November 2003
Environmental Technology
Verification Report
SEVERN TRENT SERVICES
ECLOX
RAPID TOXICITY TESTING SYSTEM
Prepared by
Battelle
Batteiie
. . . Putting Technology To Work
Under a cooperative agreement with
U.S. Environmental Protection Agency
ETV ETV ET
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November 2003
Environmental Technology Verification
Report
ETV Advanced Monitoring Systems Center
Severn Trent Services
Eclox
Rapid Toxicity Testing System
by
Ryan James
Amy Dindal
Zachary Willenberg
Karen Riggs
Battelle
Columbus, Ohio 43201
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Notice
The U.S. Environmental Protection Agency (EPA), through its Office of Research and
Development, has financially supported and collaborated in the extramural program described
here. This document has been peer reviewed by the Agency. Mention of trade names or
commercial products does not constitute endorsement or recommendation by the EPA for use.
11
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Foreword
The U.S. Environmental Protection Agency (EPA) is charged by Congress with protecting the
nation's air, water, and land resources. Under a mandate of national environmental laws, the
Agency strives to formulate and implement actions leading to a compatible balance between
human activities and the ability of natural systems to support and nurture life. To meet this
mandate, the EPA's Office of Research and Development provides data and science support that
can be used to solve environmental problems and to build the scientific knowledge base needed
to manage our ecological resources wisely, to understand how pollutants affect our health, and to
prevent or reduce environmental risks.
The Environmental Technology Verification (ETV) Program has been established by the EPA to
verify the performance characteristics of innovative environmental technology across all media
and to report this objective information to permitters, buyers, and users of the technology, thus
substantially accelerating the entrance of new environmental technologies into the marketplace.
Verification organizations oversee and report verification activities based on testing and quality
assurance protocols developed with input from major stakeholders and customer groups
associated with the technology area. ETV consists of seven environmental technology centers.
Information about each of these centers can be found on the Internet at http://www.epa.gov/etv/.
Effective verifications of monitoring technologies are needed to assess environmental quality
and to supply cost and performance data to select the most appropriate technology for that
assessment. Under a cooperative agreement, Battelle has received EPA funding to plan,
coordinate, and conduct such verification tests for "Advanced Monitoring Systems for Air,
Water, and Soil" and report the results to the community at large. Information concerning this
specific environmental technology area can be found on the Internet at
http: //www. ep a. gov/etv/centers/center 1. html.
in
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Acknowledgments
The authors wish to acknowledge the support of all those who helped plan and conduct the
verification test, analyze the data, and prepare this report. Many thanks go to Battelle's Medical
Research and Evaluation Facility for providing the facilities for and personnel capable of
working with chemical warfare agents and biotoxins. We would also like to thank
Karen Bradham, U.S. EPA National Exposure Research Laboratory; Steve Allgeier, U.S. EPA
Office of Water; Ricardo DeLeon, Metropolitan Water District of Southern California; Yves
Mikol, New York City Department of Environmental Protection; and Stanley States, Pittsburgh
Water and Sewer Authority, for their careful review of the test/QA plan and this verification
report.
IV
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Contents
Page
Notice ii
Foreword iii
Acknowledgments iv
List of Abbreviations viii
1 Background 1
2 Technology Description 2
3 Test Design and Procedures 4
3.1 Introduction 4
3.2 Test Design 5
3.3 Test Samples 6
3.3.1 Quality Control Samples 6
3.3.2 Drinking Water Fortified with Contaminants 8
3.3.3 Drinking Water Fortified with Potential Interferences 8
3.4 Test Procedure 8
3.4.1 Test Sample Preparation and Storage 8
3.4.2 Test Sample Analysis Procedures 9
3.4.3 Stock Solution Confirmation Analysis 9
4 Quality Assurance/Quality Control 12
4.1 Quality Control of Stock Solution Confirmation Methods 12
4.2 Quality Control of Drinking Water Samples 12
4.3 Audits 13
4.3.1 Performance Evaluation Audit 13
4.3.2 Technical Systems Audit 14
4.3.3 Audit of Data Quality 14
4.4 QA/QC Reporting 14
4.5 Data Review 15
5 Statistical Methods and Reported Parameters 16
5.1 Endpoints and Precision 16
5.2 Toxicity Threshold 16
5.3 False Positive/Negative Responses 17
5.4 Field Portability 17
5.5 Other Performance Factors 17
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6 Test Results 18
6.1 Endpoints and Precision 18
6.1.1 Contaminants 18
6.1.2 Potential Interferences 18
6.1.3 Precision 23
6.2 Toxicity Threshold 24
6.3 False Positive/Negative Responses 25
6.4 Field Portability 25
6.5 Other Performance Factors 26
7 Performance Summary 27
8 References 28
Figures
Figure 2-1. Eclox Rapid Toxicity Testing System 2
Tables
Table 3-1. Contaminants and Potential Interferences 5
Table 3-2. Summary of Quality Control and Contaminant Test Samples 7
Table 3-3. Dose Confirmation Results 10
Table 3-4. Water Quality Parameters 11
Table 4-1. Summary of Performance Evaluation Audit 13
Table 4-2. Summary of Data Recording Process 15
Table 6-la. Aldicarb Percent Inhibition Results 19
Table 6-lb. Colchicine Percent Inhibition Results 19
Table 6-lc. Cyanide Percent Inhibition Results 20
Table 6-ld. Dicrotophos Percent Inhibition Results 20
VI
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Table 6-le. Thallium Sulfate Percent Inhibition Results 21
Table 6-lf. Botulinum Toxin Percent Inhibition Results 21
Table 6-lg. Ricin Percent Inhibition Results 22
Table 6-lh. Soman Percent Inhibition Results 22
Table 6-li. VX Percent Inhibition Results 23
Table 6-2. Potential Interferences Results 24
Table 6-3. Toxicity Thresholds 25
Table 6-4. False Negative Responses 26
VI1
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List of Abbreviations
AMS
ASTM
ATEL
DI
DDW
EPA
ETV
HOPE
HRP
ID
LD
\iL
mL
NSDWR
%D
PE
QA
QC
QMP
SOP
ISA
Advanced Monitoring Systems
American Society for Testing and Materials
Aqua Tech Environmental Laboratories
deionized water
dechlorinated drinking water from Columbus, Ohio
U.S. Environmental Protection Agency
Environmental Technology Verification
high-density polyethylene
horseradish peroxidase
identification
lethal dose
microliter
milliliter
National Secondary Drinking Water Regulations
percent difference
performance evaluation
quality assurance
quality control
quality management plan
standard operating procedure
technical systems audit
Vlll
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Chapter 1
Background
The U.S. Environmental Protection Agency (EPA) supports the Environmental Technology
Verification (ETV) Program to facilitate the deployment of innovative environmental tech-
nologies 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.
