AQX1MAN RevD Orion AquaMate 7100_8100_User_Manual
AQX1MAN RevD Orion AquaMate 7100_8100_User_Manual
AQX1MAN RevD Orion AquaMate 7100_8100_User_Manual
Thermo Scientific
Orion AquaMate
User Manual
All other trademarks are the property of Thermo Fisher Scientific Inc. and its subsidiaries.
Thermo Fisher Scientific Inc. provides this document to its customers with a product purchase to use in the
product operation. This document is copyright protected and any reproduction of the whole or any part of
this document is strictly prohibited, except with the written authorization of Thermo Fisher Scientific Inc.
The contents of this document are subject to change without notice. All technical information in this
document is for reference purposes only. System configurations and specifications in this document
supersede all previous information received by the purchaser.
Thermo Fisher Scientific Inc. makes no representations that this document is complete, accurate or error
free and assumes no responsibility and will not be liable for any errors, omissions, damage or loss that
might result from any use of this document, even if the information in the document is followed properly.
This document is not part of any sales contract between Thermo Fisher Scientific Inc. and a purchaser. This
document shall in no way govern or modify any Terms and Conditions of Sale, which Terms and
Conditions of Sale shall govern all conflicting information between the two documents.
For Research Use Only. This instrument or accessory is not a medical device and is not intended to be used
for the prevention, diagnosis, treatment or cure of disease.
Table of Contents
Chapter 1 .................................................................................................................................... 7
Spectrophotometer Introduction .............................................................................................. 7
Spectrophotometer Overview ...................................................................................................... 7
Orion AquaMate 7100 Vis Spectrophotometer ............................................................................ 8
Orion AquaMate 8100 UV-Vis Spectrophotometer ...................................................................... 8
Packing Lists................................................................................................................................ 8
Orion AquaMate 7100 Vis Spectrophotometer Packing List .............................................. 8
Orion AquaMate 8100 UV-Vis Spectrophotometer Packing List ........................................ 9
Orion AquaMate User Documentation on USB.......................................................................... 10
Intended Use ............................................................................................................................. 10
Operating Precautions ............................................................................................................... 10
Safety and Special Notices ........................................................................................................ 11
Chapter 2 .................................................................................................................................. 12
Spectrophotometer Basics ..................................................................................................... 12
Spectrophotometer Components ............................................................................................... 12
Instrument Touchscreen................................................................................................... 13
Instrument Connections ................................................................................................... 14
Optional Accessories........................................................................................................ 14
Sample Compartment ................................................................................................................ 15
Sample Compartment for AQ7100 and AQ8100 .............................................................. 15
Single Cell Holder ............................................................................................................ 16
Tray Features ................................................................................................................... 16
Removal – grasp cell holder and lift up and forward ........................................................ 16
Optional Sample Holders ................................................................................................. 17
Cell Holder Replacement ................................................................................................. 18
Optional Printer ................................................................................................................ 19
Selecting and Positioning Vials and Cuvettes .................................................................. 22
Z-dimension ..................................................................................................................... 22
Chapter 3 .................................................................................................................................. 23
Orion AquaMate Instrument Setup and Touchscreen and Features ................................... 23
Instrument Screen Navigation ................................................................................................... 23
User Interface Familiarity ................................................................................................. 25
User Interface Content ............................................................................................................... 27
Screen 1 Startup Screen .................................................................................................. 27
Screen 2 – Method Development, Diagnostics, and Data ................................................ 34
Screen 3 – Multi-wavelength and OD600 ......................................................................... 44
Instrument Settings .................................................................................................................... 45
Settings ............................................................................................................................ 45
SmartStart ........................................................................................................................ 46
Ending and Exporting Experiments .................................................................................. 54
Exporting Data ................................................................................................................. 55
Chapter 4 .................................................................................................................................. 57
Water Analysis Test Menu ...................................................................................................... 57
Preprogrammed Methods .......................................................................................................... 57
Method Selection and Experiment ............................................................................................. 58
CHAPTER 1
1
Spectrophotometer Introduction
Spectrophotometer Overview
Thermo Scientific™ Orion™ AquaMate™ Vis and UV-Vis spectrophotometers offer the following
features and benefits:
• Easy operation using the 260+ preprogrammed methods for common colorimetric reagents
• Easy access to approved regulatory methods for wastewater and drinking water.
• Secured Smart Method selection for frequently used methods
• Glove-friendly touchscreen user interface.
• Flexibility to create new methods for additional reagents or samples – create new methods
using calibration standards or update methods using published wavelengths and equations
• Use a variety of circular and rectangular vial sizes with a variety of vial holder options.
• Performance verification tests ensure wavelength accuracy and instrument functionality,
plus built-in filters allow for wavelength verification with no additional equipment required
• Additional functions include standard curve concentration measurements, wavelength
scanning, multiple fixed-wavelength measurements, absorbance ratio and difference
• One year instrument warranty
Packing Lists
Orion AquaMate 7100 Vis Spectrophotometer Packing List
• Vis Spectrophotometer with 7-inch color touchscreen and Tungsten-Halogen Lamp
• 12-25 mm Round Vial Holder (P/N: AQX1LWLVH)
• 24 mm Round vials, 12-pack quantity (P/N: AC2V24)
• External AC to DC Universal Power Supply, 100–240 volts, 50–60 Hz (AQX1PWRSUP)
• Standard power cord bundle (North American, EU, and UK) w/AQ7100
o NA Cord (P/N: AQX1NACBL)
o EU Cord (P/N: AQX1EUCBL)
o UK Cord (P/N: AQX1UKCBL)
• Optional APAC power cord bundle (China, Australia, and India) w/AQ7100APAC
o China Cord (P/N: AQX1CNCBL)
o Australia Cord (P/N: AQX1AUCBL)
o India Cord (P/N: AQX1INCBL)
• AquaMate User Guide, Methods List, and Reagent Instructions on USB
• AquaMate Getting Started Guide
• Read Me First Warning Guide (Multilanguage)
• AquaMate Site and Safety Guide
• CE Declaration of Conformity
• Printed instrument test verification report
• Dust cover
• USB cable
Note: The APAC (China, Australia, and India) power cord bundle must be specified by either
AQ7100APAC or AQ8100APAC upon ordering.
o Warranty Information
Intended Use
Please read this user manual thoroughly. Any use outside of these instructions may invalidate
the instrument warranty and cause permanent damage to the instrument.
Operating Precautions
Warning: Do not operate this system without following the safety precautions described in this
manual and the documentation that came with your system.
The spectrophotometer contains precise optical components. Handle it carefully and follow
these precautions.
• Allow instrument to come to room temperature after unboxing before power on.
Important: Instructions that must be followed to avoid damaging system hardware or data loss
Caution: Statements that indicate a hazardous situation that, if not avoided, could result in
minor or moderate injury
Warning: A hazardous situation that, if not avoided, could result in death or serious injury
Thermo Fisher Scientific provides this document to its customers with a product purchase to
use in the product operation. The contents of this document are subject to change without
notice. All technical information in this document is for reference purposes only. System
configurations and specifications in this document supersede all previous information received
by the purchaser.
Thermo Fisher Scientific makes no representations that this document is complete, accurate or
error free and assumes no responsibility and will not be liable for any errors, omissions,
damage or loss that might result from any use of this document, even if the information in the
document is followed properly.
This document is not part of any sales contract between Thermo Fisher Scientific Inc. and a
purchaser. This document shall in no way govern or modify any terms and conditions of sale,
which terms and conditions of sale shall govern all conflicting information between the two
documents.
For Research Use Only. This instrument is not a medical device and is not intended for the
prevention, diagnosis, treatment or cure of disease.
Warning: Avoid an explosion or fire hazard. This instrument is not designed for use in an
explosive atmosphere.
CHAPTER 2
2
Spectrophotometer Basics
Spectrophotometer Components
The following are some of the major components visible on the outside of the instrument:
Touchscreen
Printer
Housing
Sample Compartment
Electrical
Connections
USB Ports
Instrument Touchscreen
The instrument Touchscreen has 3-screen to navigate and can be swiped from left-to-right.
Below are the three user interface screens to navigate through by swiping left or right. Within
each screen there appear a series if application icons that are briefly described below. When
any application is selected, the user is directed to an “Application Home” screen. Methods can
be selected and adjusted. New methods can be created. Diagnostics and experiment data can
be reviewed. By tapping on the gear in the upper right corner, the Settings menu can be
accessed from any screen.
METHODS: AquaMate water method libraries C-MODE: 1-point cal. @ select λ and units. MULTI-WAVELENGTH: multi-λ scanning
SCAN: ABS or %T, min, max, interval, and speed. QUANT: Multi-point cal. curve development. OD600: reserved
LIVE DISPLAY: ABS or %T at specified wavelength. FIXED: Dual - λ and factor measurements.
KINETICS - time based experiments
DIAGNOSTICS: review verification test reports.
PERFORMANCE VERIFICATION REPORTS
DATA VIEWER: Logged experimental data.
Instrument Connections
Electrical Connections
• Single USB-A supports flash memory devices for method and data storage
• Export data to network or PC via Ethernet or Wi-Fi USB adaptor (not shown)
Warning: Avoid shock hazard. Always turn off the instrument and unplug it from the
wall outlet or power strip before you unplug the power cord from the instrument
connector.
Optional Accessories
The USB ports support the following peripheral devices:
• Printer
Sample Compartment
Remove all tape from the exterior of the instrument and inside the sample compartment.
Tray Features
• Able to contain spills up to 150 mL
NOTICE Clean the tray with water and mild detergent. Ethanol and iso-propyl alcohol can be
used if necessary but do not soak the tray in alcohols. Do not allow acetone, chlorocarbons or
other aggressive organic solvents to contact the tray. The PC-ABS plastic will soften and
discolor.
Insertion – allow front magnet to engage. Lower cell holder into place, allowing back magnet to
guide and engage
Loosen the captive screw and the base of the cell holder to remove it. Attach a new sample
holder in the same way.
Optional Printer
If the unit comes with the optional printer, follow the following steps and then reference Chapter
11 for Printer Setup through the touch screen.
4. Observing the bottom of the printer, align the guide rail on the printer with the guide rail
on the AquaMate Spectrophotometer
5. Push the printer forward until the connectors are fully connected. You will hear a snap
when the connectors have engaged properly.
Note: See the manufacturer’s specifications and work within the recommended range.
Position vials and cuvettes so that the clear sides face the light beam, one clear side facing the
front of the instrument and the other facing the back.
Note: Always place vials in the instrument in the same orientation in the light beam. An
alignment mark on the vial helps orientation the vial consistently and correctly.
Z-dimension
The figure below illustrates the position of the light beam in the instrument. Beam size
specifications are shown below.
• Distance from bottom of the vial/cuvette to center of beam (Z-dimension): 8.5 mm
• Beam dimensions: 2 mm (wide) by 7 mm (tall)
CHAPTER 3
3
Orion AquaMate Instrument Setup
and Touchscreen and Features
Instrument Screen Navigation
Below are the three user interface screens to navigate through Screen 1, Screen 2, and Screen
3 by swiping left or right. Within each screen there appears a series of application icons that
are included in a legend to briefly describe each application.
When any application is selected, the user is directed to an “Application Home” screen.
Methods can be selected and adjusted. New methods can be created. Diagnostics and
experiment data can be reviewed and performance verification reports can be viewed,
generated and exported.
METHODS: AquaMate water method libraries C-MODE: 1-point cal. @ select λ and units. MULTI-WAVELENGTH: multi-λ scanning
SCAN: ABS or %T, min, max, interval, and speed. QUANT: Multi-point cal. curve development. OD600: reserved
LIVE DISPLAY: ABS or %T at specified wavelength. FIXED: Dual - λ and factor measurements.
KINETICS - time based experiments
DIAGNOSTICS: review verification test reports.
PERFORMANCE VERIFICATION REPORTS
DATA VIEWER: Logged experimental data.
For example, in the image below when using a SCAN application, when tapping the method
name the keyboard will appear to edit. When tapping the min or max wavelength, a wavelength
keypad appears. When tapping the interval or the speed fields, respective pop keypads will
appear to edit the method/experiment. Finally, a diskette icon is available to be used to save
the method with the edited fields and name. Other areas to look for are the ellipsis icons that
will expand access to editing features and more.
This section uses the Scan application as an example. As shown, using the ellipsis will open
additional fields for the saved method allowing the following:
• Method revisions
• Method export
• DROPLETS - Five (5) droplet folders with respective water methods. Any method
identified as having a regulatory approval is duplicated in the Drinking water or
Wastewater droplet folders. Droplet folder version numbers are provided.
Droplet Folders
There are three main water method Droplet Folders and two Regulatory folders:
1. AquaMate Orion
2. Merck/Millipore
3. Chemetrics
4. Wastewater (regulatory)
Any AquaMate water method that qualifies as a regulatory Wastewater (WW) or Drinking Water
(DW) method is duplicated within the respective WW or DW folders. Within each folder you can
search either by the numerical method name or by the parameter. For example, you can
search by either AC2002 or by Alkalinity-M. You can also sort by method name, date of
creation, date last updated or date last used.
NOTE: Each method description provides the wavelength, appropriate vial size, and
measurement range. If the incorrect vial size is used, the results will not be accurate.
Note: Please refer to the method description for measurement capabilities for each
method. This instrument will report values outside of the stated range capabilities that may not
be acceptable for the user’s specific purpose or for regulatory reporting requirements.
NOTE: The regulatory method range may be limited to a different range of the AquaMate
method itself.
SCAN Application
SCAN is a multiwavelength scan, selecting absorption (ABS) or percent transmission (%T),
across a selectable wavelength range and selectable speed and interval resolution. The value
of SCAN is to evaluate the absorption or transmission characteristics of a sample or a sample
with reagent. This is valuable when determining what the best wavelength is to establish a new
method.
Custom sample scanning methods can be created for sample characterization, yet no
concentration measurements are made here.
