and incubate in a Sample solution: 10 mg/mL
shaking water bath at 40° for 4 h. Acceptance criteria: [α]D25 between +145° and +151°
Condition a C18 solid-phase extraction column5 by • WATER, Water Determination, Method I, Appendix IIB
washing with methanol (2 × 10 mL) and water (4 × 10 Acceptance criteria: NMT 11%
mL). Quantitatively transfer the digested sample
solution to the conditioned column, and slowly pass it
through the column. Wash the column with water
beta-Cyclodextrin
.
(2 × 10 mL). Gently pass nitrogen through the column

to dry it (10 min). Apply 2.5 mL of tetrahydrofuran to First Published: Prior to FCC 6
the column, let stand for 5 min, and collect the eluate.
Use the eluate as the Sample solution. β-Cyclodextrin
Chromatographic system, Appendix IIA BCD
Detector: Flame ionization (C6H10O5)7 Formula Wt 1135.0
Column: 25-m × 0.32-mm capillary column coated INS: 459 CAS: [7585-39-9]
with a 0.5-µm layer of dimethylpolysiloxane gum6 UNII: JV039JZZ3A [betadex]
Column temperature: 60° for 1 min, 20°/min to
300°, 300° for 7 min
DESCRIPTION
Beta-Cyclodextrin occurs as a white, fine, crystalline solid,
Injection port temperature: 265°
frequently a fine, crystalline powder. It is a nonreducing
Carrier gas: Helium
cyclic compound consisting of seven alpha-(1,4) linked D-
Flow rate: 1 mL/min
glucopyranosyl units. It is slightly soluble in water.
Injection syringe: Heated, gas-tight
Function: Encapsulating agent; stabilizer
Injection volume: 1 µL
Packaging and Storage: Store in tight containers in a dry
Analysis: Separately inject equal volumes of the
place.
Standard solution and Sample solution into the
chromatograph, record the chromatograms, and IDENTIFICATION
measure the peak responses. Calculate the • INFRARED ABSORPTION, Spectrophotometric Identification
concentration (mg/kg) of 1-decanol in the sample Tests, Appendix IIIC
taken using the following formula: Reference standard: USP beta-Cyclodextrin RS
Result = (RU/RS) × CS × (2.5/S) × 1000
Analysis: The spectrum of the sample exhibits maxima
at the same wavelengths as those in the spectrum of
RU = internal standard ratio (1-decanol peak the Reference standard.
response/1-octanol response) from the • CHROMATOGRAPHY, Appendix IIA
Sample solution Analysis: The retention time of the major peak in the
RS = internal standard ratio (1-decanol peak chromatogram of Sample solution corresponds to that in
response/1-octanol response) from the the chromatogram of Standard solution, obtained as
Standard solution directed in the Assay (below).
CS = concentration of 1-decanol in the Standard
solution (µg/mL)
ASSAY
• PROCEDURE
2.5 = dilution factor for the Sample solution from
Mobile phase: Acetonitrile and water [65:35], filtered
the mL of tetrahydrofuran applied to the
and degassed
solid-phase extraction column
Internal standard solution: 20 mg/mL of glycerol,
S = mg of sample taken to prepare the Sample
filtered through a 0.45-µm membrane filter [NOTE—Use
solution
fresh or store in a freezer and thaw in hot water.]
1000 = µg/mg to mg/kg conversion factor
Standard stock solution: 10 mg/mL of USP beta-
3 Gluczyme 8000 (Wacker Chemie, Munich, Germany). Cyclodextrin RS [NOTE—Use fresh or store in a freezer
4 Available from Wacker Chimie (Munich, Germany).
5 Isolute C18, 10 mL (ICT, Bad Homburg, Germany), or equivalent.
and thaw in hot water.]
6 HP-1 (Agilent Technologies), or equivalent.
332 / beta-Cyclodextrin / Monographs FCC 9
Standard solution: Mix 1.0 mL of the Standard stock Organic Impurities

solution with 1.0 mL of Internal standard solution. • REDUCING SUGARS (DEXTROSE EQUIVALENT), Reducing Sugars,
System suitability solution: 5 mg/mL each of USP Appendix X
alpha-Cyclodextrin RS and USP beta-Cyclodextrin RS, Sample: 60 to 120 mg
filtered through a 0.45-µm membrane filter Acceptance criteria: NMT 1.0%, calculated on the
Sample: 1 g anhydrous basis
Sample stock solution: Transfer the Sample into a 100- • TOLUENE
mL volumetric flask, dilute to volume with water, and Toluene standard stock solution: 1000 µg/mL (or
mix. Filter this solution through a 0.45-µm membrane 1.153 µL/mL) of toluene in methanol
filter. Toluene standard solutions: 10 µg/mL, 50 µg/mL,
Monographs
Sample solution: Mix 1.0 mL of the filtered Sample 100 µg/mL, 250 µg/mL, and 500 µg/mL of toluene in
stock solution with 1.0 mL of Internal standard solution. methanol: from Toluene standard stock solution
Chromatographic system, Appendix IIA Trifluorotoluene standard stock solution: 800 µg/mL
Mode: High-performance liquid chromatography (or 0.673 µL/mL) of α,α,α-trifluorotoluene in methanol
Detector: Refractive index Trifluorotoluene standard solutions: 80 µg/mL, 400 µg
Column: 25-cm × 4.6-mm (id) column packed with 10- /mL, and 800 µg/mL of α,α,α-trifluorotoluene in
µm porous silica gel bonded with aminopropylsilane methanol, from Trifluorotoluene standard stock solution
(Alltech 35643, or equivalent), and a guard column [NOTE—These are the surrogate standards.]
that contains the same packing Sample: 500 mg
Column temperature: 25° ± 2° Chromatographic system, Appendix IIA
Detector temperature: 25° Mode: Gas chromatography connected to a purge
Flow rate: About 2.0 mL/min and trap apparatus (below)
Injection size: 20 µL Detector: Photoionization
System suitability Column: 30-m × 0.53-mm (id) fused silica open
Sample: System suitability solution tubular column, or equivalent, with a 1.5-µm
Suitability requirement 1: The relative standard crossbonded 5% diphenyl, 95% dimethyl
deviation for replicate injections is NMT 2.0%. polysiloxane (Restek RTX-5, or equivalent) stationary
Suitability requirement 2: The alpha-cyclodextrin and phase
beta-cyclodextrin peaks exhibit baseline separation. Column temperature: Hold at 40° for 2 min, increase
[NOTE—The relative retention times for alpha- at 20°/min to 180°, hold at 180° for 2 min.
cyclodextrin and beta-cyclodextrin are about 0.8 and Flow rate: 20 mL/min
1.0, respectively.] Carrier gas: Helium (ultra-high-purity)
Analysis: Separately inject the Sample solution and the Purge and trap apparatus: The purging apparatus
Standard solution into the chromatograph, record the uses disposable 15- × 150-mm test tubes. Use ultra-
chromatograms, and measure the responses for the high-purity helium to purge the sample for 15 min at
major peaks. Calculate the quantity, in mg, of a flow rate of 60 mL/min. Maintain at 100° all lines
(C6H10O5)7 in the portion of the Sample taken by the that the sample vapor passes through in the purge
formula: module. The trap consists of a 30.5-cm × 2.7-mm
(id) stainless steel tube, with a packing of a porous
Result = C × (RU / RS) × 100 polymer based on 2,6-diphenyl-p-phenylene oxide
(Tenax, or equivalent). The length of the packing in
the tube is 24 cm. The entire void volume of the trap
C = concentration (mg/mL) of anhydrous beta-
is at the vented end of the trap column. Maintain the
cyclodextrin in the Standard solution,
trap at 100°. Recondition the trap for a subsequent
corrected for moisture content by a
run by baking it for 5 min at 190°.
titrimetric water determination
Calibration: Place an empty purge tube into the
RU = peak response ratio of the beta-cyclodextrin
purging apparatus. Fill a 5-mL syringe with 0.5 N
peak to the internal standard peak obtained
sodium hydroxide, and inject into this solution an
from the Sample solution
accurately measured 5-µL aliquot of 10 µg/mL of
RS = peak response ratio of the beta-cyclodextrin
Toluene standard solution. Introduce this solution into
peak to the internal standard peak obtained
the purge tube. Start the instrument for the run,
from the Standard solution
automated if desired, by purging for 15 min, then
Acceptance criteria: NLT 98.0% and NMT 101.0% of
heating the trap at 180° for 3 min. Repeat this
(C6H10O5)7 as beta-cyclodextrin, calculated on the
sequence for each of the Toluene standard solutions and
anhydrous basis
Trifluorotoluene standard solutions. Plot standard curves
IMPURITIES of the standard concentration (CS), in µg/mL, versus
Inorganic Impurities detector response (rS) for the Toluene standard solutions
• LEAD, Lead Limit Test, Flame Absorption Spectrophotometric and Trifluorotoluene standard solutions.
Method, Appendix IIIB Analysis: Place the Sample in a purge tube with 0.1 g
Sample: 10 g of salicylic acid. Attach the purge tube to the purging
Acceptance criteria: NMT 1 mg/kg apparatus. Add 5 µL of 400 µg/mL of Trifluorotoluene
FCC 9 Monographs / beta-Cyclodextrin / 333
standard solution to a 5-mL syringe filled with 0.5 N the trap has a total volume of NMT 15 mL. The
sodium hydroxide. Add the contents of the syringe to purge gas is passed through the water column as
the sample/salicylic acid mixture in the purge finely divided bubbles with a diameter of less than 3
apparatus, and start the run, automated if desired, by mm at the origin and is introduced NMT 5 mm from
purging for 15 min, then heating the trap at 180° for the base of the water column. Use a trap not shorter
3 min. The procedure is valid only when the detector than or narrower than 25 cm × 2.67 mm (id). Pack
response of the surrogate standard (Trifluorotoluene the trap to contain the indicated minimum lengths of
standard solution) in the sample preparation is within adsorbents in the following order, beginning at the
±15% of the value from the standard curve for trap inlet: 7.7 cm of 2,6-diphenylene oxide polymer
Trifluorotoluene standard. Calculate the concentration, (TENAX GC, or equivalent), 7.7 cm of silica gel, and
Monographs
in µg/g (numerically equivalent to mg/kg), of toluene 7.7 cm of coconut charcoal. The desorber is capable
in the Sample taken by the formula: of rapidly heating the trap to 250°, which is the
maximum temperature to be used. Condition the
Result = 5CS / WS assembled trap before use at 225° overnight with an
inert gas at a flow rate of not less than 20 mL/min.
Before daily use, condition the trap for 15 min at
CS = concentration of toluene (µg/mL) in the
225°.
sample, calculated from the standard curve
Analysis: Introduce exactly 20 µL of each Standard
for the Toluene standard solutions
solution on the inner wall of the sample purge. Desorb
WS = weight of Sample (g) taken
according to equipment instructions, and
record the peak areas. Prepare a calibration curve by
• TRICHLOROETHYLENE
plotting the peak area responses versus the weight of
Standard stock solution: 1 mg/mL of reagent-grade
trichloroethylene introduced into the purge. Introduce
trichloroethylene in methanol
the Sample on the fritted sparger of the sample purge.
Standard solutions: Transfer 0.5, 1.0, 2.0, 3.0, and 5.0
Purge and desorb according to equipment instructions.
mL of the Standard stock solution into five 50-mL
Record the peak area of trichloroethylene, and read the
volumetric flasks and dilute to volume with water.
corresponding weight (X) of trichloroethylene from the
These Standard solutions contain 10 ng/µL, 20 ng/µL,
calibration curve. Calculate the amount of
40 ng/µL, 60 ng/µL, and 100 ng/µL of
trichloroethylene, in mg/kg, by the formula:
trichloroethylene.
Sample: 250 mg Result = X/W
Detector: Flame-ionization X = weight of trichloroethylene (ng) in the
Column: 30-m × 0.32-mm (id) capillary column sample, determined from the calibration
coated with a 1-µm film thickness of curve
dimethylpolysiloxane oil (such as DB-1, OV-1, or W = weight of Sample (mg) taken
equivalent) Acceptance criteria: NMT 1 mg/kg
Column temperature: Hold at 40° for 3 min, increase
at 4°/min to 220°. SPECIFIC TESTS
Detector temperature: 280° • OPTICAL (SPECIFIC) ROTATION, Appendix IIB
Carrier gas: Helium Sample solution: 1 g in 100 mL of water
Purge gas: Nitrogen Acceptance criteria: [α]D20 between +160° and +164°,
Flow rate: 40 mL/min calculated on the anhydrous basis
Injection size: 20 µL • RESIDUE ON IGNITION (SULFATED ASH), Appendix IIC
Purge and trap apparatus: The apparatus1 comprises Sample: 1 to 2 g
three sections: the sample purge, the trap, and the Acceptance criteria: NMT 0.1%
desorber. The sample purge accepts 5-mL samples • WATER, Water Determination, Appendix IIB
with a water column NLT 3 cm deep, and the Acceptance criteria: NMT 14.0%
gaseous headspace between the water column and
1 The apparatus used is based on that described in the U.S. Environmental
Protection Agency Test Method for Purgeable Halocarbons—Method 601.