ETV works in partnership with recognized testing organizations; with stakeholder groups
consisting of buyers, vendor organizations, and permitters; and with the full participation of
individual technology developers. The program evaluates the performance of innovative tech-
nologies 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 EPA's National Exposure Research Laboratory and its verification organization partner,
Battelle, operate the Advanced Monitoring Systems (AMS) Center under ETV. The AMS Center
recently evaluated the performance of the Severn Trent Services Eel ox rapid toxicity testing
system. Rapid toxicity testing systems were identified as a priority technology verification
category through the AMS Center stakeholder process.
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Chapter 2
Technology Description
The objective of the ETV AMS Center is to verify the performance characteristics of environ-
mental monitoring technologies for air, water, and soil. This verification report provides results
for the verification testing of Eclox. Following is a description of Eclox, based on information
provided by the vendor. The information provided below was not subjected to verification in this
test.
Eclox (Figure 2-1) is a broadband chemiluminescence test that qualitatively assesses a water
sample to determine whether it has been contaminated. The technique, used extensively in the
medical field as an immunodiagnostic tool, is based upon the reaction of luminol and an oxidant
in the presence of a catalyst enzyme—horseradish peroxidase (FtRP). This reaction produces a
flash of light (chemiluminescence) that is measured by a luminometer. An enhancer is added
prior to the HRP so that the light output produced is of a steady measurable level. Free radical
scavengers or antioxidants such as those contained in feces or urine interfere with the reaction,
thus reducing the light emission. Substances such as phenols, amines, heavy metals, or
compounds that interact with the enzyme also reduce the light output.
Figure 2-1. Eclox Rapid Toxicity
Testing System
To analyze a water sample, 100 microliters (\iL) of three
reagents are added to 1 milliliter (mL) of the sample, and
the sample cuvette is placed in the luminometer for four
I minutes. Results are compared with a contaminant-free
reference, i.e., deionized water, which gives a high light
output. Samples containing pollution give lower light
levels. Comparing the light output from sample water to
that obtained from the reference indicates the contamina-
tion levels in the sample water. This test gives a measure
of the relative toxicity of a water sample with respect to a
control sample. It is up to the user to define the response
protocols to activate, based on the level of inhibition
exhibited by a water sample.
The Eclox includes a luminometer, a 100-[J,L and a
1,000-[J,L pipette and pipette tips, cuvettes, reagent, a
pre-conditioner, a cuvette holder, and a CD-ROM with
software to download results.
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The luminometer stores a total of 60 measurements, and the data can be downloaded to a
personal computer using the supplied software. The stored values are downloaded to a Microsoft
Access database file and can be exported to a Microsoft Excel spreadsheet.
The complete Eclox weighs approximately 20 pounds. Overall dimensions for the kit are
20-V2 inches x 17-V2 inches x 8 inches. The luminometer contained in the system weighs a few
pounds and is approximately 9 inches x 5 inches x 3 inches. The cost of the full Eclox kit is
$7,900.
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Chapter 3
Test Design and Procedures
3.1 Introduction
The objective of this verification test of rapid toxicity technologies was to evaluate their ability
to detect certain toxins and to determine their susceptibility to interfering chemicals in a
controlled experimental matrix. Rapid toxicity technologies do not identify or determine the
concentration of specific contaminants, but serve as a screening tool to quickly determine
whether water is potentially toxic. Rapid toxicity technologies use bacteria (e.g., Vibrio
Jischeri), enzymes (e.g., luciferase), or small crustaceans (e.g., Daphnia magna) that either
directly, or in combination with reagents, produce a background level of light or use dissolved
oxygen at a steady rate in the absence of toxic contaminants. Toxic contaminants in water are
indicated by a change in the color or intensity of light produced or by a decrease in the dissolved
oxygen uptake rate in the presence of the contaminants.
As part of this verification test, Eclox 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 Eclox can detect the toxicity caused by each contaminant, its
response to interfering compounds in clean drinking water, such as water treatment chemicals
and by-products, was evaluated. Table 3-1 shows the contaminants and potential interferences
that were evaluated during this verification test.
This verification test was conducted according to procedures specified in the Test/QA Plan for
Verification of Rapid Toxicity Technologies .(1) Eclox was verified by analyzing a dechlorinated
drinking water (DDW) sample from Columbus, Ohio, fortified with various concentrations of
the contaminants and interferences shown in Table 3-1. Hereafter in this report, DDW will refer
to dechlorinated drinking water from Columbus, Ohio. Where possible, the concentration of
each contaminant or potential interference was confirmed independently by Aqua Tech
Environmental Laboratories (ATEL), Marion, Ohio, or by Battelle, depending on the analyte.
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Table 3-1. Contaminants and Potential Interferences
Category Contaminant
Carbamate pesticide aldicarb
Pharmaceutical colchicine
Industrial chemical cyanide
Organophosphate pesticide dicrotophos
Rodenticide thallium sulfate
Biological toxins botulinum toxin, ricin
Nerve agents soman, VX
Potential interferences aluminum, copper, iron, manganese, zinc,
chloramination by-products, and chlorination
by-products
Eclox was evaluated by
• Endpoint and precision—percent inhibition for all concentration levels of contaminants and
potential interfering compounds and precision of replicate analyses
• Toxicity threshold for each contaminant
• False negative responses—contaminants that were reported as producing inhibition results
similar to the negative control when the contaminant was present at lethal concentrations
• False positive responses—occurrence of inhibition significantly greater than the inhibition
reported for unspiked American Society for Testing and Materials (ASTM) Type n deionized
(DI) water samples (zero inhibition)
• Field portability
• Ease of use
• Throughput.
3.2 Test Design
Eclox was used to analyze the DDW sample fortified with contaminants at concentrations
typically ranging from lethal levels to concentrations several orders of magnitude less than the
lethal dose. The lethal dose of each contaminant was determined by calculating the concentration
at which 250 mL of water would probably cause the death of a 154-pound person. These calcula-
tions were based on toxicological data available for each contaminant. For soman, the stock
solution confirmation showed degradation in the water; therefore, the concentrations analyzed
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were less than anticipated. Whether the concentration is still a lethal dose, as is the case for all
contaminants, depends on the characteristics of each individual person and the amount of
contaminant ingested. Inhibition results (endpoints) from four replicates of each contaminant at
each concentration level were evaluated to assess the ability of Eclox to detect toxicity at various
concentrations of contaminants, as well as to measure the precision of Eclox results.