The wavelength can be edited by using the sliding scale adjustment bar or by tapping the blue
wavelength to edit the value directly. Either ABS or %T can be selected. Each time a change
is made, the system should be re-blanked before allowing the Live Display readings to
continue.
Once the instrument is blanked, the system will provide automatically real-time and continuous
measurements until the X in the upper left corner is tapped.
• C-Mode – select a wavelength, enter a known standard concentration, and select the
units of measure. By blanking and measuring of the standard - a single point
calibration report of ABS and calculated Cal Factor are calculated and reported.
• Quant – is a calibration curve development application. The user can select the
expiration date, wavelength, reference wavelength, equation, units of concentration,
and enter the known standards that will be used to develop the curve. Methods can
be saved by name and used to measure subsequent sample concentration.
• Fixed – permits the user to enter a single or multiwavelength method base on 3rd party
documented data; such as ABS or %T, wavelength_1, wavelength_2, units of
measure, and the respective wavelength factors, and either a direct, additive,
differential, or ratiometric equation. Methods can be saved by name and used to
measure subsequent samples concentration.
• Data Viewer – lists the date and number of experiments conducted on that date. By
selecting that day, the reports are unfolded and a report can be selected, viewed, and
printed and/or exported.
Below is an image of Screen 2 that can be associated with the descriptions above. A summary
legend is provided at the bottom.
C-Mode
C-Mode permits the user to select a wavelength, introduce a known standard concentration,
select from the units of measure. By blanking and measuring, the user will have developed a
single point calibration and report what the absorbance is and the factor necessary to correlate
to the known concentration value that was entered.
Quant
Quant is a calibration curve development application. The user can select the expiration date,
wavelength, reference wavelength, equation, units of concentration, and enter the known
standards that will be used to develop the curve. Methods can be saved by name and used to
measure subsequent sample concentration
Quant Options
By tapping on the equation that is listed to the right of the wavelength, a selection of equations
will appear. As the complexity of the equation increases, so does the number of points
required. Choose the equation that you feel is best suitable. When the multi-point calibration is
completed, the resulting equation and the correlation results (r2) will appear. These results are
based on the quality of your blank and the accuracy of the standards prepared and entered.
Be sure to save your results by tapping the blue diskette icon in the upper right-hand corner.
Tap to
expand field
Diagnostics Menu
Diagnostic opens a stored list of performance verifications that can be completed on the
instrument. If a scheduled interval is overdue, a red clock icon will appear. These verification
intervals are dependent on lab protocol. By selecting one of the available tests and
performance verification can be run. Tests that require no additional accessories are:
• Wavelength Accuracy
• Drift at 500 nm
• Baseline Flatness
Data Viewer
When selected, Data Viewer will list the date and number of experiments conducted on that
date. By selecting that day, the reports are unfolded and a report can be selected, viewed, and
printed and/or exported. You can see the experiment and any graphical data as it appeared on
the day of the experiment.
• OD600 – reserved
Instrument Settings
Settings
The instrument settings are accessible by tapping the gear in the upper right corner from any
main screen. From the settings window the user can:
• Smart Start – activation will only show Smart Methods on the Home Screen.
• Language will permit the user to select English, Deutsch, Italiano, Espanol, Francais,
Portuguese, and multiple Asian languages
• Disk space
• Software Update
SmartStart
The following are SmartStart features:
• Locks SmartStart mode with optional password protection; thus limiting the user to
only the methods made available
Unlocking SmartStart
The user must have the password of a USB Password Reset Key to unlock the instruments. If
the password cannot be remembered, you will have to contact your technical support team to
request a key be sent to you.
Language
Several languages are available for selection. Please select the language that is suitable to
meet your needs. If there are any issues with the language accuracy, please contact technical
support at wlp.techsupport@thermofisher.com.
Printer Settings
Below are the printer settings window. If you have a thermal printer option, please reference
Section 2 of this manual.
The user can select either a Thermal printer or a USB/Network printer for configuration. The
report header can be configured to reflect the use case for the spectrophotometer.
NOTE: For the 7100 AquaMate, the tungsten lamp should be placed in a lamp saver
mode and will shut off after 15-minutes of non-use. Please remember that the Tungsten
lamp will need to be warmed-up to get accurate results.
Software Update
The Software Updates allow two updates to be made. The first is the firmware update and the
second is specific to updating the water method library. Water Methods are only specific to
AquaMate Spectrophotometers and are typical of any aqueous method that utilizes reagent
chemistry for results. If for any reason you are unable to update your library methods, please
contact technical support with your specific model and serial number.
Software Update
For a software update, contact wlp.techsupport@thermofisher.com for the latest firmware and
library methods and place these on your USB stick. Insert the stick into the front USB port.
Tap on Update Software. Tap on the most recent release date version and tap on Update. The
instrument will self-guide you through the process and will automatically reboot itself.
• Go Back
Go Back to experiment
Send to printer
Export to file
Confirm experiment is
ended.
Exporting Data
If you choose to export your data, be sure to have setup a network path via Ethernet or via
WiFi. A typical method is to save the data directly to a USB drive. The report can be saved as
both a CSV file and/or a JPEG file.
Below is an example of the USB file directory and a JPEG file image of the experiment.
CHAPTER 4
4
Water Analysis Test Menu
Preprogrammed Methods
Thermo Scientific Orion AquaMate 8100 UV-Vis and AquaMate 7100 Vis spectrophotometers
include over 260 preprogrammed methods for use with Thermo Scientific™ Orion™ AQUAfast™,
Merck, and CHEMetrics reagent chemistries. Preprogrammed methods provide values for the
test parameters required to run specific reagent chemistries on the instrument, including
wavelength, vial path length, concentration factors/curves and measurement units. Any
methods that have a regulatory approved for wastewater or drinking water have are
conveniently duplicated within their respective droplet folders.
All preprogrammed methods and documentation are stored on the instrument as a methods
library package and is also available for dowload at thermofisher.com/aquamate. The
pre-programmed methods are specific to AquaMate Spectrophotometers only. Operators can
modify preprogrammed methods or create their own custom methods, so additional parameters
and test methods can be added at any time.
The following instructions are for using Orion AQUAfast reagent, Merck or CHEMetrics
chemistries with the AquaMate spectrophotometer. Preprogrammed methods use a specific vial
size (path length) in the formula and the vial size specified in these instructions must be used
for accurate analysis. The majority of AQUAfast reagent methods use a 24mm round vial, Cat.
No. AC2V24 or 16mm round vial, Cat. No. AC2V16. Other vial sizes are noted in the individual
reagent chemistry instructions.
Method descriptions provided detail the method number, parameter, wavelength, vial type and
size, as well as detailed information about the method itself. Follow the method directions per
reagent instructions in Chapter 5 or per vendor instructions.
AQUAMATE
Note: Please refer to the method description for measurement capabilities for each
method. This instrument will report values outside of the stated range capabilities that may not
be acceptable for the user’s specific purpose or for regulatory reporting requirements.
Method Options
Within the method selected, the fields that appear in blue are normally editable fields. In the
example below, the Sample Base Name (Sample), can be customized by using the
alphanumeric touchscreen field that appears when tapped. When the field being edited is
complete, press the Done key on the touchscreen keypad.
• If you are working with a new batch or reagents, you have the option of preparing a
known and traceable standard, edit the Standard Concentration Value, and tap Re-
calibrate. This will update the Calibration Factor. Otherwise, move onto Measuring.
• Insert the prepared sample and tap Measure to get the results.
End experiment
Note: Ideally, the same vial should be used or one that has been matched to the sample vial.
Note: The blank measurement is stored and used while working in the sample test method. The
blank measurement will be cleared automatically if any test method settings are changed, if the
test method is saved, or if a new test method is loaded.
Note: When running a reverse color test method, a reagent blank measurement is required
after the standard blank. Insert the vial containing the reagent blank and press the Blank
function key. Open the lid and remove the reagent blank. See the Using the Reverse Color
Feature section for detailed instructions.
Note: If the Statistics option is set to Off, statistics will not be displayed.
Below appears an example screen for a COD method and the typical fields highlighted in blue
that may require attention. Furthermore, the final screen that addresses ending and
experiment, naming the experiment, printing and exporting experiment results.
1. Load the test method in the Water Analysis test menu. The Reverse Color (Negative ABS)
option should be set to ON for the method.
2. Prepare the sample reagent blank into the vial defined by the method.
3. Place the vial into the holder in the sample chamber and close the chamber door.
4. Press the Blank function key to measure the reagent blank.
5. Open the sample chamber door and remove the vial from the holder.
6. Follow the method directions and prepare the sample to be measured.
7. Place the vial into the holder in the sample chamber and close the chamber door.
8. Press the Measure function key to display the results.
9. Continue to run additional samples as needed.
10. When complete, end the experiment and export or print the data.
CHAPTER 5
5
Orion AQUAfast Reagent Chemistry
Instructions for Orion AquaMate
Orion AQUAfast Colorimetric Reagents
Compatible with Orion AquaMate Instruments
Use the information in the following table to identify the Orion AQUAfast reagent method file
name on AQ7100 or AQ8100 and the test parameters associated with each method. This
information is also included on the Orion AquaMate user documentation CD or on our website
at www.thermoscientific.com/water.
Test methods and ranges are subject to change without notice. For a list of the most up-to-date
test methods, visit www.thermoscientific.com/water.
Notes:
• The terms total alkalinity, alkalinity-m, m-value and alkalinity to pH 4.3 are identical.
• For accurate results, exactly 10 ml of water sample must be taken for the test.
Notes
• The terms alkalinity-p, p-value and alkalinity to pH 8.2 are identical.
• For accurate test results, exactly 10 ml of water sample must be taken for the test.
• This method was developed from a volumetric procedure for the determination of
alkalinity-p. Due to undefined conditions, the deviations from the standardized method may
be greater.
Notes:
• Before use, clean the vials and the measuring beaker with hydrochloric acid (approximately
20%). Rinse them thoroughly with deionized water.
• To get accurate results the sample temperature must be between 20 °C and 25 °C.
• A low test result may be given in the presence of fluorides and polyphosphates. The effect
of this is generally insignificant unless the water has fluoride added artificially.
Notes:
• Before use, clean the vials and the measuring beaker with hydrochloric acid (approximately
20%). Rinse them thoroughly with deionized water.
• To get accurate results the sample temperature must be between 20 °C and 25 °C.
• A low test result may be given in the presence of fluorides and polyphosphates. The effect
of this is generally insignificant unless the water has fluoride added artificially.
Notes:
• The tablets must be added in the correct sequence.
• The Ammonia No. 1 tablet will only dissolve completely after the Ammonia No. 2 tablet has
been added.
• The temperature of the sample is important for full color development. At a temperature
below 20 °C, the reaction period is 15 minutes.
• Conversion: mg/l NH4 = mg/l N x 1.29
mg/l NH3 = mg/l N x 1.22
Notes:
• Extremely basic or acidic water samples should be adjusted to pH 7 with a 0.5 mol/l (1 N)
sulfuric acid solution or 1 mol/l (1 N) sodium hydroxide solution.
Notes:
• Strong alkaline or acidic water samples must be adjusted to approximately pH 7 before
analysis (use 1 mol/l hydrochloric acid or 1 mol/l sodium hydroxide).
• If chlorine is known to be present, add one drop of 0.1 mol/l sodium thiosulfate for each 0.3
mg/l Cl2 in a one liter water sample.
• Iron interferes with the test. The interferences can be eliminated as follows: Determine the
amount of total iron present in the water sample. To produce the blank add an iron
standard solution with the same iron concentration to the vial instead of deionized water.
• Conversion: mg/l NH4 = mg/l N x 1.29
mg/l NH3 = mg/l N x 1.22
Notes:
• Strong alkaline or acidic water samples must be adjusted to approximately pH 7 before
analysis (use 1 mol/l hydrochloric acid or 1 mol/l sodium hydroxide).
• If chlorine is known to be present, add one drop of 0.1 mol/l sodium thiosulfate for each 0.3
mg/l Cl2 in a one liter water sample.
• Iron interferes with the test. The interferences can be eliminated as follows: Determine the
amount of total iron present in the water sample. To produce the blank, add an iron
standard solution with the same iron concentration to the vial instead of deionized water.
• Conversion: mg/l NH4 = mg/l N x 1.29
mg/l NH3 = mg/l N x 1.22
Notes:
• Alternatively, the AC203510 method can be used with 10mm square vials and the
AC203550 method can be used with 50mm rectangular vials. All blank and sample
volumes must remain the same as those specified in these instructions, so samples may
need to be prepared in separate containers and then transferred into the selected vial.
• Vial cleaning: As many household cleaners (i.e. dishwasher detergent) contain reducing
substances, the subsequent determination of bromine may show lower results. To avoid
any measurement errors, only use glassware free of chlorine demand.
Preparation: Put all applicable glassware into sodium hypochlorite solution (0.1 g/l) for one
hour and then rinse all glassware thoroughly with deionized water.
• Preparing the sample: When preparing the sample, the escape of bromine gases, i.e. by
pipetting or shaking, must be avoided. The analysis must take place immediately after
taking the sample.
• The DPD color development is carried out at a pH value of 6.2 to 6.5. The reagent tablet
therefore contains a buffer for the pH adjustment. Strong alkaline or acidic water samples
must be adjusted between pH 6 and pH 7 before the reagent is added (use 0.5 mol/l
sulfuric acid or 1 mol/l sodium hydroxide).
• Exceeding the measuring range: Concentrations above 22 mg/l bromine can lead to results
showing 0 mg/l. In this event, the water sample must be diluted with water free of bromine.
10 ml of the diluted sample should be mixed with the reagent and the measurement
repeated.
• Oxidizing agents such as chlorine or ozone interfere as they react in the same way as
bromine.
Notes:
• Ensure that all particles of the tablet are dissolved – chloride causes an extremely fine
distributed turbidity with a milky appearance. Heavy shaking leads to bigger sized particles
that can cause false readings.
• High concentrations of electrolytes and organic compounds have different effects on the
precipitation reaction.
