United States
Environmental Protection
Agency
Office Of Water
(4305)
EPA 823-R-93-002
August 1993
£EPA
Guidance For Assessing
Chemical Contaminant Data
For Use In Fish
Volume 1
Fish Sampling And Analysis
-------
GUIDANCE FOR ASSESSING CHEMICAL CONTAMINANT DATA
FOR USE IN FISH ADVISORIES
VOLUME 1: FISH SAMPLING AND ANALYSIS
Contract No. 68-C3-0303
Prepared for
Work Assignment Managers
Jeffrey Bigler
Alison Greene
Office of Science and Technology
Office of Water
U.S. Environmental Protection Agency
Washington, DC
-------
TABLE OF CONTENTS
TABLE OF CONTENTS
List of Figures vi
List of Tables viii
Acknowledgments xi
1 Introduction 1-1
1.1 Historical Perspective 1-1
1.2 Purpose 1-3
1.3 Objectives 1-4
1.4 Relationship of Manual to Other Guidance Documents 1-5
1.5 Organization of this Manual 1-6
2 Monitoring Strategy 2-1
2.1 Screening Studies (Tier 1) 2-4
2.2 Intensive Studies (Tier 2) 2-14
3 Target Species 3-1
3.1 Purpose of Using Target Species 3-1
3.2 Criteria for Selecting Target Species 3-2
3.3 Freshwater Target Species 3-3
3.3.1 Bottom-Feeding Target Species 3-5
3.3.2 Predator Target Species 3-5
3.4 Estuarine/Marine Target Species 3-11
3.4.1 Selection of Target Shellfish Species 3-20
3.4.2 Selection of Target Finfish Species 3-24
4 Target Analytes 4-1
4.1 Recommended Target Analytes 4-1
4.2 Selection of Target Analytes 4-4
4.3 Target Analyte Profiles 4-4
4.3.1 Metals 4-4
4.3.2 Organochlorine Pesticides 4-10
4.3.3 Organophosphate Pesticides 4-18
4.3.4 Chlorophenoxy Herbicides 4-23
4.3.5 Polychlorinated Biphenyls (Total) 4-23
4.3.6 Dioxins and Dibenzofurans 4-27
4.4 Target Analytes under Evaluation 4-32
4.4.1 Metals 4-33
4.4.2 Organics 4-35
in
-------
TABLE OF CONTENTS
Page
5 Screening Values for Target Analytes 5-1
5.1 General Equations for Calculating Screening Values 5-2
511.1 Noncarcinogens 5-3
5.1.2 Carcinogens 5-3
5.1.3 Recommended Values for Variables in Screening
Value Equations 5-4
5.2 Recommended Screening Values for Target Analytes 5-5
5.3 Comparison of Target Analyte Concentrations
with Screening Values 5-13
5.3.1 Metals 5-13
5.3.2 Organics 5-14
6 Field Procedures 6-1
6.1 Sampling Design 6-1
6.1.1 Screening Studies (Tier 1) 6-2
6.1.2 Intensive Studies (Tier 2) 6-12
6.2 Sample Collection 6-22
6.2.1 Sampling Equipment and Use 6-22
6.2.2 Preservation of Sample Integrity 6-29
6.2.3 Field Recordkeeping 6-30
6.3 Sample Handling 6-40
6.3.1 Sample Selection 6-40
6.3.2 Sample Packaging 6-45
6.3.3 Sample Preservation 6-47
6.3.4 Sample Shipping 6-48
7 Laboratory Procedures I—Sample Handling 7-1
7.1 Sample Receipt and Chain-of-Custody 7-1
7.2 Sample Processing 7-3
7.2.1 General Considerations 7-4
7.2.2 Processing Fish Samples 7-6
7.2.3 Processing Shellfish Samples 7-16
7.3 Sample Distribution 7-21
7.3.1 Preparing Sample Aliquots 7-21
7.3.2 Sample Transfer 7-24
8 Laboratory Procedures II—Sample Analyses 8-1
8.1 Recommended Analytes 8-1
8.1.1 Target Analytes 8-1
8.1.2 Lipid 8-1
8.2 Analytical Methods 8-3
8.2.1 Lipid Method 8-3
8.2.2 Target Analyte Methods 8-3
8.3 Quality Assurance and Quality Control Considerations 8-9
8.3.1 QA Plans 8-11
IV
-------
TABLE OF CONTENTS
8.3.2 Method Documentation 8-11
8.3.3 Minimum QA and QC Requirements for Sample
Analyses 8-12
8.4 Documentation and Reporting of Data 8-46
8.4.1 Analytical Data Reports 8-46
8.4.2 Summary Reports 8-48
9 Data Analysis and Reporting 9-1
9.1 Data Analysis 9-1
9.1.1 Screening Studies 9-1
9.1.2 Intensive Studies 9-2
9.2 Data Reporting 9-3
9.2.1 State Data Reports 9-3
9.2.2 Reports to the National Fish Tissue Data Repository ... 9-3
10 Literature Cited 10-1
Appendixes
A Fish and Shellfish Species for which State Consumption Advisories
Have Been Issued A-1
B Target Analytes Analyzed in National or Regional Monitoring
Programs B-1
C Pesticides and Herbicides Recommended as Target Analytes .... C-1
D Target Analyte Dose-Response Variables and Associated
Information D-1
E Quality Assurance and Quality Control Guidance E-1
F Recommended Procedures for Preparing Whole Fish Composite
Homogenate Samples F-1
G General Procedures for Removing Edible Tissues from Shellfish . . G-1
H Comparison of Target Analyte Screening Values (SVs) with
Detection and Quantitation Limits of Current Analytical Methods . . H-1
I Sources of Recommended Reference Materials and Standards .... 1-1
-------
LIST OF FIGURES
LIST OF FIGURES
Page
2-1 Recommended strategy for State fish and shellfish contaminant
monitoring programs 2-2
3-1 Geographic distributions of three bivalve species used extensively in
national contaminant monitoring programs (based on data from
Abbott, 1974) 3-23
4-1 States issuing fish and shellfish advisories for mercury 4-9
4-2 States issuing fish and shellfish advisories for chlordane 4-12
4-3 States issuing fish and shellfish advisories for PCBs 4-26
4-4 States issuing fish and shellfish advisories for dioxin/
dibenzofurans 4-31
6-1 Example of a sample request form 6-3
6-2 Example of a field record for fish contaminant monitoring
program—screening study 6-32
6-3 Example of a field record for shellfish contaminant monitoring
program—screening study 6-33
6-4 Example of a field record for fish contaminant monitoring
program—intensive study 6-34
6-5 Example of a field record for shellfish contaminant monitoring
program—intensive study 6-36
6-6 Example of a sample identification label 6-37
6-7 Example of a chain-of-custody tag or label 6-37
6-8 Example of a chain-of-custody record form 6-39
6-9 Recommended measurements of body length and size for fish
and shellfish 6-42
7-1 Preparation of fish fillet composite homogenate samples 7-7
7-2 Example of a sample processing record for fish contaminant
monitoring program—fish fillet composites 7-9
7-3 Illustration of basic fish filleting procedure 7-13
7-4 Preparation of shellfish edible tissue composite homogenate
samples 7-17
7-5 Example of a sample processing record for shellfish contaminant
monitoring program—edible tissue composites 7-18
7-6 Example of a fish and shellfish monitoring program sample aliquot
record 7-23
vi
-------
LIST OF FIGURES
Page
7-7 Example of a fish and shellfish monitoring program sample transfer
record 7-25
8-1 Recommended contents of analytical standard operating procedures
(SOPs) 8-12
9-1 Recommended data reporting requirements for screening and
intensive studies 9-4
VII
-------
LIST OF TABLES
LIST OF TABLES
Page
2-1 Recommended Strategy for State Fish and Shellfish Contaminant
Monitoring Programs 2-5
3-1 Recommended Target Species for Inland Fresh Waters 3-4
3-2 Recommended Target Species for Great Lakes Waters 3-4
3-3 Comparison of Freshwater Finfish Species Used in Several National
Fish Contaminant Monitoring Programs .. 3-6
3-4 Average Fish Tissue Concentrations of Xenobiotics for Major Finfish
Species Sampled in the National Study of Chemical Residues in
Fish 3-7
3-5 Average Fish Tissue Concentrations of Dioxins and Furans for Major
Finfish Species Sampled in the National Study of Chemical Residues
in Fish 3-8
3-6 Principal Freshwater Fish Species Cited in State Fish Consumption
Advisories 3-9
3-7 Recommended Target Species for Northeast Atlantic Estuaries and
Marine Waters (Maine through Connecticut) 3-12
3-8 Recommended Target Species for Mid-Atlantic Estuaries and Marine
Waters (New York through Virginia) 3-13
3-9 Recommended Target Species for Southeast Atlantic Estuaries and
Marine Waters (North Carolina through Florida) 3-14
3-10 Recommended Target Species for Gulf of Mexico Estuaries and
Marine Waters (West Coast of Florida through Texas) 3-15
3-11 Recommended Target Species for Pacific Northwest Estuaries and
Marine Waters (Alaska through Oregon) 3-16
3-12 Recommended Target Species for Northern California Estuaries and
Marine Waters (Klamath River through Morro Bay) 3-17
3-13 Recommended Target Species for Southern California Estuaries and
Marine Waters (Santa Monica Bay to Tijuana Estuary) 3-18
3-14 Sources of Information on Commercial and Sportfishing Species in
Various Coastal Areas of the United States 3-19
3-15 Estuarine/Marine Species Used in Several National Fish and
Shellfish Contaminant Monitoring Programs 3-21
3-16 Principal Estuarine/Marine Fish and Shellfish Species Cited in State
Consumption Advisories 3-25
VIII
-------
LIST OF TABLES
Page
4-1 Recommended Target Analytes 4-2
4-2 Contaminants Resulting in Fish and Shellfish Advisories 4-5
4-3 Polychlorinated Biphenyl (PCS) Congeners Recommended for
Quantitation as Potential Target Analytes 4-28
4-4 Dibenzo-p-Dioxins and Dibenzofurans Recommended as Target
Analytes 4-32
5-1 Recommended Values for Mean Body Weights (BWs) and Fish
Consumption Rates (CRs) for Selected Subpopulations 5-6
5-2 Dose-Response Variables and Recommended Screening Values
(SVs) for Target Analytes 5-8
5-3 Example Screening Values (SVs) for Various Subpopulations and
Risk Levels (RLs) 5-12
5-4 Toxicity Equivalency Factors (TEFs) for Tetra- through Octa-
Chlorinated Dibenzo-p-Dioxins and Dibenzofurans 5-15
6-1 Values of [2/n2m2(n-1)]1/2 for Various Combinations of n and m . . 6-19
6-2 Estimates of Statistical Power of Hypothesis of Interest Under
Specified Assumptions 6-21
6-3 Summary of Fish Sampling Equipment . 6-24
6-4 Summary of Shellfish Sampling Equipment 6-26
6-5 Checklist of Field Sampling Equipment and Supplies for Fish and
Shellfish Contaminant Monitoring Programs 6-28
6-6 Safety Considerations for Field Sampling Using a Boat 6-29
6-7 Recommendations for Preservation of Fish and Shellfish Samples
from Time of Collection to Delivery at the Processing
Laboratory 6-46
7-1 Recommendations for Container Materials, Preservation, and Holding
Times for Fish and Shellfish Tissues from Receipt at Sample
Processing Laboratory to Analysis 7-3
7-2 Weights (g) of Individual Homogenates Required for Screening Study
Composite Homogenate Sample 7-15
7-3 Recommended Sample Aliquot Weights and Containers for Various
Analyses 7-22
8-1 Contract Laboratories Conducting Dioxin/Dibenzofuran Analyses in
Fish and Shellfish Tissues 8-2
8-2 Current References for Analytical Methods for Contaminants in Fish
and Shellfish Tissues 8-5
8-3 Recommended Analytical Techniques for Target Analytes 8-6
8-4 Range of Detection and Quantitation Limits of Current Analytical
Methods for Recommended Target Analytes 8-7
IX
-------
LIST OF TABLES
Page
8-5 Approximate Range of Costs per Sample for Analysis of
Recommended Target Analytes 8-10
8-6 Recommended Quality Assurance and Quality Control Samples .. 8-15
8-7 Minimum Recommended QA and QC Samples for Routine
Analysis of Target Analytes 8-24
8-8 Fish and Shellfish Tissue Reference Materials 8-27
-------
ACKNOWLEDGMENTS
ACKNOWLEDGMENTS
This report was prepared by the Research Triangle Institute under EPA Contract
No. 68-C3-0303 for the U.S. Environmental Protection Agency, Office of Water,
Fish Contaminant Section. The primary authors were Patricia Cunningham and
Karen Gold, with assistance from Eva Estes, Kerrie Boyle, Peter Grohse, and
Kathleen Mohar. The EPA work assignment managers for this document were
Alison Greene and Jeffrey Bigler who provided overall project coordination as
well as technical assistance and guidance. Preparation of this guidance
document required extensive effort by members of the EPA Fish Contaminant
Workgroup (listed below). These members, representing EPA Headquarters,
EPA Regions and State and Federal agencies, provided technical information,
review, and recommendations throughout the preparation of this document.
FISH CONTAMINANT WORKGROUP
EPA Headquarters Staff
Carin Bisland
Richard Hoffman
Clyde Houseknecht
Michael Kravitz
Elizabeth Southerland
Margaret Stasikowski
Irene Suzukida-Horner
Elizabeth Tarn
William Telliard
Tina Levine
Michael Metzger
Richard Whiting
Dennis Borum
Jacqueline Moya
Other EPA Office Staff
David DeVault
Brian Melzian
John Paul
EPA/Office of Water
EPA/Office of Water
EPA/Office of Water
EPA/Office of Water
EPA/Office of Water
EPA/Office of Water
EPA/Office of Water
EPA/Office of Water
EPA/Office of Water
EPA/Office of Pesticide Programs
EPA/Office of Pesticide Programs
EPA/Office of Pesticide Programs
EPA/Office of Drinking Water
EPA/Office of Health and Environmental
Assessment
EPA/Great Lakes National Program Office
EPA/Office of Reserach and Development-
Narragansett, Rl
EPA/Office of Research and Development-
Narragansett, Rl
xi
-------
ACKNOWLEDGMENTS
Dennis McMullen
Laurence Burkhard
Michael Dourson
Donald Klemm
EPA Regional Staff
Charles Kanetsky
Jerry Stober
Peter Redmon
Diane Evans
Philip Crocker
Bruce Herbold
Other Federal Agency Staff
Michael Bolger
Leon Sawyer
Lee Barclay
Frank De Luise
Donald Steffeck
Jerry Schulte
Adriana Cantillo
Maxwell Eldridge
Betty Hackley
Alicia Jarboe
Bruce Morehead
Don Dycus
J. Kent Crawford
State Agency Staff
Robert Cooner
Brian Hughes
William Keith
Thomas McChesney
Randall Mathis
Gerald Pollock
Robert McConnell
Richard Green
Eldert Hartwig
Randall Manning
Robert Flentge
C. Lee Bridges
EPA/Environmental Monitoring and
Systems Laboratory-Cincinnati, OH
EPA/Office of Research and Development-
Duluth, MN
EPA/Office of Health and Environmental
Assessment-Cincinnati, OH
EPA/Office of Health and Environmental
Assessment-Cincinnati, OH
Region 3
Region 4
Region 5
Region 6
Region 7
Region 9
FDA
FDA
FWS
FWS
FWS
ORSANCO
NOAA
NOAA
NOAA
NOAA
NOAA
TVA
USGS
Alabama
Alabama
Arkansas
Arkansas
Arkansas
California
Colorado
Delaware
Florida
Georgia
Illinois
Indiana
XII
-------
ACKNOWLEDGMENTS
Emelise Cormier
Albert Hindrichs
Elaine Sorbet
Deirdre Murphy
Jack Schwartz
John Hesse
Richard Powers
Lisa Williams
Pamela Shubat
Alan Buchanan
David Tunink
Donald Normandeau
Paul Hauge
Lawrence Skinner
Ken Eagleson
Jay Sauber
Luanne Williams
Michael Ell
Martin Schock
Abul Anisuzzaman
Gene Foster
Barbara Britton
Peter Sherertz
Ram Tripathi
Jim Amrhein
Bruce Baker
Other Organizations
James Wiener
Deborah Schwackhamer
Louisiana
Louisiana
Louisiana
Maryland
Massachusetts
Michigan
Michigan
Michigan
Minnesota
Missouri
Nebraska
New Hampshire
New Jersey
New York
North Carolina
North Carolina
North Carolina
North Dakota
North Dakota
Ohio
Oregon
Texas
Virginia
Virginia
Wisconsin
Wisconsin
American Fisheries Society
University of Minnesota
XIII
-------
1. INTRODUCTION
SECTION 1
INTRODUCTION
1.1 HISTORICAL PERSPECTIVE
Contamination of aquatic resources, including freshwater, estuarine, and marine
fish and shellfish, has been documented in the scientific literature for many
regions of the United States (NAS, 1991). Environmental concentrations of
some pollutants have decreased over the past 20 years as a result of better
water quality management practices. However, environmental concentrations of
other heavy metals, pesticides, and toxic organic compounds have increased
due to intensifying urbanization, industrial development, and use of new
agricultural chemicals. Our Nation's waterbodies are among the ultimate
repositories of pollutants released from these activities. Pollutants come from
permitted point source discharges (e.g., industrial and municipal facilities),
accidental spill events, and nonpoint sources (e.g., agricultural practices,
resource extraction, urban runoff, in-place sediment contamination, ground water
recharge, and atmospheric deposition).
Once these toxic contaminants reach surface waters, they may concentrate
through aquatic food chains and bioaccumulate in fish and shellfish tissues.