The response of Eclox to compounds used during the water treatment process (identified as
potential interferences in Table 3-1) was evaluated by analyzing separate aliquots of DDW
fortified with each potential interference at approximately one-half of the concentration limit
recommended by the EPA's National Secondary Drinking Water Regulations (NSDWR)(2)
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 from
St. Petersburg, Florida, which uses chloramination as its disinfection process, was obtained. The
samples were analyzed after residual chlorine was removed using the vendor-provided
dechlorinating reagent.
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 and the
Verification Test Coordinator. In addition to comprehensive testing in Battelle laboratories, Eclox
was operated in the basement of a Columbus, Ohio, home to test its ability to be transported and
operated in a non-laboratory setting.
3.3 Test Samples
Test samples used in the verification test included drinking water and quality control (QC)
samples. Table 3-2 shows the number and type of samples analyzed. QC samples included method
blanks and positive and negative control samples. The fortified drinking water samples were
prepared from a single drinking water sample collected from the Columbus, Ohio, system. The
water was dechlorinated using a vendor-provided dechlorination reagent and then fortified with
various concentrations of contaminants and interferences. Using this DDW (Columbus, Ohio,
dechlorinated drinking water), individual solutions containing each contaminant and potential
interference were prepared and analyzed. The DDW containing the potential interferences was
analyzed at a single concentration level, while four dilutions (made using the DDW) were
analyzed for each contaminant using Eclox. Mixtures of contaminants and interfering compounds
were not analyzed. One concentration level of cyanide was analyzed in the field setting.
3.3.1 Quality Control Samples
QC samples included method blank samples, which consisted of ASTM Type EDI water; positive
control samples, which consisted of ASTM Type n DI water or DDW (depending on vendor
preference) fortified with a contaminant and concentration selected by the vendor; and negative
control samples, which consisted of the unspiked DDW. The method blank samples were used to
help ensure that no sources of contamination were introduced in the sample handling and analysis
procedures.
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Table 3-2. Summary of Quality Control and Contaminant Test Samples
Type of Sample
Quality control
DDW fortified
with contaminants
Field location
DDW fortified
with potential
interferences
Disinfectant
by-products
Sample Characteristics
Method blank
Positive control (Phenol)
Negative control (unspiked
DDW)
Aldicarb
Colchicine
Cyanide
Dicrotophos
Thallium sulfate
Botulinum toxin*-1
Ricin(c)
Soman
VX
Cyanide
Aluminum
Copper
Iron
Manganese
Zinc
Chloramination by-
products
Chlorination by-products
Concentration
Levels (mg/L)
NS(a)
115
NS
280; 28; 2.8; 0.28
240; 24; 2.4; 0.24
250; 0.25; 0.05;
0.025
1,400; 140; 14; 1.4
2,400; 240; 24; 2.4
0.30; 0.030; 0.0030;
0.0030
15; 1.5; 0.15; 0.015
0.068;(d) 0.0068;
0.00068; 0.000068
0.49; 0.049; 0.0049;
0.00049
0.05
0.36
0.65
0.069
0.26
3.5
NS
NS
No. of Sample Analyses
12
14
47
4 per concentration level
4 per concentration level
4 per concentration level
4 per concentration level
4 per concentration level
4 per concentration level
4 per concentration level
4 per concentration level
4 per concentration level
4
4
4
4
4
4
4
4
W NS = Samples not fortified with any contaminant or potential interference.
^ Lethal dose solution also contained 3 mg/L phosphate and 1 mg/L sodium chloride.
(c) Lethal dose solution also contained 3 mg/L phosphate, 26 mg/L sodium chloride, and 2 mg/L sodium azide.
(d) Due to the degradation of soman in water, the stock solution confirmation analysis confirmed that the concentration of the
lethal dose was 23% of the expected concentration of 0.30 mg/L.
Phenol was suggested by the vendor for use as the positive control sample; and, while per-
formance limits were not placed on the results, nearly complete inhibition for this contaminant
indicated to the operator that Eclox was functioning properly. The negative control sample was
used to set a background inhibition of the DDW, the matrix in which each test sample was
prepared.
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3.3.2 Drinking Water Fortified with Contaminants
Approximately 150 liters of Columbus, Ohio, tap water were collected in a high-density
polyethylene (HDPE) container. A portion of that sample was dechlorinated with two drops of
vendor-provided dechlorinating reagent for every 50 mL of water. All subsequent test samples
were prepared from this DDW and stored in glass containers to avoid chlorine leaching from
HDPE containers.
A stock solution of each contaminant was prepared in ASTM Type n DI water at concentrations
above the lethal dose concentration level. The stock solution was diluted in DDW to obtain one
sample containing the lethal dose concentration for each contaminant and three additional
samples with concentrations 10, 100, and 1,000 times less than the lethal dose. At concentra-
tions near the lethal dose, Eclox was more sensitive to cyanide than to the other contaminants, so
more dilute solutions had to be prepared and analyzed. Table 3-2 lists each concentration level
and the number of samples analyzed at each level.
3.3.3 Drinking Water Fortified with Potential Interferences
Individual aliquots of the DDW were fortified with one-half the concentration specified by the
EPA's NSDWR for each potential interference. Table 3-2 lists the interferences, along with the
concentrations at which they were tested. Four replicates of each of these samples were
analyzed. To test the sensitivity of Eclox to by-products of the chlorination process as potential
interferences, the unspiked DDW (same as the negative control) was used since the water sample
originated from a utility that uses chlorination as its disinfectant procedure. In a similar test
involving the by-products of the chloramination process, an additional water sample was
obtained from St. Petersburg, Florida, a city that uses chloramination as its disinfectant
procedure. The residual chlorine in both of these samples was removed using the vendor-
provided dechlorination reagent, and then the samples were analyzed in replicate with no
additional fortification of contaminants.