• Ions that also form deposits with silver nitrate in acidic media, such as bromides, iodides
and thiocyanates, interfere with the analysis.
• Highly alkaline water should, if necessary, be neutralized using nitric acid before analysis.
Notes:
• Alternatively, the AC207010 method can be used with 10mm square vials and the
AC207050 method can be used with 50mm rectangular vials. All blank and sample
volumes must remain the same as those specified in these instructions, so samples may
need to be prepared in separate containers and then transferred into the selected vial.
• Vial cleaning: As many household cleaners (i.e. dishwasher detergent) contain reducing
substances, the subsequent determination of chlorine may show lower results. To avoid
any measurement errors, only use glassware free of chlorine demand.
Preparation: Put all applicable glassware into sodium hypochlorite solution (0.1 g/l) for one
hour and then rinse all glassware thoroughly with deionized water.
• For individual testing of free and total chlorine, the use of different sets of glassware is
recommended (EN ISO 7393-2, 5.3).
• Preparing the sample: When preparing the sample, the escape of chlorine gases, i.e. by
pipetting or shaking, must be avoided. The analysis must take place immediately after
taking the sample.
• The DPD color development is carried out at a pH value of 6.2 to 6.5. The reagents
therefore contain a buffer for the pH adjustment. Strong alkaline or acidic water samples
must be adjusted between pH 6 and pH 7 before the reagent is added (use 0.5 mol/l
sulfuric acid or 1 mol/l sodium hydroxide).
• Exceeding the measuring range: Concentrations above 10 mg/l chlorine using tablets can
lead to results showing 0 mg/l. In this event, the water sample must be diluted with water
free of chlorine. 10 ml of the diluted sample should be mixed with the reagent and the
measurement repeated.
• Turbidity can lead to errors. The use of the DPD No. 1 tablet in samples with high calcium
ion contents and/or high conductivity can lead to turbidity of the sample and therefore
incorrect measurements.
• Oxidizing agents such as bromine or ozone interfere as they react in the same way as
chlorine.
Notes:
• Alternatively, the AC207110 method can be used with 10mm square vials and the
AC207150 method can be used with 50mm rectangular vials. All blank and sample
volumes must remain the same as those specified in these instructions, so samples may
need to be prepared in separate containers and then transferred into the selected vial.
• Vial cleaning: As many household cleaners (i.e. dishwasher detergent) contain reducing
substances, the subsequent determination of chlorine may show lower results. To avoid
any measurement errors, only use glassware free of chlorine demand.
Preparation: Put all applicable glassware into sodium hypochlorite solution (0.1 g/l) for one
hour and then rinse all glassware thoroughly with deionized water.
• For individual testing of free and total chlorine, the use of different sets of glassware is
recommended (EN ISO 7393-2, 5.3).
• Preparing the sample: When preparing the sample, the escape of chlorine gases, i.e. by
pipetting or shaking, must be avoided. The analysis must take place immediately after
taking the sample.
• The DPD color development is carried out at a pH value of 6.2 to 6.5. The reagents
therefore contain a buffer for the pH adjustment. Strong alkaline or acidic water samples
must be adjusted between pH 6 and pH 7 before the reagent is added (use 0.5 mol/l
sulfuric acid or 1 mol/l sodium hydroxide).
• Exceeding the measuring range: Concentrations above 10 mg/l chlorine using tablets can
lead to results showing 0 mg/l. In this event, the water sample must be diluted with water
free of chlorine. 10 ml of the diluted sample should be mixed with the reagent and the
measurement repeated.
• Turbidity can lead to errors. The use of the DPD No. 1 tablet in samples with high calcium
ion contents and/or high conductivity can lead to turbidity of the sample and therefore
incorrect measurements.
• Oxidizing agents such as bromine or ozone interfere as they react in the same way as
chlorine.
Notes:
• Alternatively, the AC207210 method can be used with 10mm square vials and the
AC207250 method can be used with 50mm rectangular vials. All blank and sample
volumes must remain the same as those specified in these instructions, so samples may
need to be prepared in separate containers and then transferred into the selected vial.
• Vial cleaning: As many household cleaners (i.e. dishwasher detergent) contain reducing
substances, the subsequent determination of chlorine may show lower results. To avoid
any measurement errors, only use glassware free of chlorine demand.
Preparation: Put all applicable glassware into sodium hypochlorite solution (0.1 g/l) for one
hour and then rinse all glassware thoroughly with deionized water.
• For individual testing of free and total chlorine, the use of different sets of glassware is
recommended (EN ISO 7393-2, 5.3).
• Preparing the sample: When preparing the sample, the escape of chlorine gases, i.e. by
pipetting or shaking, must be avoided. The analysis must take place immediately after
taking the sample.
• The DPD color development is carried out at a pH value of 6.2 to 6.5. The reagents
therefore contain a buffer for the pH adjustment. Strong alkaline or acidic water samples
must be adjusted between pH 6 and pH 7 before the reagent is added (use 0.5 mol/l
sulfuric acid or 1 mol/l sodium hydroxide).
• Exceeding the measuring range: Concentrations above 10 mg/l chlorine using tablets can
lead to results showing 0 mg/l. In this event, the water sample must be diluted with water
free of chlorine. 10 ml of the diluted sample should be mixed with the reagent and the
measurement repeated.
• Turbidity can lead to errors. The use of the DPD No. 1 tablet in samples with high calcium
ion contents and/or high conductivity can lead to turbidity of the sample and therefore
incorrect measurements.
• Oxidizing agents such as bromine or ozone interfere as they react in the same way as
chlorine.
Notes:
• Vial cleaning: As many household cleaners (i.e. dishwasher detergent) contain reducing
substances, the subsequent determination of chlorine may show lower results. To avoid
any measurement errors, only use glassware free of chlorine demand.
Preparation: Put all applicable glassware into sodium hypochlorite solution (0.1 g/l) for one
hour and then rinse all glassware thoroughly with deionized water.
• For individual testing of free and total chlorine, the use of different sets of glassware is
recommended (EN ISO 7393-2, 5.3).
• Preparing the sample: When preparing the sample, the escape of chlorine gases, i.e. by
pipetting or shaking, must be avoided. The analysis must take place immediately after
taking the sample.
• The DPD color development is carried out at a pH value of 6.2 to 6.5. The reagents
therefore contain a buffer for the pH adjustment. Strong alkaline or acidic water samples
must be adjusted between pH 6 and pH 7 before the reagent is added (use 0.5 mol/l
sulfuric acid or 1 mol/l sodium hydroxide).
• Exceeding the measuring range: Concentrations above 2 mg/l chlorine using powder packs
can lead to results showing 0 mg/l. In this event, the water sample must be diluted with
water free of chlorine. 10 ml of the diluted sample should be mixed with the reagent and
the measurement repeated.
• Oxidizing agents such as bromine or ozone interfere as they react in the same way as
chlorine.
Notes:
• Vial cleaning: As many household cleaners (i.e. dishwasher detergent) contain reducing
substances, the subsequent determination of chlorine may show lower results. To avoid
any measurement errors, only use glassware free of chlorine demand.
Preparation: Put all applicable glassware into sodium hypochlorite solution (0.1 g/l) for one
hour and then rinse all glassware thoroughly with deionized water.
• For individual testing of free and total chlorine, the use of different sets of glassware is
recommended (EN ISO 7393-2, 5.3).
• Preparing the sample: When preparing the sample, the escape of chlorine gases, i.e. by
pipetting or shaking, must be avoided. The analysis must take place immediately after
taking the sample.
• The DPD color development is carried out at a pH value of 6.2 to 6.5. The reagents
therefore contain a buffer for the pH adjustment. Strong alkaline or acidic water samples
must be adjusted between pH 6 and pH 7 before the reagent is added (use 0.5 mol/l
sulfuric acid or 1 mol/l sodium hydroxide).
• Exceeding the measuring range: Concentrations above 2 mg/l chlorine using powder packs
can lead to results showing 0 mg/l. In this event, the water sample must be diluted with
water free of chlorine. 10 ml of the diluted sample should be mixed with the reagent and
the measurement repeated.
• Oxidizing agents such as bromine or ozone interfere as they react in the same way as
chlorine.
Notes:
• Oxidizing agents interfere as they react in the same way as chlorine.
10. Close the vial tightly with the cap and swirl or invert several times until the tablets are
dissolved. Wipe the exterior of the vial.
11. Place the vial into the holder in the sample chamber and close the chamber door.
12. Tap the Sample function key to display the result in mg/l chlorine dioxide.
Notes:
• Alternatively, the AC209950 method can be used with 50mm rectangular vials. All blank
and sample volumes must remain the same as those specified in these instructions, so
samples may need to be prepared in separate containers and then transferred into the
selected vial.
• Vial cleaning: As many household cleaners (i.e. dishwasher detergent) contain reducing
substances, the subsequent determination of chlorine dioxide may show lower results. To
avoid any measurement errors, only use glassware free of chlorine demand.
Preparation: Put all applicable glassware into sodium hypochlorite solution (0.1 g/l) for one
hour and then rinse all glassware thoroughly with deionized water.
• Preparing the sample: When preparing the sample, the escape of chlorine dioxide gases,
i.e. by pipetting or shaking, must be avoided. The analysis must take place immediately
after taking the sample.
• The DPD color development is carried out at a pH value of 6.2 to 6.5. The reagent tablet
therefore contains a buffer for the pH adjustment. Strong alkaline or acidic water samples
must be adjusted between pH 6 and pH 7 before the tablet is added (use 0.5 mol/l sulfuric
acid or 1 mol/l sodium hydroxide).
• Exceeding the measuring range: Concentrations above 19 mg/l chlorine dioxide can lead to
results showing 0 mg/l. In this event, the water sample must be diluted with water free of
chlorine dioxide. 10 ml of the diluted sample should be mixed with the reagent and the
measurement repeated.
• Oxidizing agents such as chlorine or ozone interfere as they react in the same way as
chlorine dioxide.
1. Open one 16 mm COD reaction vial and add 2 ml of deionized water (this is the reagent
blank vial).
2. Open a second 16 mm reaction vial and add 2 ml of sample (this is the sample vial).
3. Close the vials tightly with the caps and gently invert the vials several times to mix the
contents. CAUTION: The vials will become hot during mixing.
4. Heat the vials for 120 minutes in the preheated reactor at a temperature of 150 °C.
5. CAUTION: The vials will be hot.
Remove the vials from the reactor and allow them to cool to 60 °C or less. Gently invert the
vials several times to mix the contents while still warm. Allow the vials to cool to room
temperature before measuring. Wipe the exteriors of the vials.
6. Load and run the CODL00 method.
7. Place the reagent blank vial into the holder in the sample chamber and close the chamber
door.
8. Press the Blank function key to measure the reagent blank.
9. Open the sample chamber door. Remove the reagent blank vial from the holder.
10. Place the sample vial into the holder in the sample chamber and close the chamber door.
11. Tap the Sample function key to display the result in mg/l oxygen.
Notes:
• Reverse color methods use a reagent that, when prepared with samples, deceases in color
as the concentration of the species being measured in the samples increases. Reverse
color methods require the use of a reagent blank. The reagent blank is a mixture of the
reagent and deionized water and provides a zero concentration point with the darkest color
(highest absorbance). The color of samples prepared with the reagent will decrease as the
concentration increases for this method.
• Run samples and blanks with the same batch of vials. The blank is stable when stored in
the dark and can be used for further measurements with vials of the same batch.
• Do not place the hot vials in the sample chamber. Cool the vials to room temperature for
final measurements.
• Suspended solids in the vial lead to incorrect measurements. For this reason, it is
important to place the vials carefully in the sample chamber. The precipitate at the bottom
of the sample should be not suspended.
• Clean the outside of the vials with a towel. Fingerprints or other marks must be removed.
• Samples can be measured when the chloride content does not exceed 1000 mg/l.
• In exceptional cases, compounds contained in the water cannot be oxidized adequately, so
results may be lower than reference methods.
1. Open one 16 mm COD reaction vial and add 2 ml of deionized water (this is the blank vial).
2. Open a second 16 mm COD reaction vial and add 2 ml of sample (this is the sample vial).
3. Close the vials tightly with the caps and gently invert the vials several times to mix the
contents. CAUTION: The vials will become hot during mixing.
4. Heat the vials for 120 minutes in the preheated reactor at a temperature of 150 °C.
5. CAUTION: The vials will be hot.
Remove the vials from the reactor and allow them to cool to 60 °C or less. Gently invert the
vials several times to mix the contents while still warm. Allow the vials to cool to room
temperature before measuring. Wipe the exteriors of the vials.
6. Load and run the CODH00 method.
7. Place the blank vial into the holder in the sample chamber and close the chamber door.
8. Tap the Blank function key to measure the blank.
9. Open the sample chamber door. Remove the blank vial from the holder.
10. Place the sample vial into the holder in the sample chamber and close the chamber door.
11. Tap the Sample function key to display the result in mg/l oxygen.
Notes:
• Run samples and blanks with the same batch of vials. The blank is stable when stored in
the dark and can be used for further measurements with vials of the same batch.
• Do not place the hot vials in the sample chamber. Cool the vials to room temperature for
final measurements.
• Suspended solids in the vial lead to incorrect measurements. For this reason, it is
important to place the vials carefully in the sample chamber. The precipitate at the bottom
of the sample should be not suspended.
• Clean the outside of the vials with a towel. Fingerprints or other marks must be removed.
• Samples can be measured when the chloride content does not exceed 1000 mg/l.
• In exceptional cases, compounds contained in the water cannot be oxidized adequately, so
results may be lower than reference methods.
• For samples under 100 mg/l it is recommended to repeat the test using the COD low range
test (CODL00).
1. Open one 16 mm COD reaction vial and add 0.2 ml of deionized water (this is the blank
vial).
2. Open a second 16 mm COD reaction vial and add 0.2 ml of sample (this is the sample
vial).