Aquatic organisms may bioaccumulate environmental contaminants to more than
1,000,000 times the concentrations detected in the water column (U.S. EPA,
1992c, 1992d). Thus, fish and shellfish tissue monitoring serves as an important
indicator of contaminated sediments and water quality problems, and many
States routinely conduct chemical contaminant analyses of fish and shellfish
tissues as part of their comprehensive water quality monitoring programs
(Cunningham and Whitaker, 1989). Tissue contaminant monitoring also enables
State agencies to detect levels of contamination in fish and shellfish tissue that
may be harmful to human consumers. If States conclude that consumption of
chemically contaminated fish and shellfish poses an unacceptable human health
risk, they may issue local fish consumption advisories or bans for specific
waterbodies and specific fish and shellfish species for specific populations.
In 1989, the American Fisheries Society (AFS), at the request of the U.S.
Environmental Protection Agency (EPA), conducted a survey of State fish and
shellfish consumption advisory practices. Questionnaires were sent to health
departments, fisheries agencies, and water quality/environmental management
departments in all 50 States and the District of Columbia. Officials in all 50
States and the District responded.
1-1
-------
1. INTRODUCTION
Respondents were asked to provide information on several issues including
Agency responsibilities
Sampling strategies
• Sample collection procedures
Chemical residue analysis procedures
• Risk assessment methodologies
Data interpretation and advisory development
• State concerns
Recommendations for Federal assistance.
Cunningham et al. (1990) summarized the survey responses and reported that
monitoring and risk assessment procedures used by States in their fish and
shellfish advisory programs varied widely. States responded to the question
concerning assistance from the Federal government by requesting that Federal
agencies
Provide a consistent approach for State agencies to use in assessing health
risks from consumption of chemically contaminated fish and shellfish
Develop guidance on sample collection procedures
Develop and/or endorse uniform, cost-effective analytical methods for
quantitation of contaminants
Establish a quality assurance (QA) program that includes use of certified
reference materials for chemical analyses.
In March 1991, the National Academy of Sciences (MAS) published a report
entitled Seafood Safety (MAS, 1991) that reviewed the nature and extent of
public health risks associated with seafood consumption and examined the
scope and adequacy of current seafood safety programs. After reviewing over
150 reports and publications on seafood contamination, the NAS Institute of
Medicine concluded that high concentrations of chemical contaminants exist in
various fish species in a number of locations in the country. The report noted
that the fish monitoring data available in national and regional studies had two
major shortcomings that affected their usefulness in assessing human health
risks:
In some of the more extensive studies, analyses were performed on
nonedible portions of finfish (e.g., liver tissue) or on whole fish, which
precludes accurate determination of human exposures.
Studies did not use consistent methods of data reporting (e.g., both
geometric and arithmetic means were reported in different studies) or failed
to report crucial information on sample size, percent lipid, mean values of
contaminant concentrations, or fish size, thus precluding direct comparison
1-2
-------
1. INTRODUCTION
of the data from different studies and complicating further statistical analysis
and risk assessment.
As a result of these MAS concerns and State concerns expressed in the AFS
survey, the EPA Office of Water established a Fish Contaminant Workgroup. It
is composed of representatives from EPA and the following State and Federal
agencies:
Food and Drug Administration (FDA)
• Fish and Wildlife Service (FWS)
• Ohio River Valley Water Sanitation Commission (ORSANCO)
National Oceanic and Atmospheric Administration (NOAA)
Tennessee Valley Authority (TVA)
• United States Geological Survey (USGS)
and representatives from 26 States: Alabama, Arkansas, California, Colorado,
Delaware, Florida, Georgia, Illinois, Indiana, Louisiana, Maryland, Massachu-
setts, Michigan, Minnesota, Missouri, Nebraska, New Hampshire, New Jersey,
New York, North Carolina, North Dakota, Ohio, Oregon, Texas, Virginia, and
Wisconsin.
The objective of the EPA Fish Contaminant Workgroup was to formulate
guidance for States on how to sample and analyze chemical contaminants in fish
and shellfish where the primary end uses of the data included development of
fish consumption advisories. The Workgroup compiled documents describing
protocols currently used by various Federal agencies, EPA Regional offices, and
States that have extensive experience in fish contaminant monitoring. Using
these documents, they selected methods considered most cost-effective and
scientifically sound for sampling and analyzing fish and shellfish tissues. These
methods are recommended as standard procedures for use by the States and
are described in this manual.
1.2 PURPOSE
The purpose of this manual is to provide overall guidance to States on methods
for sampling and analyzing contaminants in fish and shellfish tissue that will
promote consistency in the data States use to determine the need for fish
consumption advisories. This manual provides guidance only and does not
constitute a regulatory requirement for the States. It is intended to describe
what the EPA Office of Water believes to be scientifically sound methods for
sample collection, chemical analyses, and statistical analyses of fish and
shellfish tissue contaminant data for use in fish contaminant monitoring programs
that have as their objective the protection of public health. This nonregulatory,
technical guidance manual is intended for use as a handbook by State and local
agencies that are responsible for sampling and analyzing fish and shellfish
tissue. Adherence to this guidance will enhance the comparability of fish and
shellfish contaminant data, especially in interstate waters, and thus provide more
standardized information on fish contamination problems.
1-3
-------
1. INTRODUCTION
The three-ring binder format was selected by EPA for this manual to enhance
its use as a working document and to facilitate inclusion of additional refinements
and updates of the recommended procedures. It is anticipated that additional
contaminants will be added to the list of target analytes, that screening values
may change as new toxicologic data are evaluated, and that new chemical
analysis procedures may be recommended for some target analytes as they are
developed.
The EPA Office of Water realizes that adoption of these recommended methods
requires adequate funding. In practice, funding varies among States and
resource limitations will cause States to tailor their fish and shellfish contaminant
monitoring programs to meet their own needs. States must consider tradeoffs
among the various parameters when developing their fish contaminant
monitoring programs. These parameters include
• Total number of stations sampled
Intensity of sampling at each site
Number of chemical analyses and their cost
Resources expended on data storage and analysis, QA and quality control
(QC), and sample archiving.
These tradeoffs will limit the number of sites sampled, number of target analytes
analyzed at each site, number of target species collected, and number of
replicate samples of each target species collected at each site (Crawford and
Luoma, 1993).
1.3 OBJECTIVES
The specific objectives of the manual are to
1. Recommend a tiered monitoring strategy designed to
• Screen waterbodies (Tier 1) to identify those harvested sites where
chemical contaminant concentrations in the edible portions of fish and
shellfish exceed human consumption levels of potential concern
(screening values [SVs]). SVs for contaminants with carcinogenic
effects are calculated based on selection of an acceptable cancer risk
level. SVs for contaminants with noncarcinogenic effects are
concentrations determined to be without appreciable noncancer health
risk. For a contaminant with both carcinogenic and noncarcinogenic
effects, the lower (more conservative) of the two calculated SVs is used.
• Conduct intensive followup sampling (Tier 2, Phase I) to determine the
magnitude of the contamination in edible portions of fish and shellfish
1-4
-------
1. INTRODUCTION
species commonly consumed by humans in water-bodies identified in the
screening process.
• Conduct intensive sampling at additional sites (Tier 2, Phase II) in a
waterbody where screening values were exceeded to determine the
geographic extent of contamination in various size classes of fish and
shellfish.
2. Recommend target species and criteria for selecting additional species if the
recommended target species are not present at a site.
3. Recommend target analytes to be analyzed in fish and shellfish tissue and
criteria for selecting additional analytes.
4. Recommend risk-based procedures for calculating target analyte screening
values.
5. Recommend standard field procedures including
Site selection
Sampling time
Sample type and number of replicates
Sample collection procedures including sampling equipment
Field recordkeeping and chain of custody
Sample processing, preservation, and shipping.
6. Recommend cost-effective, technically sound analytical methods and
associated QA and QC procedures, including identification of
• Analytical methods for target analytes with detection limits capable of
measuring tissue concentrations at or below SVs
• Sources of recommended certified reference materials
• Federal agencies currently conducting QA interlaboratory comparison
programs.
7. Recommend procedures for data analysis and reporting of fish and shellfish
contaminant data.
8. Recommend QA and QC procedures for all phases of the monitoring
program and provide guidance for documenting QA and QC requirements
in a QA plan or in a combined work/QA project plan.
1.4 RELATIONSHIP OF MANUAL TO OTHER GUIDANCE DOCUMENTS
This manual is the first in a series of four documents to be prepared by the EPA
Office of Water as part of a Federal Assistance Plan to help States standardize
_
-------
1. INTRODUCTION
fish consumption advisories. The remaining three documents will provide
guidance on risk assessment, risk management, and risk communication.
This sampling and analysis manual is not intended to be an exhaustive guide to
all aspects of sampling, statistical design, development of risk-based screening
values, laboratory analyses, and QA and QC considerations for fish and shellfish
contaminant monitoring programs. Key references are provided that detail
various aspects of these topics. In addition, States may obtain a list of related
documents relevant to fish and shellfish contaminant monitoring by accessing the
EPA Nonpoint Source Bulletin Board System (NPS BBS). The phone number
of the BBS is (301) 589-0205.
1.5 ORGANIZATION OF THIS MANUAL
This manual provides specific guidance on sampling, chemical analysis, and
data reporting and analysis procedures for State fish and shellfish contaminant
monitoring programs. Appropriate QA and QC considerations are integral parts
of each of the recommended procedures.
Monitoring Strategy: Section 2 outlines the recommended strategy for State
fish and shellfish contaminant monitoring programs. This strategy is designed
to (1) routinely screen waterbodies to identify those locations where chemical
contaminants in edible portions of fish and shellfish exceed human health
screening values and (2) sample more intensively those waterbodies where
exceedances of these SVs have been found in order to assess the magnitude
and the geographic extent of the contamination.
Target Species: Section 3 discusses the purpose of using target species and
criteria for selection of target species for both screening and intensive studies.
Lists of recommended target species are provided for inland fresh waters, Great
Lakes waters, and seven distinct estuarine and coastal marine regions of the
United States.
Target Analytes: Section 4 presents a list of recommended target analytes to
be considered for inclusion in screening studies and discusses criteria used in
selecting these analytes.
Screening Values: Section 5 describes the EPA risk-based procedure for
calculating screening values for target analytes.
Field Procedures: Section 6 recommends field procedures to be followed from
the time fish or shellfish samples are collected until they are delivered to the
laboratory for processing and analysis. Guidance is provided on site selection
and sample collection procedures; the guidance addresses material and
equipment requirements, time of sampling, size of animals to be collected,
sample type, and number of samples. Sample identification, handling,
preservation, shipping, and storage procedures are also described.
1-6
-------
1. INTRODUCTION
Laboratory Procedures: Section 7 describes recommended laboratory
procedures for sample handling including: sample measurements, sample
processing procedures, and sample preservation and storage procedures.
Section 8 presents recommended laboratory procedures for sample analyses,
including cost-effective analytical methods and associated QC procedures, and
information on sources of certified reference materials and Federal agencies
currently conducting interlaboratory comparison programs.
Data Analysis and Reporting: Section 9 includes procedures for data analysis
to determine the need for additional monitoring and risk assessment and for data
reporting. This section also describes the National Fish Tissue Data Repository
(NFTDR), a national database of fish and shellfish contaminant monitoring data.
Supporting documentation for this guidance is provided in Section 10, Literature
Cited, and in Appendixes A through I.
1-7
-------
2. MONITORING STRATEGY
SECTION 2
MONITORING STRATEGY
The objective of this section is to describe the strategy recommended by the
EPA Office of Water for use by States in their fish and shellfish contaminant
monitoring programs. A two-tiered strategy is recommended as the most cost-
effective approach for State contaminant monitoring programs to obtain data
necessary to evaluate the need to issue fish or shellfish consumption advisories.
This monitoring strategy is shown schematically in Figure 2-1 and consists of
Tier 1—Screening studies of a large number of sites for chemical
contamination where sport, subsistence, and/or commercial fishing is
conducted. This screening will help States identify those sites where
concentrations of chemical contaminants in edible portions of commonly
consumed fish and shellfish indicate the potential for significant health risks
to human consumers.
Tier 2—Two-phase Intensive studies of problem areas identified in
screening studies to determine the magnitude of contamination in edible
portions of commonly consumed fish and shellfish species (Phase I), to
determine size-specific levels of contamination, and to assess the
geographic extent of the contamination (Phase II).
This basic approach of using relatively low-cost, nonintensive screening studies
to identify areas for more intensive followup sampling is used in a variety of
water quality programs involving public health protection (California
Environmental Protection Agency, 1991; Oregon Department of Environmental
Quality, 1990; TVA, 1991; U.S. EPA, 1989d).
One key objective in the recommendation of this approach is to improve the data
used by States for issuing fish and shellfish consumption advisories. Other
specific aims of the recommended strategy are
To ensure that resources for fish contaminant monitoring programs are
allocated in the most cost-effective way. By limiting the number of sites
targeted for intensive studies, as well as the number of target analytes at
each intensive sampling site, screening studies help to reduce overall
program costs while still allowing public health protection objectives to be
met.
2-1
-------
Tierl
Send fish contaminant residue
data to the National Fish
Tissue Data Repository
(NFTDR)
Conduct Waterbody Screening Study *
1. Collect composite sample for each target species
2. Determine concentrations of selected target anarytes in
composite sample for each target species
Any
Target Analyte
Concentration Exceeds
Appropriate Screening Value
(SV) for Any Target
Species
No
Additional
Monitoring
Needed
until Next
Screening
Study
i YES
Tier 2, Phase I
Tier 2, Phase II
Conduct Phase I Intensive Study to
Assess Magnitude of Contamination **
1. Collect replicate composite samples for each target
2. Determine arithmetic mean concentrations of target
anarytes that exceed SVs for each target species
ro
ro
Conduct Phase II Intensive Study to Determine
Geographic Extent of Contamination
1. Select number of additional sites in the waterbody to be sampled
2. At each Phase II site, collect replicate composite samples for three
size classes of each target species
3. Determine arithmetic mean concentrations of target anarytes
exceeding SVs for three size classes of each target species
No
Additional
Monitoring
Needed
until Next
Screening
Study
Arithmetic
Mean Concentration of
Any Target Analyte
Exceeds SV for Any
Target Species
Perform Risk
Assessment to
Evaluate Need for
Issuance of a Fish
sumption Advi
ro
O
O
2
O
w
-------
Arithmetic
Mean Concentration of
Any Target Analyte at
Any Site Exceeds SV
for Any Target
Species
Perform Risk
Assessment to
Evaluate Need for
Issuance of a Rsh
umption Advlso
"i
Risk Management — Evaluate Options to Protect Public Health
1. Identify nature of the advisory for the general adult population and/or subpopulalions
2. Identify contaminated fish/shellfish species and size classes to be cited in the advisory (as appropriate)
3. Identify geographic extent of the advisory
No
Additional «
Monitoring f-,
Needed {'$
until Next 'JT
Screening f,
Study ;*
• ' '*-ff:~ -% ^
Risk Communication
t. Issue final fish consumption advisory SSSKSS? ••"
2. Send fish advisory information to EPA Fish
Contamination Section so that information can
be entered into the National Fish Consumption
Advisory Database
3. Report all new advisories issued in the biennial
State 305(b) report
ro
U
Revisit walerbodies as funding permits to assess
changes in target analyte concentrations
* Tier 1 Screening Study Options — If resources are limited for conducting Tier 2 intensive studies.
States may pursue other options during screening studies (see discussion in Section 6.1).
** Tier 2, Phase I, Intensive Study Options — If resources are limited for conducting Tier 2 Phase II
studies. States may pursue other options during Phase I (see discussion in Section 6.2).
O
i
3J
O
CO
Figure 2-1. Recommended strategy for State fish/shellfish contaminant monitoring programs.
-------
2. MONITORING STRATEGY
To ensure that sampling data are appropriate for developing risk-based
consumption advisories.
• To ensure that sampling data are appropriate for determining contaminant
concentrations in various size (age) classes of each target species so that
States can give size-specific advice on contaminant concentrations (as
appropriate).
• To ensure that sampling designs are appropriate to allow statistical
hypothesis testing. Such sampling designs permit the use of statistical tests
to detect a difference between the average tissue contaminant concentration
at a site and the human health screening value for any analyte.
The following elements must be considered when planning either screening
studies or more intensive followup sampling studies:
Study objective
Target species (and size classes)
• Target analytes
• Target analyte screening values
Sampling locations
Sampling times
• Sample type
Sample replicates
• Sample analysis
Data analysis and reporting.
Detailed guidance for each of these elements, for screening studies (Tier 1) and
for both Phase I and Phase II of intensive studies (Tier 2), is provided in this
document. The key elements of the monitoring strategy are summarized in
Table 2-1, with reference to the section number of this document where each
element is discussed.
2.1 SCREENING STUDIES (TIER 1)
The primary aim of screening studies is to identify frequently fished sites where
concentrations of chemical contaminants in edible fish and shellfish composite
samples exceed specified human health screening values and thus require more
intensive followup sampling. Ideally, screening studies should include all
waterbodies where commercial, recreational, or subsistence fishing is practiced;
specific sampling sites should include areas where various types of fishing are
conducted routinely (e.g., from a pier, from shore, or from private and
commercial boats), thereby exposing a significant number of individuals to
potentially adverse health effects. Composites of skin-on fillets (except for
catfish and other scaleless species, which are usually prepared as skin-off fillets)
and edible portions of shellfish are recommended for contaminant analyses in
screening studies to provide conservative estimates of typical exposures for the
general population. Note: If consumers remove the skin and fatty areas from
_
-------
Table 2-1. Recommended Strategy for State Fish and Shellfish Contaminant Monitoring Programs
Program element
Tier 1 Screening study
Tier 2 Intensive study (Phase I)
Tier 2 Intensive study (Phase II)
Objective
(see Section 2)
Target species and
size classes
(see Sections 3
and 6)
Identify frequently fished sites where
commonly consumed fish and shellfish target
species are contaminated and may pose
potential human health risk.
Select target species from commonly
consumed species using the following
additional criteria: known to bioaccumulate
high concentrations of contaminants and
distributed over a wide geographic area.
Recommended types of target species:
Inland fresh 1 bottom-feeder
waters: 1 predator
Great Lakes: 1 bottom-feeder
1 predator
Estuarine/ 1 shellfish and
marine: 1 fish species
or
2 fish species (one species
should be bottom-feeder).