3.4 Test Procedure
3.4.1 Test Sample Preparation and Storage
A drinking water sample was collected as described in Section 3.3.2 and, because free chlorine
inhibits the chemiluminescent reaction that generates the light production within the Eclox
reagent and can degrade the contaminants during storage, was immediately dechlorinated with
the dechlorinating reagent provided by the vendor. All the contaminant samples, potential inter-
ference samples, and negative control QC samples were made from this DDW, while the method
blank sample was prepared from ASTM Type n DI water. The positive control samples were
made using ASTM Type n DI water in Class A volumetric glassware. All QC samples were
prepared prior to the start of the testing and stored at room temperature for a maximum of
60 days. The aliquots of DDW containing the contaminants were prepared within seven days of
testing and stored in the dark at room temperature without chemical preservation. Aliquots to be
analyzed by each technology were placed in uniquely labeled sample containers. The sample
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containers were assigned an identification (ID) number. A master log of the samples and sample
ID numbers for each technology was kept by Battelle.
3.4.2 Test Sample Analysis Procedure
To analyze DDW samples, 100 \iL of three reagents were added to 1 mL of the water sample to
be analyzed, and the sample cuvette was placed in the Eclox immediately. The sample was
analyzed for four minutes. Software within the Eclox automatically calculated the result (percent
inhibition) for each sample. For each contaminant, Eclox analyzed the lethal dose concentration
and three additional concentration levels four times. Only one concentration of potential inter-
ference was analyzed. To test the field portability of Eclox, a single concentration level of
cyanide, prepared in the same way as the other DDW samples, was analyzed in replicate by
Eclox in the basement of a Columbus, Ohio, home. Sample analysis procedures were performed
in the same way as during testing in the laboratory. Two operators performed all the analyses
using Eclox. Both held bachelor's degrees in the sciences and spent approximately four hours
with the vendor to become familiar with using Eclox.
3.4.3 Stock Solution Confirmation Analysis
The concentrations of the contaminant and interfering compound stock solutions were verified
with standard analytical methods, with the exception of colchicine, ricin, and botulinum toxin—
contaminants without standard analytical methods. Aliquots to be analyzed by standard methods
were preserved as prescribed by the method. In addition, the same standard methods were used
to measure the concentrations of each contaminant/potential interference in the unspiked DDW
so that background concentrations of contaminants or potential interferences were accounted for
within the displayed concentration of each contaminant/potential interference sample. Table 3-3
lists the standard methods used to measure each analyte; the results from the stock solution
confirmation analyses (obtained by reporting the correct lethal dose concentration for the
contaminants and the single concentration that was analyzed for the potential interferences); and
the background levels of the contaminants and potential interferences measured in the DDW
sample, which were all non-detect or negligible.
Standard methods were also used to characterize several water quality parameters such as the
concentration of trihalomethanes, haloacetic acids, and total organic halides; turbidity; dissolved
organic carbon content; pH; alkalinity; specific conductivity; and hardness. Table 3-4 lists these
measured water quality parameters for both the water sample collected in Columbus, Ohio,
representing a water system using chlorination as the disinfecting process, and the water sample
collected in St. Petersburg, Florida, representing a water system using chloramination as the
disinfecting process.
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Table 3-3. Dose Confirmation Results
Contaminant
Aldicarb
Colchicine
Cyanide
Dicrotophos
Thallium sulfate
Botulinum toxin
Ricin
Soman
VX
Method
EPA531.1(3)
(a)
EPA335.1(4)
EPASW846(8141A)(5)
EPA 200.8(6)
(a)
(a)
(<0
(<0
Average Concentration
± Standard Deviation N
= 4 (mg/L)
280 ±28
NA(b)
250 ±15
1,400 ±140
2,400 ± 24
NA
NA
0.068(d)± 0.001
0.49 ±0.01
Background in
DDW (mg/L)
<0.0007
NA
0.008
<0.002
<0.001
NA
NA
<0.05
<0.05
Potential Interference
Aluminum
Copper
Iron
Manganese
Zinc
EPA 200. 8
EPA 200.8
EPA 200.8
EPA 200. 8
EPA 200. 8
0.36 ±0.01
0.65 ±0.01
0.069 ±0.08
0.26 ±0.01
3.5 ±0.35
<0.10
0.011
<0.04
<0.01
0.3
(a) No standard method available. QA audits and balance calibration assured accurately prepared solutions.
^ NA = Not applicable.
(G:I Purity analyses performed on chemical and biological agent materials using Battelle standard operating
procedures.
^ The result of the dose confirmation analysis for soman was 23% of the expected concentration of 0.30 mg/L.
10
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Table 3-4. Water Quality Parameters
Parameter
Turbidity
Organic carbon
Specific conductivity
Alkalinity
pH
Hardness
Total organic halides
Total trihalomethanes
Total haloacetic acids
Dechlorinated Columbus,
Ohio, Tap Water (disinfected
Method by chlorination)
EPA180.1(7)
SM5310(8)
SM2510(8)
SM 2320(8)
EPA150.1(9)
EPA 130.2(9)
SM 5320B(8)
EPA 524.2(10)
EPA 552.2(11)
0.1NTU(a)
2.5 mg/L
364 [irnho
42 mg/L
7.65
112 mg/L
190 |ig/L
52.8 |ig/L
75.7 |ig/L
Dechlorinated
St. Petersburg, Florida,
Tap Water (disinfected by
chloramination)
0.3 NTU
2.9 mg/L
460 [irnho
97 mg/L
7.95
160 mg/L
83 |ig/L
2.4 fig/L
13.5 |ig/L
(a) NTU = nephelometric turbidity units
11
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Chapter 4
Quality Assurance/Quality Control
QA/QC procedures were performed in accordance with the quality management plan (QMP) for
the AMS Center^ and the test/QA plan for this verification test.(1)
4.1 Quality Control of Stock Solution Confirmation Methods
The stock solutions for aldicarb, cyanide, dicrotophos, and thallium sulfate were analyzed using a
standard reference method at ATEL. As part of ATEL's standard operating procedures (SOPs),
various QC samples were analyzed with each sample set. These included matrix spike, laboratory
control spike, and method blank samples. According to the standard methods used for the
analyses, recoveries of the QC spike samples analyzed with samples from this verification test
were within acceptable limits of 75% to 125%, and the method blank samples were below the
detectable levels for each analyte. For VX and soman, the confirmation analyses were performed
at Battelle using a Battelle SOP. Calibration standard recoveries of VX and soman were always
between 69% and 130%, and most of the time were between 90% and 100%. Standard analytical
methods for colchicine, ricin, and botulinum toxin were not available and, therefore, were not per-
formed. QA audits and balance calibrations assured that solutions for these compounds were
accurately prepared.