3. Close the vials tightly with the caps and gently invert the vials several times to mix the
contents. CAUTION: The vials will become hot during mixing.
4. Heat the vials for 120 minutes in the preheated reactor at a temperature of 150 °C.
5. CAUTION: The vials will be hot.
Remove the vials from the reactor and allow them to cool to 60 °C or less. Gently invert the
vials several times to mix the contents while still warm. Allow the vials to cool to room
temperature before measuring. Wipe the exteriors of the vials.
6. Load and run the CODHP0 method.
7. Place the blank vial into the holder in the sample chamber and close the chamber door.
8. Tap the Blank function key to measure the blank.
9. Open the sample chamber door. Remove the blank vial from the holder.
10. Place the sample vial into the holder in the sample chamber and close the chamber door.
11. Tap the Sample function key to display the result in mg/l oxygen.
Notes:
• Run samples and blanks with the same batch of vials. The blank is stable when stored in
the dark and can be used for further measurements with vials of the same batch.
• Do not place the hot vials in the sample chamber. Cool the vials to room temperature for
final measurements.
• Suspended solids in the vial lead to incorrect measurements. For this reason, it is
important to place the vials carefully in the sample chamber. The precipitate at the bottom
of the sample should be not suspended.
• Clean the outside of the vials with a towel. Finger prints or other marks must be removed.
• Samples can be measured when the chloride content does not exceed 1000 mg/l.
• In exceptional cases, compounds contained in the water cannot be oxidized adequately, so
results may be lower than reference methods.
• For samples under 1000 mg/l it is recommended to repeat the test using the COD mid
range test (CODH00) or for samples under 100 mg/l it is recommended to repeat the test
using the COD low range test (CODL00).
Notes:
• Alternatively, the AC202950 method can be used with 50mm rectangular vials. All blank
and sample volumes must remain the same as those specified in these instructions, so
samples may need to be prepared in separate containers and then transferred into the
selected vial.
Notes:
• For determination of total copper, a digestion is required.
• Extremely acid water samples (pH 2 or less) must be adjusted between pH 4 and pH 6
before the reagent is added (with 8 mol/l potassium hydroxide solution, KOH).
• Accuracy is not affected by undissolved powder.
• Interferences:
Cyanide (CN- ) Cyanide prevents full color development. Add 0.2 ml formaldehyde to 10 ml
water sample and wait for a reaction time of 4 minutes (cyanide is masked).
After this perform test as described. Multiply the result by 1.02 to correct the
sample dilution by formaldehyde.
Silver (Ag+) If turbidity remains and turns black, silver interference is likely. Add 10 drops
of saturated potassium chloride solution to 75 ml of water sample. Filtrate
through a fine filter. Use 10 ml of the filtered water sample to perform test.
Notes:
• If cyanuric acid is present, a cloudy solution will occur. Small single particles are not
necessarily caused by cyanuric acid.
• Dissolve the tablet completely (swirl the vial for approximately 1 minute). Undissolved
particles of the tablet can cause results that are too high.
• Exceeding the measurement range: samples with concentration above 90 mg/l must be
diluted with water free of cyanuric acid. 10 ml of the diluted sample should be tested as
described above and the displayed results calculated using the dilution factor.
Notes:
• For the best accuracy, just one vial (the same vial) should be used for the reagent blank
and all sample measurements. Different vials may exhibit minor variations that can reduce
the accuracy of the readings.
• Reverse color methods use a reagent that, when prepared with samples, decreases in color
as the concentration of the species being measured in the samples increases. Reverse
color methods require the use of a reagent blank. The reagent blank is a mixture of the
reagent and deionized water and provides a zero concentration point with the darkest color
(highest absorbance). The color of samples prepared with the reagent will decrease as the
concentration increases for this method.
• The same batch of SPADNS reagent solution must be used for testing (reagent blank and
sample measurement)
• The calibration solution and water samples should have the same temperature (+/- 1 °C).
• As the test result is highly dependent on exact sample and reagent volumes, the sample
and reagent volumes should always be measured using a 10 ml or 2 ml volumetric pipette
(class A).
• The accuracy of the test methods decreases above a level of 1.2 mg/l fluoride. Although
the results are sufficiently accurate for most applications, more exact results can be
achieved using a 1:1 dilution of the sample prior to use and subsequent multiplication of
the result by 2.
• SPADNS reagent solution contains arsenite. Chlorine concentrations up to 5 mg/l do not
interfere.
• Seawater and wastewater samples must be distilled.
Notes:
• Strong alkaline or acidic water samples must be adjusted between pH 4 and pH 10 before
the tablet is added (use 1 mol/l hydrochloric acid or 1 mol/l sodium hydroxide).
Notes:
• If the water sample is cloudy, you must filter it before performing the blank measurement.
• The temperature of the water sample should not exceed 21 °C.
• Using the hydrazine spoon: 1 g is equivalent to one level spoon.
• Qualitative folded filter papers for medium precipitates are recommended.
• To check whether the reagent has aged (if it has been stored for a lengthy period), perform
the test as described above using tap water. If the result is above the detection limit of 0.05
mg/l, you should only use the reagent with reservations, as there may be a major deviation
in results.
Notes:
• Alternatively, the AC207850 method can be used with 50mm rectangular vials. All blank
and sample volumes must remain the same as those specified in these instructions, so
samples may need to be prepared in separate containers and then transferred into the
selected vial.
• This method determines the total dissolved iron as Fe2+ and Fe3+.
• For the determination of total dissolved and undissolved iron (soluble and insoluble iron),
digestion is required. An example is described here:
i. Add 1 ml of concentrated sulfuric acid to 100 ml water sample. Heat and boil for 10
minutes or until all particles are dissolved. After cooling down, the sample is set to
a pH value of 3 to 6 by using ammonia solution. Refill with deionized water to the
previous volume of 100 ml and mix well. 10 ml of this pre-treated solution is used
for analysis (perform as described by the selected test method).
ii. Water that has been treated with organic compounds like corrosion inhibitors must
be oxidized as necessary to break down the iron. Therefore add 1 ml concentrated
sulfuric acid and 1 ml concentrated nitric acid to 100 ml water sample and boil to
approximately half volume. After cooling down, proceed as described above.
Notes:
• The reagent reacts with all soluble iron and most insoluble forms of iron in the water
sample.
• Iron oxide requires prior digestion: use mild, vigorous or Digesdahl digestion. An example
of digestion with acid is described here:
i. Add 1 ml of concentrated sulfuric acid to 100 ml water sample. Heat and boil for 10
minutes or until all particles are dissolved. After cooling down, the sample is set to
a pH value of 3 to 6 by using ammonia solution. Refill with deionized water to the
previous volume of 100 ml and mix well. 10 ml of this pre-treated solution is used
for analysis (perform as described by the selected test method).
ii. Water that has been treated with organic compounds like corrosion inhibitors must
be oxidized as necessary to break down the iron. Therefore add 1 ml concentrated
sulfuric acid and 1 ml concentrated nitric acid to 100 ml water sample and boil to
approximately half volume. After cooling down, proceed as described above.
• Very strong alkaline or acidic samples must be adjusted to a pH value between 3 and 5
before analysis.
• Accuracy is not affected by undissolved powder.
• Water samples containing visible rust should be allowed to react for at least 5 minutes.
Notes:
• For determination of total iron, digestion is required. TPTZ reagent recovers most insoluble
iron oxides without digestion.
• Rinse all glassware with 1:1 hydrochloric acid solution first and then rinse with deionized
water to remove iron deposits that can cause slightly high results.
• Strong alkaline or acidic water samples must be adjusted between pH 3 and pH 8 before
the reagent is added (use 0.5 mol/l sulfuric acid or 1 mol/l sodium hydroxide).
• Interferences: When interferences occur, color development is inhibited or a precipitate is
formed. The values below refer to a standard with an iron concentration of 0.5 mg/l. The
following substances do not interfere when present up to the levels given:
Substance No Interference To Substance No Interference To
Cadmium 4.0 mg/l Manganese 50 mg/l
Chromium (3+) 0.25 mg/l Mercury 0.4 mg/l
Chromium (6+) 1.2 mg/l Molybdenum 4.0 mg/l
Cobalt 0.05 mg/l Nickel 1.0 mg/l
Copper 0.6 mg/l Nitrite Ion 0.8 mg/l
Cyanide 2.8 mg/l
Notes:
• Rinse all glassware with 1:1 nitric acid solution first and then rinse with deionized water.
• Water samples that contain more than 300 mg/l CaCO3 hardness: after adding the ascorbic
acid powder pack, add 10 drops of Rochelle salt solution.
• After addition of the alkaline cyanide reagent solution, a cloudy or turbid solution may form
in some water samples. The turbidity should disappear after the PAN indicator solution is
added.
• Water samples containing more than 5 mg/l iron should be allowed to react for at least 10
minutes.
Notes:
• This test is applicable for the determination of soluble manganese in water and
wastewater.
• Highly buffered water samples or extreme pH values may exceed the buffering capacity of
the reagents and requires sample pre-treatment. If samples were acidified for storing,
adjust the pH between 4 and 5 with 5 mol/l (5 N) sodium hydroxide before test. Do not
exceed pH 5, as manganese may precipitate.
• Interferences:
Interfering Substance Interference Level
Calcium Greater than 700 mg/l
Chloride Greater than 70,000 mg/l
Iron Greater than 5 mg/l
Magnesium Greater than 100,000 mg/l
Notes:
• Filter turbid water samples using filter paper and funnel before analysis.
• Highly buffered water samples or extreme pH values should be adjusted to a pH of nearly 7
with 1 mol/l nitric acid or 1 mol/l sodium hydroxide.
• Concentrations above 10 mg/l Cu causes too high test values if the reaction time of 5
minutes is increased – so it is very important to perform the test procedure as described.
• Substances which may interfere when present in concentrations at:
Aluminum 50 mg/l
Chromium 1000 mg/l
Iron 50 mg/l
Nickel 50 mg/l
Nitrite All Levels
Notes:
• Some solids may not dissolve.
• Conversion: mg/l NO3 = mg/l N x 4.43
Notes:
• The following ions can produce interferences since under the reaction conditions they
cause precipitation: antimony (III), iron (III), lead, mercury (I), silver, chloroplatinate,
metavanadate and bismuth. Copper (II) ions may cause lower test results as they
accelerate the decomposition of the diazonium salt. It is unlikely in practice that these
interfering ions will occur in such high concentrations that they cause significant reading
errors.
• Conversion: mg/l NO2 = mg/l N x 3.29
Notes:
• Interferences:
o Strong oxidizing and reducing substances interfere.
o Cupric and ferrous ions cause low results.
o Antimonous, auric, bismuth, chloroplatinate, ferric, lead, mercurous, metavanadate
and silver ions interfere by causing precipitation.
o In samples with very high concentrations of nitrate (> 100 mg/l N) a small amount of
nitrite will be found. Such high levels of nitrate appear to undergo a slight amount of
reduction to nitrite, either spontaneously or during the reaction time of the test.
1. Open two TN Hydroxide LR Digestion Vials and add the contents of one TN Persulfate
Reagent Power Pack straight from the foil into each vial. Use a funnel to add the reagent.
Wipe off any persulfate reagent that may get on the lid or tube threads.
2. Add 2 ml of deionized water to the first digestion vial (this is the blank vial).
3. Add 2 ml of sample to the second digestion vial (this is the sample vial).
4. Close the vials tightly with the caps and shake the vials for at least 30 seconds to mix the
contents. The reagent many not dissolve completely.
5. Heat the digestion vials for 30 minutes in the preheated reactor at a temperature of 100 °C.
6. CAUTION: The vials will be hot. Remove the digestion vials from the reactor and allow
them to cool to room temperature.
7. Open the cooled digestion vials and add the contents of one TN Reagent A Power Pack
straight from the foil into each vial. Use a funnel to add the reagent.
8. Close the vials tightly with the caps and shake the vials for at least 15 seconds to mix
contents.
9. Wait for a reaction period of 3 minutes.
10. Open the digestion vials and add the contents of one TN Reagent B Power Pack straight
from the foil into each vial. Use a funnel to add the reagent.
11. Close the vials tightly with the caps and shake the vials for at least 15 seconds to mix
contents. The reagent will not completely dissolve.
12. Wait for a reaction period of 2 minutes.
13. Open two TN Acid LR/HR (Reagent C) Vials and add 2 ml of the digested, treated blank to
the first vial (this is the blank vial).
14. Add 2 ml of the digested, treated sample to the second vial (this is the sample vial).
15. Close the vials tightly with the caps and gently invert the vials at least 10 times to mix
contents. Hold the vial in a vertical position with the cap pointing up. Turn the vial upside-
down. Wait for all of the solution to flow down to the cap. Return the vial to the upright
position. Wait for all the solution to flow to the bottom of the vial. This process is one
inversion; 10 inversions equal about 30 seconds. Wipe the exteriors of the vials.
CAUTION: The vials will become warm during mixing.
16. Wait for a reaction period of 5 minutes.
17. Load and run the ACD004 method.
18. Place the blank vial into the holder in the sample chamber and close the chamber door.
Notes:
• Appropriate safety precautions and a good lab technique should be used during the whole
procedure.
• Volumes for samples and blank should always be measured using 2 ml volumetric pipettes
(class A).
• One blank is sufficient for each set of samples. After taking the blank measurement, it is
possible to measure several samples.
• It is very important to remove the vials from the reactor after exactly 30 minutes.
• Large quantities of nitrogen free, organic compounds that are included in some water
samples may reduce the effectiveness of the digestion by reacting with the persulfate
reagent. Samples that are well known to contain large quantities of organic compounds
must be diluted and digestion and measurement must be repeated for checking the
effectiveness of the digestion.
• Application: for water, wastewater and seawater.
• Interfering substances that resulted in a concentration change of 10%: Bromide more than
60 mg/l and chloride more than 1000 mg/l produce positive interferences.
1. Open two TN Hydroxide HR Digestion Vials and add the contents of one TN Persulfate
Reagent Power Pack straight from the foil into each vial. Use a funnel to add the reagent.