Assess and verify magnitude of
tissue contamination at screening site
for commonly consumed target
species.
Resample target species at sites
where they were found to be
contaminated in screening study.
Assess geographic extent of
contamination in selected size classes
of commonly consumed target
species.
Resample at additional sites in the
waterbody three size classes of the
target species found to be
contaminated in Phase I study.
o
a
z
o
o
10
61
See notes at end of table:
(continued)
-------
Table 2-1 (continued)
Program element
Tier 1 Screening study
Tier 2 Intensive study (Phase I)
Tier 2 Intensive study (Phase II)
Target species and
size classes
(continued)
Target analytes
(see Section 4)
OPTIONAL: If resources are limited and a
State cannot conduct Tier 2 intensive
studies, the State may find it more cost-
effective to collect additional samples during
the Tier 1 screening study. States may
collect (1) one composite sample of each of
three size classes for each target species,
(2) replicate composite samples for each
target species, or (3) replicate composite
samples of each of three size classes for
each target species.
Consider all target analytes listed in
Table 4-1 for analysis as resources
allow. Include additional site-specific
target analytes as appropriate based
on historic data.
OPTIONAL: If resources are limited
and a State cannot conduct Tier 2,
Phase II, intensive studies, the State
may find it more cost-effective to
collect additional samples during the
Tier 2, Phase I, intensive study.
States may collect replicate
composite samples of three size
classes of the target species found to
be contaminated to assess size-
specific contaminant concentrations.
Other commonly consumed target
species may also be sampled if
resources allow.
Analyze only for those target analytes
from Tier 1 screening study that
exceeded SVs.
OPTIONAL: If resources allow, select
additional commonly consumed target
species using same criteria as in
Phase I study.
Analyze only for those target analytes
from Tier 2, Phase I, study that
exceeded SVs.
10
i
2
o
0>
See notes at end of table.
(continued)
-------
Table 2-1 (continued)
Program element
Tier 1 Screening study
Tier 2 Intensive study (Phase I)
Tier 2 Intensive study (Phase II)
Screening values
(see Section 5)
Calculate SVs using oral RfDs for
noncarcinogens and using oral slope factors
and an appropriate risk level (1O"4 to 10~7)
for carcinogens, for adults consuming 6.5
g/d to 140 g/d or more of fish and shellfish
(based on site-specific dietary data).
Note: In this guidance document, EPA's
Office of Water used a 6.5-g/d consumption
rate, 70-kg adult body weight, and, for
carcinogens, used a 10~5 risk level, 70-year
exposure, and assumed no loss of
contaminants during preparation or cooking.
States may use other SVs for site-specific
exposure scenarios by adjusting values for
consumption rate, body weight, risk level,
exposure period, and contaminant loss
during preparation or cooking.
Use same SVs as in screening study. Use same SVs as in screening study.
Sampling sites
(see Section 6)
Sample target species at sites in each
harvest area that have a high probability of
contamination and at presumed clean sites
as resources allow.
Sample target species at each site
identified in the screening study
where fish/shellfish tissue
concentrations exceed SVs to assess
the magnitude of contamination.
Sample at additional sites in the
harvest area three size classes of the
target species found to be
contaminated in Phase I study to
assess the geographic extent of the
contamination in the waterbody.
O
o
2
o
i
See notes at end of table.
(continued)
m
O
-------
Table 2-1 (continued)
Program element
Her 1 Screening study
Tier 2 Intensive study (Phase I)
Tier 2 Intensive study (Phase II)
Sampling times
(see Section 6)
Sample during legal harvest season
when target species are most available to
consumers. Ideally, sampling time should not
include the spawning period for target
species unless the target species can be
legally harvested during this period.
Same as screening study.
Same as screening study.
Sample type
(see Sections 6
and?)
Collect composite fillet samples (skin on,
belly flap included) for each target fish
species and composite samples of edible
portions of target shellfish species. The
exceptions to the "skin on, belly flap
included" recommendation is to use skin-off
fillets for catfish and other scaleless species.
OPTIONAL States may use whole fish or
other sample types, if necessary, to improve
exposure estimates of local fish-consuming
populations.
Same as screening study.
Same as screening study but collect
composite samples for three size
classes of each target species.
Same as screening study.
Same as screening study.
Sample replicates
(see Section 6)
Collect one composite sample for each
target species. Collection of replicate
composite samples Is encouraged but Is
optional. If resources allow, collect a
minimum of one replicate composite sample
for each target species at 10% of the
screening sites for QC.
Collect replicate composites for each
target species at each Phase I site.
Collect replicate composites of three
size classes for each target species
at each Phase II site.
ISJ
§
5
o
CO
ro
oo
See notes at end of table.
(continued)
-------
Table 2-1 (continued)
Program element
Tier 1 Screening study
Tier 2 Intensive study (Phase I)
Tier 2 Intensive study (Phase II)
Sample analysis
(see Section 8)
Data analysis and
reporting
(see Sections 6,
7, 8, and 9)
Use standardized and quantitative analytical
methods with limits of detection adequate to
allow reliable quantitation of selected target
analytes at or below SVs.
For each target species, compare target
analyte concentrations of composite sample
with SVs to determine which sites require
Tier 2, Phase I, intensive study.
The following information should be reported
for each target species at each site:
Site location (e.g., sample site name,
waterbody name, type of waterbody, and
latitude/longitude)
Scientific and common name of target
species
Use same analytical methods as in Use same analytical methods as in
screening study. screening study.
For each target species, compare
target analyte arithmetic mean
concentrations of replicate composite
samples with SVs to determine which
sites require Phase II intensive study.
If resources are insufficient to
conduct Phase II intensive study,
conduct a risk assessment and
assess the need for issuing a
preliminary fish or shellfish
consumption advisory.
The following information should be
reported for each target species
at each site:
Same as screening study.
Same as screening study
For each of three size classes within
each target species, compare target
analyte arithmetic mean
concentrations of replicate composite
samples at each Phase II site with
SVs to determine geographic extent
of fish or shellfish contamination.
Assess the need for issuing a final
fish or shellfish consumption advisory.
The following information should be
reported for each of three size
classes within each target species at
each site:
• Same as screening study.
Same as screening study
ro
O
I
3J
O
m
o
to
to
See notes at end of table.
(continued)
-------
Table 2-1 (continued)
Program element
Tier 1 Screening study
Tier 2 Intensive study (Phase I)
Tier 2 Intensive study (Phase II)
Data analysis and
reporting
(continued)
Sampling date and time
Sampling gear type used
Sampling depth
Number of QC replicates (optional)
Number of individual organisms used in
the composite sample and in the QC
replicate composite sample if applicable
Predominant characteristics of specimens
used in the composite sample and in the
QC replicate if applicable (e.g., life stage,
age, sex, total length or body size) and
description of fish fillet or edible parts of
shellfish (tissue type) used
Analytical methods used (including a
method for lipid analysis) and method
detection and quantitation limits for each
target analyte.
Sample cleanup procedures
Data qualifiers
Percent lipid in each composite sample.
Same as screening study
Same as screening study
Sampling depth
Number of replicates
Number of individual organisms
used in each replicate composite
sample
Predominant characteristics of
specimens used in each replicate
composite sample (e.g., life stage,
age, sex, total length or body size)
and description of fish fillet or
edible parts of shellfish (tissue
type) used
Same as screening study
Same as screening study.
Same as screening study.
Same as screening study.
Same as screening study
Same as screening study
Sampling depth
Same as Phase I study
' Same as Phase I study
1 Same as Phase I study
Same as screening study
Same as screening study.
Same as screening study.
Same as screening study.
10
3
O
O
3
o
m
o
ro
See notes at end of table.
(continued)
-------
Table 2-1 (continued)
Program element
Tier 1 Screening study
Tier 2 Intensive study (Phase I)
Her 2 Intensive study (Phase II)
Data analysis and
reporting
(continued)
For each target analyte:
- Total wet weight of composite sample (g)
used in analysis
Measured concentration (wet weight) in
composite sample including units of
measurement for target analyte
Measured concentration (wet weight) in
the QC replicate, if applicable.
Evaluation of laboratory performance
(i.e., description of all QA and QC
samples associated with the sample(s)
and results of all QA and QC analyses)
Comparison of measured concentration
of composite sample with SV and clear
indication of whether SV was exceeded
For each target analyte:
- Total wet weight of each replicate
composite sample (g) used in
analysis
- Measured concentration (wet
weight) in each replicate
composite sample and units of
measurement for target analyte
- Range of concentrations (wet
weight) for each set of replicate
composite samples
- Mean (arithmetic) concentration
(wet weight) for each set of
replicate composite samples
- Standard deviation of mean
concentration (wet weight)
- Same as screening study
- Comparison of target analyte
arithmetic mean concentration of
replicate composite samples with
SV using hypothesis testing and
clear indication of whether the SV
was exceeded
For each target analyte:
- Same as Phase I study
- Same as Phase I study
- Same as Phase I study
- Same as Phase I study
- Same as Phase I study
- Same as screening study
- Same as Phase I study
JO
O
I
a
z
o
ro
QA = Quality assurance.
QC = Quality control.
RfDs = Reference doses.
SVs = Screening values.
m
O
-------
2. MONITORING STRATEGY
a fish before preparing it for eating, exposures to some contaminants can be
reduced (Armbruster et al., 1987,1989; Cichy, Zabik, and Weaver, 1979; Foran,
Cox, andCroxton, 1989; Gall and Voiland, 1990; Reinert, Stewart, and Seagram,
1972; Sanders and Haynes, 1988; Skea et al., 1979; Smith, Funk, and Zabik,
1973; Voiland et al., 1991; Wanderstock et al., 1971; Zabik, Hoojjat, and Weaver,
1979).
Because the sampling sites in screening studies are focused primarily on the
most likely problem areas and the numbers of commonly consumed target
species and samples collected are limited, relatively little detailed information is
obtained on the magnitude and geographic extent of contamination in a wide
variety of harvestable fish and shellfish species of concern to consumers. More
information is obtained through additional intensive followup studies (Tier 2,
Phases I and II) conducted at potentially contaminated sites identified in
screening studies.
Although the EPA Office of Water recommends that screening study results not
be used as the sole basis for conducting a risk assessment, the Agency
recognizes that this practice may be unavoidable if monitoring resources are
limited or if the State must issue an advisory based on detection of elevated
concentrations in one composite sample. States have several options for
collecting samples during the Tier 1 screening study (see Figure 2-1), which can
provide additional information on contamination without necessitating additional
field monitoring expenditures as part of the Tier 2 intensive studies.
The following assumptions are made in this guidance document for sampling fish
and shellfish and for calculating human health SVs:
• Use of commonly consumed target species that are dominant in the catch
and have high bioaccumulation potential
Use of fish fillets (with skin on and belly flap tissue included) for scaled
finfish species, use of skinless fillets for scaleless finfish species, and use
of edible portions of shellfish
Use of fish and shellfish above legal size to maximum size in the target
species
Use of a 10~5 risk level, a human body weight of 70 kg (average adult), a
consumption rate of 6.5 g/d for the general population, and a 70-yr lifetime
exposure period to calculate SVs for carcinogens. Note: The EPA is
currently reviewing the 6.5-g/d consumption rate for the general population.
Use of a human body weight of 70 kg (average adult) and a consumption
rate of 6.5 g/d for the general population to calculate SVs for noncar-
cinogens.
2-12
-------
2. MONITORING STRATEGY
Use of no contaminant loss during preparation and cooking or from
incomplete absorption in the intestines.
For certain site-specific situations, States may wish to use one or more of the
following exposure assumptions to protect the health of subpopulations at
potentially greater risk:
Use of commonly consumed target species that are dominant in the catch
and have the highest bioaccumulation potential
Use of whole fish or whole body of shellfish (excluding shell of bivalves),
which may provide a better estimate of contaminant exposures in sub-
populations that consume whole fish or shellfish
Use of the largest (oldest) individuals in the target species to represent the
highest likely exposure levels
Use of a 10"6 or 10"7 risk level, body weights less than 70 kg for women and
children, site-specific consumption rates (i.e., 30 g/d for sport fisherman or
140 g/d for subsistence fishermen or other consumption rates based on
dietary studies of local fish-consuming populations), and a 70-yr exposure
period to calculate SVs for carcinogens. Note: The EPA is currently
reviewing the consumption rate for sport and subsistence fishermen.
Use of body weights less than 70 kg for women and children and site-
specific consumption rates (i.e., 30 g/d for sport fishermen or 140 g/d for
subsistence fishermen or other consumption rates based on dietary studies
of local fish-consuming populations) to calculate SVs for noncarcinogens.
There are additional aspects of the screening study design that States should
review because they affect the statistical analysis and interpretation of the data.
These include
Use of composite samples, which results in loss of information on the
distribution of contaminant concentrations in the individual sampled fish and
shellfish. Maximum contaminant concentrations in individual sampled fish,
which can be used as an indicator of potentially harmful levels of
contamination (U.S. EPA, 1989d), are not available when composite
sampling is used.
Use of a single sample per screening site for each target species, which
precludes estimating the variability of the contamination level at that site and,
consequently, of conducting valid statistical comparisons to the target
analyte SVs.
2-13
-------
2. MONITORING STRATEGY
Uncertainty factors affecting the numerical calculation of quantitative health
risk information (i.e., references doses and cancer slope factors) as well as
human health SVs.
States should consider the potential effects of these study design features when
evaluating screening study results.
2.2 INTENSIVE STUDIES (TIER 2)
The primary aims of intensive studies are to assess the magnitude of tissue
contamination at screening sites, to determine the size class or classes of fish
within a target species whose contaminant concentrations exceed the SVs, and
to assess the geographic extent of the contamination for the target species in the
waterbody under investigation. With respect to the design of intensive studies,
EPA recommends a sampling strategy that may not be feasible for some site-
specific environments. Specifically, EPA recognizes that some waterbodies
cannot sustain the same intensity of sampling (i.e., number of replicate
composite samples per site and number of individuals per composite sample)
that others (i.e., those used for commercial harvesting) can sustain. In such
cases, State fisheries personnel may consider modifying the sampling strategy
(e.g., analyzing individual fish) for intensive studies to protect the fishery
resource. Although one strategy cannot cover all situations, these sampling
guidelines are reasonable for the majority of environmental conditions, are
scientifically defensible, and provide information that can be used to assess the
risk to public health. Regardless of the final study design and protocol chosen
for a fish contaminant monitoring program, State fisheries, environmental, and
health personnel should always evaluate and document the procedures used to
ensure that results obtained meet State objectives for protecting human health.
The allocation of limited funds to screening studies or to intensive studies should
always be guided by the goal of conducting adequate sampling of State fish and
shellfish resources to ensure the protection of the public's health. The amount
of sampling that can be performed by a State will be determined by available
economic resources. Ideally, State agencies will allocate funds for screening as
many sites as is deemed necessary while reserving adequate resources to
conduct subsequent intensive studies at sites where excessive fish tissue
contamination is detected. State environmental and health personnel should use
all information collected in both screening and intensive studies to (1) conduct
a risk assessment to determine whether the issuance of an advisory is
warranted, (2) use risk management to determine the nature and extent of the
advisory, and then (3) effectively communicate this risk to the public. Additional
information on risk assessment, risk management, and risk communication
procedures will be provided in subsequent volumes in this series.
2-14
-------
3. TARGET SPECIES
SECTION 3
TARGET SPECIES
The primary objectives of this section are to: (1) discuss the purpose of using
target species, (2) describe the criteria used to select target species, and (3)
provide lists of recommended target species. Target species recommended for
freshwater and estuarine/marine ecosystems are discussed in Sections 3.3 and
3.4, respectively.
3.1 PURPOSE OF USING TARGET SPECIES
The use of target species allows comparison of fish and shellfish tissue
contaminant monitoring data among sites over a wide geographic area.
Differences in habitat, food preferences, and rate of contaminant uptake among
various fish and shellfish species make comparison of contaminant monitoring
results within a State or among States difficult unless the contaminant data are
from the same species. It is virtually impossible to sample the same species at
every site, within a State or region or nationally, due to the varying geographic
distributions and environmental requirements of each species. However, a
limited number of species can be identified that are distributed widely enough to
allow for collection and comparison of contaminant data from many sites.
Three aims are achieved by using target species in screening studies. First,
States can cost-effectively compare contaminant concentrations in their State
waters and then prioritize sites where tissue contaminants exceed human health
screening values. In this way, limited monitoring resources can be used to
conduct intensive studies at sites exhibiting the highest degree of tissue
contamination in screening studies. By resampling target species used in the
screening study in Phase I intensive studies and sampling additional size classes
and additional target species in Phase II intensive studies as resources allow,
States can assess the magnitude and geographic extent of contamination in
species of commercial, recreational, or subsistence value. Second, the use of
common target species among States allows for more reliable comparison of
sampling information from various sites within a State to information from sites
in adjacent States with which they share waters. Such information allows States
to design and evaluate their own contaminant monitoring programs more
efficiently, which should further minimize overall monitoring costs. For example,
monitoring by one State of fish tissue contamination levels in the upper reaches
of a particular river can provide useful information to an adjacent State on tissue
contamination levels that might be anticipated in the same target species at
sampling sites downstream. Third, the use of a select group of target fish and
_
-------
3. TARGET SPECIES
shellfish species will allow for the development of a national database for
tracking the magnitude and geographic extent of pollutant contamination in these
target species nationwide and will permit analyses of trends in fish/shellfish
contamination over time.
3.2 CRITERIA FOR SELECTING TARGET SPECIES
The appropriate choice of target species is a key element of any chemical
contaminant monitoring program. Criteria for selecting target species used in the
following national fish and shellfish contaminant monitoring programs were
reviewed by the EPA Fish Contaminant Workgroup to assess their applicability
for use in selecting target species for State fish contaminant monitoring
programs:
National Study of Chemical Residues in Fish (U.S. EPA)
• National Dioxin Study (U.S. EPA)
• 301 (h) Monitoring Program (U.S. EPA)
National Pesticide Monitoring Program (U.S. FWS)
National Contaminant Biomonitoring Program (U.S. FWS)
• National Status and Trends Program (NOAA).