4.2 Quality Control of Drinking Water Samples
A method blank sample consisting of ASTM Type n DI water was analyzed once by Eclox for
approximately every 20 drinking water samples that were analyzed. According to the Eclox
procedure, the first sample of each analysis set is treated as the control sample that is used to
correct the response of the instrument with respect to a clean water sample. For this verification
test, this sample was the method blank. When the method blank sample (ASTM Type n DI water)
was analyzed, Eclox did not report a percent inhibition. Toward the end of testing, it was
ascertained that, to obtain inhibition data about the method blank samples, ASTM Type n DI
water should have been analyzed as a sample in some position other than the first in the analysis
set. Two method blank samples were analyzed in this manner, producing small inhibitions of 3%
and 2%. A negative control sample (unspiked DDW) was analyzed with approximately every four
samples. The absolute inhibitions of the negative controls were small, indicating that they caused
inhibition similar to the ASTM Type n DI water, which was used as the zero control sample (i.e.,
set to zero inhibition). A positive control sample also was analyzed once for approximately every
20 DDW samples. While performance limits were not placed on the results of the positive control
12
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sample, the vendor informed Battelle that, if the positive control samples did not cause almost
complete inhibition, it would indicate to the operator that Eclox was operating incorrectly. For 14
positive control samples of phenol, the average inhibition was 99% ± 6%.
4.3 Audits
4.3.1 Performance Evaluation Audit
The concentration of the standards used to prepare the contaminant and potential interferences
was confirmed by analyzing solutions of each analyte prepared in ASTM Type n DI water from
two separate commercial vendors using the confirmation methods. The standards from one source
were used to prepare the stock solution during the verification test, while the standards from a
second source were used exclusively to confirm the accuracy of the measured concentration of the
first source. The percent difference (%D) between the measured concentration of the performance
evaluation (PE) sample and the prepared concentration of that sample was calculated using the
following equation:
%D = — x 100%
A
(1)
whereMis the absolute value of the difference between the measured and the prepared concen-
tration and A is the prepared concentration. The %D between the measured concentration of the
PE standard and the prepared concentration had to be less than 25% for the measurements to be
considered acceptable. Table 4-1 shows the results of the PE audit for each compound. All %D
values were less than 25.
Table 4-1. Summary of Performance Evaluation Audit
Contaminant
Potential
interference
Aldicarb
Cyanide
Dicrotophos
Thallium sulfate
Aluminum
Copper
Iron
Manganese
Zinc
Average Measured
Concentration ±
Standard Deviation
(mg/L)
0.00448 ±0.000320
0.207 ±0.026
0.00728 ±0.000699
0.090 ±0.004
0.512 ±0.013
0.106 ±0.002
0.399 ±0.004
0.079 ±0.003
0.106 ±0.016
Actual
Concentration
(mg/L)
0.00500
0.200
0.00748
0.100
0.500
0.100
0.400
0.100
0.100
Percent
Difference
11
4
3
10
2
6
0.30
21
6
13
-------
Given the lack of confirmation methodology for some of the contaminants in this verification
test, PE audits were not performed for all of the contaminants. PE audits were performed when
more than one source of the contaminant or potential interference was commercially available
and when methods were available to perform the confirmation. To assure the purity of the other
standards, documentation, such as certificates of analysis, was obtained for colchicine,
botulinum toxin, and ricin. In the case of VX and soman, which were obtained from the U.S.
Army, the reputation of the source, combined with the confirmation analysis data, provided
assurance of the concentration analyzed.
4.3.2 Technical Systems Audit
The Battelle Quality Manager conducted a technical systems audit (TSA) to ensure that the
verification test was performed in accordance with the test/QA plan(1) and the AMS Center
QMP.(12) As part of the audit, the Battelle Quality Manager reviewed the contaminant standard
and stock solution confirmation methods, compared actual test procedures with those specified
in the test/QA plan, and reviewed data acquisition and handling procedures. Observations and
findings from this audit were documented and submitted to the Battelle verification test
coordinator for response. No findings were documented that required any significant action. The
records concerning the TSA are permanently stored with the Battelle Quality Manager.
The EPA Quality Manager also conducted a TSA to ensure that the verification test was
performed in accordance with the test/QA plan(1) and the AMS Center QMP.(12) As part of the
audit, the EPA Quality Manager compared actual test procedures with those specified in the
test/QA plan and reviewed data acquisition and sample preparation records and procedures. No
significant findings were observed during the EPA TSA. The records concerning the TSA are
permanently stored with the EPA Quality Manager.
4.3.3 Audit of Data Quality
At least 10% of the data acquired during the verification test were audited. Battelle's Quality
Manager traced the data from the initial acquisition, through reduction and statistical analysis,
to final reporting, to ensure the integrity of the reported results. All calculations performed on
the data undergoing the audit were checked.
4.4 QA/QC Reporting
Each internal assessment and audit was documented in accordance with Sections 3.3.4 and 3.3.5
of the QMP for the ETV AMS Center.(12) Once the assessment report was prepared, the Battelle
verification test coordinator ensured that a response was provided for each adverse finding or
potential problem and implemented any necessary follow-up corrective action. The Battelle
Quality Manager ensured that follow-up corrective action was taken. The results of the TSA
were sent to the EPA.
14
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4.5 Data Review
Records generated in the verification test were reviewed before these records were used to
calculate, evaluate, or report verification results. Table 4-2 summarizes the types of data
recorded. The review was performed by a technical staff member involved in the verification
test, but not the staff member who originally generated the record. The person performing the
review added his/her initials and the date to a hard copy of the record being reviewed.
Table 4-2. Summary of Data Recording Process
Data to be
Recorded
Responsible
Party
Where
Recorded
How Often
Recorded
Disposition of Data(a)
Dates, times of test Battelle
events
Sample preparation Battelle
(dates, procedures,
concentrations)
Test parameters Battelle
(contaminant
concentrations,
location, etc.)
Laboratory
record books
Laboratory
record books
Start/end of test, and
at each change of a
test parameter
When each sample
was prepared
Laboratory
record books
When set or
changed
Used to organize/check
test results; manually
incorporated in data
spreadsheets as
necessary
Used to confirm the
concentration and
integrity of the samples
analyzed, procedures
entered into laboratory
record books
Used to organize/check
test results, manually
incorporated in data
spreadsheets as
necessary
Stock solution
confirmation
analysis, sample
analysis, chain of
custody, and
results
Battelle or
contracted
laboratory
Laboratory
record books,
data sheets, or
data acquisition
system, as
appropriate
Throughout sample
handling and
analysis process
Transferred to
spreadsheets/agreed
upon report
(a) All activities subsequent to data recording were carried out by Battelle.