Wipe off any persulfate reagent that may get on the lid or tube threads.
2. Add 0.5 ml of deionized water to the first digestion vial (this is the blank vial).
3. Add 0.5 ml of sample to the second digestion vial (this is the sample vial).
4. Close the vials tightly with the caps and shake the vials for at least 30 seconds to mix
contents. The reagent many not dissolve completely.
5. Heat the digestion vials for 30 minutes in the preheated reactor at a temperature of 100 °C.
6. CAUTION: The vials will be hot. Remove the digestion vials from the reactor and allow
them to cool to room temperature.
7. Open the cooled digestion vials and add the contents of one TN Reagent A Power Pack
straight from the foil into each vial. Use a funnel to add the reagent.
8. Close the vials tightly with the caps and shake the vials for at least 15 seconds to mix
contents.
9. Wait for a reaction period of 3 minutes.
10. Open the digestion vials and add the contents of one TN Reagent B Power Pack straight
from the foil into each vial. Use a funnel to add the reagent.
11. Close the vials tightly with the caps and shake the vials for at least 15 seconds to mix
contents. The reagent will not completely dissolve.
12. Wait for a reaction period of 2 minutes.
13. Open two TN Acid LR/HR (Reagent C) Vials and add 2 ml of the digested, treated blank to
the first vial (this is the blank vial).
14. Add 2 ml of the digested, treated sample to the second vial (this is the sample vial).
15. Close the vials tightly with the caps and gently invert the vials at least 10 times to mix
contents. Hold the vial in a vertical position with the cap pointing up. Turn the vial upside-
down. Wait for all of the solution to flow down to the cap. Return the vial to the upright
position. Wait for all the solution to flow to the bottom of the vial. This process is one
inversion; 10 inversions equal about 30 seconds. Wipe the exteriors of the vials.
CAUTION: The vials will become warm during mixing.
16. Wait for a reaction period of 5 minutes.
17. Load and run the ACD007 method.
18. Place the blank vial into the holder in the sample chamber and close the chamber door.
Notes:
• Appropriate safety precautions and a good lab technique should be used during the whole
procedure.
• Volumes for samples and blank should always be measured using 2 ml volumetric pipettes
(class A).
• One blank is sufficient for each set of samples. After taking the blank measurement, it is
possible to measure several samples.
• It is very important to remove the vials from the reactor after exactly 30 minutes.
• Large quantities of nitrogen free, organic compounds that are included in some water
samples may reduce the effectiveness of the digestion by reacting with the persulfate
reagent. Samples that are well known to contain large quantities of organic compounds
must be diluted and digestion and measurement must be repeated for checking the
effectiveness of the digestion.
• Application: for water, wastewater and seawater.
• Interfering substances that resulted in a concentration change of 10%: Bromide more than
60 mg/l and chloride more than 1000 mg/l produce positive interferences.
9. Transfer the contents of the vial with glycine solution into the vial containing the DPD No. 1
and DPD No. 3 tablets.
10. Close the vial tightly with the cap and swirl or invert several times until the tablets are
dissolved. Wipe the exterior of the vial.
11. Place the vial into the holder in the sample chamber. Close the sample chamber door.
12. Wait for a reaction period of 2 minutes.
13. Tap the Sample function key to display the result in mg/L ozone.
Notes:
• Vial cleaning: As many household cleaners (i.e. dishwasher detergent) contain reducing
substances, the subsequent determination of ozone may show lower results. To avoid any
measurement errors, only use glassware free of chlorine demand.
Preparation: Put all applicable glassware into a sodium hypochlorite solution (0.1 g/l) for
one hour and then rinse all glassware thoroughly with deionized water.
• Preparing the sample: When preparing the sample, the loss of ozone, i.e. by pipetting or
shaking, must be avoided. The analysis must take place immediately after taking the
sample.
• The DPD color development is carried out at a pH value of 6.2 to 6.5. The reagent tablet
therefore contains a buffer for the pH adjustment. Strong alkaline or acidic water samples
must be adjusted between pH 6 and pH 7 before the tablet is added (use 0.5 mol/l sulfuric
acid or 1 mol/l sodium hydroxide).
• Exceeding the measuring range: Concentrations above 6 mg/l ozone can lead to results
showing 0 mg/l. In this case, the water sample must be diluted with water free of ozone. 10
ml of the diluted sample should be mixed with the reagent and the measurement repeated.
• Oxidizing agents such as bromine and chlorine interfere as they react in the same way as
ozone.
Notes:
• Water samples with low values of alkalinity-m (below 35 mg/l CaCO3) may give wrong pH
readings.
• pH values below 6.5 and above 8.4 can produce results inside the measuring range. A
plausibility test (pH meter) is recommended.
• The accuracy of the colorimetric determination of pH values depends on various boundary
conditions (buffer capacity of the sample, salt contents, etc.).
• Salt error: Correction of test results (average values) for samples with salt contents of:
Indicator Salt Content
1 molar 2 molar 3 molar
Phenol Red
– 0.21 – 0.26 – 0.29
• The values of Parson and Douglas (1926) are based on the use of Clark and Lubs buffers.
1 Mol NaCl = 58.4 g/l = 5.8 %
Notes:
• When testing chlorinated water, the residual chlorine contents can influence the color
reaction of the liquid reagent. This can be avoided (without interfering with the pH
measurement) by adding a small crystal of sodium thiosulfate (Na2S2O3 • 5 H2O) to the
sample before adding the phenol red solution. Phenol red tablets already contain
thiosulfate.
• Due to differing drop sizes results can show a discrepancy in accuracy by comparison with
tablets. This can be minimized by using a pipette (0.18 ml phenol red solution is equivalent
to 6 drops).
• After use, replace the bottle cap securely.
• Store the phenol red solution in a cool, dry place ideally at between 6 °C and 10 °C.
Notes:
• Only ortho-phosphate ions react.
• The tablets must be added in the correct sequence.
• The test sample should have a pH value between 6 and 7.
• Interferences: Higher concentrations of Cu, Ni, Cr (III), V (V) and W (VI) interfere due to
their color. Silicates do not interfere (masked by citric acid in the tablets).
• Ortho-phosphate ions react with the reagent to form an intense blue color.
• Phosphate in organic and condensed inorganic forms (meta-, pyro- and polyphosphates)
must be converted to ortho-phosphate ions before analysis. Pretreatment of the sample
with acid and heat provides the conditions for hydrolysis of the condensed inorganic forms.
Organically combined phosphates are converted to ortho-phosphate ions by heating with
acid and persulfate. The amount of organically combined phosphates can be calculated:
mg/l phosphate (organic) = mg/l phosphate (total) - mg/l phosphate (acid hydrolysable)
• Phosphate, ortho = Phosphorus, reactive
Notes:
• For samples under 5 mg/l PO4, it is recommended to analyze the sample using the AC2095
phosphate (ortho) low range tablet test method.
• Only ortho-phosphate ions react.
• Phosphate in organic and condensed inorganic forms (meta-, pyro- and polyphosphates)
must be converted to ortho-phosphate ions before analysis. Pretreatment of the sample
with acid and heat provides the conditions for hydrolysis of the condensed inorganic forms.
Organically combined phosphates are converted to ortho-phosphate ions by heating with
acid and persulfate. The amount of organically combined phosphates can be calculated:
mg/l phosphate (organic) = mg/l phosphate (total) - mg/l phosphate (acid hydrolysable)
• The ortho-phosphate ions react with the vanadate-molybdate reagent under acid conditions
to form a yellow colored product.
• Phosphate, ortho = Phosphorus, reactive
Notes:
• Ortho-phosphate ions react with the reagent to form an intense blue color.
• Phosphate in organic and condensed inorganic forms (meta-, pyro- and polyphosphates)
must be converted to ortho-phosphate ions before analysis. Pretreatment of the sample
with acid and heat provides the conditions for hydrolysis of the condensed inorganic forms.
Organically combined phosphates are converted to ortho-phosphate ions by heating with
acid and persulfate. The amount of organically combined phosphates can be calculated:
mg/l phosphate (organic) = mg/l phosphate (total) - mg/l phosphate (acid hydrolysable)
• Application: for water, wastewater and seawater.
• Highly buffered samples or samples with extreme pH values should be adjusted between
pH 2 and pH 10 before analysis with 1 mol/l hydrochloric acid or 1 mol/l sodium hydroxide.
• Phosphate, ortho = Phosphorus, reactive
• Interferences: Large amounts of turbidity may cause inconsistent results.
Interference Interference Level Interference Interference Level
Aluminum greater than 200 mg/l Nickel greater than 300 mg/l
Arsenate at any level Silica (Silicium dioxide) greater than 50 mg/l
Chromium greater than 100 mg/l Silicate greater than 10 mg/l
Copper greater than 10 mg/l Sulfide at any level
Iron greater than 100 mg/l Zinc greater than 80 mg/l
Notes:
• Ortho-phosphate ions react with the reagent to form an intense blue color.
• Phosphate in organic and condensed inorganic forms (meta-, pyro- and polyphosphates)
must be converted to ortho-phosphate ions before analysis. Pretreatment of the sample
with acid and heat provides the conditions for hydrolysis of the condensed inorganic forms.
Organically combined phosphates are converted to ortho-phosphate ions by heating with
acid and persulfate. The amount of organically combined phosphates can be calculated:
mg/l phosphate (organic) = mg/l phosphate (total) - mg/l phosphate (acid hydrolysable)
• Application: for water, wastewater and seawater.
• Highly buffered samples or samples with extreme pH values should be adjusted between
pH 2 and pH 10 before analysis with 1 mol/l hydrochloric acid or 1 mol/l sodium hydroxide.
• Phosphate, ortho = Phosphorus, reactive
• Interferences: Large amounts of turbidity may cause inconsistent results.
Interference Interference Level Interference Interference Level
Aluminum greater than 200 mg/l Nickel greater than 300 mg/l
Arsenate at any level Silica (Silicium dioxide) greater than 50 mg/l
Chromium greater than 100 mg/l Silicate greater than 10 mg/l
Copper greater than 10 mg/l Sulfide at any level
Iron greater than 100 mg/l Zinc greater than 80 mg/l
1. Open one 16 mm PO4-P Acid Reagent Digestion Tube and add 5 ml of sample.
2. Add the contents of one Potassium Persulfate F10 Power Pack straight from the foil into
the vial. Use a funnel to add the reagent.
3. Close the vial tightly with the cap and invert the vial several times to mix the contents.
4. Heat the vial for 30 minutes in the preheated reactor at a temperature of 100 °C.
5. CAUTION: The vial will be hot. Remove the vial from the reactor and allow it to cool to
room temperature.
6. Open the cooled digestion vial and add 2 ml of 1.54 N Sodium Hydroxide Solution to the
vial.
7. Close the vial tightly with the cap and gently invert the vial several times to mix the
contents. Wipe the exterior of the vial.
8. Load and run the ACD095 method.
9. Place the vial into the holder in the sample chamber and close the chamber door.
10. Tap the Blank function key to measure the blank.
11. Open the sample chamber door. Remove the vial from the holder.
12. Add the contents of one Phosphate Rgt. F10 Power Pack straight from the foil into the vial.
Use a funnel to add the reagent.
13. Close the vial tightly with the cap and swirl the vial for 10-15 seconds to mix the contents.
The reagent will not completely dissolve. Wipe the exterior of the vial.
14. Wait for a reaction period of 2 minutes.
15. Place the vial into the holder in the sample chamber and close the chamber door.
16. Tap the Sample function key to display the result in mg/l total phosphate as phosphorus
(P).
Notes:
• Appropriate safety precautions and good lab technique should be used during the whole
procedure.
• Ortho-phosphate ions react with the reagent to form an intense blue color.
• Phosphate in organic and condensed inorganic forms (meta-, pyro- and polyphosphates)
must be converted to ortho-phosphate ions before analysis. Pretreatment of the sample
with acid and heat provides the conditions for hydrolysis of the condensed inorganic forms.
Organically combined phosphates are converted to ortho-phosphate ions by heating with
acid and persulfate. The amount of organically combined phosphates can be calculated:
mg/l phosphate (organic) = mg/l phosphate (total) - mg/l phosphate (acid hydrolysable)
• Application: for water, wastewater and seawater.
• Highly buffered samples or samples with extreme pH values should be adjusted between
pH 2 and pH 10 before analysis with 1 mol/l hydrochloric acid or 1 mol/l sodium hydroxide.
• Phosphate, ortho = Phosphorus, reactive
• Interferences: Large amounts of turbidity may cause inconsistent results.
Interfering Substance Interference Level
Aluminum greater than 200 mg/l
Arsenate at any level
Chromium greater than 100 mg/l
Copper greater than 10 mg/l
Iron greater than 100 mg/l
Nickel greater than 300 mg/l
Silica (Silicium dioxide) greater than 50 mg/l
Silicate greater than 10 mg/l
Sulfide at any level
Zinc greater than 80 mg/l
1. Open one 16 mm PO4-P Acid Reagent Digestion Tube and add 5 ml of sample.
2. Close the vial tightly with the cap and gently invert the vial several times to mix the
contents.
3. Heat the vial for 30 minutes in the preheated reactor at a temperature of 100 °C.
4. CAUTION: The vial will be hot. Remove the vial from the reactor and allow it to cool to
room temperature.
5. Open the cooled digestion vial and add 2 ml of 1.00 N Sodium Hydroxide Solution to the
vial.
6. Close the vial tightly with the cap and gently invert the vial several times to mix the
contents. Wipe the exterior of the vial.
7. Load and run the ACD095AH method.
8. Place the vial into the holder in the sample chamber and close the chamber door.
9. Tap the Blank function key to measure the blank.
10. Open the sample chamber door. Remove the vial from the holder.
11. Add the contents of one Phosphate Rgt. F10 Power Pack straight from the foil into the vial.
Use a funnel to add the reagent.