National Water-Quality Assessment Program (USGS).
The criteria used to select target species in many of these programs are similar
although the priority given each criterion may vary depending on program aims.
The EPA Fish Contaminant Workgroup believes the most important criterion for
selecting target species for State fish and shellfish contaminant monitoring
programs assessing human consumption concerns is that the species are
commonly consumed in the study area and are of commercial, recreational, or
subsistence fishing value. Two other criteria of major importance are that the
species have the potential to bioaccumulate high concentrations of chemical
contaminants and have a wide geographic distribution. EPA recommends that
States use the same criteria to select species for both screening and intensive
site-specific studies.
In addition to the three primary criteria for target species selection, it is also
important that the target species be easy to identify taxonomically because there
are significant species-specific differences in bioaccumulation potential. Because
many closely related species can be similar in appearance, reliable taxonomic
identification is essential to prevent mixing of closely related species with the
target species. Note: Under no circumstance should individuals of more than
one species be mixed to create a composite sample (U.S. EPA, 1991e). It is
also both practical and cost-effective to sample target species that are abundant,
easy to capture, and large enough to provide adequate tissue samples for
chemical analyses.
3-2
-------
3. TARGET SPECIES
It cannot be overemphasized that final selection of target species will require the
expertise of State fisheries biologists with knowledge of local fish/shellfish
species that best meet the selection criteria and knowledge of local human
consumption patterns. Although, ideally, all fish or shellfish species consumed
from a given waterbody by the local population should be monitored, resource
constraints may dictate that only a few of the most frequently consumed species
be sampled.
In the next two sections, lists of recommended target species are provided for
freshwater ecosystems (inland fresh waters and the Great Lakes) and
estuarine/marine ecosystems (Atlantic, Gulf, and Pacific waters), and the
methods used in developing each list are discussed.
3.3 FRESHWATER TARGET SPECIES
As part of the two-tiered sampling strategy proposed for State fish contaminant
monitoring programs, EPA recommends that States collect one bottom-feeding
fish species and one predator fish species at each freshwater screening study
site. Some suggested target species for use in State fish contaminant
monitoring programs are shown in Table 3-1 for inland fresh waters and in Table
3-2 for Great Lakes waters.
The lists of target species recommended by the EPA Fish Contaminant
Workgroup for freshwater ecosystems were developed based on a review of
species used in the following national monitoring programs:
• National Study of Chemical Residues in Fish (U.S. EPA)
• National Dioxin Study (U.S. EPA)
National Pesticide Monitoring Program (U.S. FWS)
• National Contaminant Biomonitoring Program (U.S. FWS)
• National Water-Quality Assessment Program (USGS)
and on a review of fish species cited in State fish consumption advisories or
bans (RTI, 1993). Separate target species lists were developed for inland fresh
waters (Table 3-1) and Great Lakes waters (Table 3-2) because of the distinct
ecological characteristics of these waters and their fisheries. Each target
species list has been reviewed by regional and State fisheries experts.
Use of two distinct ecological groups of finfish (i.e., bottom-feeders and
predators) as target species in freshwater systems is recommended. This
permits monitoring of a wide variety of habitats, feeding strategies, and
physiological factors that might result in differences in bioaccumulation of
contaminants. Bottom-feeding species may accumulate high contaminant
concentrations from direct physical contact with contaminated sediment and/or
by consuming benthic invertebrates and epibenthic organisms that live in
contaminated sediment. Predator species are also good indicators of persistent
pollutants (e.g., mercury or DDT and its metabolites) that may be biomagnified
through several trophic levels of the food web. Species used in several Federal
_
-------
3. TARGET SPECIES
Table 3-1. Recommended Target Species for Inland Fresh Waters
Family name
Percichthyidae
Centrarchidae
Percidae
Cyprinidae
Catostomidae
Ictaluridae
Esocidae
Salmonidae
Common name
White bass
Largemouth bass
Smallmouth bass
Black crappie
White crappie
Walleye
Yellow perch
Common carp
White sucker
Channel catfish
Flathead catfish
Northern pike
Lake trout
Brown trout
Rainbow trout
Scientific name
Morone chrysops
Micropterus salmoides
Micropterus dolomieui
Pomoxis nigromaculatus
Pomoxis annularis
Stizostedion vitreum
Perca flavescens
Cyprinus carpio
Catostomus commersoni
Ictalurus punctatus
Pylodictis olivaris
Esox lucius
Salvelinus namaycush
Salmo trutta
Oncorhynchus mykiss*
'Formerly Salmo gairdneri.
Table 3-2. Recommended Target Species for Great Lakes Waters
Family name
Percichthyidae
Centrarchidae
Percidae
Cyprinidae
Catostomidae
Ictaluridae
Esocidae
Salmonidae
Common name
White bass
Smallmouth bass
Walleye
Common carp
White sucker
Channel catfish
Muskellunge
Chinook salmon
Lake trout
Brown trout
Rainbow trout
Scientific name
Morone chrysops
Micropterus dolomieui
Stizostedion vitreum
Cyprinus carpio
Catostomus commersoni
Ictalurus punctatus
Esox masquinongy
Oncorhynchus tschawytscha
Salvelinus namaycush
Salmo trutta
Oncorhynchus mykiss*
aFormeriy Salmo gairdneri.
3-4
-------
3. TARGET SPECIES
programs to assess the extent of freshwater fish tissue contamination nationwide
are compared in Table 3-3.
3.3.1 Bottom-Feeding Target Species
EPA recommends that, whenever practical, States use common carp (Cyprinus
carpio), channel catfish (Ictalurus punctatus), and white sucker (Catostomus
commersoni) in that order as bottom-feeding target species in both inland fresh
waters (Table 3-1) and in Great Lakes waters (Table 3-2). These bottom-feeders
have been used consistently for monitoring a wide variety of contaminants
including dioxins/dibenzofurans (Crawford and Luoma, 1993; U.S. EPA, 1992c,
1992d; Versar Inc., 1984), organochlorine pesticides (Crawford and Luoma,
1993; Schmitt et al., 1983, 1985, 1990; U.S. EPA, 1992c, 1992d), and heavy
metals (Crawford and Luoma, 1993; Lowe et al., 1985; May and McKinney,
1981; Schmitt and Brumbaugh, 1990; U.S. EPA, 1992c, 1992d). These three
species are commonly consumed in the areas in which they occur and have also
demonstrated an ability to accumulate high concentrations of environmental
contaminants in their tissues as shown in Tables 3-4 and 3-5. Note: The
average contaminant concentrations shown in Tables 3-4 and 3-5 for fish
collected for the National Study of Chemical Residues in Fish (U.S. EPA, 1992c,
1992d) were derived from concentrations in fish from undisturbed areas and from
areas expected to have elevated tissue contaminant concentrations. The mean
contaminant concentrations shown, therefore, may be higher than or lower than
those found in the ambient environment because of site selection criteria used
in this study.
In addition, these three species are relatively widely distributed throughout the
continental United States, and numerous States are already sampling these
species in their contaminant monitoring programs. A review of the database
National Listing of State Fish and Shellfish Consumption Advisories and Bans
(RTI, 1993) indicated that the largest number of States issuing advisories for
specific bottom-feeding species did so for carp (21 States) and channel catfish
(22 States), with eight States issuing advisories for white suckers (see Table 3-
6). Appendix A lists the freshwater fish species cited in consumption advisories
for each State.
3.3.2 Predator Target Species
EPA recommends that, whenever practical, States use predator target species
listed in Tables 3-1 and 3-2 for inland fresh waters and Great Lakes waters,
respectively. Predator species, because of their more definitive habitat and
water temperature preferences, generally have a more limited geographic
distribution. Thus, a greater number of predator species than of bottom feeders
have been used in national contaminant monitoring programs (Table 3-3) and
these are recommended for use as target species in freshwater ecosystems.
Predator fish that prefer relatively cold freshwater habitats include many
members of the following families: Salmonidae (trout and salmon), Percidae
(walleye and yellow perch), and Esocidae (northern pike and muskellunge).
_
-------
3. TARGET SPECIES
Table 3-3. Comparison of Freshwater Finfish Species Used in Several
National Fish Contaminant Monitoring Programs
u.s. EPA
National
Dloxin Study
U.S. FWS
NPMP"and
NCBP"
U.S. EPA
NSCRF6
USGS
NAWOA"
BOTTOM FEEDERS
Family Cyprinidae
Carp (Cyprinus carpio)
Family Ictaluridae
Channel catfish (Ictalurus punctetus)
Family Catostomidae
White sucker (Catastomus commersoni)
Longnose sucker (C. catostomus)
Largescale sucker (C, macrocheilus)
Spotted sucker (Minytrema melanops)
Redhorse sucker (Moxostoma sp.)
included variety of species:
Silver redhorse (M. anisurum)
Grey redhorse (M. congestion)
Black redhorse (M. duquesnei)
Golden redhorse (M. erythrurum)
Shorthead redhorse (M. macrolepidotum)
Blacktail redhorse (M. poedlurum)
Or other ictalurid
Or other catostomid
PREDATORS
Family Salmonidae
Rainbow trout (Oncorhynchus mytiss)
[formerly Salmo gairdneri]
Brown trout (Salmo trutta)
Brook trout (Salvelinus fontinalis)
Lake trout (Salmo namaycush)
Family Percidae
Walleye (Stizostedion vitreum)
Sauger (Stizostedion canadense)
Yellow perch (Perca flavescens)
Family Percichthyidae
White bass (Morone chtysops)
Family Centrarchidae
Largemouth bass (Micropterus salmoidos)
Smallmouth bass (Micropterus dolomieui)
Black crappie (Pomoxis nigromaculatus)
White crappie (Pomoxis annularis)
Bluegill sunfish (Lepomis macrochirus)
Family Esocidae
Northern pike (Esox lucius)
Family Ictaluridae
Flathead catfish (Pylodictis olivaris)
Or other percid
O
O
Or other centrarchid
O
O
O
Or other percid
O
O
Or other centrarchid
O
O
O
• Recommended target species
O Alternate target species
"National Pesticide Monitoring Program
''National Contaminant Biomonitoring Program
"National Study of Chemical Residues in Rsh
dNational Water Quality Assessment Program
Sources: Versar, Inc., 1984; Schmitt et al., 1990; Schmitt el al., 1983; May and McKinney, 1981; U.S. EPA, 1992c. 1992d;
Crawford and Luoma, 1993.
3-6
-------
Table 3-4. Average Fish Tissue Concentrations of Xenoblotlcs for Major Flnflsh Species
Sampled In the National Study of Chemical Residues In Fish8
Fish Species
Bottom Feeders'3
Carp
White Sucker
Channel Cat
Redhorse Sucker
Spotted Sucker
Predators b
Largemouth Bass
Smaflmouth Bass
Walleye
Brown Trout
White Bass
Northern Pike
Flathead Cat
White Crappie
Bluefish
Alpha BHC
3.10
3.31
2.87
0.82
1.45
0.15
0.36
ND
1.59
0.34
0.55
0.92
0.23
0.38
Gamma-BHC
4.34
1.66
3.17
0.41
2.63
0.07
0.15
NO
NO
0.79
ND
0.58
NO
0.12
Biphenyl
4.38
1.28
1.24
1.25
3.35
0.38
0.33
0.40
0.81
0.62
0.59
0.60
0.21
0.20
Chlorpyrilos
8.23
1.75
6.97
0.35
0.56
0.23
0.08
0.04
ND
1.32
11.43
2257
ND
ND
Oicolol
0.88
0.48
0.59
ND
0.05
0.20
ND
ND
0.94
ND
0.31
1.28
ND
ND
Dieldnn
44.75
22.75
15.44
5.35
5.52
5.01
2.34
3.73
20.13
9.35
9.04
37.38
ND
2.87
Endrin
1.40
0.24
9.07
0.97
ND
ND
ND
ND
ND
ND
ND
3.45
ND
ND
Heptachlor
Epoxida
4.00
1.09
0.50
ND
ND
0.30
0.07
021
2.08
1.40
ND
0.57
ND
ND
Mercury
(ppm)
0.11
0.11
0.09
027
0.12
0.46
0.34
0.51
0.14
0.35
0.34
0.27
022
0.22
Mirex
3.70
4.35
14.59
0.57
1.79
0.21
1.99
0.08
43.98
0.11
2.39
ND
ND
0.13
Oxychlordane
8.20
310
6.41
237
0.05
0.47
0.54
1.11
5.38
0.84
4.00
063
ND
ND
PCBs
2941.13
1697.81
1300.52
487.72
13390
232.26
496.22
368.65
2434.07
288.35
788.40
521.19
22.34
368.06
aThese average fish tissue concentrations may be higher than or lower than those found in the ambient environment because of site selection criteria used in this study.
bValues were calculated using whole-body samples for bottom-feeders and fillet samples for predators.
Individual values below detection were set at zero. Units = ppb, unless noted. ND = Not detected
Source: U.S. EPA, 1991 h.
Fish Species
Bottom Feeders^
Carp
White Sucker
Channel Cat
Redhorse Sucker
Spotted Sucker
Predators^
Largemouth Bass
Smallmouth Bass
Walleye
Brown Trout
White Bass
Northern Pike
Flalhead Cat
White Crappie
Bluefish
Pentachloro-
anisole
16.50
9.06
3960
2.87
17.68
0.57
0.23
0.76
0.09
0.93
1.51
0.31
0.33
0.05
Pentachloro-
benzene
1.04
0.39
1.32
0.02
0.02
0.02
0.02
ND
0.60
ND
0.09
ND
ND
ND
DDE
415.43
78.39
62777
87.25
75.31
55.72
33.63
34.00
158.90
17.44
59.50
755.18
10.04
29.13
Total
Chlordane
67.15
16.42
54.39
16.48
12.33
2.89
4.01
3.62
7.25
10.67
5.45
16.07
0.34
7.74
Total
Nonachlor
63.15
20.83
66.28
30.73
15.00
4.21
7.82
8.04
32.60
16.00
13.88
14.04
0.28
7.56
123TCB
1.54
0.16
0.14
0.55
3.34
0.22
0.70
0.29
1.10
0.21
0.30
0.10
0.08
6.25
124TCB
4.77
0.30
0.37
6.48
12.00
0.19
0.59
0.38
0.98
0.10
0.23
0.18
0.08
4.66
135 TCB
0.08
0.14
ND
0.08
1.00
0.03
0.04
ND
ND
ND
ND
ND
ND
0.57
1234TECB
0.30
0.15
0.88
0.09
0.09
0.01
004
0.004
0.09
0.01
0.01
ND
ND
ND
Trifluralin
12.55
ND
1.00
ND
ND
ND
ND
ND
NO
ND
ND
44.37
ND
NO
Hexachloro-
benzene
3.58
362
2.36
0.58
0.02
0.20
036
0.11
3.06
0.83
0.20
0.85
ND
ND
CO
3D
O
m
TJ
m
o
m
-------
Table 3-5. Average Fish Tissue Concentrations of Dloxlns and Furans for Major Flnflsh Species
Sampled In the National Study of Chemical Residues In Fish8
Fish Species
Bottom Feeders'3
Carp
White Sucker
Channel Catfish
Redhorse Sucker
Spotted Sucker
Predators*3
Largemouth Bass
Smallmouth Bass
Walleye
Brown Trout
White Bass
Northern Pike
Flathead Cattish
White Crapple
Blueltsh
2378
TCDD
7.76
B.08
11.56
4.65
1.73
1.73
0.72
0.88
2.52
3.00
0.77
0.78
2.13
0.85
12378
PeCDD
3.63
2.05
2.37
1.50
2.34
0.59
0.50'
0.54*
1.01
0.66
0.46*
0.43
0.60
0.56
123478
HxCDD
2.16
1.03
1.61
1.40
1.70
1.12
1.13*
0.99*
1.07*
1.05*
1.23*
0.90
1.29*
1.23*
123678
HxCDD
6.81
1.96
5.62
2.36
12.08
1.28
0.79
0.73
0.98
0.78
0.91
1.06
1.03*
0.98*
123789
HxCDD
1.54
0.88
1.29
0.84
1.14
0.64
0.64*
0.62*
0.68*
0.61*
0.69'
0.50
0.83*
0.69'
1234678
HpCDD
22.29
3.72
9.40
4.94
17.48
2.48
0.67
0.88
1.18
1.01
0.73
1.67
1.33
0.65
2378
TCDF
10.15
22.89
2.22
30.09
7.49
2.18
1.93
1.83
3.74
5.07
1.01
1.63
10.46
2.11
12378
PeCDF
1.31
1.10
0.52
0.75
2.12
0.37
0.36'
0.35'
0.60
0.40
0.44
0.40
0.54
0.41
23478
PeCDF
4.01
2.64
2.91
1.28
2.06
0.47
0.51
0.38
1.36
0.49
0.66
0.56
0.67
0.59
123478
HxCDF
2.54
2.21
2.41
2.10
2.22
1.24
.28
.04
.47
.04
.41*
.05
1.33*
1.42*
123678
HxCDF
1.91
1.29
1.41
1.16
1.79
.23
.23
.09*
.12*
.16*
.42*
1.20*
1.33'
1.42'
123789
HxCDF
1.16
.06
.38'
.19*
.28*
.21*
.26'
.07'
.09*
.13"
.38*
.17*
1.30*
1.39*
234678
HxCDF
1.20
1.09
1.62
1.50
1.78
0.88
0.89*
0.75
0.94'
o.er
0.98"
0.61*
0.95*
0.98*
1234678
HpCDF
2.49
1.23
2.55
1.57
1.77
0.82'
0.69
0.74
0.67'
0.63
0.56
0.56
0.96*
0.72'
1234789
HpCDF
.22
.13
.26
.36*
.08
1.21*
1.30*
1.21*
1.16*
1.17*
1.30*
1.10*
1.34*
1.31'
TEC
13.06
12.79
14.80
9.22
6.23
1.91
0.65*
0.79"
3.31
3.44
0.66
0.99
3.80
1.41
aThese average fish tissue concentrations may be higher than or lower than those found in the ambient environment because of site selection criteria used in this study.
bValues were calculated using whole-body samples for bottom-feeders and fillet samples for predators. Values below
detection have been replaced by one-half detection limit for the given sample. Asterisk indicates all values below detection.
