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:
LuminoTox PECs
Lab Bell Inc.
2263, avenue du College PHONE: (819) 539-8508, ext. 107
Shawinigan, Quebec FAX: (819) 539-8880
CANADA G9N 6V8
www.lab-bell.com
info(S>lab-bell.com
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 Lab_Bell Inc. LuminoTox photosynthetic enzymatic complexes (PECs) 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, LuminoTox PECs 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 LuminoTox PECs 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.
LuminoTox PECs 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
• 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 similar to the
negative control when present at lethal concentrations (the concentration at which 250 milliliters of
water would probably cause the death of a 154-pound person) or a negative background inhibition that
caused falsely low inhibition
• Other performance factors (sample throughput, ease of use, reliability).
The LuminoTox PECs 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 inhibits the
photosynthetic process that the LuminoTox PECs Test Kit depends on to indicate toxicity and can degrade the
contaminants during storage. Inhibition results (endpoints) from four replicates of each contaminant at each
concentration level were evaluated to assess the ability of the LuminoTox PECs Test Kit to detect toxicity, as
well as to measure the precision of the LuminoTox PECs Test Kit results. The response of the LuminoTox
PECs 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 atrazine); 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 LuminoTox PECs Test Kit is based on information provided by the vendor.
This technology description was not verified in this test.
The LuminoTox PECs Test Kit is a portable biosensor that indicates the presence of toxic chemicals in water.
It uses PECs that have been stabilized through a method patented by Lab_Bell Inc. The PECs are membranes
isolated from chloroplasts that are as simple to use as a chemical, but react more rapidly than a living
organism because toxic compounds do not have to penetrate the cell wall of an organism. The photosynthetic
electron chain is what is inhibited by contamination. When stimulated by light, the PECs emit fluorescence.
The LuminoTox PECs Test Kit measures the fluorescence parameters produced both in background water and
samples containing contaminants. Decreases in fluorescence parameters as a result of the presence of toxic
contamination are expressed as percent inhibition.
The LuminoTox PECs Test Kit consists of the LuminoTox analyzer, a bottle of PECs for 50 tests, reaction
buffer, a blank water control, and a positive control. Also provided are disposable syringes in which the test is
performed and fabric syringe covers to protect the reaction from light. Aluminum foil can be used as a light
protector.
The LuminoTox analyzer is 21.6 by 12.7 by 7.6 centimeters and weighs 1 kilogram. It is battery-operated and
is portable. The analyzer has a built-in RS-232 serial port outlet, which can also be used for transferring data
to a spreadsheet (which was not done during this test), and is compatible with a printer. A total of 100
measurements can be stored in the internal memory. The rechargeable battery operates for eight hours. Each
kit costs $89, and the analyzer costs approximately $7,500.
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VERIFICATION RESULTS
Parameter
Contaminants
inDDW
Potential
interferences
inDDW
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
26
0
2
47
3
77
2
-5
63
-5
LD/10
2
-5
-3
31
10
80
5
-6
19
-8
LD/100
0
-8
-2
-8
8
6
-7
0
-3
-2
Average Inhibition
(%)
0
70
7
5
12
LD/1,000
-2
-12
-6
-7
4
9
-10
-1
-12
3
Standard
Deviation (%)
4
1
6
6
5
Range of
Standard
Deviations
(%)
1-3
2-6
1-5
1-7
2-4
1-6
4-9
4-6
2-7
3-5
Toxicity
Thresh.
(mg/L)
260
ND
ND
25
ND
280
ND
ND
280
ND
Both the chlorinated and chloraminated disinfection by-product samples produced an inhibition
significantly greater than the negative control and, therefore, were considered false positive
responses. However, the disinfectant by-product samples produced an inhibition of less than 15%,
leaving enough fluorescence available for subsequent inhibition due to contamination.
Botulinum toxin complex B, colchicine, dicrotophos, ricin, soman, and VX exhibited non-detectable
responses at the lethal dose concentration.
The LuminoTox PECs Test Kit contained detailed instructions and clear illustrations. The contents
of the LuminoTox PECs Test Kit were well identified with labels on the vials. Storage requirements
were stated in the instructions and on the reagent vials. Preparation of the test samples for analysis
was straightforward. However, the PECs had to be stored in ice between every sample analysis to
keep them from coming to room temperature, which was somewhat inconvenient because the
melting ice caused the lab bench and operators' hands to be wet most of the time. The necessity to
record four numbers as raw data was somewhat burdensome; however, Lab_Bell has indicated that it
is modifying this. No formal scientific education would be required to use the LuminoTox PECs
Test Kit.
The LuminoTox PECs Test Kit was transported from a laboratory setting to a storage room for the
field portability evaluation. The LuminoTox PECs Test Kit was tested with one contaminant,
cyanide, at the lethal dose concentration. The results of the test were very similar to the laboratory
results. Inhibition in the laboratory was 47% ± 1%, and in the non-laboratory location, 51% ± 1%.
Approximately 20 analyses were completed per hour, and approximately 50 samples could be
analyzed with the supplies contained in one LuminoTox PECS Test Kit.
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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