334 / gamma-Cyclodextrin / Monographs FCC 9
measure the responses for all peaks. Calculate the

gamma-Cyclodextrin
.
content of gamma-cyclodextrin in the Sample taken by

First Published: Prior to FCC 6 the peak area percentage method using the following
equation:
γ-Cyclodextrin A = (B/C) × 100
gamma-CD
Cyclooctaamylose
Cyclomaltooctaose A = percentage of gamma-cyclodextrin in the
sample
B = peak area of gamma-cyclodextrin in the
Monographs
chromatogram
C = sum of the areas of all peaks recorded in
the chromatogram
Acceptance criteria: NLT 98.0% as (C6H10O5)8,
calculated on the anhydrous basis
IMPURITIES
• LEAD, Atomic Absorption Spectrophotometric Graphite
Furnace Method, Method I, Appendix IIIB
Sample preparation: Reflux 5 g of sample with 30 mL
nitric acid for 1 h. Remove the reflux condenser, and
(C6H10O5)8 Formula wt 1297.14 attach a condenser to the flask. Continue to heat and
CAS: [17465-86-0]
collect the distilled nitric acid. Allow the residue to
UNII: KZJ0BYZ5VA [gamma cyclodextrin]
cool, add 20 mL of water, and allow it to cool again.
DESCRIPTION Add 2 mL of orthophosphoric acid, and dilute to 100
Gamma-Cyclodextrin occurs as a white or almost white mL with water.
crystalline solid. It is a nonreducing cyclic saccharide Acceptance criteria: NMT 1 mg/kg
consisting of eight α-1,4-linked D-glucopyranosyl units Organic Impurities
manufactured by the action of cyclomaltodextrin • REDUCING SUGARS (AS GLUCOSE)
glucanotransferase on hydrolyzed starch followed by Sample: 1 g
purification of the gamma-Cyclodextrin. It is freely soluble Analysis: Transfer the Sample into a 250-mL Erlenmeyer
in water and is very slightly soluble in ethanol. flask, dissolve in 10 mL of water, add 25 mL of alkaline
Function: Stabilizer; emulsifier; carrier cupric citrate TS, and cover the flask with a small
Packaging and Storage: Store in tight containers in a dry beaker. Boil gently for exactly 5 min, and cool rapidly
place. to room temperature. Add 25 mL of 10% acetic acid
solution, 10.0 mL of 0.1 N iodine, 10 mL of dilute
IDENTIFICATION hydrochloric acid TS, and 3 mL of starch TS, and titrate
• INFRARED SPECTRA, Spectrophotometric Identification Tests, with 0.1 N sodium thiosulfate to the disappearance of
Appendix IIIC the blue color. Calculate the content of reducing
Sample preparation: Mineral oil mull substances, %R, (as D-glucose) by the equation:
Analysis: The spectrum of the sample exhibits relative
maxima at the same wavelengths as those of the %R = [(V1N1 − V2N2) × FE]/W
spectrum below.
ASSAY V1 = volume of the iodine solution (mL)

• PROCEDURE N1 = normality of the iodine solution
Mobile phase: Deionized water V2 = volume of the sodium thiosulfate solution
Sample solution: 10 mg/mL (mL)
Chromatographic system, Appendix IIA N2 = normality of the sodium thiosulfate solution
Mode: High-performance liquid chromatography FE = equivalence factor for D-glucose, determined
Detector: Differential refractometer empirically, 2.7
Column: 30-cm × 7.8-mm (id); column packed with W = weight of the sample taken (g)
25-µm diameter beads of silver bonded to sulfonated Acceptance criteria: NMT 0.5%
divinyl benzene–styrene copolymer (Aminex HPX-42A, • VOLATILE ORGANIC COMPOUNDS
Bio-Rad Laboratories, or equivalent) Standard solutions: Prepare solutions of 8-
Column temperature: 65° ± 10° cyclohexadecen-1-one in hexane in the range of 1 µg/
Flow rate: 0.3 to 1.0 mL/min mL to 60 µg/mL.
Injection size: About 20 µL Sample solution: Dissolve 50 g of sample in 700 mL of
Analysis: Inject the Sample solution into the water in a 1-L round bottom flask and add a magnetic
chromatograph, record the chromatogram, and stirrer. Attach the flask to the lower part of a Bleidner
FCC 9 Monographs / gamma-Cyclodextrin / 335
apparatus (see Figure 1, below), and connect a 100-mL Analysis: Separately inject the Standard solutions and
round-bottom flask containing about 70 mL of hexane the Sample solution into the chromatograph and record
and a few boiling stones to the other side of the the resulting chromatograms. Using the chromato-
apparatus. Fill the Bleidner apparatus with equal grams from the Standard solutions, create a calibration
amounts of water and hexane, and place a reflux curve of the concentration of 8-cyclohexadecen-1-one
condenser on the top. Heat both flasks with heating versus the response factor. Calculate the area(s) under
mantels to boiling. Using the magnetic stirrer, stir the the peak for each volatile organic compound in the
contents of the 1-L flask well. Keep the contents of the Sample solution, and convert it to mg per kg of
two flasks boiling for 8 h. After cooling, remove the gamma-cyclodextrin using the calibration curve.
100-mL flask, transfer the contents to a 100-mL Acceptance criteria: NMT 20 mg/kg
Monographs
volumetric flask, and dilute to volume with hexane.
Chromatographic system, Appendix IIA SPECIFIC TESTS
Mode: Gas chromatography • IODINE REACTION
Detector: Flame-ionization Sample: 0.2 g
Column: 30-m × 0.32-mm (id) column with a Analysis: Place the Sample in a test tube and add 2 mL
stationary phase consisting of 0.25-µm, cross-bonded, of a 0.1 N iodine solution. Heat the mixture in a water
95% dimethyl 5% diphenyl polysiloxane (JBW bath, and allow to cool at room temperature.
Scientific DB-5.625, or equivalent) Acceptance criteria: A clear, brown solution forms.
Temperature • OPTICAL (SPECIFIC) ROTATION, Appendix IIB
Injector: 280° Sample: 10 mg/mL
Detector: 280° Acceptance criteria: [α]D25 between +174° and +180°
Column: Hold at 70° for 4 min, increase at 10°/min • RESIDUE ON IGNITION (SULFATED ASH), Method I (for Solids),
to 250°. Appendix IIC
Carrier gas: Nitrogen Acceptance criteria: NMT 0.1%
Flow rate: 70 mL/min
Figure 1. Bleidner Apparatus

336 / gamma-Cyclodextrin / Monographs FCC 9
• WATER, Water Determination, Appendix IIB

Monographs
γ-Cyclodextrin (Mineral Oil Mull)
IDENTIFICATION
Cyclohexane
.

First Published: FCC 7 General Provisions).
Hexahydrobenzene ASSAY
Hexamethylene • PROCEDURE
Hexanaphthene Analysis: Proceed as directed under M-1b, Appendix XI.
Acceptance criteria: NLT 99.5% (w/w)
IMPURITIES
C6H12 Formula wt 84.16 • LEAD, Lead Limit Test, Flame Atomic Absorption
CAS: [110-82-7] Spectrophotometric Method, Appendix IIIB
UNII: 48K5MKG32S [cyclohexane] Sample: 5 g
Analysis: Proceed as directed using the Diluted Standard
DESCRIPTION Lead Solutions for the 1 mg/kg Lead Limit.
Cyclohexane occurs as a clear, colorless, flammable liquid Acceptance criteria: NMT 2 mg/kg
with a faint, characteristic odor that is ether-like. It is • SULFUR, Appendix IIIC
produced by reacting benzene with hydrogen. Acceptance criteria: NMT 10 mg/kg
Cyclohexane is insoluble in water and miscible with Organic Impurities
ethanol and ether. • BENZENE, Appendix IIIC
Function: Extraction solvent Analysis: Proceed as directed using the conditions
Packaging and Storage: Store in tight containers in a described under Column No. 6.
cool place. Store remote from fire. [CAUTION—Cyclohexane Acceptance criteria: NMT 0.1%
is highly flammable.]
FCC 9 Monographs / Cyclohexane / 337
• POLYCYCLIC AROMATIC HYDROCARBONS cm; also, for checking spectrophotometer