15
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Chapter 5
Statistical Methods and Reported Parameters
The statistical methods presented in this chapter were used to verify the performance parameters
listed in Section 3.1.
5.1 Endpoints and Precision
Each DDW sample containing contaminants was compared with a negative control sample that,
for this verification test, was unspiked DDW. This comparison was made by subtracting the
percent inhibition of the negative control within a sample set from the inhibition produced by
each sample in the sample set. Therefore, the percent inhibition of the negative control sample
within each sample set was zero percent.
The standard deviation (S) of the results for the replicate samples was calculated, as follows, and
used as a measure of technology precision at each concentration.
where n is the number of replicate samples, Ik is the percent inhibition measured for the A*h
sample, and / is the average percent inhibition of the replicate samples. Because the average
inhibitions were frequently near zero for this data set, relative standard deviations often would
have greatly exceeded 100%, making the results difficult to interpret. Therefore, the precision
results were left in the form of standard deviations so the reader could easily view the
uncertainty around the average for results that were both near zero and significantly larger than
zero.
5.2 Toxicity Threshold
The toxicity threshold was defined as the lowest concentration of contaminant to exhibit a
percent inhibition significantly greater than the negative control. Also, each concentration level
higher than the toxicity threshold had to be significantly greater than the negative control, and
the inhibition produced by each lower concentration level had to be significantly less than that
produced by the toxicity threshold concentration. Since the inhibition of the negative control
16
-------
sample was subtracted from the inhibition of each sample, the percent inhibition of the negative
control was always zero. An inhibition was significantly greater than the negative control if the
average, plus or minus the standard deviation, did not include zero.
5.3 False Positive/Negative Responses
A response would be considered false positive if an unspiked drinking water sample produced
an inhibition such that the subsequent addition of toxic contaminants could not be detected.
Depending on the degree of inhibition in the sample, toxicity due to subsequent contamination
of that sample may not be detectable or could be exaggerated as a result of the baseline
inhibition. To test for this possibility, the percent inhibition of the unspiked drinking water was
determined with respect to ASTM Type n DI water. Therefore, the result of the negative control
was not subtracted from the result for these samples. The percent inhibition of drinking water
samples collected from water systems using chlorination and chloramination as the disinfecting
process were reported as determined by Eclox with no further correction. For Eclox, a result
would be considered false positive if the drinking water samples produced inhibitions
significantly greater than zero.
A response was considered false negative when Eclox was subjected to a lethal concentration of
some contaminant in the DDW and did not indicate inhibition significantly greater than the
negative control and the other concentration levels analyzed. Requiring the inhibition of the
lethal dose sample to be significantly greater than the negative control and the other concentra-
tion levels more thoroughly incorporated uncertainty for Eclox when determining a false
negative response. For any result to be significantly different from the negative control, the
inhibition needed to be significantly greater than zero.
5.4 Field Portability
The results obtained from the measurements made on DDW samples in the laboratory and field
setting were compiled independently and compared to assess the performance of the Eclox under
different analysis conditions. Means and standard deviations of the endpoints generated in both
locations were used to make the comparison. Also, qualitative observations of Eclox in a non-
laboratory setting were made by the verification test coordinator and operators. Factors such as
the ease of transport and set-up, demand for electrical power, and space requirement were
documented.
5.5 Other Performance Factors
Ease of use (including clarity of the instruction manual, user-friendliness of software, and
overall convenience) was qualitatively assessed throughout the verification test through
observations of the operators and verification test coordinator. Sample throughput was evaluated
quantitatively based on the number of samples that could be analyzed per hour.
17
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Chapter 6
Test Results
6.1 Endpoints and Precision
Tables 6-1 a-i present the percent inhibition data for nine contaminants, and Table 6-2 presents
data for five potential interferences and drinking water samples disinfected by both chlorination
and chloramination. Given in each table are the concentrations analyzed, the percent inhibition
results for each replicate at each concentration, and the average and standard deviation of the
inhibition of the four replicates at each concentration. Samples that produced negative percent
inhibition values indicated an increase in light production by the enzyme relative to the negative
control.
6.1.1 Contaminants
The contaminants that were analyzed by Eclox during this verification test resulted in percent
inhibition data that varied considerably among contaminants. The percent inhibitions for
aldicarb, dicrotophos, thallium sulfate, ricin, and VX were significantly different from the
negative control and the lower concentration levels for only the highest concentration level
(lethal dose). For colchicine, the percent inhibition increased steadily in proportion to the
concentration in the sample. Eclox was especially sensitive to cyanide at concentrations near the
lethal dose. Complete inhibition was produced for cyanide concentrations from the lethal dose to
at least as low as 0.25 mg/L, one thousand times less concentrated than the lethal dose. No
detectable inhibition was produced by botulinum toxin or soman.
6.1.2 Potential Interferences
Table 6-2 presents the results from the samples that were analyzed to test the effect of potential
interferences on Eclox. Aluminum, copper, and iron exhibited percent inhibitions near zero,
indicating little or no response to these compounds, while manganese and zinc exhibited higher
inhibitions of 62% and 10%, respectively.
18
-------
Table 6-1 a. Aldicarb Percent Inhibition Results
Concentration
(mg/L)
0.28
2.8
28
280
(Lethal Dose)
Inhibition Average
h
8
-4
15 7
8
7
1
4 4
6
31
39
32
36
Table 6-1 b. Colchicine Percent Inhibition
Concentration
(mg/L)
0.24
2.4
24
240
(Lethal Dose)
Inhibition Average
5
14 9
15
13
V
17
40
50 43
37
44
87
100
84
96
Standard
Deviation
10
8
2
4
Results
Standard
Deviation
7
6
6
8
19
-------
Table 6-1 c. Cyanide Percent Inhibition Results
Concentration
(mg/L)
0.025
0.05
0.25
Inhibition Average
(%) (%)
-7
-,'
11
22
/9 13
4
95
109 103
98
Standard
Deviation
8
9
7
96
250 ^ 97
97
4
0.05 25 ^ 1Q
(Field Location) 5
19
Table 6-ld. Dicrotophos Percent Inhibition Results
Concentration
(mg/L)
1.4
14
140
1,400
(Lethal Dose)
Inhibition Average
(%) (%)
-14
;'
7
5
\
3
5
75 4
8
27
27 29
28
34
Standard
Deviation
10
6
6
3
20
-------
Table 6-1 e. Thallium Sulfate Percent Inhibition Results
Concentration
(mg/L)
2.4
24
240
2,400
(Lethal Dose)
Inhibition
(%)
-4
-2
-7
0
-8
-1
-5
3
-3
12
-3
13
43
49
46
45
Table 6-lf. Botulinum Toxin
Concentration
(mg/L)
0.0003
0.003
0.03
0.30
(Lethal Dose)
Inhibition
(%)
-3
-1
1
4
1
-1
-6
-2
-3
-3
-2
-3
1
0
-4
-4
Standard
Average Deviation
(%) (%)
-3 3
-3 5
5 9
46 3
Percent Inhibition Results
Standard
Average Deviation
(%) (%)
1 3
-2 3
-3 1
-2 3
21
-------
Table 6-1 g. Ricin Percent Inhibition Results
Concentration
(mg/L)
0.015
0.15
1.5
Inhibition Average
(%) (%)
-2
-5,
1
0
I
3
1
!