12. Close the vial tightly with the cap and swirl the vial for 10-15 seconds to mix contents. The
reagent will not completely dissolve. Wipe the exterior of the vial.
13. Wait for a reaction period of 2 minutes.
14. Place the vial into the holder in the sample chamber and close the chamber door.
15. Tap the Sample function key to display the result in mg/l acid hydrolysable phosphate as
phosphorus (P).
Notes:
• Appropriate safety precautions and good lab technique should be used during the whole
procedure.
• Ortho-phosphate ions react with the reagent to form an intense blue color.
• Phosphate in organic and condensed inorganic forms (meta-, pyro- and polyphosphates)
must be converted to ortho-phosphate ions before analysis. Pretreatment of the sample
with acid and heat provides the conditions for hydrolysis of the condensed inorganic forms.
Organically combined phosphates are converted to ortho-phosphate ions by heating with
acid and persulfate. The amount of organically combined phosphates can be calculated:
mg/l phosphate (organic) = mg/l phosphate (total) - mg/l phosphate (acid hydrolysable)
• Application: for water, wastewater and seawater.
• Highly buffered samples or samples with extreme pH values should be adjusted between
pH 2 and pH 10 before analysis with 1 mol/l hydrochloric acid or 1 mol/l sodium hydroxide.
• Phosphate, ortho = Phosphorus, reactive
• Interferences: Large amounts of turbidity may cause inconsistent results.
Interfering Substance Interference Level
Aluminum greater than 200 mg/l
Arsenate at any level
Chromium greater than 100 mg/l
Copper greater than 10 mg/l
Iron greater than 100 mg/l
Nickel greater than 300 mg/l
Silica (Silicium dioxide) greater than 50 mg/l
Silicate greater than 10 mg/l
Sulfide at any level
Zinc greater than 80 mg/l
Notes:
• The tablets must be added in the correct sequence.
Notes:
• The tablets must be added in the correct sequence.
• Phosphate ions do not interfere under the given reaction conditions.
• If phosphate is known to be absent, the addition of the Silica PR Tablet may be omitted.
Notes:
• Occasionally water samples contain forms of silica which reacts very slowly with
molybdate. The nature of these forms is not known.
• A pre-treatment with sodium hydrogen carbonate and then with sulfuric acid will make
these forms reactive to molybdate (pre-treatment is given in “Standard Methods for the
Examination of Water and Wastewater” under “Silica Digestion with Sodium Bicarbonate”).
Substance Interference
Iron Large amounts interfere
Phosphate Does not interfere at concentrations less than 50 mg/l PO4
At 60 mg/l PO4 the interference is approximately 2%
At 75 mg/l PO4 the interference is approximately 11 %
Sulfide Interferes at all levels
Notes:
• If sulfate ions are present a cloudy solution will appear.
Notes:
• The tablets must be added in the correct sequence.
• Chlorine and other oxidizing agents that react with DPD do not interfere with the test.
• To avoid loss of sulfide, collect the sample carefully with a minimum of aeration. It is
essential to test the sample immediately after collection.
• The sample temperature should be 20 °C. A different temperature can lead to higher or
lower results.
Notes:
• The tablets must be added in the correct sequence.
• In the case of high levels of residual chlorine, perform the analysis with a dechlorinated
water sample.
Health Organization recommends that drinking water color does not to exceed 15 true color
units.
• Spectrophotometers allow for more accurate and precise results than colorimeters, but are
more expensive. Lowest detection limits and better accuracy at low levels can be achieved
on a spectrophotometer by choosing a sample cell with a longer cell path length.
• Colorimeters are useful for testing outside the laboratory and indicate relative color
intensity. Color values determined on a colorimeter may differ from color values determined
on a spectrophotometer, especially when the colorimeter wavelength does not match the
wavelength used on the spectrophotometer.
• Sample cells for color testing are made of glass. Glass is suitable for testing in the visible
wavelength range. A round glass 24 mm or 25 mm cell gives satisfactory results. For best
accuracy and precision at color levels that are lower than 15 color units, a rectangular
glass 50 mm cell is recommended. The 10 mm sample cell size is not recommended for
color testing.
Preprogrammed
Sample
Color Method name or user Wavelength(s) Scope of the method
Cell Size
defined
Platinum–Cobalt Color CLRPT50 50 mm 455 nm Water and wastewater
Method: absorbance CLRPT25 25 mm samples of color similar
measured at one CLRPT24 24 mm to Pt-Co color
wavelength. User Method 10-50 mm standards
Range: 0 -500 PCU
Adaptation of NCASI CLRPTP50 50 mm 465 nm Color in pulp mill
253/803: absorbance CLRPTP25 25 mm wastewaters
measured at one CLRPTP24 24 mm
wavelength. User Method 10-50 mm
Range: 0 -500 PCU
ISO 7887; GB 11903: User Method 50 nm 436 nm Raw and potable water,
Water Quality - 525 nm industrial water of low
Examination and 620 nm color
Determination of Color
Tristimulus Values User Method 10 mm or Multiple Water and wastewater
Method: transmittance is 50 mm wavelengths samples of color
measured at multiple 400 to 700nm different from, but not
wavelength in the 400 – range excluding, Pt-Co
700 nm range standards
Application Notes
Application notes for color testing are available through your local technical sales
representative, our technical support service, our website and other Thermo Scientific web
locations. These notes give detailed information on how to test for color on our Thermo
Scientific Orion AquaMate spectrophotometers: AQ7100 visible range and AQ8100 UV/Vis
range. Application Notes available include:
• Log 133, Color of water and wastewater by Pt-Co method at 455 nm
• Log 134, Color of pulp mill wastewater by Pt-Co method at 465 nm (NCASI method)
References
1. Standard Methods for the Examination of Water and Wastewater, Method 2120B.
www.standardmethods.org
2. Standard Methods for the Examination of Water and Wastewater, Method 2120.
www.standardmethods.org
3. Available through Fisher Scientific at www.fishersci.com
4. NCASI, Technical Bulletin No. 253, Dec. 1971. See 40CFR Part 136, Table IB, footnote 18.
www.ecfr.gov
5. EPA Color Method 110.1. http://www.umass.edu/tei/mwwp/acrobat/epa110.1colorspec.pdf
References
1. EPA Method 415.3 Rev 1.1. UV254 for SUVA. http://www.epa.gov/microbes/ordmeth.htm
2. Standard Methods 5910B, UV-Absorbing Organic Constituents. www.standardmethods.org
3. Orion AquaMate 8100 UV-Vis Instrument User Guide
Recommended Equipment
• Thermo Scientific Orion AquaMate AQ8100 spectrophotometer
• Filtration apparatus (e.g., Fisher XX1004707); 0.45-µm filters, 47 mm (e.g., Fisher
HNWP04700)
• Vacuum source-aspirator, air flow or water flow, hand-operated or low pressure electric
vacuum pump
• UV disposable cuvettes 1 cm (Fisher 14-377-009) or quartz sample cells
• Carousel for 10 cm cell, if needed (Orion AQ100C)
• Orion pH meter and electrode
Solutions
1. Organic-free deionized water (DI).
2. Reagents for pH adjustment. Sodium hydroxide, 0.1N; Hydrochloric acid, 0.1N; Orion pH
4.01 and 7.00 Buffers (Orion 910104, Orion 910107).
3. Spectrophotometer Check Solution (SCS), optional: Organic Carbon, KHP in pH 7
phosphate buffer, see Reference 2 for preparation; or, Commercial SCS, PN 222-234700
(www.unitylabservices.com).
Instrument Setup
Turn on the spectrophotometer. Choose a suitable cell size and method for the organic carbon
content expected in your samples. Refer to the following chart. Select the appropriate cell
holder. Select the method and load it. Press the Run Test function to start the analysis. Refer
to the following chart for choosing the method.
Sample Storage
Samples are not preserved. Analyze as soon as possible after collection. Samples may be
stored for up to 48 hours at <6°C prior to analysis. See EPA Method 415.3 for SUVA storage.
For UVA, UV254: Set up the filtration apparatus with a 0.45 um filter. Wash the filter with 50 mL
DI and discard the rinse water. Filter 50 mL of the sample. Test the filtrate.
Sample Measurement
Ensure the instrument has been zeroed properly. Touching only the frosted sides of the cell,
rinse the clean cell with a portion of the filtered sample, then fill with the filtered sample. Wipe
dry. Insert the sample cell into the holder, close the lid, and Tap the Sample key. Record the
displayed result. The results can be saved to the USB stick, if desired. If the reading is >0.900
absorbance, dilute and retest. Multiply the reading by the dilution factor. If the results are
<0.010 absorbance, consider using a larger cell. Load the appropriate method, and re-zero the
instrument.
pH Adjustment of Sample
1. Note: Do not adjust the pH of a sample which will be used for a SUVA calculation. Proceed
to the filtration step.
2. Calibrate the pH probe in pH 4.01 and 7.00 buffers.
3. Warm the sample up to room temperature.
4. Shake the sample to insure homogeneity.
5. Measure 50 mL of the sample to a 100-mL beaker using a graduated cylinder.
6. Immerse the pH probe in the sample and record the initial pH.
7. Adjust the sample into the range of pH 4 to 10, by adding drop-wise 0.1N sodium hydroxide
to raise the pH or by adding drop-wise 0.1N hydrochloric acid to lower the pH. Different
strength acid or base can be used, if needed.
8. Note that the overall volume change should not be greater than 1% (0.5 mL). Discard and
re-prepare with stronger acid or base if the volume changes more than 1%.
9. Record the adjusted pH. Proceed to the filtration step (p. 1).
Bias: readings of a KHP organic carbon standard at 25.0 mg/L in phosphate buffer (prepared
per SM 5910B) tested in a 1 cm cell demonstrate good accuracy:
• The average AQ8100 absorbance result is within 0.002 absorbance units of the average
reading expected per SM 5910B; 0.6% difference from the expected absorbance.
• The average AQ8100 organic carbon concentration result (calculated per SM 5910B) is
within 0.1 mg/L of the expected value; 0.4% difference (99.6% recovery) from the expected
value of 25.0 mg/L organic carbon.
Precision # of Samples Maximum %RSD
Result (AQ8100) Evaluation
Method UV254_1 Tested (per SM 5910B)
Absorbance 14 < 10.7% RSD 0.3% RSD Good
Precision: readings of a KHP organic carbon standard at 25.0 mg/L in phosphate buffer
(prepared per SM 5910B) tested in a 1 cm cell demonstrate good precision:
• The relative standard deviation (RSD) of 14 test results on the AQ8100 is 0.3% RSD, well
within the maximum 10.7% limit expected per SM 5910B.
Notes
1. If the preprogrammed method is not on the memory stick, download the method file from
the Online Library at www.thermoscientific.com/waterlibrary, or call Technical Support.
2. Alternately, use disposable cuvettes formulated for UV measurements, available in the 1
cm (10 mm) cell path.
CHAPTER 6
6
Standard Curve Test Menu
Concentration Measurements using the Quant
Standard Curve Application (Custom Method)
The Standard Curve test technique is used to perform a quantitative analysis experiment using
a multipoint calibration curve via the Quant application. A calibration curve is composed of
standards of well-known concentration. A fit of this standard curve is used to measure the
concentration of samples.
This test technique is ideal when using colorimetric reagents for which the test kit manufacturer
does not specify a factor or equation for obtaining the concentration of the test kit. Always follow
the reagent instructions provided by the test kit manufacturer when creating a custom method.
Note: If the manufacturer does not specify the wavelength for the test kit, the Scan application
should be used with a prepared standard as a technique to determine the wavelength
parameter for the reagent before creating a standard curve.
Accessing Quant
From Screen 1, swipe the to the left to have Screen 2 appear. The Quant application is
selected by tapping the respective icon. Quant is a calibration curve development application.
The user can select the expiration date, wavelength, reference wavelength, equation, units of
concentration, and enter the known standards that will be used to develop the curve. Methods
can be saved by name and used to measure subsequent sample concentration
End
experiment
Ending An Experiment
When your measurements are completed, select the “X” in the upper left hand corner and it will
terminate. You will need to confirm that the experiments will end and the file will be saved.
The data may be also be printed as a report (if network printer or tape printer are installed) or
the date may be exported to a network folder of to a USB drive.
Go Back to experiment
Send to printer
Export to file
Confirm experiment is
ended
Exporting Data
If you choose to export your data, be sure to have setup a network path via Ethernet or via
WiFi. A typical method is to save the data directly to a USB drive. The report can be saved as
both a CSV file and/or a JPEG file. Below is an example of the JPEG file image.
From the Quant home page, select the listed method curve that you would like to review and
edit. In the example below, the Chlorine Curve_C is selected. Once selected, tap the ellipsis to
bring up the curve options to Edit, select Smart Method, Duplicate, Export, or Delete.
Once the curve options window appears, select the Edit option. In the edit mode, you can
rename the method, change the expiration date of the method, or choose a different equation to
represent the calibration curve. Tap on the following more common parameters highlighted in
blue to revise and update:
• Method name
• Calibration expiration
• Calibration equation
If any changes are made, please be sure to save the changes with the new name. Standard
points can be deleted or added, however the curve will have to be re-ran.
Once the method has been revised, sample measurements can continue.
CHAPTER 7
7
Wavelength Scanning Test Menu
The Wavelength Scanning test technique lets you measure the absorption or percent
transmission spectrum of a sample. You can use scans to determine peak wavelengths or to
evaluate the quality of a material.
This test technique is useful when using colorimetric reagents for which the test kit
manufacturer does not specify a wavelength parameter for the test kit. Always follow the
reagent instructions provided by the test kit manufacturer when creating a custom method.
Note: If the manufacturer does not specify the factor or equation for obtaining the concentration
of the test kit, the Standard Curve test technique should be used with prepared standards to
determine the factor or equation for the reagent.
Once the parameters have been set for the scan you can continue.
1. Press Continue to run a scan test
2. Install a blank sample and Blank the system.
3. Install the sample of interest and tap Measure. This can be repeated for multiple samples.
4. Once the Scan is completed, up to three (3) reference wavelengths can be selected to
display results specifically at those wavelengths. In the example below, 622, 663, and 700
nm have been selected.