[NOTE—Because of the sensitivity of the test, the performance only, use cells with an optical path
possibility of errors arising from contamination is great. length in the range of 1.000 ± 0.005 cm. With
It is of the greatest importance, therefore, that all distilled water in the cells, determine any absorbance
glassware be scrupulously cleaned to remove all organic differences.
matter such as oil, grease, detergent residues, etc. Spectrophotometer: Use an instrument with a
Examine all glassware, including stoppers and spectral range of 250–400 nm with a spectral slit
stopcocks, under ultraviolet light to detect any residual width of 2 nm or less. Under instrument operating
fluorescent contamination. As a precautionary measure conditions for these absorbance measurements, the
it is a recommended practice to rinse all glassware with spectrophotometer shall also meet the performance
Monographs
purified isooctane immediately before use. No grease is requirements in Table 1.
to be used on stopcocks or joints. Great care to avoid
contamination of samples in handling and to assure Table 1
absence of any extraneous material arising from Parameter Value
inadequate packaging is essential. Because some of the Absorbance repeatability ± 0.01 at 0.4 absorbance
polynuclear hydrocarbons sought in this test are very
Absorbance accuracy ± 0.05 at 0.4 absorbance
susceptible to photo-oxidation, the entire procedure is
Wavelength repeatability ± 0.2 nm
to be carried out under subdued light.]
Apparatus Wavelength accuracy ± 1.0 nm
Separatory funnels: 250-, 500-, 1000-, and preferably
2000-mL capacity, equipped with tetrafluoroethylene Organic solvents: All solvents used throughout this
polymer stopcocks procedure shall meet the specifications and tests
Reservoir: 500-mL capacity, equipped with a 24/40- described below. The Isooctane, Benzene, Acetone, and
standard taper male fitting at the bottom and a Methanol designated below shall meet the following
suitable ball joint at the top for connecting to the test: To the specified quantity of solvent in a 250-mL
nitrogen supply. The male fitting should be equipped Erlenmeyer flask, add 1 mL of purified n-Hexadecane
with glass hooks. and evaporate on the steam bath under a stream of
Chromatographic tube: 180 mm × 15.7 ± 0.1 mm nitrogen. [NOTE—A loose aluminum foil jacket around
(i.d.), equipped with a coarse, fritted-glass disc, a the flask will speed evaporation.] Discontinue
tetrafluoroethylene polymer stopcock, and a female evaporation when NMT 1 mL of residue remains. To
24/40-standard tapered fitting at the opposite end. the residue from Benzene add a 10-mL portion of
Overall length of the column with the female joint is purified Isooctane, re-evaporate, and repeat once to
235 mm. ensure complete removal of Benzene.
Disc: Tetrafluoroethylene polymer 2-in diameter disc Alternatively, the evaporation time can be reduced by
approximately 3/16-in thick with a hole bored in the using the optional evaporation flask. In this case the
center to closely fit the stem of the chromatographic solvent and n-Hexadecane are placed into the flask on
tube the steam bath, the tube assembly is inserted, and a
Heating jacket: Conical, suitable for a 500-mL stream of nitrogen is fed through the inlet tube while
separatory funnel; used with variable transformer heat the outlet tube is connected to a solvent trap and
control vacuum line in such a way as to prevent any flow-back
Suction flask: 250- or 500-mL filter flask of condensate into the flask.
Condenser: 24/40 joints, fitted with a drying tube, Dissolve the 1 mL of n-Hexadecane residue in Isooctane
length optional and dilute to 25 mL. Determine the absorbance in the
Evaporation flask (optional): 250- or 500-mL 5-cm path length cells compared to Isooctane as
capacity all-glass flask equipped with a standard taper reference. The absorbance of the solution of the
stopper having inlet and outlet tubes permitting solvent residue (except for Methanol) is NMT 0.01 per
passage of nitrogen across the surface of the liquid to cm path length between 280 and 400 nm. For
be evaporated Methanol this absorbance value is 0.00.
Vacuum distillation assembly: Use an all glass (for Isooctane (2,2,4-trimethylpentane): Use 180 mL for
purification of dimethyl sulfoxide) 2-L distillation flask the test described under Organic solvents. Purify, if
with a heating mantle; a Vigreaux, or equivalent, necessary, by passage through a column of activated
vacuum-jacketed condenser about 45-cm in length; silica gel, grade 12, or equivalent, about 90-cm in
and a distilling head with a separable cold finger length and 5- to 8-cm in diameter.
condenser. Use of tetrafluoroethylene polymer sleeves Benzene, reagent grade: Use 150 mL for the test
on the glass joints will prevent freezing. Do not use described under Organic solvents. Purify, if necessary,
grease on stopcocks or joints. by distillation or otherwise.
Nitrogen cylinder: Water-pumped or equivalent Acetone, reagent grade: Use 200 mL for the test
purity nitrogen in cylinder equipped with regulator described under Organic solvents. Purify, if necessary,
and valve to control flow at 5 psig by distillation.
Spectrophotometric cells: Use fused quartz cells with
an optical path length in the range of 5.000 ± 0.005
338 / Cyclohexane / Monographs FCC 9
Eluting mixtures 85% phosphoric acid and 50 g of norit A (decolorizing

10% Benzene in isooctane: Pipet 50 mL of Benzene carbon), or equivalent. Stopper the flask and stir with a
into a 500-mL glass-stoppered volumetric flask and magnetic stirrer (tetrafluoroethylene polymer coated
adjust with Isooctane to volume. bar) for 15 min. Filter the Dimethyl sulfoxide through
20% Benzene in isooctane: Pipet 50 mL of Benzene four thicknesses of fluted paper (18.5 cm, Schleicher &
into a 250-mL glass-stoppered volumetric flask and Schuell, No. 597, or equivalent). If the initial filtrate
adjust with Isooctane to volume. contains carbon fines, refilter through the same filter
Acetone–benzene–water mixture: Add 20 mL of until a clear filtrate is obtained. Protect the sulfoxide
water to 390 mL of Acetone and 200 mL of Benzene. from air and moisture during this operation by
n-Hexadecane, 99% olefin-free: Dilute 1.0 mL of n- covering the solvent in the funnel and collection flask
Monographs
hexadecane with Isooctane to 25 mL and determine with a layer of Isooctane. Transfer the filtrate to a 2-L
the absorbance in a 5-cm cell, compared to Isooctane separatory funnel and draw off the dimethyl sulfoxide
as a reference, between 280 and 400 nm. The into the 2-L distillation flask of the Vacuum distillation
absorbance per cm path length shall not exceed 0.00 assembly and distill at or below 3 mm Hg. Discard the
in this range. Purify, if necessary, by percolation first 200-mL fraction of the distillate and replace the
through activated silica gel or by distillation. distillate collection flask with a clean one. Continue the
Methanol, reagent grade: Use 10.0 mL of methanol. distillation until 1 L of the sulfoxide has been collected.
Purify, if necessary, by distillation. At completion of the distillation, the reagent should be
Dimethyl sulfoxide: Use a pure grade, clear, water- stored in glass-stoppered bottles because it is very
white product with a melting point of 18°, minimum. hygroscopic and will react with some metal containers
Dilute 120 mL with 240 mL of distilled water in a 500- in the presence of air.
mL separatory funnel, mix, and allow to cool for 5–10 Magnesium oxide: Use Sea Sorb 43, Food Machinery
min. Add 40 mL of Isooctane to the solution and Company, Westvaco Division, or equivalent. Place
extract by shaking the funnel vigorously for 2 min. 100 g in a large beaker, add 700 mL of distilled water
Draw off the lower aqueous layer into a second 500- to make a thin slurry, and heat on a steam bath for 30
mL separatory funnel, and repeat the extraction with min with intermittent stirring. Stir well initially to
40 mL of Isooctane. Draw off and discard the aqueous ensure that all of the adsorbent is completely wetted.
layer. Wash each of the 40-mL isooctane portions three Using a Buchner funnel and a filter paper (Schleicher &
times with 50-mL portions of distilled water. The Schuell No. 597, or equivalent) of suitable diameter,
shaking time for each wash is 1 min. Discard the filter with suction. Continue suction until water no
aqueous layers. Filter the first isooctane portion longer drips from the funnel. Transfer the adsorbent to
through Sodium sulfate, anhydrous prewashed with a glass trough lined with aluminum foil free from
Isooctane (see Sodium sulfate below for preparation of rolling oil. Break up the magnesia with a clean spatula
filter), into a 250-mL Erlenmeyer flask, or optionally and spread out the adsorbent on the foil in a layer 1–2
into the Evaporation flask. Wash the first separatory cm thick. Dry for 24 h at 160 ± 1°. Pulverize the
funnel with the second 40-mL isooctane portion, and magnesia with a mortar and pestle. Sieve the
pass through the sodium sulfate into the flask. Then pulverized adsorbent between 60 and 180 mesh. Use
wash the second and first separatory funnels the magnesia retained on the 180-mesh sieve.
successively with a 10-mL portion of Isooctane, and Celite 545: Johns-Manville Company, diatomaceous
pass the solvent through the Sodium sulfate into the earth, or equivalent
flask. Add 1 mL of n-Hexadecane and evaporate the Magnesium oxide–celite 545 mixture: Place the
isooctane on the steam bath under nitrogen. Magnesium oxide (60–180 mesh) and Celite 545 in 2 to
Discontinue evaporation when NMT 1 mL of residue 1 proportions, respectively by weight, in a glass-
remains. To the residue, add a 10-mL portion of stoppered flask large enough for mixing. Shake
Isooctane and re-evaporate to 1 mL of n-Hexadecane. vigorously for 10 min, then transfer the mixture to a
Again, add 10 mL of Isooctane to the residue and glass trough lined with aluminum foil free from rolling
evaporate to 1 mL of hexadecane to ensure complete oil and spread it out in a layer 1–2 cm thick. Reheat
removal of all volatile materials. Dissolve the 1 mL of n- the mixture at 160 ± 1° for 2 h, and store in a tightly
Hexadecane in Isooctane and dilute to 25 mL. closed flask
Determine the absorbance in 5-cm path length cells, Sodium sulfate, anhydrous, reagent grade: (It is
compared to Isooctane as the reference. The preferable to use a granular form.) For each bottle of
absorbance of the solution should not exceed 0.02 per Sodium sulfate, anhydrous, reagent grade used, establish
cm path length in the 280–400 nm range. [NOTE— as follows the necessary prewash to provide such filters
Difficulty in meeting this absorbance specification may required in the method: Place 35 g of Sodium sulfate,
be due to organic impurities in the distilled water. anhydrous in a 30-mL coarse, fritted-glass funnel or in a
Repetition of the test omitting the Dimethyl sulfoxide 65-mL filter funnel with a glass wool plug; wash with
will disclose their presence. If necessary to meet the successive 15-mL portions of the indicated solvent until
specification, purify the water by re-distillation, passage a 15-mL portion of the wash shows 0.00 absorbance
through an ion-exchange resin, or otherwise.] per cm path length between 280 and 400 nm when
Purify, if necessary, as follows: To 1500 mL of Dimethyl tested as prescribed under Organic solvents above.
sulfoxide in a 2-L glass-stoppered flask, add 6.0 mL of Usually three portions of wash solvent are sufficient.
FCC 9 Monographs / Cyclohexane / 339
Sulfoxide–phosphoric acid mixture: Place 300 mL of residue remains. To the residue, add a 10-mL portion
Dimethyl sulfoxide in a 1-L separatory funnel and add of Isooctane, re-evaporate to 1 mL of n-Hexadecane,
75 mL of 85% phosphoric acid. Mix the contents of and repeat this operation once more.
the funnel and allow to stand for 10 min. [CAUTION— Transfer the residue with Isooctane to a 25-mL
The reaction between the sulfoxide and the acid is volumetric flask and dilute to volume. Determine the
exothermic. Release pressure after mixing, then keep absorbance of the solution in the 5-cm path length
the funnel stoppered.] Add 150 mL of Isooctane and cells compared to Isooctane as reference between 280
shake to pre-equilibrate the solvents. Draw off the and 400 nm. Take care to lose none of the solution in
individual layers and store in glass-stoppered flasks. The filling the sample cell. Correct the absorbance values
layers are Pre-equilibrated sulfoxide–phosphoric acid for any absorbance derived from the reagents as
Monographs
mixture and Pre-equilibrated isooctane. determined by carrying out the preceding Analysis
Sample: 25 g without the Sample. If the corrected absorbance does
Analysis: Weigh the Sample in a beaker and transfer the not exceed the limits prescribed under Acceptance
Sample to a 500-mL separatory funnel containing 100 criteria, the Sample meets the ultraviolet absorbance
mL of the Pre-equilibrated sulfoxide–phosphoric acid specifications.
mixture. Promptly complete the transfer of the Sample If the corrected absorbance per cm path length exceeds
to the funnel with portions of the Pre-equilibrated the limits prescribed in the Acceptance criteria, proceed
isooctane, using a total volume of 50 mL. as follows: Transfer the isooctane solution to a 125-mL
When the Sample is in solution, shake it vigorously for 2 flask equipped with a 24/40 joint and evaporate the
min. Set up three 250-mL separatory funnels with each isooctane on the steam bath under a stream of
containing 30 mL of Pre-equilibrated isooctane. After nitrogen to a volume of 1 mL of hexadecane. Add 10
separation of the liquid phases, allow to cool until the mL of Methanol and 0.3 g of 98% sodium
main portion of the sample–isooctane solution begins borohydride. Minimize exposure of the borohydride to
to show a precipitate. Gently swirl the funnel when the atmosphere; a measuring dipper may be used.
precipitation first occurs on the inside surface of the Immediately fit a water-cooled condenser equipped
funnel to accelerate this process. Carefully draw off the with a 24/40 joint and with a drying tube into the
lower layer, filter it slowly through a thin layer of glass flask, mix until the borohydride is dissolved, and allow
wool fitted loosely in a filter funnel into the first 250- to stand for 30 min at room temperature, with
mL separatory funnel, and wash in tandem with the intermittent swirling. At the end of this period,
30-mL portions of isooctane contained in the 250-mL disconnect the flask and evaporate the methanol on
separatory funnels. Shaking time for each wash is 1 the steam bath under nitrogen until the sodium
min. Repeat the extraction operation with two borohydride begins to come out of solution. Add 10
additional portions of the Sulfoxide–phosphoric acid mL of Isooctane and evaporate to a volume of 2–3 mL.
mixture, replacing the funnel after each extraction to Again, add 10 mL of Isooctane and concentrate to a
keep the sample in solution and washing each volume of 5 mL. Swirl the flask repeatedly to assure
extractive in tandem through the same three portions adequate washing of the sodium borohydride residues.
of isooctane. Fit the Disc on the upper part of the stem of the
Collect the successive extractives (300 mL total) in a Chromatographic tube, then place the tube with the
separatory funnel (2-L), containing 480 mL of distilled disc on the Suction flask and apply the vacuum (135
water, mix, and allow to cool for a few min after the mm Hg). Weigh out 14 g of the Magnesium
last extractive has been added. Add 80 mL of Isooctane oxide–Celite 545 mixture and pour the adsorbent
to the solution and extract by shaking the funnel mixture into the Chromatographic tube in
vigorously for 2 min. Draw off the lower aqueous layer approximately 3-cm layers. After the addition of each
into a second 2-L separatory funnel, and repeat the layer, level off the top of the adsorbent with a flat glass
extraction with 80 mL of Isooctane. Draw off and rod or metal plunger by pressing down firmly until the
discard the aqueous layer. Wash each of the 80-mL adsorbent is well-packed. Loosen the topmost few mm
extractives three times with 100-mL portions of distilled of each adsorbent layer with the end of a metal rod
water. Shaking time for each wash is 1 min. Discard before the addition of the next layer. Continue packing
the aqueous layers. in this manner until all of the 14 g of adsorbent is
Filter the first extractive through Sodium sulfate, added to the tube. Level off the top of the adsorbent
anhydrous prewashed with Isooctane (see Sodium sulfate by pressing down firmly with a flat glass rod or metal
above for preparation of the filter) into a 250-mL plunger to make the depth of the adsorbent bed
Erlenmeyer flask (or, optionally, into the Evaporation approximately 12.5 cm in depth. Turn off the vacuum
flask). Wash the first separatory funnel with the second and remove the Suction flask. Fit the 500-mL Reservoir
80-mL isooctane extractive and pass through the onto the top of the chromatographic column and pre-
sodium sulfate. Then wash the second and first wet the column by passing 100 mL of Isooctane
separatory funnels successively with a 20-mL portion of through the column. Adjust the nitrogen pressure so
Isooctane and pass the solvent through the sodium that the rate of descent of the isooctane coming off of
sulfate into the flask. Add 1 mL of n-Hexadecane and the column is between 2 and 3 mL/min. Discontinue
evaporate the isooctane on the steam bath under pressure just before the last of the isooctane reaches
nitrogen. Discontinue evaporation when NMT 1 mL of the level of the adsorbent. [CAUTION—Do not allow the
340 / Cyclohexane / Monographs FCC 9
liquid level to recede below the adsorbent level at any Acceptance criteria: See Table 2.
time.]
Remove the Reservoir and decant the 5 mL isooctane Table 2
concentrate solution onto the column, and, with slight Wavelength
Absorbance
pressure, again allow the liquid level to recede to (nm)
barely above the adsorbent level. Rapidly complete the 280–289 0.15
transfer similarly with two 5-mL portions of isooctane, 290–299 0.12
swirling the flask repeatedly each time to assure
300–359 0.08
adequate washing of the residue. Just before the final
360–400 0.02
5-mL wash reaches the top of the adsorbent, add 100
Monographs
mL of Isooctane to the Reservoir and continue the