Units = ppt (parts per trillion).
Source: U.S. EPA, 1991 h.
to
W
00
JJ
o
m
w
•o
m
o
m
-------
3. TARGET SPECIES
Table 3-6. Principal Freshwater Fish Species Cited In State Fish
Consumption Advisories9
Family name
Perdchthyidae
Centrarchidae
Common name
White bass
Striped bass
White perch
Largemouth bass
Smallmouth bass
Black crappie
White crappie
Bluegill
Rock bass
Number of States
Scientific name with advisories'*
Morone chrysops
Morone saxatilis
Morone americana
Micropterus salmoides
Micropterus dolomieui
Pomoxis nigromaculatus
Pomoxis annularis
Lepomis macrochims
Ambloplites rupestris
10
6
4
15
9
5
2
5
3
Percidae
Cyprinidae
Acipenseridae
Catostomidae
Ictaluridae
Sciaenidae
Esocidae
Salmonidae
Yellow perch
Sauger
Walleye
Common carp
Shovelnose sturgeon
Lake sturgeon
Smallmouth buffalo
Bigmouth buffalo
Shorthead redhorse
White sucker
Quillback carpsucker
White catfish
Channel catfish
Flathead catfish
Black bullhead
Brown bullhead
Yellow bullhead
Freshwater drum
Northern pike
Muskellunge
Coho salmon
Chinook salmon
Brown trout
Lake trout
Rainbow trout
Brook trout
Lake whitefish
Perca flavescens 8
Stizostedion canadense 4
Stizostedion vitreum 9
Cyprinus carpio 21
Scaphirhynchus platorynchus 1
Acipenser fulvescens 2
Ictiobus bubalus 4
Ictiobus cyprinellus 4
Moxostoma macrolepidotum 2
Catostomus commersoni 8
Carpiodes cyprinus 2
Ictalurus catus 5
Ictalurus punctatus 22
Pylodictis olivaris 4
Ictalurus melas 2
Ictalurus nebulosus 7
Ictalurus natalis 2
Aplodinotus grunniens 3
Esox lucius 7
Esox masquinongy 4
Oncorhynchus kisutch 6
Oncorhynchus tschawytscha 7
Salmo trutta 9
Salvelinus namaycush 10
Oncorhynchus mykiss* 8
Salvelinus fontinalis 3
Coregonus clupea formis 2
Anguillidae
American eel
Anguilla rostrata
aSpecies in boldface are EPA-recommended target species for inland fresh waters (see Table 3-1) and the
Great Lakes waters (Table 3-2).
bMany States did not identify individual species of finfish in their advisories.
cFormerly Salmo gairdneri.
Source: RTI, 1993.
3-9
-------
3. TARGET SPECIES
Members of the Centrarchidae (large- and smallmouth bass, crappie, and
sunfish), Percichthyidae (white bass), and Ictaluridae (flathead catfish) families
prefer relatively warm water habitats. Only two predator species (brown trout
and largemouth bass) have been used in all four of the national monitoring
programs reviewed (Table 3-3). However, most of the other predator species
recommended as target species have been used in at least one national
monitoring program.
To identify those predator species with a known ability to bioaccumulate
contaminants in their tissues, the EPA Workgroup reviewed average tissue
concentrations of xenobiotic contaminants for major predator fish species
sampled in the National Study of Chemical Residues in Fish. Unlike the bottom-
feeders (common carp, channel catfish, and white suckers), no single predator
species or group of predator species consistently exhibited the highest tissue
concentrations for the contaminants analyzed (Tables 3-4 and 3-5). However,
average fish tissue concentrations for some contaminants (i.e., mercury, mirex,
chlorpyrifos, DDE, 1,2,3-trichlorobenzene [123-TCB], and trifluralin) were higher
for some predator species than for the bottom-feeders despite the fact that only
the fillet portion rather than the whole body was analyzed for predator species.
This finding emphasizes the need for using two types of fish (i.e., bottom-feeders
and predators) with different habitat and feeding strategies as target species.
The current fish consumption advisories for these predator target species are
further justification for their recommended use. As was shown for the bottom-
feeder target species, States are already sampling the recommended predator
target species listed in Table 3-6. The largest number of States issuing
advisories for specific predator species did so for largemouth bass (15), lake
trout (10), white bass (10), smallmouth bass (9), brown trout (9), walleye (9),
rainbow trout (8), yellow perch (8), Chinook salmon (7), northern pike (7), black
crappie (5), flathead catfish (4), and muskellunge (4) (RTI, 1993).
Because some freshwater finfish species (e.g., several Great Lake salmonids)
are highly migratory, harvesting of these species may be restricted to certain
seasons because sexually mature adult fish (i.e., the recommended size for
sampling) may make spawning runs from the Great Lakes into tributary streams.
EPA recommends that spawning populations not be sampled in fish contaminant
monitoring programs. Sampling of target finfish species during their spawning
period should be avoided because contaminant tissue concentrations may
decrease during this time (Phillips, 1980) and because the spawning period is
generally outside the legal harvest period. Note: Target finfish may be sampled
during their spawning period, however, if the species can be legally harvested
at this time.
State personnel, with their knowledge of site-specific fisheries and human
consumption patterns, must be the ultimate judge of the species selected for use
in freshwater fish contaminant monitoring programs within their jurisdiction.
3-10
-------
3. TARGET SPECIES
3.4 ESTUARINE/MARINE TARGET SPECIES
EPA recommends that States collect either one shellfish species (preferably a
bivalve mollusc) and one finfish species or two finfish species at each
estuarine/marine screening site. In all cases, the primary criterion for selecting
the target species is that it is commonly consumed. Ideally, one shellfish
species and one finfish species should be sampled; however, if no shellfish
species from the recommended target species list meets the primary criterion,
EPA recommends that States use two finfish species selected from the
appropriate regional estuarine/marine target species lists. If two finfish are
selected as the target species, one should be a bottom-feeding species.
EPA recommends that, whenever practical, States use target species selected
from fish and shellfish species identified in Tables 3-7 through 3-13 for the
following specific estuarine/marine coastal areas:
Northeast Atlantic region (Maine through Connecticut)—Table 3-7
Mid-Atlantic region (New York through Virginia)—Table 3-8
Southeast Atlantic region (North Carolina through Florida)—Table 3-9
• Gulf Coast region (west coast of Florida through Texas)—Table 3-10
Pacific Northwest region (Alaska through Oregon)—Table 3-11
Northern California waters (Klamath River through Morro Bay)—Table 3-12
• Southern California waters (Santa Monica Bay to Tijuana Estuary)—Table
3-13.
The seven separate regional lists of target species recommended by the EPA
Workgroup for estuarine/marine ecosystems were developed because of
differences in species' geographic distribution and abundance and the nature of
the regional fisheries and were developed based on a review of species used in
the following national monitoring programs:
• National Dioxin Study (U.S. EPA)
Section 301 (h) Monitoring Program (U.S. EPA)
• National Status and Trends Program (NOAA)
National Study of Chemical Residues in Fish (U.S. EPA).
Because some of these programs identified some fish and shellfish species that
are not of commercial, sportfishing, or subsistence value, several recent
literature sources identifying commercial and sportfishing species were also
reviewed (Table 3-14). Some sources included information on seasonal
distribution and abundance of various life stages (i.e., adults, spawning adults,
juveniles) of fish and shellfish species. This information was useful in delineating
seven regional estuarine/marine areas nationwide. The EPA Workgroup also
reviewed fish and shellfish species cited in State consumption advisories for
estuarine/marine waters (Appendix A). Each of the final regional lists of target
species has been reviewed by State, regional, and national fisheries experts.
3-11
-------
3. TARGET SPECIES
Table 3-7. Recommended Target Species for Northeast Atlantic
Estuaries and Marine Waters (Maine through Connecticut)
Family name
FInflsh Species
Anguillidae
Percichthyidae
Pomatomidae
Sparidae
Sciaenidae
Bothidae
Common name
American eel
Striped bass
Bluefish
Scup
Weakfish
Summer flounder
Scientific name
Anguilla rostrata
Morone saxatilis
Pomatomus saltatrix
Stenotomus chrysops
Cynoscion regalis
Paralichthys dentatus
Pleuronectidae
Shellfish Species
Bivalves
Crustaceans
Four-spotted flounder
Winter flounder
Yellowtail flounder
American dab
Soft-shell clam
Hard clam
Ocean quahog
Surf clam
Blue mussel
American lobster
Eastern rock crab
Paralichthys oblongus
Pseudopleuronectes
americanus
Limanda ferruginea
Hippoglossoides
platessoides
Mya arenaria
Mercenaria mercenaria
Arctica islandica
Spisula solidissima
Mytilus edulis
Homarus americanus
Cancer irroratus
3-12
-------
3. TARGET SPECIES
Table 3-8. Recommended Target Species for Mid-Atlantic
Estuaries and Marine Waters (New York through Virginia)
Family name
Finfish Species
Anguillidae
Ictaluridae
Percichthyidae
Pomatomidae
Sparidae
Sciaenidae
Bothidae
Pleuronectidae
Common name
American eel
Channel catfish
White catfish
White perch
Striped bass
Bluefish
Scup
Weakfish
Spot
Atlantic croaker
Red drum
Summer flounder
Winter flounder
Scientific name
Anguilla rostrata
Ictalurus punctatus
Ictalurus catus
Morone americana
Morone saxatilis
Pomatomus saltatrix
Stenotomus chrysops
Cynoscion regalis
Leistomus xanthurus
Micropogonias undulatus
Sciaenops ocellatus
Paralichthys dentatus
Pseudopleuronectes
americanus
Shellfish Species
Bivalves
Crustaceans
Hard clam
Soft-shell clam
Ocean quahog
Surf clam
Blue mussel
American oyster
Blue crab
American lobster
Eastern rock crab
Mercenaria mercenaria
Mya arenaria
Arctica islandica
Spisula solidissima
Mytilus edulis
Crassostrea virginica
Callinectes sapidus
Homarus americanus
Cancer irroratus
3-13
-------
3. TARGET SPECIES
Table 3-9. Recommended Target Species for Southeast Atlantic
Estuaries and Marine Waters (North Carolina through Florida)
Family name
FJrtflsn Sp0cle$
Anguillidae
Ictaluridae
Percichthyidae
Sciaenidae
Bothidae
Shellfish Species
Bivalves
Crustaceans
Common name
American eel
Channel catfish
White catfish
White perch
Striped bass
Spot
Atlantic croaker
Red drum
Southern flounder
Summer flounder
Hard clam
American oyster
West Indies spiny lobster
Blue crab
Scientific name
Anguilla rostrata
Ictalurus punctatus
Ictalurus catus
Morone americana
Morone saxatilis
Leistomus xanthurus
Micropogonias undulatus
Sciaenops ocellatus
Paralichthys lethostigma
Paralichthys dentatus
Mercenaria mercenaria
Crassostrea virginica
Panulirus argus
Callinectes sapidus
3-14
-------
3. TARGET SPECIES
Table 3-10. Recommended Target Species for Gulf of Mexico
Estuaries and Marine Waters (West Coast of Florida through Texas)
Family name
FJnfteh Species
Ictaluridae
Ariidae
Sciaenidae
Bothidae
Shellfish Species
Bivalves
Crustaceans
Common name
Blue catfish
Channel catfish
Hardhead catfish
Spotted seatrout
Spot
Atlantic croaker
Red drum
Gulf flounder
Southern flounder
American oyster
Hard clam
White shrimp
Blue crab
Gulf stone crab
West Indies spiny lobster
Scientific name
Ictalurus furcatus
Ictalurus punctatus
Arius felis
Cynoscion nebulosus
Leistomus xanthurus
Micropogonias undulatus
Sciaenops ocellatus
Paralichthys albigutta
Paralichthys lethostigma
Crassostrea virginica
Mercenaria mercenaria
Penaeus setiferus
Callinectes sapidus
Menippe adina
Panulirus argus
3-15
-------
3. TARGET SPECIES
Table 3-11. Recommended Target Species for Pacific Northwest
Estuaries and Marine Waters (Alaska through Oregon)
Family name
Flnflsh Species
Embiotocidae
Scorpaenidae
Bothidae
Pleuronectidae
Salmonidae
Common name
Redtail Surfperch
Copper rockfish
Black rockfish
Speckled sanddab
Pacific sanddab
Starry flounder
English sole
Coho salmon
Chinook salmon
Scientific name
Amphistichus rhodoterus
Sebastes caurinus
Sebastes melanops
Citharichthys stigmaeus
Citharichthys sordidus
Platichthys stellatus
Parophrys vetulus
Onchorhynchus kisutch
Onchorhynchus tshawytscha
Shellfish Species
Bivalves
Crustaceans
Blue mussel
California mussel
Pacific oyster
Horseneck clam
Pacific littleneck clam
Soft-shell clam
Manila clam
Dungeness crab
Red crab
Mytilus edulis
Mytilus califomianus
Crassostrea gigas
Tresus capax
Protothaca staminea
Mya arenaria
Venerupis japonica
Cancer magister
Cancer productus
3-16
-------
3. TARGET SPECIES
Table 3-12. Recommended Target Species for Northern California
Estuaries and Marine Waters (Klamath River through Morro Bay)
Family name
Flnflsh Species
Triakidae
Sciaenidae
Embiotocidae
Scorpaenidae
Bothidae
Pleuronectidae
Salmonidae
Shellfish Species
Bivalves
Crustaceans
Common name
Leopard shark
White croaker
Redtailed surfperch
Striped seaperch
Black rockfish
Yellowtail rockfish
Bocaccio
Pacific sanddab
Speckled sanddab
Starry flounder
English sole
Coho salmon
Chinook salmon
Blue mussel
California mussel
Pacific littleneck clam
Soft-shell clam
Dungeness crab
Red crab
Pacific rock crab
Scientific name
Triakis semifasciata
Genyonemus lineatus
Amphistichus rhodoterus
Embiotoca lateralis
Sebastes melanops
Sebastes flavidus
Sebastes paucispinis
Citharichthys sordidus
Citharichthys stigmaeus
Platichthys stellatus
Parophrys vetulus
Onchorhynchus kisutch
Onchorhynchus tshawytscha
Mytilus edulis
Mytilus californianus
Protothaca staminea
Mya arenaria
Cancer magister
Cancer productus
Cancer antennarius
3-17
-------
3. TARGET SPECIES
Table 3-13. Recommended Target Species for Southern California
Estuaries and Marine Waters (Santa Monica Bay to Tijuana Estuary)
Family name
Finfish Species
Serranidae
Sciaenidae
Embiotocidae
Scorpaenidae
Pleuronectidae
Shellfish Species
Bivalves
Crustaceans
Common name
Kelp bass
Barred sand bass
White croaker
Corbina
Black perch
Walleye surf perch
Barred surfperch
California scorpionfish
Widow rockfish
Blue rockfish
Bocaccio
Diamond turbot
Dover sole
Blue mussel
California mussel
Pacific littleneck clam
Pacific rock crab
Red crab
California rock lobster
Scientific name
Paralabrax clathratus
Paralabrax nebulifer
Genyonemus lineatus
Menticirrhus undulatus
Embiotoca jacksoni
Hyperprosopan argenteum
Amphistichus argenteus
Scorpaena guttata
Sebastes entomelas
Sebastes mystinus
Sebastes paucispinis
Hypsopetta guttulata
Microstomus pacificus
Mytilus edulis
Mytilus californianus
Protothaca staminea
Cancer antennarius
Cancer productus
Panulirus interruptus
3-18
-------
3. TARGET SPECIES
Table 3-14. Sources of Information on Commercial and Sportflshlng
Species In Various Coastal Areas of the United States
Geographic
area Source
Atlantic Coast National Marine Fisheries Service. 1987. Marine Recreational Fishery Statistics Survey,
Atlantic and Gulf Coasts, 1986. Current Fishery Statistics Number 8392. National Oceanic
and Atmospheric Administration, U.S. Department of Commerce, Rockville, MO.
Leonard, D.L., M.A. Broutman, and K.E. Harkness. 1989. The Quality of Shellfish Growing
Waters on the East Coast of the United States. Strategic Assessment Branch, National
Oceanic and Atmospheric Administration, U.S. Department of Commerce, Rockville, MD.
Nelson, D.M., M.E. Monaco, E.A. Irlandi, L.R. Settle, and L Coston-Clements. 1991.
Distribution and Abundance of Fishes and Invertebrates in Southeast Estuaries. ELMR Report
No. 9. Strategic Assessment Division. National Oceanic and Atmospheric Administration, U.S.
Department of Commerce, Rockville, MD.
Gulf Coast National Marine Fisheries Service. 1987. Marine Recreational Fishery Statistics Survey,
Atlantic and Gulf Coasts, 1986. Current Fishery Statistics Number 8392. National Oceanic
and Atmospheric Administration, U.S. Department of Commerce, Rockville, MD.
Broutman, M.A., and D.L. Leonard. 1988. The Quality of Shellfish Growing Waters in the Gulf
of Mexico. Strategic Assessment Branch, National Oceanic and Atmospheric Administration,
Rockville, MD.
Monaco, M.E., D.M. Nelson, T.C. Czapla, and M.E. Patillo. 1989. Distribution and Abundance
of Fishes and Invertebrates in Texas Estuaries. ELMR Report No. 3. Strategic Assessment
Branch, National Oceanic and Atmospheric Administration, U.S. Department of Commerce,
Rockville, MD.
Williams, C.D., D.M. Nelson, M.E. Monaco, S.L. Stone, C. lancu, L. Coston-Clements, L.R.
Settle, and E.A. Irlandi. 1990. Distribution and Abundance of Fishes and Invertebrates in
Eastern Gulf of Mexico Estuaries. ELMR Report No. 6. Strategic Assessment Branch,
National Oceanic and Atmospheric Administration, U.S. Department of Commerce, Rockville,
MD.