0
Standard
Deviation
3
2
2
8
15 11
(Lethal Dose) 7
5
Table 6-lh. Soman Percent Inhibition Results
Concentration
(mg/L)
0.000068
0.00068
0.0068
Inhibition Average
(%) (%)
5
I
4
-2
2
4
5
S
1
Standard
Deviation
2
3
2
8
0.068(a) -3
(Lethal Dose) -3
0
^a' NS = Samples not fortified with any contaminant or potential interference.
^ Due to the degradation of soman in water, the stock solution confirmation
analysis confirmed that the concentration of the lethal dose was 23% of the
expected concentration of 0.30 mg/L.
22
-------
Table 6-1 i. VX Percent Inhibition Results
Concentration
(mg/L)
0.00049
0.0049
0.049
0.49
(Lethal Dose)
Inhibition
(%)
-7
-10
-5
-7
-7
-3
-5
-6
-6
-5
1
-4
13
7
11
6
Standard
Average Deviation
(%) (%)
-7 2
-5 2
-4 3
9 3
All of the contaminant and potential interference samples were prepared in the DDW sample and
compared with an unspiked DDW sample. Therefore, any background inhibition in the DDW
sample was corrected by subtracting the inhibition caused by the negative control sample. To
investigate whether Eclox is sensitive to by-products of disinfecting processes, dechlorinated
drinking water samples from water systems that use chlorination and chloramination were
analyzed and compared with ASTM Type n DI water as the baseline sample. This determination
is crucial because the ability of Eclox to detect toxicity is dependent on the light production of
the Eclox reagent in a clean drinking water matrix. If clean drinking water produces 100%
inhibition of light, the detection of subsequently added contaminants would not be possible. On
average, the chlorinated sample exhibited inhibitions of 6% ± 5%, while the chloraminated
sample exhibited inhibitions of 0% ± 2%. This suggests that by-products of either disinfection
process that may be present in drinking water do not interfere with Eclox results.
6.1.3 Precision
Across all the contaminants and potential interferences, the standard deviation was measured
and reported for each set of four replicates to evaluate the Eclox precision. The standard devia-
tion of the four replicate measurements was never greater than 10%.
23
-------
Table 6-2. Potential Interferences Results
Concen-
tration Inhibition Average Standard Deviation
Compound (mg/L) (%) (%) (%)
Aluminum 0.36
Copper 0.65
Iron 0.069
Manganese 0.26
ZmC 3.5
Chlorination NA(a)
by-products
Chloramination .
U J 4. NA
by-products
-8
-2
8
-5
0
0 4
17
0
2
° 2
10 Z
-3
62
70 69
58 62
57
8
?
6
(b) g
2
J
-2
7
9
6
6
4
5
2
A) NA = Not applicable.
Chlorination by-product data averaged over the negative control results with respect to the inhibition
of ASTM Type IIDI water.
6.2 Toxicity Threshold
Table 6-3 gives the toxicity thresholds as defined in Section 5.2 for each contaminant. The
lowest toxicity threshold concentration was for cyanide at 0.25 mg/L, indicating that Eclox was
most sensitive to cyanide. For botulinum toxin and soman, no inhibition significantly greater
than the negative control was detected regardless of the concentration level, indicating that the
technology was not highly responsive to these contaminants.
24
-------
Table 6-3. Toxicity Thresholds
Contaminant
Aldicarb
Colchicine
Cyanide
Dicrotophos
Thallium sulfate
Botulinum toxin
Ricin
Soman
VX
Concentration (mg/L)
280
24
0.25
1,400
2,400
ND(a)
15
ND
0.49
= Significant inhibition was not detected.
6.3 False Positive/Negative Responses
No false positive responses were generated by Eclox. High background light production (low
inhibitions with respect to ASTM Type n DI water) in both chlorinated and chloraminated
drinking water samples allowed for the possibility of detection of contaminants.
A false negative response is when a lethal dose of contaminant is present in the water sample
and no inhibition is detected. Table 6-4 gives each contaminant's lethal dose concentration and
shows whether or not the inhibition was also significantly different from zero at that concentra-
tion level. The inhibition induced by lethal doses of aldicarb, colchicine, cyanide, dicrotophos,
thallium sulfate, ricin, and VX was significantly different from zero, while botulinum toxin and
soman were not detected at the lethal dose, indicating false negative responses. Nerve agent test
strips supplied with the Eclox kit were not tested, only the chemiluminescent toxicity test was
conducted. The vendor states that the nerve agent test strip will detect soman.
6.4 Field Portability
A single concentration of cyanide was prepared and analyzed in replicate at a field location to
examine the ability of Eclox to be used in a non-laboratory setting. Eclox and necessary
accessories were conveniently transported to the field in the hard plastic carrying case provided
by the vendor. The carrying case was equipped with holders for each reagent and needed
accessories and a waste container to store the small amount of waste generated until it could be
disposed of properly. Also, detailed instructions on performing the test were permanently
attached to the lid of the case. Fully loaded, the case weighed about 20 pounds. At the field
location, Eclox was operated with four "AA" batteries on a small table in the basement of a
house. Table 6-lc shows the results of the cyanide samples analyzed at the field location, along
with the results of the cyanide samples analyzed in the laboratory. The concentration of the
25
-------
Table 6-4. False Negative Responses
Lethal Dose
Concentration False Negative
Contaminant (mg/L) Response
Aldicarb
Colchicine
Cyanide
Dicrotophos
Thallium sulfate
Botulinum toxin
Ricin
Soman
VX
280
240
250
1,400
2,400
0.30
15
0.068
0.49
no
no
no
no
no
yes
no
yes
no
solution analyzed in the field was 0.05 mg/L. The inhibition produced in the field was 13% ±
10%, and the inhibition produced in the laboratory at the same concentration was 13% ± 9%,
indicating that Eclox functioned similarly at the laboratory and non-laboratory locations. The
Eclox reagent was easy to prepare and will last up to a year as long as it is kept at approximately
4°C, making it ideal for field portability if coolers are available for overnight storage.