percolation at the 2–3 mL/min rate. Just before the last SPECIFIC TESTS
of the isooctane reaches the adsorbent level, add 100 • DISTILLATION RANGE, Appendix IIB
mL of 10% Benzene in isooctane to the Reservoir and Acceptance criteria: Between 80° and 82°
continue the percolation at the 2–3 mL/min rate. Just • NONVOLATILE RESIDUE
before the solvent mixture reaches the adsorbent level, Sample: 100 mL
add 25 mL of 20% Benzene in isooctane to the Reservoir Analysis: Evaporate the Sample to dryness in a tared
and continue the percolation at 2–3 mL/min until all platinum dish on a steam bath, heat at 105° for 30 min
this solvent mixture has been removed from the or to constant weight, cool in a desiccator, and weigh.
column. Discard all the elution solvents collected up to Acceptance criteria: NMT 1 mg/100 mL
this point.
Add 300 mL of the Acetone–benzene–water mixture to
the Reservoir and percolate through the column to
elute the polynuclear compounds. Collect the eluate in
Cyclohexanecarboxylic Acid
.
a clean 1-L separatory funnel. Allow the column to

drain until most of the solvent mixture is removed. First Published: Second Supplement, FCC 8
Wash the eluate three times with 300-mL portions of
distilled water, shaking well for each wash. The Benzoic Acid, Hexahydro
addition of small amounts of sodium chloride facilitates Carboxycyclohexane
separation. Discard the aqueous layer after each wash. Cyclohexanoic Acid
After the final separation, filter the residual benzene Cyclohexylcarboxylic Acid
through Sodium sulfate, anhydrous prewashed with Cyclohexylmethanoic Acid
Benzene (see Sodium sulfate above for preparation of Hexahydrobenzoic Acid
the filter) into a 250-mL Erlenmeyer flask (or optionally Cyclohexanecarboxylic Acid
into the Evaporation flask). Wash the separatory funnel
with two additional 20-mL portions of Benzene which
are also filtered through the Sodium sulfate. Add 1 mL
of n-Hexadecane and completely remove the benzene
by evaporation under nitrogen, using the special
procedure to eliminate benzene as previously described C7H12O2 Formula wt 128.17
under Organic solvents. Transfer the residue with FEMA: 3531
Isooctane to a 25-mL volumetric flask and adjust the CAS: [98-89-5]
volume. Determine the absorbance of the solution in UNII: H9VKD9VL18 [cyclohexanecarboxylic acid]
the 5-cm path length cells compared to Isooctane as
reference between 250 and 400 nm. Correct for any DESCRIPTION
absorbance derived from the reagents as determined Cyclohexanecarboxylic Acid occurs as a white solid under
by carrying out the procedure without a wash sample. ambient temperature. At higher temperatures, it melts and
If either spectrum shows the characteristic benzene occurs as a colorless, viscous liquid.
peaks in the 250–260 nm region, evaporate the Odor: Rum, raisins, fruity, fatty sweet
solution to remove benzene by the procedure under Solubility: Slightly soluble in water; miscible in fat;
Organic solvents. Dissolve the residue, transfer with miscible in ethanol at room temperature
Isooctane to a 25-mL volumetric flask, and dilute to Boiling Point: ~232°–233°
volume. Record the absorbance again. If the corrected Function: Flavoring agent
absorbance does not exceed the limits prescribed in
the Acceptance criteria, the Sample meets the ultraviolet
IDENTIFICATION
absorbance specifications.
Reference standard: USP Cyclohexanecarboxylic Acid
RS
Sample and standard preparation: F
FCC 9 Monographs / p-Cymene / 341
Acceptance criteria: The spectrum of the sample Acceptance criteria: NMT 1.0
exhibits maxima at the same wavelengths as those in • REFRACTIVE INDEX, Appendix II: At 20°
the spectrum of the Reference standard. Acceptance criteria: Between 1.436 and 1.441
ASSAY method (see General Provisions).
• PROCEDURE: Proceed as directed under M-1b, Appendix Acceptance criteria: Between 0.966 and 0.970
XI.
Reference standard: USP Cyclohexanecarboxylic Acid
RS
Acceptance criteria: NLT 98%
p-Cymene
.
Monographs
SPECIFIC TESTS First Published: Prior to FCC 6
Acceptance criteria: 1.516–1.520
IIB
C10H14 Formula wt 134.22
Acceptance criteria: 28°–32°
FEMA: 2356
UNII: 1G1C8T1N7Q [p-cymene]
DESCRIPTION
Cyclohexyl Acetate
.
p-Cymene occurs as a colorless to pale yellow liquid.

First Published: Prior to FCC 6 Odor: Kerosene
Solubility: Soluble in vegetable oils; insoluble or practically
insoluble in water, propylene glycol
mL of 95% alcohol.
FEMA: 2349 IDENTIFICATION
UNII: UL0RS4H1UE [cyclohexyl acetate] • INFRARED SPECTRA, Spectrophotometric Identification Tests,
Appendix IIIC
DESCRIPTION Acceptance criteria: The spectrum of the sample
Cyclohexyl Acetate occurs as a colorless to pale yellow exhibits relative maxima at the same wavelengths as
liquid. those of the spectrum below.
Odor: Green, fruity
Solubility: Soluble in alcohol ASSAY
Boiling Point: ∼174° • PROCEDURE: Proceed as directed under M-1a, Appendix
Function: Flavoring agent XI.
Acceptance criteria: NLT 97.0% of C10H14
ASSAY
• PROCEDURE: Proceed as directed under M-1b, Appendix SPECIFIC TESTS
XI. • REFRACTIVE INDEX, Appendix II: At 20°
Acceptance criteria: NLT 98.0% of C8H14O2 Acceptance criteria: Between 1.489 and 1.491
SPECIFIC TESTS method (see General Provisions).
• ACID VALUE, FLAVOR CHEMICALS (OTHER THAN ESSENTIAL Acceptance criteria: Between 0.853 and 0.855
342 / p-Cymene / Monographs FCC 9
Monographs
p-Cymene

L-Cysteine Monohydrochloride
.