Czapla, T.C., M.E. Patillo, D.M. Nelson, and M.E. Monaco. 1991. Distribution and Abundance
of Fishes and Invertebrates in Central Gulf of Mexico Estuaries. ELMR Report No. 7.
Strategic Assessment Branch, National Oceanic and Atmospheric Administration, U.S.
Department of Commerce, Rockville, MD.
West Coast National Marine Fisheries Service. 1987. Marine Recreational Fishery Statistics Survey,
Pacific Coast, 1986. Current Fishery Statistics Number 8393. National Oceanic and
Atmospheric Administration, U.S. Department of Commerce, Rockville, MD.
Leonard, D.L., and E.A. Slaughter. 1990. The Quality of Shellfish Growing Waters on the
West Coast of the United States. Strategic Assessment Branch, National Oceanic and
Atmospheric Administration, U.S. Department of Commerce, Rockville, MD.
Monaco, M.E., D.M. Nelson, R.L. Emmett, and S.A. Hinton. 1990. Distribution and Abundance
of Fishes and Invertebrates in West Coast Estuaries. Volume I: Data Summaries. ELMR
Report No. 4. Strategic Assessment Branch, National Oceanic and Atmospheric
Administration, Rockville, MD.
Emmett, R.L., S.A. Hinton, S.L. Stone, and M.E. Monaco. 1991. Distribution and Abundance
of Fishes and Invertebrates in West Coast Estuaries. Volume II: Life History Summaries.
ELMR Report No. 8. Strategic Environmental Assessment Division, Rockville, MD.
3-19
-------
3. TARGET SPECIES
Use of two distinct ecological groups of organisms (shellfish and finfish) as target
species in estuarine/marine systems is recommended. This permits monitoring
of a wide variety of habitats, feeding strategies, and physiological factors that
might result in differences in bioaccumulation of contaminants. Estuarine/marine
species used in several national contaminant monitoring programs are compared
in Table 3-15.
3.4.1 Selection of Target Shellfish Species
Selection of shellfish species (particularly bivalve molluscs) as target species
received primary consideration by the EPA Workgroup because of the
commercial, recreational, and subsistence value of shellfish in many coastal
areas of the United States. Bivalve molluscs (e.g., oysters, mussels, and clams)
are filter feeders that accumulate contaminants directly from the water column
or via ingestion of contaminants adsorbed to phytoplankton, detritus, and
sediment particles. Bivalves are good bioaccumulators of heavy metals
(Cunningham, 1979) and polycyclic aromatic hydrocarbons (PAHs) and other
organic compounds (Phillips, 1980; NOAA, 1987) and, because they are sessile,
they may reflect local contaminant concentrations more accurately than more
mobile crustacean or finfish species.
Three bivalve species—the blue mussel (Mytilus edulis), the California mussel
(Mytilus californianus), and the American oyster (Crassostrea mj/n/ca)—were
recommended and/or used in three of the national monitoring programs. Two
other bivalve species—the soft-shell clam (Mya arenaria) and the Pacific oyster
(Crassostrea gigas)—were also recommended and/or used in two national pro-
grams. Although no bivalve species was identified by name in State fish and
shellfish consumption advisories (Appendix A), seven coastal States issued
advisories for unspecified bivalves or shellfish species that may have included
these and other bivalve species. All three species are known to bioaccumulate
a variety of environmental contaminants (Phillips, 1988). The wide distribution
of these three species makes them useful for comparisons within a State or
between States sharing coastal waters (Figure 3-1). Because these three
species meet all of the selection criteria, they are recommended as target
species for use in geographic areas in which they occur.
In addition, several species of edible clams were added to the various
estuarine/marine target species lists based on recommendations received from
specific State and regional fisheries experts.
Crustaceans are also recommended as target species for estuarine/marine
sampling sites. Many crustaceans are bottom-dwelling and bottom-feeding
predator and/or scavenger species that are good indicators of contaminants that
may be biomagnified through several trophic levels of the food web. Several
species of lobsters and crabs have been recommended in one national
monitoring program, and the Dungeness crab has been recommended in two
national monitoring programs (Table 3-15). These crustaceans, although of
fishery value in many areas, are not as widely distributed nationally as the three
3-20
-------
3. TARGET SPECIES
Table 3-15. Estuarine/Marine Species Used in Several National
Fish and Shellfish Contaminant Monitoring Programs
FINFISH
Family Aciponseridae
White sturgeon (Acipenser transmontanus)
Family Ariidae
Hardhead catfish (Arius felis)
Family Percichthyidae
White perch (Morone americana)
Family Pomatomidae
Bluet ish (Pomatomus saltatrix)
Family Lutjanidae
Red snapper (Lutjanus campechanus)
Family Sparidae
Sheepshead (Archosargus probatocephalus)
Family Sciaenidae
Spotted seatrout (Cynoscion nebulosus)
Weakfish (Cynoscion regalis)
Spot (Leiostomus xanthurus)
White croaker (Genyonemus lineatus)
Atlantic croaker (Micropogonias undulatus)
Black drum (Pogonias cromis)
Red drum (Sciaenops ocellatus)
Family Serranidae
Barred sand bass (Paralabrax nebulifer)
Family Mugilidae
Striped mullet (Mugil cephalus)
Family Bothidae
Southern flounder (Paralichthys lethostigma)
Windowpane flounder (Scophthatmus aquosus)
Family Pleuronectidae
Pacific sanddab (Citharichthys sordidus)
Flathead sole (Hippoglossoides elassodon)
Diamond turbot (Hypsopsetta guttulata)
Starry flounder (Platichthys stellatus)
Hornyhead turbot (Pleuronichthys verticalis)
Winter flounder (Pseudopleuronectes americanus)
English sole (Parophrys vetulus)
Dover sole (Microstomus pacificus)
U.S. EPA
National
Dioxln
Study*
NOAA
Status
and
Trends
•
•
•
•
•
U.S. EPA
301 (h)
Program
•
•
•
•
•
U.S. EPA
NSCRFb-c
•
•
•
•
•
•
•
•
•
•
•
See notes at end of table.
(continued)
3-21
-------
3. TARGET SPECIES
Table 3-15. (continued)
SHELLFISH
Bivalves
Hard clam (Mercenaria mercenaria)
Soft-shell clam (Mya arenaria)
Ocean quahog (Arctica islandia)
Surf clam (Spisula solidissima)
Blue mussel (Mytilus edulis)
California mussel (Mytilus californianus)
American oyster (Crassostrea virginica)
Hawaiian oyster (Ostrea sandwichensis)
Pacific oyster (Crassostrea gigas)
Bent-nosed macoma (Macoma nasuta)
Baltic macoma (Macoma baltica)
White sand macoma (Macoma secta)
Crustaceans
American lobster (Homarus americanus)
West Indies spiny lobster (Panulirus argus)
California rock lobster (Panulirus interrupts)
Hawaiian spiny lobster (Panulirus penicillatus)
Eastern rock crab (Cancer irroratus)
Dungeness crab (Cancer magister)
Pacific rock crab (Cancer antennarius)
Yellow crab (Cancer anthonyi)
Red crab (Cancer productus)
U.S. EPA
National
Dloxln
Study*
•
•
•
NOAA
Status
and
Trends
•
•
•
•
U.S. EPA
301 (h)
Program
U.S. EPA
NSCRFb-c
•
•
•
"Only freshwater finfish were identified as target species; bivalves were identified as estuarin e/marine target species.
bSpecies listed were those collected at more than one site nationally; Salmonidae were not listed because they were
included on freshwater lists.
cNational Study of Chemical Residues in Fish.
3-22
-------
Mytilus edulis
Crassostrea virginica
Mytilus californianus
CO
M
CO
Figure 3-1. Geographic distributions of three bivalve species used extensively in national contaminant monitoring
programs (based on data from Abbott, 1974).
CO
a
o
m
W
•O
m
o
m
-------
3. TARGET SPECIES
bivalve species (Figure 3-1). However, they should be considered for selection
as target species in States where they are commonly consumed.
Only two crustaceans—the American lobster (Homarus americanus) and the blue
crab (Callinectes sapidus)—were specifically identified in State advisories (RTI,
1993). However, seven coastal States reported advisories in estuarine/marine
waters for unspecified shellfish species that may have included these and other
crustacean species (Table 3-16). All of the shellfish species cited in State
advisories are included as EPA-recommended target species on the appropriate
estuarine/marine regional lists.
3.4.2 Selection of Target Finfish Species
Two problems are encountered in the selection of target finfish species for
monitoring fish tissue contamination at estuarine/marine sites regionally and
nationally. First is the lack of finfish species common to both Atlantic and Gulf
Coast waters as well as Pacific Coast waters. Species used in several Federal
fish contaminant monitoring programs are compared in Table 3-15. Members
of the families Sciaenidae (seven species), Bothidae (two species), and
Pleuronectidae (eight species) were used extensively in these programs.
Bottom-dwelling finfish species (e.g., flounders in the families Bothidae and
Pleuronectidae) may accumulate high concentrations of contaminants from direct
physical contact with contaminated bottom sediments. In addition, these finfish
feed on sedentary infaunal or epifaunal organisms and are at additional risk of
accumulating contaminants via ingestion of these contaminated prey species
(U.S. EPA, 1987a). For finfish species, two Atlantic coast species, spot
(Leiostomus xanthurus) and winter flounder (Pseudopleuronectes americanus),
are recommended and/or used in three of the national monitoring programs, and
the Atlantic croaker (Micropogonias undulatus) is recommended and/or used in
two national monitoring programs. Three Pacific coast species, Starry flounder
(Platichthys stellatus), English sole (Parophrys vetulus), and Dover sole
(Microstomus pacificus), are recommended or used in two of the national
monitoring programs.
Second, because some estuarine/marine finfish species are highly migratory,
harvesting of these species may be restricted to certain seasons because
sexually mature adult fish (i.e., the recommended size for sampling) may enter
the estuaries only to spawn. EPA recommends that neither spawning
populations nor undersized juvenile stages be sampled in fish contaminant
monitoring programs. Sampling of target finfish species during their spawning
period should be avoided as contaminant tissue concentrations may decrease
during this time (Phillips, 1980) and because the spawning period is generally
outside the legal harvest period. Note: Target finfish species may be sampled
during their spawning period, however, if the species can be legally harvested
at this time. Sampling of undersized juveniles of species that use estuaries as
nursery areas is precluded by EPA's recommended monitoring strategy because
juveniles may not have had sufficient time to bioaccumulate contaminants or
attain harvestable size.
__
-------
3. TARGET SPECIES
Table 3-16. Principal Estuarine/Marine Fish and Shellfish Species Cited In State
Consumption Advisories8'6
Species
group name
FinfJsh
Percichthyidae
Ictaluridae
Anguillidae
Pomatomidae
Belonidae
Serranidae
Sciaenidae
Shellfish
Crustacean^
Common name
Striped bass
White perch
White catfish
Channel catfish
American eel
Bluefish
Atlantic needlefish
Kelp bass
Black croaker
White croaker
Queenfish
Corbina
American lobster
Blue crab
Number of States
Scientific name with advisories
Morone saxatilis
Morone americana
Ictalurus catus
Ictalurus punctatus
Anguilla rostrata
Pomatomus saltatrix
Strongylura marina
Paralabrax clathratus
Cheilotrema saturnum
Genyonemus Hneatus
Seriphus politus
Menticirrhus undulatus
s
Homarus americanus
Callinectes sapidus
5
3
4
5
6
4
1
1
1
1
1
1
•••. w. \ %
1
3
a Species in boldface are EPA-recommended target species for regional estuarine/marine waters (see
Tables 3-7 through 3-13).
b Many coastal States issued advisories for fish and shellfish species and thus did not identify specific
finfish and shellfish species in their advisories.
c Seven coastal States (American Samoa, California, Louisiana, Massachusetts, New Jersey, South
Carolina, and Texas) report advisories for unspecified shellfish or bivalve species.
Source: RTI, 1993.
3-25
-------
3. TARGET SPECIES
Because of these problems, the EPA Workgroup consulted with regional and
State fisheries experts and reviewed the list of current State fish consumption
advisories and bans to determine which estuarine/marine finfish species should
be recommended as target species. As shown in Table 3-16, the largest number
of States issuing advisories for specific estuarine and marine waters did so for
the American eel (6), channel catfish (5), striped bass (5), bluefish (4), white
catfish (4), and white perch (3). Several other estuarine/marine species were
cited in advisories for one State each (Table 3-16). Many coastal States did not
identify individual finfish species by name in their advisories (see Appendix A);
however, almost all of the species that have been cited in State advisories are
recommended as target species by EPA (see Tables 3-7 through 3-13).
These seven regional lists of recommended estuarine/marine target species are
provided to give guidance to States on species commonly consumed by the
general population. State personnel, with their knowledge of site-specific
fisheries and human consumption patterns, must be the ultimate judge of the
species selected for use in estuarine/marine fish contaminant monitoring
programs within their jurisdiction.
3-26
-------
4. TARGET ANALYTES
SECTION 4
TARGET ANALYTES
The selection of appropriate target analytes in fish and shellfish contaminant
monitoring programs is essential to the adequate protection of the health of fish
and shellfish consumers. The procedures used for selecting target analytes for
screening studies and a list of recommended target analytes are presented in
this section.
4.1 RECOMMENDED TARGET ANALYTES
Recommended target analytes for screening studies in fish and shellfish
contaminant monitoring programs are listed in Table 4-1. This list was
developed by the EPA Fish Contaminant Workgroup from a review of the
following information:
1. Pollutants analyzed in several national or regional fish contaminant
monitoring programs—The monitoring programs reviewed included
National Study of Chemical Residues in Fish (U.S. EPA)
National Dioxin Study (U.S. EPA)
• 301 (h) Monitoring Program (U.S. EPA)
National Pollutant Discharge Elimination System (U.S. EPA)
• National Pesticide Monitoring Program (U.S. FWS)
National Contaminant Biomonitoring Program (U.S. FWS)
• National Status and Trends Program (NOAA)
Great Lakes Sportfish Consumption Advisory Program
U.S. Food and Drug Administration (FDA) recommendations
• National Water-Quality Assessment Program (USGS).
Criteria for selection of the target analytes in these programs varied widely
depending on specific program objectives. The target analytes used in these
major fish contaminant monitoring programs are compared in Appendix B.
Over 200 potential contaminants are listed, including metals, pesticides,
base/neutral organic compounds, dioxins, dibenzofurans, acidic organic
compounds, and volatile organic compounds.
2. Pesticides with active registrations—The EPA Office of Pesticide
Programs (OPP's) Fate One Liners Database (U.S. EPA, 1993a) containing
information for more than 900 registered pesticides was reviewed to identify
4-1
-------
4. TARGET ANALYTES
Table 4-1. Recommended Target Analytes3
Metals Organophosphate Pesticides
Cadmium Carbophenothion
Mercury Chlorpyrifos
Selenium Diazinon
Disulfoton
Orqanochlorlne Pesticides Ethion
Terbufos
Chlordane, total (cis- and trans-chlordane,
cis- and trans-nonachlor, oxychlordane) Chlorophenoxy Herbicides
DDT, total (2,4'-DDD, 4,4'-DDD. 2,4'-DDE,
4,4'-DDE, 2,4'-DDT, 4,4'-DDT) Oxyfluorfen
Dicofol
Dieldrin PCBs
Endosulfan (I and II)
Endrin Total Aroclors9
Heptachldr epoxideb
Hexachlorobenzene Dloxlns/dlbenzofurans -°
Lindane (y-hexachlorocyclohexane; y-HCH)0
Mirexd
Tpxaphene
PAHs - Polycyclic aromatic hydrocarbons.
PCBs = Polychlorihated biphenyls.
a States should include all recommended target analytes in screening studies, if resources allow, unless historic
tissue or sediment data indicate that an analyte is not present at a level of concern for human health.
Additional target analytes should be included in screening studies if States have site-specific information (e.g.,
historic tissue or sediment data, discharge monitoring reports from municipal and industrial sources, pesticide
use application information) that these chemicals may be present at levels of concern for human health.
b Heptachlor epoxide is not a pesticide but is a metabolite of the pesticide heptachlor.
c Also known as ybenzene hexachloride (y-BHC).
d Mirex should be regarded primarily as a regional target analyte in the southeast and Great Lakes States,
unless historic tissue, sediment, or discharge data indicate the likelihood of its presence in other areas.
6 Analysis of total PCBs, as the sum of Aroclor equivalents, is recommended in both screening and intensive
studies because of the lack of adequate toxicologic data to develop screening values (SVs) for individual PCB
congeners (see Section 4.3.5). However, because of the wide range of toxicities among different PCB
congeners and the effects of metabolism and degradation on Aroclor composition in the environment,
congener analysis is deemed to be a more scientifically sound and accurate method for determining total PCB
concentrations. Consequently, States that currently do congener-specific PCB analyses should continue to do
so. Other States are encouraged to develop the capability to conduct PCB congener analysis.
' Note: The EPA Office of Research and Development is currently reassessing the human health effects of
dioxins/dibenzofurans.
8 Dioxins/dibenzofurans should be considered for analysis primarily at sites of pulp and paper mills using a
chlorine bleaching process and at industrial sites where the following organic compounds are formulated:
herbicides (containing 2,4,5-trichlorophenoxy acids and 2,4,5-trichlorophenol), hexachlorophene,
pentachlorophenol, and PCBs (U.S. EPA, 1987d). It is recommended that the 2,3,7,8-substituted tetra-
through octa-chlorinated dibenzo-p-dioxins (PCDDs) and dibenzofurans (PCDFs) be determined and a toxicily-
weighted total concentration calculated for each sample (Barnes and Bellin, 1989; U.S. EPA, 1987d) (see
Section 5.3.2.4). If resources are limited, 2,3,7,8-TCDD and 2,3,7,8-TCDF should be determined at a
minimum.
4-2
-------
4. TARGET ANALYTES
pesticides and herbicides with active registrations that met four criteria. The
screening criteria used were
Oral toxicity
Biological concentration factor (BCF) greater than 300
Half-life value of 30 days or more
Initial use application profile.