6.5 Other Performance Factors
The analysis procedure for Eclox was very straightforward. The instructions on the lid of the
case were detailed and easy to understand. Although the ETV operators had scientific
backgrounds, based on observations of the test coordinator, operators with little technical
training would probably be able to operate Eclox successfully with no instruction other than the
in-case manual. All reagents and pipettes were color-coded to assist operators in identifying the
correct items. The carrying case was used as a sample and reagent holder during testing in the
laboratory, as well as in the field, because of the convenient way in which it was designed. Eclox
must be operated on batteries because there is no electrical power option. The operators analyzed
15 samples per hour.
26
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Chapter 7
Performance Summary
Parameter
Contaminants in
DDW
Potential
interferences in
DDW
False positive
response
False negative
response
Compound
Aldicarb
Colchicine
Cyanide
Dicrotophos
Thallium
sulfate
Botulinum
toxm(c)
Ricm(e)
Soman
VX
Interference
Aluminum
Copper
Iron
Manganese
Zinc
Lethal
Dose (LD)
Cone.
(mg/L)
280
240
250(b)
1,400
2,400
0.30
15.0
0.068®
0.49
Cone.
(mg/L)
0.36
0.65
0.069
0.26
3.5
Average Inhibitions at Concentrations
Relative to the LD Concentration (%)
LD
35
92
97
29
46
-2
8
0
9
LD/10
4
43
103
4
5
o
-J
2
2
-4
LD/100
7
14
13
2
-3
-2
2
2
-5
LD/1,000
10
9
3
-1
-3
1
1
o
J
-7
Average Inhibitions at a
Single Concentration (%)
-2
4
2
62
10
Range of
Standard
Deviations
(%)
2-10
6-8
1-9
3-10
3-9
1-3
2-3
2-5
2-3
Standard
Deviation
(%)
7
9
6
6
4
Chlorinated (6% ± 5%) and chloraminated (0% ± 2%) drinking water samples were not
inhibitory with respect to ASTM Type II DI water. This shows that there were no false ]
responses.
Toxicity
Thresh.
(mg/L)(a)
280
24
0.25
1,400
2,400
ND(d)
15
ND
0.49
positive
At the lethal concentration level, inhibitions produced by botulinum toxin and soman were not
significantly different from the negative control or inhibitions generated by lower concentrations
of the same contaminant, indicating false negative responses.
Field portability
Inhibitions for cyanide at 0.05 mg/L at the field location were 13% ± 10%, while laboratory
testing of the same concentration produced an inhibition of 13% ± 9%. Eclox was easily
transported and operated in the field. Detailed instructions in the carrying case and organized
packaging made field analysis convenient.
Other
performance
factors
Although the operators had scientific backgrounds, upon observation of the test procedures, it
seems likely that operators with little technical training would probably be able to operate Eclox
by following the detailed instructions provided with Eclox. Reagents and pipettes were color-
coded to ensure mistake-free analysis. Waste container was included. Operators were able to
analyze 15 samples per hour in this test.
See Tables 6-la-I in the report for the precision around each individual inhibition result.
Cyanide LD/10, LD/100, and LD/1,000 concentrations are 0.25, 0.05, and 0.025 mg/L.
Lethal dose solution also contained 3 mg/L phosphate and 1 mg/L sodium chloride.
ND = Not detectable.
Lethal dose solution also contained 3 mg/L phosphate, 26 mg/L sodium chloride, and 2 mg/L sodium azide.
Due to the degradation of soman in water, the stock solution confirmation analysis confirmed that the concentration of the
lethal dose was 23% of the expected concentration of 0.30 mg/L.
27
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Chapter 8
References
1. Test/QA Plan for Verification of Rapid Toxicity Technologies, Battelle, Columbus, Ohio,
June 2003.
2. United States Environmental Protection Agency, National Secondary Drinking Water
Regulations: Guidance for Nuisance Chemicals, EPA/810/K-92/001, July 1992.
3. U.S. EPAMethod 531.1, "Measurement of n-Methylcarbamoyloximes and
n-Methylcarbamates in Water by Direct Aqueous Injection HPLC with Post Column
Derivatization," in Methods for the Determination of Organic Compounds in Drinking
Water—Supplement III, EPA/600/R-95/131, 1995.
4. U.S. EPA Method 335.1, "Cyanides, Amenable to Chlorination," in Methods for the
Chemical Analysis of Water and Wastes, EPA/600/4-79/020, March 1983.
5. SW846 Method 8141 A, "Organophosphorous Compounds by Gas Chromatography:
Capillary Column Technique," Revision 1, September 1994.
6. U.S. EPA Method 200.8, "Determination of Trace Elements in Waters and Wastes by
Inductively-Coupled Plasma Mass Spectrometry," in Methods for the Determination of
Metals in Environmental Samples, Supplement I, EPA/600/R-94/111, 1994.
7. U.S. EPA Method 180.1, "Turbidity (Nephelometric)," Methods for the Determination of
Inorganic Substances in Environmental Samples, EPA/600/R-93/100, 1993.
8. American Public Health Association, et al. Standard Methods for the Examination of Water
and Wastewater. 19th Edition, 1997. Washington, DC.
9. U.S. EPA, Methods for Chemical Analysis of Water and Wastes, EPA/600/4-79/020.
10. U.S. EPA Method 524.2, "Purgeable Organic Compounds by Capillary Column GC/Mass
Spectrometry," Methods for the Determination of Organic Compounds in Drinking
Water—Supplement III, EPA/600/R-95/131.
28
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11. U.S. EPA Method 552.2, "Haloacetic Acids and Dalapon by Liquid-Liquid Extraction,
Derivatization and GC with Electron Capture Detector," Methods for the Determination of
Organic Compounds in Drinking Water—Supplement IIIEPA/600/R-95/131.
12. Quality Management Plan (QMP) for the ETV Advanced Monitoring Systems Center,
Version 4.0, U.S. EPA Environmental Technology Verification Program, Battelle,
Columbus, Ohio, December 2002.
29
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