First Published: Prior to FCC 6 the spectrum of the Reference standard.
Last Revision: FCC 6
ASSAY
• PROCEDURE
L-2-Amino-3-mercaptopropanoic Acid Monohydrochloride Sample: 300 mg, previously dried
Analysis: Transfer the Sample into a 250-mL glass-
stoppered flask. Add 20 mL of water, 4 g of potassium
iodide, 5 mL of 2.7 N hydrochloric acid, and 25.0 mL
of 0.1 N iodine. Stopper the flask, and allow the
mixture to stand for 30 min in a dark place. Titrate the
C3H7NO2S · HCl · H2O Formula wt, monohydrate 175.63 excess iodine with 0.1 N sodium thiosulfate. Perform a
C3H7NO2S · HCl Formula wt, anhydrous 157.62 blank determination (see General Provisions), and make
INS: 920 CAS: monohydrate [7048-04-6] any necessary correction. Each mL of 0.1 N iodine is
anhydrous [52-89-1] equivalent to 15.76 mg of C3H7NO2S · HCl.
UNII: ZT934N0X4W [cysteine hydrochloride] Acceptance criteria: NLT 98.0% and NMT 101.5%
C3H7NO2S · HCl, on the dried basis
DESCRIPTION
L-Cysteine Monohydrochloride occurs as a white, crystalline IMPURITIES
powder. It is freely soluble in water and in alcohol. The Inorganic Impurities
anhydrous form melts with decomposition at about 175°. • LEAD, Lead Limit Test, Appendix IIIB
Function: Nutrient Sample solution: Prepare as directed for organic
Packaging and Storage Store in well-closed, light- compounds.
resistant containers. Control: 5 µg Pb (5 mL of Diluted Standard Lead
Solution)
IDENTIFICATION Acceptance criteria: NMT 5 mg/kg
Tests, Appendix IIIC. SPECIFIC TESTS
Reference standard: USP L-Cysteine Hydrochloride RS • LOSS ON DRYING, Appendix IIC: room temperature for 24
Sample and Standard preparation: K h in a vacuum desiccator using a suitable desiccant and
maintaining a pressure of NMT 5 mm Hg
Acceptance criteria: NLT 8.0% and NMT 12.0%
FCC 9 Monographs / L-Cystine / 343
• OPTICAL (SPECIFIC) ROTATION, Appendix IIB Sample preparation: Mineral oil mull
Sample : 8 g, undried Acceptance criteria: The spectrum of the sample
Analysis: Dissolve the Sample in sufficient 1 N exhibits relative maxima at the same wavelengths as
hydrochloric acid to make 100 mL. those of the spectrum below.
Acceptance criteria
[α]D20 between +5.0° and +8.0°, calculated on the dried ASSAY
basis • NITROGEN DETERMINATION, Appendix IIIC
[α]D25 between +4.9° and +7.9°, calculated on the dried Sample: 200 mg
basis Analysis: Calculate the percent L-Cystine using the
• RESIDUE ON IGNITION (SULFATED ASH), Appendix IIC equation:
Monographs
Sample: 1 g
% L-Cystine = N × 8.58
N = percent nitrogen
Acceptance criteria: NLT 98.5% and NMT 101.5%
L-Cystine
.
C6H12N2O4S2, calculated on the dried basis

IMPURITIES
3,3′-Dithiobis(2-aminopropanoic acid) • LEAD, Lead Limit Test, Appendix IIIB
Sample solution: Prepare as directed for organic
compounds.
Solution)
C6H12N2O4S2 Formula wt 240.30
INS: 921 CAS: [56-89-3] SPECIFIC TESTS
UNII: 48TCX9A1VT [cystine] • LOSS ON DRYING, Appendix IIC: 105° for 3 h
DESCRIPTION • OPTICAL (SPECIFIC) ROTATION, Appendix IIB
L-Cystine occurs as colorless to white crystals. It is soluble in Sample solution: 2 g of previously dried sample in
diluted mineral acids and in alkaline solutions. It is very sufficient 1 N hydrochloric acid to make 100 mL
slightly soluble in water and in alcohol. Acceptance criteria: [α]D20 between −215° and
Function: Nutrient −225°, on the dried basis
Packaging and Storage: Store in well-closed containers. • RESIDUE ON IGNITION (SULFATED ASH), Appendix IIC
Sample: 2 g
IDENTIFICATION Acceptance criteria: NMT 0.1%
Appendix IIIC
344 / L-Cystine / Monographs FCC 9
Monographs
L-Cystine (Mineral Oil Mull)
IDENTIFICATION
5′-Cytidylic Acid
.
• A. INFRARED ABSORPTION, Spectrophotometric Identification

First Published: First Supplement, FCC 7 Tests, Appendix IIIC
Reference standard: USP 5′-Cytidylic Acid RS
Cytidine 5′-monophosphate Sample and standard preparation: A
Cytidylic acid Acceptance criteria: The spectrum of the sample
CMP exhibits maxima at the same wavelengths as those in
Cytidine 5′-phosphoric acid the spectrum of the Reference standard.
• B. PROCEDURE
Acceptance criteria: The retention time of the major
peak (excluding the solvent peak) in the chromatogram
of the Sample solution corresponds to that of the
Standard solution in the Assay.
ASSAY
C9H14N3O8P Formula wt 323.20
CAS: [63-37-6] • PROCEDURE
Mobile phase: 0.1 M potassium dihydrogen phosphate
UNII: F469818O25 [5′-cytidylic acid]
(KH2PO4) in degassed water, adjusted with 0.1 M
DESCRIPTION dipotassium hydrogen phosphate (K2HP04) to a pH of
5′-Cytidylic Acid occurs as colorless or white crystals, or as a 5.6
white, crystalline powder. It is very slightly soluble in Standard solution: 0.02 mg/mL of USP 5′-Cytidylic Acid
water, and practically insoluble in alcohol. It is produced RS in Mobile phase. [NOTE—Ultrasonication for 15 min
by enzymatic cleavage of yeast riboncleic acid (RNA) with at 30° may be necessary to aid in complete dissolution.]
a 5′-phosphodiesterase followed by heat treatment, further Sample solution: 0.02 mg/mL in Mobile phase. [NOTE—
purification steps, and washing of crystals with ethanol. Ultrasonication for 15 min at 30° may be necessary to
Function: Source of 5′-Cytidylic Acid aid in complete dissolution.]
Packaging and Storage: Store in tight containers Chromatographic system, Appendix IIA
protected from light and moisture. Mode: High-performance liquid chromatography
Detector: UV 254 nm
FCC 9 Monographs / 5′-Cytidylic Acid / 345
Column: 25-cm × 4.6-mm; packed with 5-µm reversed developed using a Varian Vista MPX ICP OES unit.]
phase C18 silica gel1 The instrument parameters are as follows: set the
Column temperature: Ambient ultraviolet detector to scan arsenic at 188.980 nm.
Flow rate: About 1.0 mL/min Set the sample read time to 20 s. Set the forward
Injection size: 50 µL power from the RF generator to 1150 watts. Use an
System suitability argon plasma feed gas flow of 13.5 L/min with the
Sample: Standard solution auxiliary gas set to flow at 2.25 L/min. The sample is
Suitability requirements delivered to the spray chamber by a multi-channel
Suitability requirement 1: The relative standard peristaltic pump set to deliver sample at a rate of 20
deviation of the 5′-cytidylic acid peak area responses rpm. Samples are flushed through the system for 20 s
Monographs
from replicate injections is NMT 2.0%. prior to analysis. A 40-s read delay is also
Suitability requirement 2: The resolution, R, programmed into the sampling routine to allow for
between the 5′-cytidylic acid peak and all other fluid flow equilibration after the high-speed flush,
peaks is NLT 2.0. prior to the first analytical read of the sample.
Analysis: Separately inject equal volumes of the Standard Between samples, the pumping system is washed by
solution and Sample solution into the chromatograph, flushing the Diluent for 20 s.
and measure the responses for the major peaks on the Analysis: Generate a standard curve using Diluent as a
resulting chromatograms. [NOTE—The approximate Blank and the Standard solutions. [NOTE—The
retention time for 5′-cytidylic acid is 4.6 min.] Calculate correlation coefficient for the best-fit line should not be
the percentage of disodium 5′-cytidylic acid, less than 0.999.]
C9H14N3O8P, in the sample taken: Similarly, analyze the Sample solution on the ICP.
Calculate the concentration (mg/kg) of arsenic in the
Result = (rU/rS) × (CS/CU) × 100 Sample taken:
rU = peak area response of 5′-cytidylic acid in the Result = (C/W) × F
Sample solution
rS = peak area response of 5′-cytidylic acid in the C = concentration of arsenic in the Sample
Standard solution solution determined from the standard
CS = concentration of 5′-cytidylic acid in the curve (µg/mL)
Standard solution (mg/mL) W = weight of Sample taken (g)
CU = concentration of sample in the Sample F = final volume of the Sample solution, 100 mL
solution (mg/mL) Acceptance criteria: NMT 2 mg/kg
Acceptance criteria: 98.0%–103.0%, calculated on the • CADMIUM
anhydrous basis [NOTE—When water is specified as a diluent, use
deionized ultra-filtered water. When nitric acid is
IMPURITIES specified, use nitric acid of a grade suitable for trace
Inorganic Impurities element analysis with as low a content of cadmium as
• ARSENIC practical.]
[NOTE—When water is specified as a diluent, use Diluent: 4% nitric acid in water
deionized ultra-filtered water. When nitric acid is Standard stock solution: 100 µg/mL of cadmium
specified, use nitric acid of a grade suitable for trace prepared by diluting a commercially available 1000 mg
element analysis with as low a content of arsenic as /kg cadmium ICP standard solution
practical.] Standard solutions: 0.005, 0.05, 0.1, 0.2, 0.5, 1, and 2
Diluent: 4% nitric acid in water µg/mL of cadmium: from Standard stock solution
Standard stock solution: 100 µg/mL of arsenic diluted with Diluent
prepared by diluting a commercially available 1000 mg Sample: 5 g
/kg arsenic ICP standard solution Sample solution: Dissolve the Sample in 40 mL of 10%
Standard solutions: 0.05, 0.1, 0.2, 0.5, 1, and 2 µg/mL nitric acid in a 100-mL volumetric flask, and dilute with
of arsenic: from Standard stock solution diluted with water to volume.
Diluent Spectrophotometric system, Plasma Spectrochemistry,
Sample: 5 g Appendix IIC
Sample solution: Dissolve the Sample in 40 mL of 10% Mode: Inductively coupled plasma–optical emission
nitric acid in a 100-mL volumetric flask, and dilute with spectroscopy (ICP–OES)
water to volume. Setup: Same as that described in the test for Arsenic,
Spectrophotometric system, Plasma Spectrochemistry, but set to scan for cadmium at 228.802 nm
Appendix IIC Analysis: Generate a standard curve using Diluent as a
Mode: Inductively coupled plasma–optical emission Blank and the Standard solutions. [NOTE—The
spectroscopy (ICP–OES) correlation coefficient for the best-fit line should not be
Setup: Use a suitable ICP–OES configured in a radial less than 0.999.]
optical alignment. [NOTE—This method was
1 YMC-Pack ODS-AQ (YMC Europe GmbH, Dinslaken, Germany), or
equivalent.
346 / 5′-Cytidylic Acid / Monographs FCC 9
Similarly, analyze the Sample solution on the ICP. Standard solutions: 0.025, 0.05, 0.1, 0.2, 0.5, 1, and 2
Calculate the concentration (mg/kg) of cadmium in the µg/mL of mercury: from Standard stock solution diluted
Sample taken: with Diluent
Sample: 5 g
Result = (C/W) × F Sample solution: Dissolve the Sample in 40 mL of 10%
nitric acid in a 100-mL volumetric flask, and dilute with
C = concentration of cadmium in the Sample
water to volume.
solution determined from the standard
Spectrophotometric system, Plasma Spectrochemistry,
curve (µg/mL)
Appendix IIC
W = weight of Sample taken (g)
Mode: Inductively coupled plasma–optical emission
F = final volume of the Sample solution, 100 mL
Monographs
spectroscopy (ICP–OES)
Setup: Same as that described in the test for Arsenic,
• LEAD
but set to scan for mercury at 194.164 nm
[NOTE—When water is specified as a diluent, use
Analysis: Generate a standard curve using Diluent as a
Blank and the Standard solutions. [NOTE—The
specified, use nitric acid of a grade suitable for trace
correlation coefficient for the best-fit line should not be
element analysis with as low a content of lead as
less than 0.999.]
practical.]
Similarly, analyze the Sample solution on the ICP.
Diluent: 4% nitric acid in water
Calculate the concentration (mg/kg) of mercury in the
Standard stock solution: 100 µg/mL of lead prepared
Sample taken:
by diluting a commercially available 1000 mg/kg lead
ICP standard solution Result = (C/W) × F
Standard solutions: 0.05, 0.1, 0.2, 0.5, 1, and 2 µg/mL
of lead: from Standard stock solution diluted with C = concentration of mercury in the Sample
Diluent solution determined from the standard
Sample: 5 g curve (µg/mL)
Sample solution: Dissolve the Sample in 40 mL of 10% W = weight of Sample taken (g)
nitric acid in a 100-mL volumetric flask, and dilute with F = final volume of the Sample solution, 100 mL
water to volume. Acceptance criteria: NMT 0.5 mg/kg
Spectrophotometric system, Plasma Spectrochemistry, Organic Impurities
Appendix IIC • ETHANOL
Mode: Inductively coupled plasma–optical emission Standard solution: 20 mg/kg of ethanol in 1 N
spectroscopy (ICP–OES) sodium hydroxide. Add 10 mL of this solution to a 20-
Setup: Same as that described in the test for Arsenic, mL headspace vial, and cap tightly.
but set to scan for lead at 220.353 nm Sample solution: 100 mg/g in 1 N sodium hydroxide.
Analysis: Generate a standard curve using Diluent as a Add 10 mL of this solution to a 20-mL headspace vial,
Blank and the Standard solutions. [NOTE—The and cap tightly.
correlation coefficient for the best-fit line should not be Chromatographic system, Appendix IIA
less than 0.999.] Mode: Gas chromatography equipped with pressure-
Similarly, analyze the Sample solution on the ICP. loop headspace autosampler
Calculate the concentration (mg/kg) of lead in the Detector: Flame ionization
Sample taken: Column: 30-m × 0.53-mm (id) capillary column with
a 6% cyanopropylphenyl–94% dimethylpolysiloxane
Result = (C/W) × F stationary phase and a 3.00-µm film thickness2
Temperature
C = concentration of lead in the Sample solution
Column: 20 min at 40°; increase to 240° at 10°/min;
determined from the standard curve (µg/
maintain at 240° for 10 min
mL)
Injection port: 140°
W = weight of Sample taken (g)
Detector: 250°
F = final volume of the Sample solution, 100 mL
Carrier gas: Nitrogen
Flow rate: 2.5 mL/min
• MERCURY
Headspace unit: 2.5 mL/min
[NOTE—When water is specified as a diluent, use
Equilibration temperature: 80°
Equilibration time: 60 min
specified, use nitric acid of a grade suitable for trace
Loop temperature: 85°
element analysis with as low a content of mercury as
Transfer temperature: 90°
practical.]
Pressurization time: 0.5 min
Diluent: 4% nitric acid in water
Loop fill time: 0.1 min
Standard stock solution: 100 µg/mL of mercury
Injection time: 1 min
prepared by diluting a commercially available 1000 mg
Injection size: 1 mL of headspace
/kg mercury ICP standard solution
2 CP-Select 624 CB (Varian-Chrompack, Palo Alto, CA), or equivalent.
FCC 9 Monographs / 5′-Cytidylic Acid / 347
System suitability 5′-inosinic acid, and 5′-adenylic acid) in the sample