At the time of this review, complete environmental fate information was
available for only about half of the registered pesticides. As more data
become available, additional pesticides will be evaluated for possible
inclusion on the target analyte list.
Use of the OPP Database was necessary because many pesticides and
herbicides with active registrations have not been monitored extensively
either in national or State fish contaminant monitoring programs.
3. Contaminants that have triggered States to issue fish and shellfish
consumption advisories or bans—The database, National Listing of State
Fish and Shellfish Consumption Advisories and Bans (RTI, 1993), was
reviewed to identify specific chemical contaminants that have triggered
issuance of consumption advisories by the States. As shown in Table 4-2,
four contaminants have triggered advisories in the largest number of States:
polychlorinated biphenyls (PCBs), mercury, chlordane, and
dioxins/dibenzofurans.
4. Published literature on the chemistry and health effects of potential
contaminants—The physical, chemical, and toxicologic factors considered
to be of particular importance in developing the recommended target analyte
list were
Oral toxicity
• Potential of the analyte to bioaccumulate
Prevalence and persistence of the analyte in the environment
Biochemical fate of the analyte in fish and shellfish
Human health risk of exposure to the analyte via consumption of
contaminated fish and shellfish
Analytical feasibility.
Final selection of contaminants for the recommended target analyte list (Table
4-1) was based on their frequency of inclusion in national monitoring programs,
on the number of States issuing consumption advisories for them, and on their
origins, chemistry, potential to bioaccumulate, estimated human health risk, and
4-3
-------
4. TARGET ANALYTES
feasibility of analysis. Primary consideration was given to the recommendations
of the Committee on Evaluation of the Safety of Fishery Products, published in
Seafood Safety (NAS, 1991), and to the recommendations of the EPA Fish
Contaminant Workgroup.
4.2 SELECTION OF TARGET ANALYTES
States should include all recommended target analytes (Table 4-1) in screening
studies, if resources allow, unless historic tissue or sediment or pollutant source
data indicate that an analyte is not present at a level of concern (see Section 5).
For the pesticides with active registrations, use and rate application information
maintained by the State's Department of Agriculture should be reviewed to
identify watersheds where these pesticides have been used historically or are
currently used and are likely to be present in aquatic systems as a result of
agricultural runoff or drift.
It is important to note that pesticide uses and labels may change over time. The
State agency responsible for designing the fish contaminant monitoring program
should be aware of all historic and current uses of each pesticide within its State,
including the locations, application rates, and acreage where the pesticide has
been or currently is applied to ensure that all potentially contaminated sites are
included in the sampling plan.
Additional target analytes should be included in screening programs if States
have site-specific chemical information (e.g., historic tissue or sediment data,
discharge monitoring reports from municipal and industrial sources, or pesticide
use data) that these contaminants may be present at levels of concern for
human health. Compounds that are currently under review by the EPA Office
of Water for inclusion as recommended target analytes are discussed in Section
4.4. Specific factors that were considered in the selection of currently
recommended target analytes are summarized in the following sections.
4.3 TARGET ANALYTE PROFILES
4.3.1 Metals
Three metals—cadmium, mercury, and selenium—are recommended as target
analytes in screening studies. Cadmium and mercury have been included in six
major fish contaminant monitoring programs (see Appendix B). Selenium has
been monitored in five national programs. Consumption advisories are currently
in effect for cadmium, mercury, and selenium in two, twenty-seven, and five
States, respectively (Table 4-2). Also, these three metals have been identified
as having the greatest potential toxicity resulting from ingestion of contaminated
fish and shellfish (NAS, 1991).
4-4
-------
4. TARGET ANALYTES
Table 4-2. Contaminants Resulting In Fish and Shellfish Advisories
Number of States
Contaminant Issuing advisories
Metals
Arsenic 1
Cadmium 2
Chromium 1
Copper 1
Lead 4
Mercury 27
Selenium 5
Zinc 1
Organometallics 1
Unidentified metals 3
Pesticides
Chlordane 24
DDT and metabolites 9
Dieldrin 3
Heptachlor epoxide 1
Hexachlorobenzene 2
Kepone 1
Mirex 3
Photomirex 1
Toxaphene 2
Unidentified pesticides 2
Polycycllc aromatic hydrocarbons (PAHs) 3
Polychlorlnated blphenyls (PCBs) 31
Dloxlns/dlbenzofurans 22
Other chlorinated organlcs
Dichlorobenzene 1
Hexachlorobutadiene 1
Pentachlorobenzene 1
Pentachlorophenol 1
Polybrominated biphenyls (PBBs) 1
Tetrachlorobenzene 2
Tetrachloroethane 1
Others
Gasoline 1
Creosote 2
Phthalate esters 1
Multiple pollutants 2
Unspecified pollutants 3
Source: RTI, 1993.
4-5
-------
4. TARGET ANALYTES
4.3.1.1 Cadmium-
Cadmium is commonly found in zinc, lead, and copper deposits (May and
McKinney, 1981). It is released into the environment from several anthropogenic
sources: smelting and refining of ores, electroplating, application of phosphate
fertilizers, surface mine drainage (U.S. EPA, 1978), and waste disposal
operations (municipal incineration and land application) (U.S. EPA, 1979a, U.S.
EPA, 1987c). Cadmium is also used in the manufacture of paints, alloys,
batteries, and plastics and has been used in the control of moles and plant
diseases in lawns.
Cadmium is a cumulative human toxicant; it has been shown to cause renal
dysfunction and a degenerative bone disease, itai-itai, in Japanese populations
exposed via consumption of contaminated rice, fish, and water. Because
cadmium is retained in the kidney, older individuals (over 40-50 years of age)
typically have both the highest renal concentrations of cadmium and the highest
prevalence of renal dysfunction (U.S. EPA, 1979a). Cadmium is a known
carcinogen in animals, and there is limited evidence of the carcinogenicity of
cadmium or cadmium compounds in humans. It has been classified as a
probable human carcinogen by inhalation (B1) by EPA (IRIS, 1992).
Cadmium has been found to bioaccumulate in fish and shellfish tissues in fresh
water (Schmitt and Brumbaugh, 1990) and in estuarine/marine waters (NOAA,
1987, 1989a) nationwide. In the National Contaminant Biomonitoring Program
(NCBP), geometric mean concentrations of cadmium in freshwater fish were
found to have declined from 0.07 ppm in 1976 to 0.03 ppm in 1984 (Schmitt and
Brumbaugh, 1990). This trend contradicts the general trend of increasing
cadmium concentrations in surface waters, which Smith et al. (1987) attribute to
increasing U.S. coal combustion (Schmitt and Brumbaugh, 1990). Two States
(New York and Ohio) have issued advisories for cadmium contamination (RTI,
1993).
Cadmium should be considered for inclusion in all State fish and shellfish
contaminant monitoring programs.
4.3.1.2 Mercury—
The major source of atmospheric mercury is the natural degassing of the earth's
crust, amounting to 2,700 to 6,000 tons per year (WHO, 1990). Primary points
of entry of mercury into the environment from anthropogenic sources are
industrial discharges and wastes (e.g., the chlorine-alkali industry) and
atmospheric deposition resulting from combustion of coal and municipal refuse
incinerators (Glass et al., 1990). Primary industrial uses of mercury are in the
manufacture of batteries, vapor discharge lamps, rectifiers, fluorescent bulbs,
switches, thermometers, and industrial control instruments (May and McKinney,
1981), and these products ultimately end up in landfills or incinerators. Mercury
has also been used as a slimicide in the pulp and paper industry, as an
antifouling and mildew-proofing agent in paints, and as an antifungal seed
__
-------
4. TARGET ANALYTES
dressing and in chlor-alkali production facilities (Farm Chemicals Handbook,
1989; Friberg and Vostal, 1972).
Although mercury use and losses from industrial processes in the United States
have been reduced significantly since the 1970s, mercury contamination
associated with increased fossil fuel combustion is of concern in some areas and
may pose more widespread contamination problems in the future. An estimated
5,000 tons of mercury per year are released into the environment from fossil fuel
burning (Klaassen et al., 1986). There is also increasing evidence of elevated
mercury concentrations in areas where acid rain is believed to be a factor,
although the extent of this problem has not been documented with certainty
(Sheffy, 1987; Wiener, 1987). Volatilization from surfaces painted with mercury-
containing paints, both indoors and outdoors, may have been a significant
source in the past (Agocs et al., 1990; Sherry, 1987). The United States
estimated that 480,000 pounds of mercuric fungicides were used in paints and
coatings in 1987 (NPCA, 1988). In July 1990, EPA announced an agreement
with the National Paint and Coatings Association to cancel all registrations for
use of mercury or mercury compounds in interior paints and coatings. In May
1991, the paint industry voluntarily canceled all remaining registrations for
mercury in exterior paints.
Cycling of mercury in the environment is facilitated by the volatile character of
its metallic form and by bacterial transformation of metallic and inorganic forms
to stable alkyl mercury compounds, particularly in bottom sediments, which leads
to bioaccumulation of mercury (Wood, 1974). Practically all mercury in fish
tissue is in the form of methylmercury, which is toxic to humans (NAS, 1991;
Tollefson, 1989).
The EPA has determined that the evidence of carcinogenicity of mercury in both
animals and humans is inadequate and has assigned this metal a D,
carcinogenicity classification (IRIS, 1992). Both inorganic and organic forms of
mercury are neurotoxicants. Fetuses chronically exposed to organic mercury
have been found to be born mentally retarded and with symptoms similar to
those of cerebral palsy (Marsh, 1987). Individuals chronically exposed to
mercury via long-term ingestion of mercury-contaminated fish have been found
to exhibit a wide range of symptoms, including numbness of the extremities,
tremors, spasms, personality and behavior changes, difficulty in walking,
deafness, blindness, and death (U.S. EPA, 1981 a). Organomercury compounds
were the causative agents of Minamata Disease, a neurological disorder reported
in Japan during the 1950s among individuals consuming contaminated fish and
shellfish (Kurland et al., 1960), with infants exposed prenatally found to be at
significantly higher risk than adults.
The EPA is especially concerned about evidence that the fetus and possibly
pregnant women are at increased risk of adverse neurological effects from
exposure to methylmercury (e.g., Marsh et al., 1987; Piotrowski and Inskip, 1981;
Skerfving, 1988; WHO, 1976, 1990).
4-7
-------
4. TARGET ANALVTES
Mercury has been found in both fish and shellfish from estuarine/marine (NOAA,
1987, 1989a) and fresh waters (Schmitt and Brumbaugh, 1990) at diverse
locations nationwide. In contrast to cadmium and selenium, concentrations of
mercury in freshwater fish tissue did not change between 1976 and 1984
(Schmitt and Brumbaugh, 1990). Mercury, the only metal analyzed in the
National Study of Chemical Residues in Fish, was detected at 92.2 percent of
374 sites surveyed. Maximum, arithmetic mean, and median concentrations in
fish tissue were 1.80,0.26, and 0.17 ppm, respectively (U.S. EPA, 1991 h, 1992c,
1992d). Fish consumption advisories have been issued in 27 States as a result
of mercury contamination (see Figure 4-1). In particular, mercury is responsible
for a large number of the fish advisories currently in effect for lakes in Wisconsin,
Michigan, and Minnesota and for rivers and lakes in Florida (RTI, 1993).
Mercury should be considered for inclusion in all State fish and shellfish
contaminant monitoring programs. Only two national programs (301 (h) and the
FDA) currently analyze specifically for methylmercury; however, six programs
analyze for total mercury (Appendix B). Because of the higher cost of methyl-
mercury analysis, EPA recommends that total mercury be determined in State
fish contaminant monitoring programs and the conservative assumption be made
that all mercury is present as methylmercury in order to be most protective of
human health.
It should be noted that Bache et al. (1971) analyzed methylmercury
concentrations in lake trout of known ages and found that methylmercury
concentration and the ratio of methylmercury to total mercury increased with age.
Relative proportions of methylmercury in fish varied between 30 and 100
percent, with methylmercury concentrations lower than 80 percent occurring in
fish 3 years of age or younger. Thus, when high concentrations of total mercury
are detected, and if resources are sufficient, States may wish to repeat sampling
and obtain more specific information on actual concentrations of methylmercury
in various age or size classes of fish.
4.3.1.3 Selenium—
Selenium is a natural component of many soils, particularly in the west and
southwest regions of the United States (NAS, 1991). It enters the environment
primarily via emissions from oil and coal combustion (May and McKinney, 1981;
Pillay et al., 1969). Selenium is an essential nutrient but is toxic to both humans
and animals at high concentrations and has been shown to act as a mutagen in
animals (NAS, 1991). Long-term adverse effects from ingestion by humans have
not been studied thoroughly. The EPA has determined that the evidence of
carcinogenicity of selenium in both humans and animals is inadequate and,
therefore, has assigned this metal a D carcinogenicity classification. However,
selenium sulfide has been classified as a probable human carcinogen (B2) (IRIS,
1992).
Selenium is frequently detected in ground and surface waters in most regions of
the United States and has been detected in marine fish and shellfish (NOAA,
-------
H American Samoa
D Guam
Source: RTI, 1993.
States issuing advisory (27)
Virgin Islands
D Puerto Rico
3J
ft
m
Current 7/15/93
<
Figure 4-1. States issuing fish and shellfish advisories for mercury.
-------
4. TARGET ANALYTES
1987,1989a) and in freshwater fish (Schmitt and Brumbaugh, 1990) from several
areas nationwide. Selenium has been monitored in five national fish contaminant
monitoring programs (Appendix B). Definitive information concerning the
chemical forms of selenium found in fish and shellfish is not available (NAS,
1976,1991). Five States (California, Colorado, North Carolina, Texas, and Utah)
have issued advisories for selenium contamination in fish (RTI, 1993).
Selenium should be considered for inclusion in all State fish and shellfish
monitoring programs.
4.3.2 Organochlorlne Pesticides
The following organochlorine pesticides and metabolites are recommended as
target analytes in screening studies: total chlordane (sum of cis- and trans-
chlordane, cis- and trans-nonachlor, and oxychlordane), total DDT (sum of 2,4'-
and 4,4'-isomers of DDT, ODD, and DDE), dicofol, dieldrin, endosulfan I and II,
endrin, heptachlor epoxide, hexachlorobenzene, lindane (y-hexachlorocyclo-
hexane), mirex, and toxaphene (see Appendix C). Mirex is of particular concern
in the Great Lakes States and the southeast States (NAS, 1991). All of these
compounds are neurotoxins and most are known or suspected human carcino-
gens (IRIS, 1992; Sax, 1984).
With the exception of endosulfan I and II, dicofol and total DDT, each of the
pesticides on the recommended target analyte list (Table 4-1) has been included
in at least five major fish contaminant monitoring programs, and seven of the
compounds have triggered at least one State fish consumption advisory
(Table 4-2). Although use of some of these pesticides has been terminated or
suspended within the United States for as long as 20 years (Appendix C), these
compounds still require long-term monitoring. Many of the organochlorine
pesticides were used in large quantities for over a decade and are present in
sediments at high concentrations. Organochlorine pesticides are not easily
degraded or metabolized and, therefore, persist in the environment. These
compounds are either insoluble or have relatively low solubility in water but are
quite lipid soluble. Because these compounds are not readily metabolized or
excreted from the body and are readily stored in fatty tissues, they can
bioaccumulate to high concentrations through aquatic food chains to secondary
consumers (e.g., fish, piscivorous birds, and mammals including humans).
Pesticides may enter aquatic ecosystems from point source industrial discharges
or from nonpoint sources such as aerial drift and/or runoff from agricultural use
areas, leaching from landfills, or accidental spills or releases. Agricultural runoff
from crop and grazing lands is considered to be the major source of pesticides
in water, with industrial waste (effluents) from pesticide manufacturing the next
most common source (Li, 1975). Significant atmospheric transport of pesticides
to aquatic ecosystems can also result from aerial drift of pesticides, volatilization
from applications in terrestrial environments, and wind erosion of treated soil (Li,
1975). Once in water, pesticide residues may become adsorbed to suspended
4-10
-------
4. TARGET ANALYTES
material, deposited in bottom sediment, or absorbed by organisms in which they
are detoxified and eliminated or accumulated (Nimmo, 1985).
The reader should note that two of the organochlorine pesticides have active
registrations: endosulfan and dicofol. States should contact their appropriate
State agencies to obtain information on both the historic and current uses of
these pesticides.
4.3.2.1 Chlordane (Total)—
Chlordane is a multipurpose insecticide that has been used extensively in home
and agricultural applications in the United States for the control of termites and
many other insects. This pesticide is similar in chemical structure to dieldrin,
although less toxic (Toxicity Class II), and has been classified as a probable
human carcinogen (B2) by EPA (IRIS, 1992; Worthing, 1991).
Although the last labeled use of Chlordane as a termiticide was phased out in the
United States beginning in 1975, it has been monitored in eight national fish
contaminant programs (Appendix B) and has been widely detected in freshwater
fish (Schmitt et al., 1990) and in both estuarine/marine finfish (NOAA, 1987) and
marine bivalves (NOAA, 1989a) at concentrations of human health concern. The
cis- and trans-isomers of Chlordane and nonachlor, which are primary
constituents of technical-grade Chlordane, and oxychlordane, the major
metabolite of Chlordane, were monitored as part of the National Study of
Chemical Residues in Fish. These compounds were detected in fish tissue at
the following percentage of the 362 sites surveyed: cis-chlordane (64 percent),
trans-chlordane (61 percent), cis-nonachlor (35 percent), trans-nonachlor (77
percent), and oxychlordane (27 percent) (U.S. EPA, 1992c, 1992d). Chlordane's
presence in fish tissue has resulted in consumption advisories in 24 States (RTI,
1993) (see Figure 4-2).
Total Chlordane (i.e., sum of cis- and trans-chlordane, cis- and trans-nonachlor,
and oxychlordane) should be considered for inclusion in all State fish and
shellfish contaminant monitoring programs (NAS, 1991).
