THE ENVIRONMENTAL TECHNOLOGY VERIFICATION
PROGRAM
U.S. Environmental Protection Agency
Batreiie
Trtc Business o/ Innovation
ETV Joint Verification Statement
TECHNOLOGY TYPE: Rapid Toxicity Testing System
APPLICATION: Detecting Toxicity in Drinking Water
TECHNOLOGY
NAME:
COMPANY:
ADDRESS:
WEB SITE:
E-MAIL:
Chem-IQ Tox™
Aqua Survey, Inc.
469 Point Breeze Road
P.O. Box 72
Flemington, NJ 08822
www.aquasurvey.com
mail@aquasurvey.com
PHONE: (908) 788-8700
FAX: (908) 788-9165
The U.S. Environmental Protection Agency (EPA) has established the Environmental Technology Verification
(ETV) Program to facilitate the deployment of innovative or improved environmental technologies through
performance verification and dissemination of information. The goal of the ETV Program is to further
environmental protection by accelerating the acceptance and use of improved and cost-effective technologies.
ETV seeks to achieve this goal by providing high-quality, peer-reviewed data on technology performance to
those involved in the design, distribution, financing, permitting, purchase, and use of environmental
technologies. Information and ETV documents are available at www.epa.gov/etv.
ETV works in partnership with recognized standards and testing organizations, with stakeholder groups
(consisting of buyers, vendor organizations, and permitters), and with individual technology developers. The
program evaluates the performance of innovative technologies by developing test plans that are responsive to
the needs of stakeholders, conducting field or laboratory tests (as appropriate), collecting and analyzing data,
and preparing peer-reviewed reports. All evaluations are conducted in accordance with rigorous quality
assurance (QA) protocols to ensure that data of known and adequate quality are generated and that the results
are defensible.
The Advanced Monitoring Systems (AMS) Center, one of six technology areas under ETV, is operated by
Battelle in cooperation with EPA's National Exposure Research Laboratory. The AMS Center evaluated the
performance of the Aqua Survey, Inc. Chem-IQ Tox™ Test Kit. This verification statement provides a
summary of the test results.
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VERIFICATION TEST DESCRIPTION
Rapid toxicity technologies use various biological organisms and chemical reactions to indicate the presence
of toxic contaminants. The toxic contaminants are indicated by a change or appearance of color or a change in
intensity. As part of this verification test, the Chem-IQ Tox™ Test Kit was subjected to various
concentrations of contaminants such as industrial chemicals, pesticides, rodenticides, pharmaceuticals, nerve
agents, and biological toxins. Each contaminant was added to separate drinking water samples and analyzed.
In addition to determining whether the Chem-IQ Tox™ Test Kit could detect the toxicity caused by each
contaminant, its response to interfering compounds, such as water treatment chemicals and by-products in
clean drinking water, was evaluated.
The Chem-IQ Tox™ Test Kit was evaluated by
• Endpoints and precision—percent inhibition for all concentration levels of contaminants and potential
interfering compounds and precision of replicate analyses
• Toxicity threshold for each contaminant—contaminant level at which higher concentrations generate
inhibition significantly greater than the negative control and lower concentrations do not. Note that Aqua
Survey, Inc. recommends that a 20% inhibition is required for a conclusive indication of toxicity. During
this test, a thorough evaluation of the toxicity threshold was performed. Therefore, the toxicity threshold
was determined with respect to the negative control rather than the 20% inhibition threshold
• False positive responses—chlorination and chloramination by-product inhibition with respect to
unspiked American Society for Testing and Materials Type II deionized water samples
• False negative responses—contaminants that were reported as producing inhibition less than 20% when
present at lethal concentrations (the concentration at which 250 milliliters of water would probably cause
the death of a 154-pound person) or negative background inhibition that caused falsely low inhibition
• Other performance factors (sample throughput, ease of use, reliability).
The Chem-IQ Tox™ Test Kit was verified by analyzing a dechlorinated drinking water sample from
Columbus, Ohio (DDW), fortified with contaminants (at concentrations ranging from lethal levels to
concentrations up to 1,000 times less than the lethal dose) and interferences (metals possibly present as a
result of the water treatment processes). Dechlorinated water was used because free chlorine can interfere
with the performance of the test and can degrade the contaminants during storage. Inhibition (endpoints) from
four replicates of each contaminant at each concentration level were evaluated to assess the ability of the
Chem-IQ Tox™ Test Kit to detect toxicity, as well as to measure the precision of the Chem-IQ Tox™ Test
Kit results. The response of the Chem-IQ Tox™ Test Kit to possible interferents was evaluated by analyzing
them at one-half of the concentration limit recommended by the EPA's National Secondary Drinking Water
Regulations guidance. For analysis of by-products of the chlorination process, the unspiked DDW was
analyzed because Columbus, Ohio, uses chlorination as its disinfectant procedure. For the analysis of by-
products of the chloramination process, a separate drinking water sample was obtained from the Metropolitan
Water District of Southern California (LaVerne, California), which uses chloramination as its disinfection
process. The samples were analyzed after residual chlorine was removed using sodium thiosulfate. Sample
throughput was measured based on the number of samples analyzed per hour. Ease of use and reliability were
determined based on documented observations of the operators.
Quality control samples included method blank samples, which consisted of American Society for Testing
and Materials Type II deionized water; positive control samples (fortified with copper chloride); and negative
control samples, which consisted of the unspiked DDW.