Sample: Standard solution taken:
Suitability requirement: The relative standard
deviation of the ethanol peak area responses from Result = (rU/rS) × (CS/CU) × 100
replicate injections is NMT 5.0%.
rU = peak area of the analyte from the Sample
Analysis: Separately inject equal volumes of the
solution
Standard solution and Sample solution into the
rS = peak area of the analyte from the Standard
chromatograph, record the chromatograms, and
solution
measure the peak responses. [NOTE—The approximate
CS = concentration of analyte in the Standard
retention time for ethanol is 11 min.]
solution (mg/mL)
Monographs
Acceptance criteria: The peak area from the Sample
CU = concentration of analyte in the Sample
solution does not exceed that from the Standard
solution (mg/mL)
solution (NMT 200 mg/kg).
Acceptance criteria: The sum of the percentages for all
• OTHER RIBONUCLEOTIDES
nucleotide impurities is NMT 0.5%, calculated on the
Mobile phase and Chromatographic system: Prepare
anhydrous basis.
as directed in the Assay.
Standard solution: Mixture of USP Disodium 5′- SPECIFIC TESTS
Uridylate RS, USP 5′-Adenylic Acid RS, USP 5′-Cytidylic • PH, pH Determination, Appendix IIB
Acid RS, USP Disodium Guanylate RS, and USP Sample solution: 5 mg/mL
Disodium Inosinate RS each at 0.02 mg/mL in Mobile Acceptance criteria: 2.7–3.7
phase • WATER, Water Determination, Method I, Appendix IIB
Sample solution: 1.0 mg/mL. [NOTE—Ultrasonication Acceptance criteria: NMT 6.0%
for 15 min at 30° may be necessary to aid in complete • BILE-TOLERANT GRAM-NEGATIVE BACTERIA, Appendix XIIC
dissolution.] Sample preparation: Proceed as directed using a 10-g
Suitability requirements sample and incubating at 30°–35° for 18–24 h.
Sample: Standard solution Acceptance criteria: Negative in 10 g
Suitability requirement 1: The relative standard • ENTEROBACTER SAKAZAKII (Cronobacter spp.), Appendix XIIC
deviation of the 5′-cytidylic acid peak area responses Sample preparation: Proceed as directed using a 10-g
from replicate injections is NMT 2.0%. sample and incubating at 30°–35° for 18–24 h.
Suitability requirement 2: The resolution, R, between Acceptance criteria: Negative in 10 g
the 5′-cytidylic acid peak and all other nucleotide • SALMONELLA SPP., Appendix XIIC
peaks is NLT 2.0. Sample preparation: Dissolve 25 g of sample at a
Analysis: Separately inject equal volumes of the sample/broth ratio of 1/8, and proceed as directed.
Standard solution and Sample solution into the Acceptance criteria: Negative in 25 g
chromatograph, and measure the responses for all • TOTAL AEROBIC MICROBIAL COUNT, Method I (Plate Count
nucleotide peaks on the resulting chromatograms, Method), Appendix XIIB
except the peak from 5′-cytidylic acid. [NOTE—The Acceptance criteria: NMT 1,000 cfu/g
approximate retention times are 4.6 min (5′-cytidylic • TOTAL YEASTS AND MOLDS COUNT, Method I (Plate Count
acid), 6.2 min (5′-uridylic acid), 10.3 min (5′-guanylic Method), Appendix XIIB
acid), 11.5 min (5′-inosinic acid), and 27.5 min (5′- Acceptance criteria: NMT 100 cfu/g
adenylic acid).] Separately calculate the percentage of
each analyte (disodium 5′-uridylate, 5′-guanylic acid,
348 / α-Damascone / Monographs FCC 9
1-(2,6,6-Trimethyl-1-cyclohexenyl)-2-buten-1-one
α-Damascone
.
First Published: Third Supplement, FCC 8
4-(2,6,6-Trimethyl-2-cyclohexen-1-yl)-but-2-en-4-one
4-(2,6,6-Trimethyl-2-cyclohexenyl)-2-butene-4-one
1-(2,6,6-Trimethyl-2-cyclohexenyl)-2-buten-1-one C13H20O Formula wt 192.3
FEMA: 3243
CAS: [23726-92-3]
UNII: J131X1Y3RE [beta-damascone]
Monographs
DESCRIPTION
β-Damascone occurs as a colorless to pale yellow liquid.
C13H20O Formula wt 192.3 Odor: Fruity; floral
FEMA: 3659 Boiling Point: ∼67° to 70°
CAS: [43052-87-5] Solubility in Alcohol, Appendix VI: One mL dissolves in
UNII: UP5JTZ9821 [alpha-damascone] 10 mL of 95% alcohol.
DESCRIPTION
α-Damascone occurs as a colorless to pale yellow liquid. IDENTIFICATION
Odor: Fruity floral • INFRARED ABSORPTION, Spectrophotometric Identification
Boiling Point: ∼90°–100° Tests, Appendix IIIC
Solubility in Alcohol, Appendix VI: One mL dissolves in Reference standard: USP Beta-Damascone RS
10 mL of 95% alcohol. Sample and standard preparation: F
Function: Flavoring agent Acceptance criteria: The spectrum of the sample
IDENTIFICATION the spectrum of the Reference standard.
Tests, Appendix IIIC ASSAY
Reference standard: USP Alpha-Damascone RS • PROCEDURE: Proceed as directed under M-1b, Appendix
Sample and standard preparation: F XI.
Acceptance criteria: The spectrum of the sample Reference standard: USP Beta-Damascone RS
exhibits maxima at the same wavelengths as those in Acceptance criteria: NLT 90% (sum of cis- and trans-
the spectrum of the Reference standard. isomers)
ASSAY IMPURITIES
• PROCEDURE: Proceed as directed under M-1b, Appendix Organic Impurities
XI. • RELATED COMPOUNDS
Reference standard: USP Alpha-Damascone RS Procedure: Proceed as directed under M-1b, Appendix
Acceptance criteria: NLT 98% (sum of cis- and trans- XI.
isomers); among the sum, 92%–96% trans-, 4%–8% Reference standards: USP Alpha-Damascone RS; USP
cis-isomer Delta-Damascone RS
Acceptance criteria: NMT 1% α-damascone; NMT 1%
SPECIFIC TESTS δ-damascone
• REFRACTIVE INDEX, Appendix IIB: At 20°
Acceptance criteria: 1.492–1.499 SPECIFIC TESTS
• SPECIFIC GRAVITY: Determine at 25° by any reliable • REFRACTIVE INDEX, Appendix IIB: At 20°
method (see General Provisions). Acceptance criteria: 1.496–1.501
Acceptance criteria: 0.928–0.938 • SPECIFIC GRAVITY: Determine at 20° by any reliable
β-Damascone
.
First Published: Third Supplement, FCC 8
Damasione
Dorinone
4-(2,6,6-Trimethylcyclohex-1-enyl)but-2-en-4-one
FCC 9 Monographs / Dammar Gum / 349
DESCRIPTION
δ-Damascone
.
Crude Dammar Gum occurs as irregular, white to yellow to