5. Either tap the wavelength number to enter a new specific reference wavelength or drag the
vertical wavelength line to a new reference value.
6. If additional features for a sample scan are of interest, tap the ellipsis to the right of the
sample to open other options
Once the scan is completed, follow the save and export steps. The graphical data can be
manipulated through a two-finger stretch or compress touchscreen method.
Choosing
Area Function
To have the area under the curve calculated, only two wavelengths are to be referenced. By
toggling the 3Pt Net off, automatically the wavelengths 1 and 2 are selected and the area under
the curve can be calculated by toggling that feature on.
CHAPTER 8
8
AquaMate On-board Software
Performance Verification Tests and Reports
AquaMate instruments are preconfigured with default
Performance Verification Tests. These tests cannot be deleted
or edited.
Can
Performance Verification Test Description
Duplicate?
Wavelength Accuracy Xenon This test verifies the wavelength axis performance of the No
spectrophotometer.
Can
Performance Verification Test Description
Duplicate?
Baseline Flatness 1000 to 200 nm Measures any systematic deviation from perfect zero when No
scanning across the common wavelength range. Data is
smoothed to remove the impact of noise (noise can be
measured separately). Result is the maximum deviation from
zero and is compared to the specification for the instrument.
Stray Light SRE 220 filter (UV-Vis models) Measures stray light at the specified wavelength. Yes
Stray Light SRE 400 Filter (Vis models) The filter is a long pass filter that cuts on slightly above the
test wavelength. At the test wavelength it should be entirely
dark – i.e. 0%T. Longer wavelengths pass through the filter,
therefore any transmittance measured at 220 nm is actually
photons of longer wavelength that are “stray light”. Sources of
stray light include second order effects imperfections in the
grating, and imperfections or dirt on mirrors.
Wavelength Accuracy This test verifies the wavelength axis performance of the Yes
spectrophotometer.
Can
Performance Verification Test Description
Duplicate?
Photometric Accuracy This test verifies the photometric (absorbance) performance of the Yes
spectrophotometer.
User enters
• Calibration wavelengths
• Certificate absorbance values
• Calibration uncertainty value
from the calibration certificate. User also enters the
performance specification for the absorbance value being
tested from the specification sheet for the instrument .b
a We recommend against use of didymium glass “dual standard” filters calibrated for both wavelength peaks and
photometric accuracy. Customers have reported difficulty in reproducing calibration values for this type of standard
although their instruments do reproduce the calibration values for other, more widely accepted and recognized
standards. Customers who call for support because an instrument fails a photometric accuracy test with a didymium
photometric standard may be required to verify photometric accuracy with a different standard before return shipment
for warranty service is authorized.
b Future releases of software may include a look-up table that populates the instrument specification based on the
user-entered certificate absorbance value.
Toggle to measure in
transmittance mode
Standard ID may be
used to identify the
corresponding filter
in a filter set
Expiration interval can be set in number of days. When set, the instrument
will display a notification indicating the results of the corresponding verification
test have expired. The time interval is relative to the currently configured
instrument date. The instrument checks for the expiration date once each day
– either at 12:00 AM or at the time the instrument is first powered up on a
certain day. Changing the system time after a verification schedule is
configured, may cause the instrument to display the expiration notification. It
is therefore suggested that the all expiration dates be verified when the system
time is changed.
CHAPTER 9
9
Absorbance, % Transmittance and
Concentration Measurements
Absorbance & % Transmittance Measurements
Using the Fixed Application for Basic A-%T-C Test Method
The Fixed Application from Screen 2 puts the instrument into an “instant measurement” mode.
The user simply walks up to the instrument, inserts a sample and measures it. Depending on
whether the mode is set to Absorbance (A), % Transmittance (%T), or Concentration, the
measurement results appear, along with the type of measurement, date and time, wavelength
and vial position.
Select a new Fixed scan window and through touchscreen tapping and window editing:
2. Select ABS or %T
Depending upon the complexity of the equation, either one or two wavelengths and Factors will
appear.
Once the method is set, follow the blanking and measuring steps. The instrument will
automatically blank and measure at both respective wavelengths and calculate the results.
Note: The variety of equations (additive, difference, and ratio) measures the absorption
at two different wavelengths. Reference wavelength correction is available to eliminate
the effects of a sample matrix. Typically used in quality control applications, these
methods provide a convenient and quick diagnostic test for sample quality.
By tapping the C-Mode, the screen below will appear. By tapping on the editable fields, the
user can change the wavelength, standard concentration value, and units.
After running the standard, a Standard Absorbance of 2.361 is measured and a factor of 0.424
is calculated. Any subsequent samples can be entered periodically to get a direct read. In the
example below, 1.545 mg/L is measured.
Multi-wavelength
The Multi-Wavelength Application is selected from Screen 3. This places the instrument into a
mode that requires a blank and subsequent samples for measurement. The Multiwavelength
application obtains multiple fixed-wavelength measurements. It is a fast alternative to scanning
if the wavelengths of interest are well known. By tapping the icon, the screen(s) below will
appear. Tap the + symbol to add a measurement.
Up to twenty (20) discrete wavelengths can be entered into the wavelength table for a
measurement in either ABS or %T mode. In the example below, only five (5) have been
selected for ABS measurement. Tap the Save icon to save the scanning method. Tap
continue to blank and measure.
3-Point Net
The 3-Point Net application determines the height of a peak based on a sloping baseline drawn
between two wavelengths on either side of the peak. This type of analysis is beneficial when
the precise peak height is needed for an assay. A factor can be multiplied by the measured
peak height to give the concentration of the measured analyte in the appropriate concentration
units. This section covers:
Kinetics
Kinetics is an active scan at a selected fixed wavelength and optional reference wavelength, over a fixed period of
time with data being streamed at a selected intervals and integration period. When the experiment time is
reached the experiment has ended. Methods can be saved by name and used to repeat a kinetics scan. This
application is meant to see the reaction or decay of a sample over time.
The Kinetics application measures the change in the sample absorbance as a function of time.
The local control software allows the determination of a linear rate over a region, which can be
defined after the data acquisition. Frequently used in enzymatic kinetics, a factor can be
multiplied by the slope of the linear rate fit to determine activity Recalling and Recalculating
Graphical Kinetics Results
You can work with graphical or tabular data and perform the same functions with either.
However, the location of the function keys depends on the display type.
Note: The Kinetics application can measure only one sample at a time.
CHAPTER 10
10
Maintenance
The spectrophotometer is durable and reliable, so routine maintenance is minimal. This section
explains:
• Routine Care
• Changing the Fuse
• Replacing the Tungsten Lamp (Orion AquaMate 7100 Vis instruments only)
Warning: Operating the instrument with the cover off exposes the operator to potentially
dangerous voltages and ultraviolet (UV) radiation. Therefore, we recommend that only
authorized service representatives perform procedures requiring removal of the instrument
cover and replacement of electrical components. To protect both yourself and the instrument,
be sure to contact an authorized service representative to perform any service procedure you
do not feel comfortable performing.
Routine Care
Routine care for the spectrophotometer does not require a lot of time. To help minimize
maintenance time and increase the life and performance of the instrument, please follow these
guidelines:
• Always replace the dust cover when the instrument is not turned on to prevent dust from
accumulating in and on the instrument.
• Do not use or store the instrument in a corrosive environment.
• Gently wipe the outside of the instrument, including the touchscreen, with a soft cloth to
remove any dust or spills. Water, isopropyl alcohol and other common laboratory cleaning
agents may be used if necessary.
• Always clean up spills as soon as they occur to prevent or minimize damage to the
instrument. If concentrated acids or bases, or any hydrocarbon materials, are spilled on the
instrument, clean up the affected area immediately.
Ensuring your vials are clean both inside and out is important to the quality of your results for
two reasons: 1) contaminating material may absorb light resulting in falsely high absorbance
readings; and 2) contaminants in the vial may react chemically with subsequent reagents or
standards introduced into the vial.
Cleaning methods depend to some extent on the nature of the contaminating material. It is
important to identify the residual material in the vial that needs to be removed. Refer to the
following table for suggestions on cleaning methods, solvents and material.
Important: Keeping the vial clean is very important for long vial life.
• Never store vials or cuvettes long term in a water or solvent bath between uses. If the
solvent you are using dries, impurities in the water or solvent may be deposited on the
inside of the vial or cuvette, causing permanent damage.
• Use only lens cleaning tissue/paper or fine soft cloth to wipe optical surfaces. Most paper
products (such as facial tissues, paper towels, etc.) contain wood fibers that can damage
the vial or cuvette material.
• At the end of the day, ensure that all vials or cuvettes are well cleaned and stored in a
suitable container after drying.
Term Definition
Dilute acid Dilute nitric acid (<10%)
Acid Hydrochloric (5M) acid or nitric acid (5M) (see the Note below)
Solvent rinse Rinse with the solvent that was originally used to solvate your analyte
Copious water rinse Use a pure water (deionized, distilled, RO) and rinse at least 10 times
Use a neutral pH detergent (Triton X-100), if available, to dilute acid wash;
Detergent
water rinse to remove residue
Note: Do not use 5M nitric acid on an anti-reflection mirror coated vials or cuvettes.
Important: Using an ultrasonic cleaning bath for your vials or cuvettes is not recommended.
Each bath generates a different frequency; therefore, if your bath operates at the resonant
frequency of a vial or cuvette, the vial or cuvette will break. If a vial or cuvette was cleaned in an
ultrasonic bath, the warranty may be void by the manufacturer.
Use the liquid and a soft, lint-free cloth to clean the windows. Do not apply too much pressure
or the surface of the windows may be damaged. Be sure to remove all fingerprints.
Warning: This lamp gets very hot during operation. Before removing the lamp, turn off the
instrument and allow the lamp to cool for 10 minutes.
This procedure is for the Orion AquaMate 8100 Vis spectrophotometer. The typical lamp
source lifetime is approximately 3 – 5 years
Warning: This lamp gets very hot during operation. Before removing the lamp, turn off the
instrument and allow the lamp to cool for 10 minutes.
3. Reinstall the new lamp and the cover in the reverse order.
a. DO NOT TOUCH the window on the new lamp.
b. Take care when handling the new lamp and its attached
bracket.
• The lamp is precision aligned to the bracket.
• Do not adjust the screws holding the lamp to the bracket.
• Ensure that the pins and holes that align the black bracket
to the base are properly engaged before tightening the
screws that hold the new lamp to the base.
• Do not overtighten screws.
CHAPTER 11
11
Customer Services
Technical Support
For any questions or if you require assistance, contact our Technical Support Specialists:
• Email wai.techservbev@thermofisher.com
• Within the United States, call 1-800-225-1480
• Outside the United States, call +1-978-232-6000 or fax +1-978-232-6031
For additional product information, contact your local authorized dealer, Thermo Scientific Orion
technical sales representative or contact us using the Water and Laboratory Products (WLP)
information on the page back of this user manual.
For the most current warranty information, refer to the Thermo Scientific Orion warranty card
included on the Thermo Scientific Orion AquaMate literature CD and available online at
www.thermoscientific.com/water.
Instrument Specifications
Orion AquaMate 8100 UV-Vis Spectrophotometer Specifications
Optical Design Dual beam
Spectral Bandwidth 2 nm
Light Source
Xenon flash lamp (5 years)
(Typical Lifetime)
Detector Dual silicon photodiodes
Wavelength Range 190 to 1100 nm
Wavelength Accuracy ±0.5 nm
Wavelength
±0.2 nm
Repeatability
Wavelength Scanning
Slow, medium and fast (up to 1600 nm/min)
Speed
Wavelength Data
0.2, 0.5, 1.0, 2.0, 3.0, 5.0 nm
Resolution
Photometric
Absorbance, % transmittance, concentration
Measurement Modes
Photometric Range –2A to +3.5A
Photometric Accuracy ±0.002A at 0.5A, ±0.004A at 1.0A, ±0.008A at 2.0A
Photometric ±0.001A at 1A
Repeatability1
≤0.00020A at 0A at 260 and 500 nm
Photometric Noise2 ≤0.00030A at 1A at 260 and 500 nm
≤0.00040A at 2A at 260 and 500 nm
Photometric Drift3 <0.0005A/Hr (At 500 nm after warm-up)
< 1.0%T 198 nm (KCl) , <0.05%T at 220 nm (NaI), <0.03%T at 340 nm
Photometric Stray Light
(NaNO2)
Display 7-inch color touchscreen, high definition, 800 × 1280 pixels
Touchscreen Glove friendly touch screen
USB type A port for USB stick (front panel), USB type B port for computer
Connectivity
(rear panel), USB type A port for printer (rear panel)
Dimensions 35.5 × 38.5 × 19.5 cm (L × W × H)
Weight 7.5 kg
Power Requirements 100 to 240 V ; 50 to 60 Hz
1
Measured at 1.0A at 546 nm 2RMS at 500 nm. 60 consecutive Measurements. 3At 500 nm after 1 hr warmup
1Measured at 1.0A at 546 nm 2RMS at 500 nm. 60 consecutive Measurements. 3At 500 nm after 1 hr
warmup
Note: We reserve the right to make product improvements and updates. Specifications are
subject to change without notice.