QA oversight of verification testing was provided by Battelle and EPA. Battelle QA staff conducted a
technical systems audit, a performance evaluation audit, and a data quality audit of 10% of the test data.
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This verification statement, the full report on which it is based, and the test/QA plan for this verification test
are all available atwww.epa.gov/etv/centers/centerl.html.
TECHNOLOGY DESCRIPTION
The following description of the Chem-IQ Tox™ Test Kit is based on information provided by the vendor.
This technology description was not verified in this test.
The Chem-IQ Tox™ Test Kit detects toxicants in drinking water using a chemical reaction that generates
fluorescence. The test can be conducted by a technician with basic laboratory skills. Sample analysis is
performed by adding two reagents to test and control water samples and measuring each sample's
fluorescence with a calibrated fluorometer. Percent inhibition values are calculated by comparing the light
production of the control with that of the test samples. If the average percent inhibition value of the replicate
test samples is greater than 20%, the test water sample is considered significantly impacted by a toxicant and
considered a positive response.
The Chem-IQ Tox™ Test Kit, which costs $250, contains 30 vials each of two reagents, 90 IQ Exposure
Chambers, disposal reagent pipettes, Chem-IQ Tox™ Test Kit score cards, and a Sharpie pen. Materials and
laboratory equipment required for the test include an Aquafluor™ hand-held fluorometer (Turner Design) or
equivalent and a supply of non-fluorescing 4 milliliter cuvettes (10 millimeter by 10 millimeter); an automatic
pipetter or equivalent with appropriate disposable tips for dispensing 10-milliliter, 250-microliter, and 50-
microliter volumes; a PCS liquid sonicator (L&R) or equivalent; a magnetic stir plate and stir bar (1/8 inch
diameter); a distilled or deionized water supply; and a digital timer that displays seconds.
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VERIFICATION RESULTS
Parameter
Contaminants in
DDW
False positive
response
False negative
response
Ease of use
Field portability
Throughput
Compound
Aldicarb
Botulinum
toxin
complex B
Colchicine
Cyanide
Dicrotophos
Nicotine
Ricin
Soman
Thallium
sulfate
VX
Interference
Aluminum
Copper
Iron
Manganese
Zinc
Lethal
Dose (LD)
Cone.
(mg/L)
260
0.3
240
250
1,400
2,800
15
1.4
2,800
2
Cone.
(mg/L)
0.5
0.6
0.15
0.25
2.5
Average Inhibition at Concentrations
Relative to the LD Concentration
LD
-16
-62
104
63
-55
50
-44
16
66
-44
LD/10
13
1
63
47
-30
84
-13
-8
16
-36
Average
Inhibition (%)
4
46
-26
11
34
LD/100
-33
4
17
-21
-38
71
10
19
-5
-14
Standard
("
LD/1,000
7
5
42
-18
-13
-3
-3
7
-25
-13
Deviation
9
3
20
9
2
Range of
Standard
Deviations
9-32
3-8
2-15
2-24
8-41
1-12
4-12
1-6
3-13
8-26
Toxicity
Thresh.
(mg/L)
ND
ND
24
25
ND
28
ND
ND
2,800
ND
Because DI water did not generate any measurable background light, the disinfection by-product
samples could not be compared with the inhibition due to DI water. Therefore only the absolute light
units produced by the chlorinated and chloraminated samples could be measured. Both of these
samples left adequate light for subsequent inhibition due to contamination and are thus not
considered to have generated false positive results.
False negative responses (inhibition less than 20%) were j
complex B, dicrotophos, ricin, soman, and VX when they
concentration.
generated for aldicarb, botulinum toxin,
were analyzed at the lethal dose
The Chem-IQ Tox™ Test Kit instructions were clearly written; but a condensed summary with only
the necessary steps may be helpful. The contents of the Chem-IQ Tox™ Test Kit were well
identified. The test was not difficult to perform, but analyzing several samples simultaneously
required practice. No formal scientific education would be required to use the Test Kit.
The Chem-IQ Tox™ Test Kit was transported from a laboratory to a storage room to simulate a non-
laboratory location. All materials were easily transported by one person in a small cardboard box.
The Test Kit was set up in less than 10 minutes, except that Reagent Two took approximately
20 minutes to thaw. A source of electricity was required for the sonicator, while the fluorometer ran
on batteries. A cooler to transport and store reagents, pipettes and tips, the sonicator and a power
source, the fluorometer, and a waste container were needed for field use. Results were obtained
within 10 minutes of starting the test.
Approximately 30 analyses were completed in one hour. The 30 analyses included method blanks,
positive controls, and test samples. Approximately 130 samples could be processed per pair of
Reagent One and Reagent Two vials.
ND = Significant inhibition was not detected.
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Original signed by Gregory A. Mack
Gregory A. Mack Date
Vice President
Energy, Transportation, and Environment Division
Battelle
6/22/06 Original signed by Andrew P. Avel
Andrew P. Avel
Acting Director
National Homeland Security Research Center
Office of Research and Development
U.S. Environmental Protection Agency
8/7/06
Date
NOTICE: ETV verifications are based on an evaluation of technology performance under specific,
predetermined criteria and the appropriate quality assurance procedures. EPA and Battelle make no expressed or
implied warranties as to the performance of the technology and do not certify that a technology will always
operate as verified. The end user is solely responsible for complying with any and all applicable federal, state,
and local requirements. Mention of commercial product names does not imply endorsement.
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