First Published: Third Supplement, FCC 8 brown tears, fragments, or powder, sometimes admixed
with fragments of bark. Refined grades are white to yellow
1-(2,6,6-Trimethyl-3-cyclohexen-1-yl)-2-buten-1-one and are free of fragments of ligneous matter. Dammar
1-(2,6,6-Trimethyl-3-cyclohexenyl)-2-buten-1-one Gum is the dried exudate from trees of the Agathis, Hopea,
or Shorea genera. It consists of a complex mixture of acidic
and neutral terpenoid compounds together with
polysaccharide material. It is insoluble in water and in
ethanol and is soluble in toluene and in limonene. A
Monographs
chloroform solution of Dammar Gum is dextrorotatory.
Function: Stabilizer; glazing agent
C13H20O Formula wt 192.3 Packaging and Storage: Store in well-closed containers.
FEMA: 3622
CAS: [57378-68-4] IDENTIFICATION
UNII: 7F4RIE5P7E [delta-damascone] • THIN-LAYER CHROMATOGRAPHY, Appendix IIA
Sample solution: 100 mg/mL in chloroform
DESCRIPTION Adsorbent: 0.2-mm layer of silica (Merck F254, or
δ-Damascone occurs as a colorless to pale yellow liquid. equivalent)
Odor: Fruity Application volume: 20 µL
Boiling Point: ∼82° at 2 mm Hg; ∼97° at 3 mm Hg Developing solvent system: Diethyl ether:heptane
Solubility in Alcohol, Appendix VI: One mL dissolves in [30:25]
10 mL of 95% alcohol. Analysis: Spray the plate with sulfuric acid, and dry it at
Function: Flavoring agent 180° for 3 min.
Acceptance criteria: Two dark spots are observed at Rf
IDENTIFICATION of 0.8 and 0.7, and the ratio of the faster-moving spot
• INFRARED ABSORPTION, Spectrophotometric Identification to the second spot is about 1.1 to 1.
Reference standard: USP Delta-Damascone RS IMPURITIES
Sample and standard preparation: F Inorganic Impurities
Acceptance criteria: The spectrum of the sample • LEAD, Lead Limit Test, Appendix IIIB,
exhibits maxima at the same wavelengths as those in Sample solution: Prepare as directed for organic
the spectrum of the Reference standard. compounds.
ASSAY Solution)
• PROCEDURE: Proceed as directed under M-1b, Appendix Acceptance criteria: NMT 5 mg/kg
XI.
Reference standard: USP Delta-Damascone RS SPECIFIC TESTS
Acceptance criteria: NLT 96.5% • ACID NUMBER, Appendix IX
Sample: 5 g
SPECIFIC TESTS Analysis: Modify the procedure in Appendix IXA by
• REFRACTIVE INDEX, Appendix IIB: At 20° adding 30 mL each of toluene and neutral ethanol to
Acceptance criteria: 1.485–1.502 the Sample and titrating with 0.5 N alcoholic potassium
• SPECIFIC GRAVITY: Determine at 25° by any reliable hydroxide, using phenolphthalein TS as indicator.
method (see General Provisions). Acceptance criteria: Between 20 and 40
Acceptance criteria: 0.920–0.940 • ASH (TOTAL), Appendix IIC
Dammar Gum
.

First Published: Prior to FCC 6 Acceptance criteria: NMT 6.0%
• MELTING RANGE, Melting Range or Temperature
Dammar Resin Determination, Appendix IIB
Damar Gum Acceptance criteria: Between 90° and 95°
Damar Resin • SOFTENING POINT, Ring-and-Ball Method, Appendix IX
Dammar Acceptance criteria: Between 86° and 90°
CAS: [9000-16-2]
350 / (E),(E)-2,4-Decadienal / Monographs FCC 9

(E),(E)-2,4-Decadienal
.
mL of 95% ethanol.
First Published: Prior to FCC 6 Function: Flavoring agent
IDENTIFICATION
trans,trans-2,4-Decadienal Appendix IIIC
Monographs
C10H16O Formula wt 152.24 ASSAY

FEMA: 3135 • PROCEDURE: Proceed as directed under M-1a, Appendix
UNII: 3G88X2RK09 [2,4-decadienal, (2e,4e)-] XI.
Acceptance criteria: NLT 89.0% of C10H16O (sum of two
DESCRIPTION isomers)
(E),(E)-2,4-Decadienal occurs as a yellow liquid. It may
contain a suitable antioxidant. SPECIFIC TESTS
Odor: Powerful, oily, chicken fat • REFRACTIVE INDEX, Appendix II: At 20°
Solubility: Soluble in alcohol, most fixed oils; insoluble or Acceptance criteria: Between 1.514 and 1.519
practically insoluble in water • SPECIFIC GRAVITY: Determine at 25° by any reliable
Boiling Point: ∼104° (7 mm Hg) method (see General Provisions).
(E),(E)-2,4-Decadienal
Solubility: Very soluble in alcohol, propylene glycol,

δ-Decalactone
.
vegetable oils; insoluble or practically insoluble in water

First Published: Prior to FCC 6 Boiling Point: ∼281°
mL of 95% ethanol.
IDENTIFICATION
C10H18O2 Formula wt 170.25 Appendix IIIC
FEMA: 2361 Acceptance criteria: The spectrum of the sample
UNII: CNA0S5T234 [δ-decalactone] exhibits relative maxima at the same wavelengths as
DESCRIPTION
δ-Decalactone occurs as a colorless liquid. ASSAY
Odor: Coconut-fruity, buttery on dilution • PROCEDURE: Proceed as directed under M-1b, Appendix
XI.
Acceptance criteria: NLT 98.0% of C10H18O2
FCC 9 Monographs / γ-Decalactone / 351
SPECIFIC TESTS • REFRACTIVE INDEX, Appendix II: At 20°

• ACID VALUE (FATS AND RELATED SUBSTANCES), Method II, Acceptance criteria: Between 1.454 and 1.459
Appendix VII • SPECIFIC GRAVITY: Determine at 25° by any reliable
Acceptance criteria: NMT 5.0 method (see General Provisions).
Monographs
δ-Decalactone
IDENTIFICATION
γ-Decalactone
.

4-Hydroxydecanoic Acid Lactone exhibits relative maxima at the same wavelengths as
ASSAY
XI.
FEMA: 2360
UNII: 7HLS05KP9O [γ-decalactone] SPECIFIC TESTS
γ-Decalactone occurs as a colorless to pale yellow liquid. Acceptance criteria: NMT 1.0
Odor: Fruity, peach • REFRACTIVE INDEX, Appendix II: At 20°
Solubility: Soluble in propylene glycol, vegetable oils; Acceptance criteria: Between 1.447 and 1.451
insoluble or practically insoluble in water • SPECIFIC GRAVITY: Determine at 25° by any reliable
mL of 95% alcohol.
352 / γ-Decalactone / Monographs FCC 9
Monographs
γ-Decalactone

Decanal
.
IDENTIFICATION
Appendix IIIC
Aldehyde C-10 exhibits relative maxima at the same wavelengths as
Capraldehyde those of the spectrum below.
ASSAY
XI.
C10H20O Formula wt 156.27 Acceptance criteria: NLT 92.0% of C10H20O
FEMA: 2362
SPECIFIC TESTS
UNII: 31Z90Q7KQJ [decanal]
Decanal occurs as a colorless to light yellow liquid. It may Acceptance criteria: NMT 10.0
contain a suitable antioxidant. • REFRACTIVE INDEX, Appendix II: At 20°
Odor: Fatty, floral-orange on dilution Acceptance criteria: Between 1.426 and 1.430
Solubility: Miscible in alcohol, most fixed oils, propylene • SPECIFIC GRAVITY: Determine at 25° by any reliable
glycol (may be turbid); insoluble or practically insoluble in method (see General Provisions).
glycerin, water Acceptance criteria: Between 0.823 and 0.832
FCC 9 Monographs / (E)-2-Decenal / 353
Monographs
Decanal
• TITER (SOLIDIFICATION POINT), Solidification Point, Appendix

Decanoic Acid
.
IIB
First Published: Prior to FCC 6 Acceptance criteria: Between 27° and 32°
Capric Acid
• WATER, Water Determination, Method Ia, Appendix IIB

(E)-2-Decenal
.
FEMA: 2364
CAS: [334-48-5] First Published: Prior to FCC 6
UNII: 4G9EDB6V73 [capric acid] Last Revision: First Supplement, FCC 6
DESCRIPTION trans-2-Decenal
Decanoic Acid occurs as white crystals having a sour, fatty,
rancid odor. It is soluble in most organic solvents and
practically insoluble in water.
Function: Component in the manufacture of other food-
grade additives; defoaming agent; flavoring agent
FEMA: 2366
SPECIFIC TESTS UNII: E93S23U2BU [2-decenal, (2e)-]
• ACID VALUE (FATS AND RELATED SUBSTANCES), Method I,
Appendix VII DESCRIPTION
Acceptance criteria: Between 320 and 329 (E)-2-Decenal occurs as a slightly yellow liquid. It may
• IODINE VALUE, Appendix VII contain a suitable antioxidant.
Acceptance criteria: NMT 0.6 Odor: Orange, wax
• RESIDUE ON IGNITION (SULFATED ASH), Appendix IIC Solubility: Soluble in alcohol, most fixed oils; insoluble or
Sample: 10.0 g practically insoluble in water
Acceptance criteria: NMT 0.1% Boiling Point: ∼229°
mL of 95% ethanol.
354 / (E)-2-Decenal / Monographs FCC 9
Function: Flavoring agent Acceptance criteria: NLT 92.0% of C10H18O (one

isomer)
IDENTIFICATION
• INFRARED SPECTRA, Spectrophotometric Identification Tests, SPECIFIC TESTS
Appendix IIIC • REFRACTIVE INDEX, Appendix II: At 20°
Acceptance criteria: The spectrum of the sample Acceptance criteria: Between 1.452 and 1.457
exhibits relative maxima at the same wavelengths as • SPECIFIC GRAVITY: Determine at 25° by any reliable
those of the spectrum below. method (see General Provisions).
ASSAY
Monographs
XI.
(E)-2-Decenal