Ordering Information
Cat. No. Description
AquaMate 8100 UV-Vis spectrophotometer with pre-loaded methods, holder for vials
AQ8100 and test tube from 12 to 25 mm OD, 10 mm square vial holder, printer, North
America, European and UK Power cords and dust cover
AquaMate 8100 UV-Vis spectrophotometer with pre-loaded methods, holder for vials
AQ8100 APAC and test tube from 12 to 25 mm OD, 10 mm square vial holder, printer, China, Indian
and Australia / NZ Power cords and dust cover
AquaMate 7100 Vis Spectrophotometer with pre-loaded methods, holder for vials and
AQ7100 test tube from 12 to 25 mm OD, North America, European and UK Power cords and
dust cover
AquaMate 7100 Vis Spectrophotometer with pre-loaded methods, holder for vials and
AQ7100 APAC test tube from 12 to 25 mm OD, China, Indian and Australia / NZ Power cords and
dust cover
Vial / Test Tube Holder for vials and test tube from 12 to 25 mm OD for Orion™
AQX1RNDVH
AquaMate 7100 & 8100 Spectrophotometers
10 mm square single-cell holder for Orion™ AquaMate 7100 & 8100
AQX1SQVH
Spectrophotometers
Long path rectangular cell holder for 20-100 mm pathlength cuvettes for Orion™
AQX1LWLVH
AquaMate 7100 & 8100 Spectrophotometers
Film/Filter holder for filters/lenses up to 50 mm long x 80 mm tall x 10 mm thick for
AQX1FLTRHDR
Orion™ AquaMate 7100 & 8100 Spectrophotometers
840-326100 AquaMate Calibration Standards Set (Requires AQX1FLTRHDR Film/Filter holder)
AQ71LMPTGST Replacement Tungsten-Halogen Lamp, Pre-aligned for Orion™ AquaMate 7100
AQ81LMPXEN Replacement Xenon Lamp Assy, Pre-aligned for Orion™ AquaMate 8100
AQX1PWRSUP Power Supply for Orion™ AquaMate 7100 & 8100 Spectrophotometers +12V 5A
Australia power cable (AS/NZS 3112) for Orion™ AquaMate 7100 & 8100
AQX1AUCBL
Spectrophotometers
AQX1CNCBL China power cable (PRC/3) for Orion™ AquaMate 7100 & 8100 Spectrophotometers
European power cable (CEE 7/7) for Orion™ AquaMate 7100 & 8100
AQX1EUCBL
Spectrophotometers
India power cable (SABS 164) for Orion™ AquaMate 7100 & 8100
AQX1INCBL
Spectrophotometers
North America power cable (NEMA 5-15) for Orion™ AquaMate 7100 & 8100
AQX1NACBL
Spectrophotometers
AQX1UKCBL UK Power cable (BS 1362) for Orion™ AquaMate 7100 & 8100 Spectrophotometers
AQX1PRNTR Snap-on Printer for Orion™ AquaMate 7100 & 8100 Spectrophotometers
AQX1PPRPSA Self-Stick Printer Paper for AquaMate Printer Accessory
AQX1PPRSTD Standard Printer Paper for AquaMate Printer Accessory
AC2V24 24 mm round vials, 12 pack
AC2V16 16 mm round vials, 10 pack
Thermoreactor for digestion methods, 100 / 120 / 150 / 160 / 165°C temperature
COD165
control
APPENDIX A
A
General Instrument Information
Parameters
Parameter Description
+-x÷ Enters math operators when in the calculator mode (Utility)
Displays the estimated percentage of lamp life used, based on a typical Xenon
% of Lamp Life Used
lamp life of five years (Utility)
3-Pt Net Calculates peak height from the tangential baseline in the graph (Scanning)
Absorbance Enters the absorbance value
Accept Name Accepts the displayed name entry (Test Name and Edit [Units])
Add Character Adds a highlighted character to the name entry (Test Name and Edit [Units])
Adds a wavelength and factor to the list in multi-wavelength tests and some
Add nm
performance verification tests
Area Calculates area under the peak in the graph (Scanning)
AutoPrint Turns the automatic printout on or off
Autoscale Rescales the graph to the original ranges of X- and Y-axes (Kinetics, Scanning)
Enters time when the baseline for scan tests needs to be collected again
Baseline Expiration
(Utility)
Beeper Turns the audible signal for key presses on and off (Utility)
Selects the zero baseline or the tangential baseline to calculate area under the
Calculation Baseline
peak in the graph (Scanning)
Calculator Enables the calculator mode (Utility)
Displays the vial or cuvette position placed in the light path (only with Auto 6 or
Cell Position #
Auto 3 sample positioner settings)
Parameter Description
Change Mode
Switches measurement modes (Basic A-%T-C and some Performance
Change to Abs
Verification tests)
Change to %T
Collect Baseline Starts the collection of the baseline (Scanning)
Concentration Sets the concentration value
Conc. of Standard Displays the entered concentration value (Adv A-%T-C)
Cursor Goes to cursor tracking mode to view data points in graph (Kinetics, Scanning)
←Cursor Moves the cursor right or left on the graph and displays the data of each point
Cursor→ (Kinetics, Scanning)
Curve Fit Selects the type of line fit calculation (Standard Curve)
Data File Name Allows entry of a name for the data file when AutoSave is on
Date Standards
Displays the date when standards were last measured (Standard Curve)
Measured
Date/Time Setup Enters the current date and time settings for the instrument (Utility)
Enters the time from test initiation to first measurement; allows for sample
Delay Time
equilibration (Adv A-%T-C and Kinetics)
Delete Character Deletes the last character of name entry (Test Name and Edit [Units])
Delete File Deletes a test or data file from the Stored Tests Directory (Utility)
Delete Name Deletes the entire name to allow a new entry (Test Name and Edit [Units])
Removes a wavelength and factor from the list (Multiwavelength and
Delete nm
Performance Verification)
Enters the volume of diluent added before measurement (Dilution Multiplier in
Diluent Volume
some Bio Tests)
Dilution Multiplier Displays the factor used to correct for sample dilution
Display Activity Indicates whether results should include protein concentration
DNA ε(260) Calculates the extinction coefficient
DNA Factor Enter the factor to calculate DNA concentration (DNA Bio Tests)
Changes a wavelength or factor in the list (Multiwavelength and Performance
Edit
Verification)
Edit Curve Manipulates the graph (Kinetics)
Selects a portion of data in a table for recalculation of result (Kinetics,
Edit Data
Scanning)
Edit Graph Manipulates the graph (Scanning)
Edit Scale Changes the graph axis scales and view individual data points (Scanning)
Enters a factor to convert a datum to a result
Abs x Factor 1 = Concentration Result
Factor Abs/min x Factor 2 = Kinetics Result
Can be entered or calculated from concentration and absorbance in Adv A-%T-
C
Enters a factor to convert a datum to a result
Factor 1 Abs(WL1) x Factor = Result (Abs Ratio, Abs Difference,
Multiwavelength)
Parameter Description
Enters a factor to convert a datum to a result
Factor 2
Abs(WL2) x Factor = Result (Abs Ratio, Abs Diff, Multiwavelength)
Enters a factor to convert a datum to a result
Factor 3-31
Abs (WL3-31) x Factor = Result (Multiwavelength)
Graph Displays the graph of collected data (Kinetics, Scanning)
Enters the numeric identifier for measurement; auto increments during the test
ID #
until turned off (set to 0)
Instrument Serial
Displays the serial number of the instrument (Utility)
Number
Intercept Enters where the line crosses the Y-axis (Abs where concentration = 0)
Interval Enters the wavelength range between data points (Scanning)
Interval Time Enters the time between repeated readings (Kinetics)
Enters a linearity value (Kinetics)
To help determine the linearity of the reaction during the measurement, the
instrument offers a linearity parameter. This is the difference between the
changes in absorbance of two measurements as shown in the following
example:
Time Abs ?A Linearity
Linearity Value 1 .1 --- ---
2 .2 .1 ---
3 .29 .09 P
4 .38 .09 P
5 .46 .08 P
6 .52 .06 F
Linearity is the ?A between ?A calculations; P=Pass and F=Fail
Loads the highlighted test from the Stored Tests Directory into active memory
Load Test
and sets the instrument to the test parameters (Utility)
Used to protect stored tests from accidental deletion or alteration; asks for a
Lock/Unlock
password to allow the user to lock or unlock the file (Utility)
Enters the lowest and highest acceptable results, outside of which the result is
Low/High Limits flagged as “Low” or “High” (Adv A-%T-C, Std Curve, Abs Ratio, Abs Diff,
Kinetics, 3-Pt Net, some Bio Tests)
Math Accesses manipulation functions of the graph (Scanning)
Measure Blank (as
Initiates measurement of the blank
function key)
Measure Blank (as Selects the frequency of zeroing the instrument as Once or Every Reading
test parameter) (Kinetics)
Selects the type of photometric data reported for a measurement (Abs, %T,
Measurement Mode
Conc) in A-%T-C, Kinetics, Scanning, Multiwavelength
Measure Samples Initiates measurement of samples
Max, X Enters a maximum X-value to manually rescale the graph (Kinetics, Scanning)
Max, Y Enters a maximum Y-value to manually rescale the graph (Kinetics, Scanning)
Min, X Enters a minimum X-value to manually rescale the graph (Kinetics, Scanning)
Min, Y Enters a minimum Y-value to manually rescale the graph (Kinetics, Scanning)
Selects a cursor point in functions using more than one cursor setting: Scan-
Next Cursor
Area and Scan-3-Pt Net calculations in the graph (Scanning)
Parameter Description
Number of Matched Enters the number of vials or cuvettes that will be run in the Correction Program
Cuvettes (maximum of 5)
Enters the number of samples to be measured in the test (Not available in
Number of Samples
Kinetics or Scanning)
Number of
Enters the number of standards to be measured for the standard curve
Standards
Printer Selects the output mode as RS-232 or Parallel (Utility)
Protein Factor Enters the factor to calculate protein concentration (DNA Bio tests)
Enters a reference wavelength value; for each reported measurement,
Ref. Wavelength measures the analytical wavelength and reference wavelength
Reported measurement = Abs @ Analytical WL – Abs @ Reference WL
Ref. Wavelength
Turns reference wavelength correction on or off
Correction
Run Standard Goes to the standards entry screen
Run Test Goes to the data collection screen
Selects the type of Sample Positioner:
1 Cell = no movement; zeros and measures sample in same position
Manual 6 = vial holder carousel moved by sample positioner touchscreen;
always zeros on position B, then returns to set position to start measurement
Sample Positioner
Auto 3 = vial holder carousel automatically moved to B, 2, 4 (always zeros on
position B, then goes to position 2 to start measurement)
Auto 6 = vial holder carousel automatically moved to B,1,2,3,4,5 (always zeros
on position B, then goes to position 1 to start measurement)
Sample Volume Enters the total volume of sample (under Dilution Multiplier in some Bio Tests)
Save Test Saves all parameters of the current test in internal memory for later recall
Scan Speed Selects the speed (nm/min) for a scan as Slow, Medium or Fast (Scanning)
Improves visibility of the display by changing the contrast between the
Screen Contrast
background and text (Utility)
Tags the highlighted test name with “>” to include the test in the SmartStart
Select Test
menu (Utility Stored Tests Directory)
Set Max. X Sets the cursor position in the graph as the minimum X and the maximum X
Set Min. X values for recalculating rate (Kinetics)
Set nms Use to enter and edit the wavelength and factor values
Selects the factor entry or the baseline to calculate area under the peak in the
Set Options
graph (Scanning)
Initiates the procedure to collect the data necessary to correct for absorbance
Setup Correction
differences between vials or cuvettes
Slope Enters abs/concentration value (Standard Curve)
Smoothing Turns data smoothing on and off (Scanning)
Software Revision Displays the version of firmware in the instrument (Utility)
SRE Tolerance Acceptable minimum stray light
Standard Enters the concentration of standards used to generate the standard curve for
Concentrations the test
Parameter Description
Selects the time since the last keystroke or instrument activity; powers down
Standby
the unit to save lamp life (Utility)
Start Wavelength Enters the beginning wavelength for a scan (Scanning)
Turns statistics on or off; calculates the average and standard deviation of
results when set on; statistics registers are cleared when set off, when
Statistics
instrument is turned off, when test parameters are changed and when test is
saved or resaved (all test types except Kinetics, Scanning, Multiwavelength)
Std Concentration Enters the concentration of the analyte in the standard solution
Stop Wavelength Enters the ending wavelength for a scan (Scanning)
Stored Tests
Displays the list of tests stored in the instrument (Utility)
Directory
Tabular Displays the list of collected data (Kinetics, Scanning)
Operator enters an alphanumeric name (maximum of 16 characters) for the
Test Name test; the name will be included on the data printout and, if the test is saved, will
be displayed in the Utility Test Directory screen (available in all tests)
Enters the time from the run initiation to the end of the test; equals Delay Time
Total Run Time
+ Interval Times + Measurement Times (Kinetics)
Selects or creates units labels for results (all stored tests except Abs Ratio,
Units
Scanning, Cell Growth)
Removes the “>” tag of the highlighted test name to remove the test from the
Unselect Test
SmartStart menu (Utility Stored Tests Directory)
Wavelength Enters values for the analytical wavelengths
𝑆𝑆𝑆𝑆 = � 𝑥𝑥𝑖𝑖
𝑆𝑆𝑆𝑆 = � 𝑦𝑦𝑖𝑖
𝑆𝑆𝑆𝑆𝑆𝑆 = � 𝑥𝑥𝑖𝑖2
𝑆𝑆𝑆𝑆𝑆𝑆 = � 𝑦𝑦𝑖𝑖2
Where:
xi = Concentration of ith standard
yi = Absorbance of ith standard
N = Number of standards
A = A(c)
Where:
A = absorbance
c = concentration
Where:
a0 = Y-axis intercept
a1...a4 = coefficients
Where:
A = absorbance
c = concentration
a1 = slope
Linear regression
through zero
The slope is calculated as: a1 = SXY/SXX
or
Where:
A = absorbance
c1, c2 = concentration
Validity of standard Graph of Valid Nonlinear Standard Curve:
curves
If this is not the case, there will be more than one solution within the specified
domain and the message “Curve cannot be used to determine sample
concentrations – it may produce ambiguous results” will appear when the
curve is viewed.
∑(𝑦𝑦𝑖𝑖 − 𝑦𝑦� )2
σ= �
N−n−1
The calculation for the correlation coefficient applies only to first order linear
regression curves (first degree polynomials)
3-Point Net
3-Point Net
(ASTM E16904)