United States
Environmental Protection
Agency
Environmental Monitoring
Systems Laboratory
Las Vegas, NV 89193-3478
Research and Development
EPA/600/S4-91/028 June 1992
or EPA Project Summary
Superfund Innovative
Technology Evaluation
Demonstration Plan for
Westinghouse Bio-Analytic
Systems Pentachlorophenol
Immunoassays
M.E. Silverstein, R.J. White, R.W. Gerlach, and J.M. Van Emon
This report has been prepared under
the Superfund Innovative Technology
Evaluation (SITE) Program and provides
a detailed description of the SITE dem-
onstration of the Westinghouse Bio-
Analytic Systems (WBAS) immunoas-
say technologies specific to the analy-
sis of pentachlorophenol. The immu-
noassays measure parts per billion con-
centrations of pentachlorophenol in
environmental water samples.
The primary objective of this dem-
onstration is to evaluate on site a
semiquantitative immunoassay field kit
for its utility as a rapid field screening
tool. This demonstration plan provides
the protocols required to obtain the
information needed for the evaluation.
Each aspect of the evaluation is de-
scribed, including test site description,
logistical and equipment consider-
ations, sample collection, quality as-
surance, and data analysis.
This plan is submitted in partial ful-
fillment of contract number 68-03-3249
and contract number 68-CO-0049 by
Lockheed Engineering & Sciences Com-
pany under the sponsorship of the U.S.
Environmental Protection Agency.
This Project Summary was develped
by EPA's Environmental Monitoring
Systems Laboratory, Las Vegas, NV, to
announce key findings of the research
project that is fully documented in a
separate report of the same title (see
Project Report ordering informtion at
back).
Introduction
Two immunoassays will be tested in
this evaluation, one formatted as a field
kit and the other as a plate immunoassay.
The field kit will be compared to a quanti-
tative high-sample-capacity plate immu-
noassay developed by WBAS that was
previously evaluated at the U.S. Environ-
mental Protection Agency (EPA), Environ-
mental Monitoring Systems Laboratory,
Las Vegas, NV (EMSL-LV). Both of these
immunoassay techniques will be compared
to results from a standard EPA gas chro-
matography/mass spectrometry method for
the analyses of pentachlorophenol in wa-
ter. The demonstration will be conducted
at the MacGillis & Gibbs Superfund Site in
New Brighton, MN. This is a National Pri-
orities List site known to have ground wa-
ter contaminated with pentachlorophenol.
The immunoassay demonstration will be
performed in tandem with a separate SITE
demonstration of a bioremediation tech-
nology (a bioreactor developed by BioTrol,
Inc., Chaska, MN) that is designed to bio-
degrade pentachlorophenol in water.
Description of Immunoassay
Technology
Immunoassays are based on receptor
molecules called antibodies which are de-
veloped in response to a particular target
analyte. Quantification of the extent of con-
tamination in an environmental sample
is based on the ability of a specific anti-
body to bind to its target analyte. Immu-
noassays are normally based on competi-
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tion for antibody binding between a known
amount of analyte labeled with an indica-
tor, such as an enzyme, and an unknown
amount of analyte from a sample. The
indicator produces a colored product that
is used for quantitation. Color intensity is
determined by the amount of analyte
present. Immunoassays can be configured
for quantitative, semiquantitative, or quali-
tative analyses.
The field kit methodology requires about
30 minutes to perform and has a detec-
tion limit of about 3 ppb. It has a linear
dynamic range from about 3 to 40 ppb
and uses no more than 2 ml of sample to
obtain analytical results. The kit immunoas-
say employs a portable spectrophotom-
eter for standard curve generation and
quantitation of pentachlorophenol concen-
trations and it requires a clean, sheltered
work area (e.g., out of the wind and direct
sunlight).
The quantitative (i.e., plate) immunoas-
say is based on a 96-well microtiter plate
format. The minimum detectable level of
pentachlorophenol is approximately 30
ppb; however, when the required sample
dilutions are considered, the minimum de-
tection level is 90 ppb, based on an 18-
ml_ sample. The method has a linear dy-
namic range of 30 to 400 ppb. Several
96-well microtiter plates can be processed
in tandem. Thus, hundreds of samples
can be analyzed during the 2.5-hour analy-
sis time required for this method. The
plate immunoassay also can be performed
under field conditions in a mobile labora-
tory. However, for this demonstration, only
the field kit will be evaluated under field
conditions because this methodology is
particularly suited for use by field person-
nel who may have limited analytical chem-
istry experience.
Sampling and Analysis Designs
The sample collection, sample analysis,
and data analysis strategies presented in
this plan are designed to address the criti-
cal issues related to assessing the gen-
eral and practical applications of immu-
noassay technology in the measurement
and monitoring aspects of the Superfund
Program. Results obtained from on site
sample analysis using the field kit immu-
noassay will be compared to results ob-
tained by analyzing splits of the same
samples by a more conventional analyti-
cal method. For this comparison, EPA
Jeanette M. Van Emon (also the EPA Project Officer) is with the Environmental
Monitoring Systems Laboratory, Las Vegas, NV 89193-3478. M.E. Silverstein,
R.J. White, and R.W. Gerlach are with Lockheed Engineering & Sciences
Company, Las Vegas, NV 89119.
The complete report, entitled "Superfund Innovative Technology Evaluation Dem-
onstration Plan for Westinghouse Bio-Analytic Systems Pentachlorophenol Im-
munoassays, " (Order No. PB92-170190/AS; Cost: $26.00; subject to change)
will be available only from:
National Technical Information Service
5285 Port Royal Road
Springfield, VA 22161
Telephone: 703-487-4650
The EPA Project Officer can be contacted at:
Environmental Monitoring Systems Laboratory
U.S. Environmental Protection Agency
Las Vegas, NV 89193-3478
Method 8270, a gas chromatography/mass
spectrometry method for semivolatile or-
ganics will be used to analyze for pen-
tachlorophenol. Split field samples will also
be analyzed off site by both immunoassay
techniques (field kit and plate) at EMSL-
LV and WBAS laboratories. A detailed
quality assurance plan for this demonstra-
tion is provided in an appendix.
Data Analysis and Management
The demonstration plan includes de-
tailed standard operating procedures for
sample analysis and data management.
The quality assurance plan is designed to
ensure that important data quality and
methodological performance criteria are
examined. A series of performance evalu-
ation (audit) samples, as well as blank
and replicate samples, are incorporated in
the analytical scheme to assess the within-
method performance parameters of the
immunoassay and to perform between-
method comparisons. The data manage-
ment system is tailored to the sample
analysis and quality assurance programs
to provide a timely means of performing
the data analysis. Data management will
also provide a mechanism for document-
ing and tracking the data generated from
the different analysis sites and by the vari-
ous methods.
Health and Safety
Pentachlorophenol is considered a toxic
substance and a suspected carcinogen.
The plan addresses health and safety as-
pects associated with handling and dis-
posing of materials contaminated with pen-
tachlorophenol.
United States
Environmental Protection
Agency
Center for Environmental
Research Information
Cincinnati, OH 45268
BULK RATE
POSTAGE & FEES PAID
EPA
PERMIT No. G-35
Official Business
Penalty for Private Use $300
EPA/600/S4-91/028
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