(Z)-4-Decenal
.
mL of 95% ethanol.
First Published: Prior to FCC 6 Function: Flavoring agent
IDENTIFICATION
cis-4-Decenal Appendix IIIC
C10H18O Formula wt 154.25 ASSAY

FEMA: 3264 • PROCEDURE: Proceed as directed under M-1a, Appendix
XI.
UNII: 5R675PGU7K [4-decenal, (4z)-]
Acceptance criteria: NLT 90.0% of C10H18O
(Z)-4-Decenal occurs as a colorless to slightly yellow liquid.
It may contain a suitable antioxidant.
Odor: Orange, fatty
Solubility: Soluble in alcohol, most fixed oils; insoluble or
practically insoluble in water
Boiling Point: ∼78° to 80° (10 mm Hg)
FCC 9 Monographs / Decyl Alcohol / 355
Monographs
(Z)-4-Decenal

Decyl Alcohol
.
IDENTIFICATION
Appendix IIIC
Alcohol C-10 Acceptance criteria: The spectrum of the sample
1-Decanol exhibits relative maxima at the same wavelengths as
ASSAY
C10H22O Formula wt 158.28 • PROCEDURE: Proceed as directed under M-1b, Appendix
FEMA: 2365 XI.
UNII: 89V4LX791F [decyl alcohol] Acceptance criteria: NLT 98.0% of C10H22O

Decyl Alcohol occurs as a colorless liquid. • ACID VALUE, FLAVOR CHEMICALS (OTHER THAN ESSENTIAL
Odor: Floral, waxy, fruity OILS), M-15, Appendix XI
Solubility: Soluble in alcohol, ether, mineral oil, propylene Acceptance criteria: NMT 1.0
glycol, most fixed oils; insoluble or practically insoluble in • REFRACTIVE INDEX, Appendix II: At 20°
glycerin, water Acceptance criteria: Between 1.435 and 1.439
356 / Decyl Alcohol / Monographs FCC 9
OTHER REQUIREMENTS
• SOLIDIFICATION POINT, Appendix IIB
Acceptance criteria: NLT 5°
Monographs
Decyl Alcohol

Dehydroacetic Acid
.

First Published: Prior to FCC 6 exhibits maxima at the same wavelengths as those in
the spectrum of the Reference standard.
3-Acetyl-6-methyl-1,2-pyran-2,4(3H)-dione ASSAY
Methylacetopyronone • DEHYDROACETIC ACID
Sample: 500 mg
Analysis: Transfer the Sample into a 250-mL Erlenmeyer
flask, dissolve it in 75 mL of neutral alcohol, add
phenolphthalein TS, and titrate with 0.1 N sodium
hydroxide to a pink endpoint that persists for at least
C8H8O4 Formula wt 168.15 30 s. Each mL of 0.1 N sodium hydroxide is equivalent
CAS: [520-45-6] to 16.82 mg of C8H8O4.
UNII: 2KAG279R6R [dehydroacetic acid] Acceptance criteria: NLT 98.0% and NMT 100.5% of
C8H8O4, calculated on the dried basis
DESCRIPTION
Dehydroacetic Acid occurs as a white or nearly white, IMPURITIES
crystalline powder. It is soluble in aqueous solutions of Inorganic Impurities
fixed alkalies, and is very slightly soluble in water. One g of • LEAD, Lead Limit Test, Atomic Absorption Spectrophotometric
sample dissolves in about 35 mL of alcohol and in 5 mL of Graphite Furnace Method, Method II, Appendix IIIB
acetone. Acceptance criteria: NMT 0.5 mg/kg
Function: Antimicrobial agent; preservative
Packaging and Storage: Store in well-closed containers. SPECIFIC TESTS
IDENTIFICATION Acceptance criteria: NMT 1%
• INFRARED ABSORPTION, Spectrophotometric Identification • MELTING RANGE, Melting Range or Temperature
Tests, Appendix IIIC Determination, Appendix IIB
Reference standard: USP Dehydroacetic Acid RS Acceptance criteria: Between 109° and 111°
Next Page
FCC 9 Monographs / Dexpanthenol / 357
• RESIDUE ON IGNITION (SULFATED ASH), Appendix IIC SPECIFIC TESTS

Sample: 2 g • LOSS ON DRYING, Appendix IIC: 140° for 4 h under a
Acceptance criteria: NMT 0.1% vacuum of NMT 5 mm Hg
Acceptance criteria: NMT 1%
• MELTING RANGE, Melting Range or Temperature
Determination, Appendix IIB
Desoxycholic Acid
.
Acceptance criteria: Between 172° and 175°

First Published: Prior to FCC 6 • RESIDUE ON IGNITION (SULFATED ASH), Appendix IIC
Sample: 1 g
Monographs
Deoxycholic Acid
3α,12α-Dihydroxycholanic Acid
Dexpanthenol
.
D(+)-Pantothenyl Alcohol
Panthenol
CAS: [83-44-3]
UNII: 005990WHZZ [deoxycholic acid]
DESCRIPTION
Desoxycholic Acid occurs as a white, crystalline powder. It is C9H19NO4 Formula wt 205.25
practically insoluble in water, slightly soluble in chloroform CAS: [81-13-0]
and in ether, soluble in acetone and in solutions of alkali UNII: 1O6C93RI7Z [dexpanthenol]
hydroxides and carbonates, and freely soluble in alcohol.
Function: Emulsifier DESCRIPTION
Packaging and Storage: Store in tight containers. Dexpanthenol occurs as a clear, viscous, somewhat
hygroscopic liquid. It is the dextrorotatory isomer of the
IDENTIFICATION alcohol analogue of pantothenic acid. Some crystallization
• PROCEDURE may occur on standing. It is freely soluble in water, in
Sample: 10 mg alcohol, in methanol, and in propylene glycol. It is soluble
Analysis: Add 2 drops of benzaldehyde and 3 drops of in chloroform and in ether, and is slightly soluble in
75% sulfuric acid to the Sample, heat at 50° for 5 min, glycerin. Its solutions are alkaline to litmus.
and then add 10 mL of glacial acetic acid. Function: Nutrient
Acceptance criteria: A green color appears (cholic acid Packaging and Storage: Store in tight containers.
produces a brown color).
IDENTIFICATION
ASSAY • A. PROCEDURE
• PROCEDURE Sample solution: 100 mg/mL
Sample: 500 mg Analysis: Add 5 mL of 1 N sodium hydroxide and 1
Analysis: Transfer the Sample into a 250-mL Erlenmeyer drop of cupric sulfate TS to 1 mL of the Sample solution,
flask, and add 20 mL of water and 40 mL of alcohol. and shake vigorously.
Cover the flask with a watch glass, heat the mixture Acceptance criteria: A deep-blue color develops.
gently on a steam bath until the sample is dissolved, • B. PROCEDURE
and allow the mixture to cool to room temperature. Sample solution: 10 mg/mL
Add a few drops of phenolphthalein TS to the solution, Analysis: Add 1 mL of 1 N hydrochloric acid to 1 mL of
and titrate with 0.1 N sodium hydroxide to a pink the Sample solution, and heat on a steam bath for
endpoint that persists for 15 s. Each mL of 0.1 N about 30 min. Cool, add 100 mg of hydroxylamine
sodium hydroxide is equivalent to 39.26 mg of hydrochloride, mix, and add 5 mL of 1 N sodium
C24H40O4. hydroxide. Allow to stand for 5 min, then adjust the pH
Acceptance criteria: NLT 98.0% and NMT 102.0% of to within a range of 2.5 to 3.0 with 1 N hydrochloric
C24H40O4, calculated on the dried basis acid, and add 1 drop of ferric chloride TS.
Acceptance criteria: A purple-red color develops.
IMPURITIES • C. INFRARED ABSORPTION, Spectrophotometric Identification
Inorganic Impurities Tests, Appendix IIIC
• LEAD, Lead Limit Test, Flame Atomic Absorption Reference standard: USP Dexpanthenol RS
Spectrophotometric Method, Appendix IIIB Sample and Standard preparation: E
Sample: 10 g

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278 / Chitosan / Monographs FCC 9

chromatogram between Va and Vb (NLT 40). [NOTE—If

n is sufficiently large, the use of area segments Ai or

manufacturer. a liquid under pressure in containers approved by the U.S.

Spectrophotometric Method, Appendix IIIB

Cholalic Acid SPECIFIC TESTS

It is very slightly soluble in water.

C9H19NO7 Formula wt 253.25 SPECIFIC TESTS

made by drying the sample in a vacuum desiccator

exhibits maxima at the same wavelengths as those in

separate 100-mL volumetric flasks, and transfer 1.5 mL

Acceptance criteria: Between 1.619 and 1.623

Standard and sample preparation: M

Acceptance criteria: The spectrum of the sample

fixed oils; 1 g dissolves in 2000 mL of water.

Cinnamon Bark Oil, Ceylon Type

and twigs of the true cinnamon shrub, Cinnamomum

zeylanicum Nees (Fam. Lauraceae). The commercial oils,

IDENTIFICATION • REFRACTIVE INDEX, Appendix IIB

with about 2% powdered tartaric acid, and filter. dilution.

Cinnamon Leaf Oil

Function: Flavoring agent

DESCRIPTION SPECIFIC TESTS

Function: Flavoring agent

C9H10O Formula wt 134.18

DESCRIPTION OTHER REQUIREMENTS

• INFRARED SPECTRA, Spectrophotometric Identification Tests,

Function: Flavoring agent

C18H16O2 Formula wt 264.32

DESCRIPTION SPECIFIC TESTS

Solubility: Miscible in alcohol, chloroform, ether, most

fixed oils; insoluble or practically insoluble in water

SPECIFIC TESTS Acceptance criteria: Between 1.077 and 1.082

• INFRARED SPECTRA, Spectrophotometric Identification Tests,

Function: Flavoring agent

• INFRARED SPECTRA, Spectrophotometric Identification Tests,

Boiling Point: ∼228°

Solubility in Alcohol, Appendix VI: One mL dissolves in 7

Acceptance criteria: Passes test

Standard solution: 4 µg/mL tridodecylamine in Acceptance criteria

Add 2.0 mL of Standard solution and 5 mL of Buffered

Add 20 mL of a spectrograde chloroform and n-heptane

IMPURITIES Odor: Intense lemon-citronella-rose

Sample: 2 g • INFRARED SPECTRA, Spectrophotometric Identification Tests,

OILS), M-15, Appendix XI

Acceptance criteria: The spectrum of the sample

exhibits relative maxima at the same wavelengths as

Acceptance criteria: The spectrum of the sample

exhibits relative maxima at the same wavelengths as

Acceptance criteria: The spectrum of the sample

exhibits relative maxima at the same wavelengths as

• INFRARED SPECTRA, Spectrophotometric Identification Tests,

Acceptance criteria: The spectrum of the sample

exhibits relative maxima at the same wavelengths as

• INFRARED SPECTRA, Spectrophotometric Identification Tests,

DESCRIPTION SPECIFIC TESTS

• ESTERS, Ester Determination, Appendix VI

DESCRIPTION SPECIFIC TESTS