THE ENVIRONMENTAL TECHNOLOGY VERIFICATION
PROGRAM
ŁEPA
U.S. Environmental Protection Agency
Balteiie
The Business of Innovation
ETV Joint Verification Statement
TECHNOLOGY TYPE:	Rapid Toxicity Testing System
APPLICATION:	Detecting Toxicity in Drinking Water
TECHNOLOGY
NAME:	LuminoTox SAPS
COMPANY:
ADDRESS:
Lab_Bell Inc.
2263, avenue du College PHONE: (819) 539-8508, ext. 107
Shawinigan, Quebec FAX: (819) 539-8880
CANADA G9N 6V8
WEB SITE:
E-MAIL:
www.lab-bell.com
info@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 stabilized aqueous photosynthetic systems (SAPS) 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 SAPS 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 SAPS 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 SAPS 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 negative background inhibition that
caused falsely low inhibition
¦	Other performance factors (sample throughput, ease of use, reliability).
The LuminoTox SAPS 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 above lppm
inhibits the photosynthetic process that the LuminoTox SAPS Test Kit depends on to indicate toxicity 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 LuminoTox SAPS Test Kit to detect
toxicity, as well as to measure the precision of the LuminoTox SAPS Test Kit results. The response of the
LuminoTox SAPS 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 at www.epa.gov/etv/centers/centerl.html.
TECHNOLOGY DESCRIPTION
The following description of the LuminoTox SAPS Test Kit Test Kit is based on information provided by the
vendor. This technology description was not verified in this test.
The LuminoTox SAPS Test Kit is a portable biosensor that uses SAPS activated by light absorption to
recognize toxic chemicals in water. SAPS are activated at a wavelength of 470 nanometers, and fluorescence
emission is read at wavelengths longer than 700 nanometers. SAPS are whole algae (Chlorella vulgaris) that
fluoresce when photosynthesis (the conversion of electromagnetic energy into stored chemical energy) is
activated by light absorption. Some of the absorbed energy is emitted as fluorescence, which is the signal
measured by the LuminoTox SAPS Test Kit. The photosynthetic electron chain is inhibited by a broad
spectrum of organic molecules (ureas, azides, phenols, quinones or amide derivatives, polyaromatic
hydrocarbons, polychlorinated biphenyls), redox species, cyanides, and metallic cations. The LuminoTox
SAPS Test Kit measures the fluorescence produced both in background water and samples containing
contaminants. Decreases in fluorescence as a result of adding toxic contamination are expressed as percent
inhibition.
Although other SAPS could be used in the LuminoTox analyzer, Lab_Bell uses Chlorella vulgaris, which is
concentrated by centrifugation in the middle of its exponential growth curve and stored at 4°C for a few
weeks. Prior to analysis, SAPS must be activated in room light for 90 minutes at ambient temperature. The
LuminoTox test is performed in the dark (in a covered syringe) by exposing 100 microliters of SAPS solution
to 2 milliliters of test sample for 10 minutes. In this short period of time, permeable molecules acting directly
on the photosynthetic electron chain are detected at low concentrations. Prolonged incubation allows the
detection of less permeable molecules.
The LuminoTox SAPS Test Kit consists of the LuminoTox analyzer, a bottle of SAPS for 50 tests, two vials
of organic standards (positive controls to ensure that the SAPs are fully functional), and one vial of distilled
water (for blank samples). Also provided are disposable syringes in which the test is performed and fabric
syringe covers to protect the reaction from light. The analyzer is 21.6 by 12.7 by 7.6 centimeters and weighs
1 kilogram. The analyzer is battery-operated, is equipped with a RS-232 serial port for transferring data, and
can be connected to a printer (not done during this test). A total of 100 measurements can be stored in the
internal memory. The rechargeable battery operates for eight hours. Reagents (including buffers and positive
and negative controls) for approximated 50 analyses cost $106, while the LuminoTox analyzer costs
approximately $7,500.

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VERIFICATION RESULTS
Parameter
Compound
Lethal
Dose (LD)
Cone.
(mg/L)
Average Inhibition at Concentrations
Relative to the LD Concentration
(%)
Range of
Standard
Deviations
(%)
Toxicity
Thresh.
(mg/L)
LD
LD/10
LD/100
LD/1,000
Contaminants in
DDW
Aldicarb
260
50
14
5
0
1-3
26
Botulinum
toxin
Complex B
0.3
-10
-6
-5
1
1-8
ND
Colchicine
240
0
4
0
3
1-5
ND
Cyanide
250
17
10
7
1
2-3
250
Dicrotophos
1,400
4
-11
-12
-10
1-2
ND
Nicotine
2,800
34
10
1
3
1-4
280
Ricin
15
0
1
-4
3
2-6
ND
Soman
1.4
-2
1
2
0
2-3
ND
Thallium
sulfate
2,800
0
1
-3
-4
2-3
ND
VX
2
5
3
-1
2
2-5
ND
Potential
interferences in
DDW
Interference
Cone.
(mg/L)
Average Inhibition
(%)
Standard
Deviation (%)
se positive. All disinfection
contamination.
Aluminum
0.5
1
4
Copper
0.6
3
1
Iron
0.15
1
2
Manganese
0.25
1
3
Zinc
2.5
-1
4
False positive
response
None of the LuminoTox SAPS Test Kit responses were considered fal
by-product test samples left enough fluorescence for inhibition due to
False negative
response
Botulinum toxin, colchicine, dicrotophos, ricin, soman, thallium sulfate, and VX exhibited non-
detectable responses at the lethal dose concentration.
Ease of use
The LuminoTox SAPS Test Kit contained detailed instructions and clear illustrations. The
contents 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. The necessity to record four numbers as raw data was somewhat burdensome;
however, Lab_Bell has indicated this procedure is being modified. No formal scientific education
would be required to use the LuminoTox SAPS Test Kit.
Field portability
The LuminoTox SAPS Test Kit was transported from a laboratory setting to a storage room for
the field portability evaluation. The limiting factor for testing in the field would be the
approximately 90 minutes required to allow the SAPS to be exposed to light prior to testing. The
LuminoTox SAPS 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 17% ± 2%, and in the non-laboratory location, 16% ± 4%.
Throughput
Approximately 20 analyses were completed per hour, and 50 samples could be analyzed with the
supplies contained in one LuminoTox SAPS Test Kit.
ND = Significant inhibition was not detected.

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Original signed by Gregory A. Mack	6/22/06
Gregory A. Mack	Date
Vice President
Energy, Transportation, and Environment Division
Battelle
Original signed by Andrew P. Avel	8/7/06
Andrew P. Avel	Date
Acting Director
National Homeland Security Research Center
Office of Research and Development
U.S. Environmental Protection Agency
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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