4.3.2.2 DDT (Total)—
Although the use of DDT was terminated in the United States in 1972, DDT and
its DDE and DDD metabolites persist in the environment and are known to
bioaccumulate (Ware, 1978). DDT, DDD, and DDE have all been classified by
EPA as probable human carcinogens (B2) (IRIS, 1992).
DDT or its metabolites have been included as target analytes in eight major fish
and shellfish monitoring programs (Appendix B) and contamination has been
found to be widespread (NOAA, 1987, 1989a; Schmitt et al., 1990). DDE, the
only DDT metabolite surveyed in fish tissue in the National Study of Chemical
Residues in Fish, was detected at more sites than any other single pollutant (99
percent of the 362 sites sampled) (U.S. EPA, 1992c, 1992d). Nine States
_
-------
D American Samoa
D Guam
Source: RTI, 1993.
States issuing advisory (24)
D Virgin Islands
D Puerto Rico
Current 7/15/93
3J
O
m
ro
Figure 4-2. States issuing fish and shellfish advisories for chlordane.
-------
4. TARGET ANALYTES
(Alabama, American Samoa, Arizona, California, Delaware, Massachusetts,
Nebraska, New York, and Texas) currently have fish consumption advisories in
effect for DDT or its metabolites (RTI, 1993).
Total DDT (i.e., sum of the 4,4'- and 2,4'- isomers of DDT and of its metabolites,
DDE and DDD) should be considered for inclusion in all State fish and shellfish
contaminant monitoring programs.
4.3.2.3 Dicofol—
This chlorinated hydrocarbon acaricide was first registered in 1957 and is
structurally similar to DDT (U.S. EPA, 1992c, 1992d). Technical-grade dicofol
may contain impurities of the p.p' and o,p' isomers of DDT, DDE, DDD, and a
compound known as extra-chlorine DDT (CI-DDT) that are inherent as a result
of the manufacturing process (U.S. EPA, 1983b). Historically, dicofol has been
used to control mites on cotton and citrus (60 percent), on apples (10 percent),
on ornamental plants and turf (10 percent), and on a variety of other agricultural
products (20 percent) including pears, apricots, and cherries (Farm Chemical
Handbook, 1989), as a seed crop soil treatment, on vegetables (e.g., beans and
corn) and on shade trees (U.S. EPA, 1992c, 1992d).
Dicofol is moderately toxic to laboratory rats and has been assigned to Toxicity
Class III based on oral exposure studies. Technical-grade dicofol induced
hepatocellular (liver) carcinomas in male mice; however, results were negative
in female mice and in rats (NCI, 1978). EPA has classified dicofol as a possible
human carcinogen (C) (U.S. EPA, 1992a). Because of concern that dicofol
would have the same effect as DDT on thinning of egg shells, the FDA required
all dicofol products to contain less than 0.1 percent DDT and related
contaminants after June 1, 1989 (51 FR 19508).
Dicofol was recommended for monitoring by the EPA Office of Water as part of
the Assessment and Control of Bioconcentratable Contaminants in Surface
Waters Program and has been included in two national monitoring programs
(see Appendix B). In the National Study of Chemical Residues in Fish, dicofol
was detected at 16 percent of the sites monitored (U.S. EPA, 1992c, 1992d).
Dicofol concentrations were greater than the quantification limit (2.5 ppb) in
samples from 7 percent of the sites. Most of the sites where dicofol was
detected were in agricultural areas where citrus and other fruits and vegetables
are grown (U.S. EPA, 1992c, 1992d). It should be noted that this national study
did not specifically target agricultural sites where this pesticide historically had
been or currently is used. Dicofol residues in fish could be much higher if
sampling were targeted for pesticide runoff. Experimental evidence indicates this
compound bioaccumulates in Bluegill sunfish (BCF from 6,600 to 17,000) (U.S.
EPA, 1993a); however, no consumption advisories are currently in effect for
dicofol (RTI, 1993).
Dicofol should be considered for inclusion in State fish and shellfish contaminant
monitoring programs, in areas where its use is or has been extensive. States
-------
4. TARGET ANALYTES
should contact their appropriate State agencies to obtain information on the
historic and current uses of this pesticide.
4.3.2.4 Dieldrin—
Dieldrin is a chlorinated cyclodiene that was widely used in the United States
from 1950 to 1974 as a broad spectrum pesticide, primarily on termites and other
soil-dwelling insects and on cotton, corn, and citrus crops. Because the toxicity
of this persistent pesticide posed an imminent danger to human health, EPA
banned the production and most major uses of dieldrin in 1974, and, in 1987, all
uses of dieldrin were voluntarily canceled by industry (see Appendix C).
Dieldrin has been classified by EPA as a probable human carcinogen (B2) (IRIS,
1992) and has been identified as a human neurotoxin (ATSDR, 1987a). Dieldrin
has been included in eight national minitoring programs (Appendix B) and is still
detected nationwide in freshwater finfish (Schmitt et al., 1990) and
estuarine/marine finfish and shellfish (NOAA, 1987, 1989a). Dieldrin was
detected in fish tissue at 60 percent of the 362 sites surveyed as part of the
National Survey of Chemical Residues in Fish (U.S. EPA, 1992c, 1992d).
Because it is a metabolite of aldrin, the environmental concentrations of dieldrin
are a cumulative result of the historic use of both aldrin and dieldrin (Schmitt et
al., 1990). Three States (Arizona, Illinois, and Nebraska) have issued advisories
for dieldrin contamination in fish (RTI, 1993).
Dieldrin should be considered for inclusion in all State fish and shellfish
contaminant monitoring programs.
4.3.2.5 Endosulfan—
Endosulfan is a chlorinated cyclodiene pesticide that is currently in wide use
primarily as a noncontact insecticide for seed and soil treatments (Appendix C).
Two stereoisomers (I and II) exist and exhibit approximately equal effectiveness
and toxicity (Worthing, 1991).
Endosulfan is highly toxic to humans and has been assigned to Toxicity Class
I. To date, no studies have been found concerning carcinogenicity in humans
after oral exposure to endosulfan (ATSDR, 1990). EPA has given endosulfan
the carcinogenicity classification E, indicating there is no evidence of
carcinogenicity for humans (U.S. EPA, 1992a).
Agricultural runoff is the primary source of this pesticide in aquatic ecosystems.
Endosulfan has been shown to be highly toxic to fish and marine invertebrates
and is readily absorbed in sediments. It therefore represents a potential hazard
in the aquatic environment (Sittig, 1980). However, data are currently insufficient
to assess nationwide endosulfan contamination (NAS, 1991). Endosulfan was
recommended for monitoring by the U.S. FDA and has been included in one
national fish contaminant monitoring program (U.S. EPA 301 (h) Program)
4-14
-------
4. TARGET ANALYTES
evaluated by the EPA Workgroup (Appendix B). No consumption advisories are
currently in effect for endosulfan I or II (RTI, 1993).
Endosulfan I and II should be considered for inclusion in all State fish and
shellfish contaminant monitoring programs in areas where its use is or has been
extensive. States should contact their appropriate State agencies to obtain
information on the historic and current uses of this pesticide.
4.3.2.6 Endrin—
Endrin is a chlorinated cyclodiene that historically was widely used as a broad
spectrum pesticide. Endrin was first registered for use in the United States in
1951. However, recognition of its long-term persistence in soil and its high levels
of mammalian toxicity led to restriction of its use beginning in 1964 {U.S. EPA,
1980a) and 1979 (44 FR 43632) and to final cancellation of its registration in
1984 (U.S. EPA, 1984a) (Appendix C).
Endrin is highly toxic to humans (Toxicity Class I), with acute exposures affecting
the central nervous system primarily (Sax, 1984). At present, evidence of both
animal and human carcinogenicity of endrin is considered inadequate (IRIS,
1992).
Although endrin has been included in six national fish contaminant monitoring
programs (Appendix B), it has not been found widely throughout the United
States. Endrin was detected in freshwater and marine species at 11 percent of
the 362 sites surveyed in the EPA National Study of Chemical Residues in Fish
(U.S. EPA, 1992c, 1992d) and was found in only 29 percent of 112 stations
sampled in the NCBP (Schmitt et al., 1990). No States have issued fish
consumption advisories for endrin (RTI, 1993).
Endrin should be considered for inclusion in all State fish and shellfish
contaminant monitoring programs.
4.3.2.7 Heptachlor Epoxidc
Heptachlor epoxide is not a formulated pesticide, but is a metabolic degradation
product of the pesticide heptachlor. It is also found as a contaminant in
heptachlor and chlordane formulations. Heptachlor has been used as a
persistent, nonsystemic contact and ingested insecticide on soils (particularly for
termite control) and seeds and as a household insecticide (Worthing, 1991).
EPA suspended the major uses of heptachlor in 1978 (ATSDR, 1987b). Acute
exposures to high doses of heptachlor epoxide in humans can cause central
nervous system effects (e.g., irritability, dizziness, muscle tremors, and
convulsions (U.S. EPA, 1986e). In animals, liver, kidney, and blood disorders
can occur (IRIS, 1989). Exposure to this compound produced an increased
incidence of liver carcinomas in rats and mice and hepatomas in female rats
(IRIS, 1989). Heptachlor epoxide has been classified by EPA as a probable
human carcinogen (B2) (IRIS, 1992).
_
-------
4. TARGET ANALYTES
Heptachlor epoxide has been included in seven national fish monitoring
programs (Appendix B) and has been detected widely in freshwater finfish
(Schmitt et al., 1990) but infrequently in bivalves and marine fish (NOAA, 1987,
1989a). Heptachlor epoxide was detected in fish tissue at 16 percent of the 362
sites where it was surveyed in the National Study of Chemical Residues in Fish
(U.S. EPA, 1992c, 1992d). One State (Nebraska) currently has fish advisories
for heptachlor epoxide contamination (RTI, 1993).
Heptachlor epoxide should be considered for inclusion in all State fish and
shellfish monitoring programs.
4.3.2.8 Hexachlorobenzene—
Hexachlorobenzene is a fungicide that was widely used as a seed protectant in
the United States until 1985 (Appendix C). The use of hexachlorobenzene and
the presence of hexachlorobenzene residues in food are banned in many
countries including the United States (Worthing, 1991). Registration of
hexachlorobenzene as a pesticide was voluntarily canceled in 1984 (Morris and
Cabral, 1986).
The toxicity of this compound is minimal; it has been given a toxicity
classification of IV (i.e., oral LD50 greater than 5,000 ppm in laboratory animals
(Farm Chemicals Handbook, 1989). However, nursing infants are particularly
susceptible to hexachlorobenzene poisoning as lactational transfer can increase
infant tissue levels to levels two to five times maternal tissue levels (ATSDR,
1989b). Hexachlorobenzene is a known animal carcinogen (ATSDR, 1989b) and
has been classified by EPA as a probable human carcinogen (B2) (IRIS, 1992)
(Appendix C).
Of the chlorinated benzenes, hexachlorobenzene is the most widely monitored
(Worthing, 1991). It was included as a target analyte in seven of the major
monitoring programs reviewed (Appendix B). Hexachlorobenzene was detected
in fish tissue at 46 percent of the 362 sites where it was surveyed in the National
Study of Chemical Residues in Fish (U.S. EPA, 1992c, 1992d). Two States
(Louisiana and Ohio) have issued advisories for hexachlorobenzene
contamination in fish and shellfish (RTI, 1993).
Hexachlorobenzene should be considered for inclusion in all State fish and
shellfish monitoring programs.
4.3.2.9 Lindane—
Lindane is a mixture of isomers of hexachlorocyclohexane (C6H6CI6), whose
major component (>99 percent) is the gamma isomer. It is commonly referred
to as either y-HCH (hexachlorocyclohexane) or y-BHC (benzene hexachloride).
Lindane is used primarily in seed treatments, soil treatments for tobacco
transplants, foliage applications on fruit and nut trees and vegetables, and wood
and timber protection. Since 1985, many uses of lindane have been banned or
_
-------
4. TARGET ANALYTES
restricted (see Appendix C). At present, its application is permitted only under
supervision of a certified applicator (U.S. EPA, 1985c).
Lindane is a neurotoxin (assigned to Toxicity Class II) and has been found to
cause aplastic anemia in humans (Worthing, 1991). Lindane has been classified
by EPA as a probable/possible human carcinogen (B2/C). Available data for this
pesticide need to be reviewed, but at a minimum the carcinogenicity
classification will be C (U.S. EPA, 1992a).
Lindane has been included in eight major fish contaminant monitoring programs
(Appendix B). This pesticide has been detected in freshwater fish (Schmitt et al.,
1990) and in marine fish and bivalves (NOAA, 1987, 1989a) nationwide.
Lindane was detected in fish tissue at 42 percent of 362 sites surveyed in the
National Study of Chemical Residues in Fish (U.S. EPA, 1992c, 1992d).
Although lindane has been widely monitored and widely detected, no
consumption advisories are currently in effect for lindane (RTI, 1993).
Lindane should be considered for inclusion in all State fish and shellfish
monitoring programs.
4.3.2.10 Mirex—
Mi rex is a chlorinated cyclodiene pesticide that was used in large quantities in
the United States from 1962 through 1975 primarily for control of fire ants in the
Southeast and, more widely, under the name Dechlorane as a fire retardant and
polymerizing agent in plastics (Kaiser, 1978).
Mirex has been assigned to Toxicity Class II and has been classified as a
probable human carcinogen by the International Agency for Research on Cancer
(IARC, 1987); however, the carcinogenicity data are currently under review by
EPA (IRIS, 1992). EPA instituted restrictions on the use of mirex in 1975, and,
shortly thereafter, the U.S. Department of Agriculture (USDA) suspended the fire
ant control program (Hodges, 1977).
Mirex has been included in eight major fish contaminant monitoring programs
(Appendix B). It has been found primarily in the Southeast and the Great Lakes
regions (NAS, 1991; Schmitt et al., 1990). Mirex was detected in fish tissue at
36 percent of 362 sites surveyed in the National Study of Chemical Residues in
Fish (U.S. EPA, 1992c, 1992d). Three States (New York, Ohio, and
Pennsylvania) currently have fish consumption advisories for mirex (RTI, 1993).
Mirex should be considered for inclusion in all State fish and shellfish monitoring
programs.
4.3.2.11 Toxaphene—
Toxaphene is a mixture of chlorinated camphenes. Historically, it was used in
the United States as an insecticide primarily on cotton (Hodges, 1977). Partly
-------
4. TARGET ANALYTES
as a consequence of the ban on the use of DDT imposed in 1972, toxaphene
was for many years the most heavily used pesticide in the United States (Eichers
et al., 1978). In 1982, toxaphene's registration for most uses was canceled (47
FR 53784).
Like many of the other organochlorine pesticides, toxaphene has been assigned
to Toxicity Class II. Unlike the other organochlorine pesticides, toxaphene is
fairly easily metabolized by mammals and is not stored in the fatty tissue to any
great extent. Toxaphene has been classified by EPA as a probable human
carcinogen (B2) (IRIS, 1992).
Toxaphene has been included in five major fish contaminant monitoring
programs (Appendix B). It has been detected frequently in both fresh (Schmitt
et al., 1990) and estuarine (NOAA, 1989a) waters but is only consistently found
in Georgia, Texas, and California (MAS, 1991). Note: A toxaphene-like
compound that is a byproduct of the paper industry has been identified in the
Great Lakes Region (J. Hesse, Michigan Department of Public Health, personal
communication, 1993). Two States (Arizona and Texas) currently have fish
advisories in effect for toxaphene (RTI, 1993).
Toxaphene should be considered for inclusion in all State fish and shellfish
monitoring programs.
4.3.3 Organophosphate Pesticides
The following organophosphate pesticides are recommended as target analytes
in screening studies: carbophenothion, chlorpyrifos, diazinon, disulfoton, ethion,
and terbufos (Appendix C). Most of these organophosphate pesticides share two
distinct features. Organophosphate pesticides are generally more toxic to
vertebrates than organochlorine pesticides and exert their toxic action by
inhibiting production of vital nervous system enzymes (e.g., cholinesterase
[ChE]). In addition, organophosphates are chemically unstable and thus are less
persistent in the environment. It is this latter feature that made them attractive
alternatives to the organochlorine pesticides that were used extensively in
agriculture from the 1940s to the early 1970s.
With the exception of chlorpyrifos, none of the organophosphates has been
included in any of the national fish contaminant monitoring programs evaluated
by the EPA Workgroup and none of these pesticides (including chlorpyrifos) has
triggered State fish consumption advisories. All of the compounds have active
pesticide registrations and have been recommended for monitoring because they
have BCFs > 300, a half-life of 30 days or more in the environment, and their
use profiles suggest they could be potential problems in some agricultural
watersheds.
The reader should note that all of the organophosphate pesticides recommended
as target analytes have active registrations. States should contact their
4-18
-------
4. TARGET ANALYTES
appropriate State agencies to obtain information on both the historic and current
uses of these pesticides. In addition, if a State determines that use of these
pesticides may be occurring in its waters, sampling should be conducted during
late spring or early summer within 1 to 2 months following pesticide application
because these compounds are degraded and metabolized relatively rapidly by
fish species. Additional discussion of appropriate sampling times for fish
contaminant monitoring programs is provided in Section 6.1.1.5.
4.3.3.1 Carbophenothlon—
Carbophenothion is a multipurpose insecticide and acaricide that is registered
for use on a wide variety of fruit, nut, vegetable, forage, ornamental, and forestry
crops; however, the majority of pesticide use is on citrus crops. There are also
limited uses of carbophenothion as a seed treatment, dip, and soil insecticide
(U.S. EPA, 1984c). Production of this organophosphate pesticide was
discontinued in 1987 by the manufacturer (Farm Chemicals Handbook, 1989)
and is not being supported by the registrant for reregistration.
Preliminary evidence suggests that carbophenothion is moderately toxic to
humans (Toxicity Class II) based on acute oral and dermal effects, and all
products warrant restricted use classification (Farm Chemicals Handbook, 1989).
EPA has assigned carbophenothion to carcinogenicity classification D—not
classifiable based on a lack of data or inadequate evidence of carcinogenicity in
at least two animal tests or in both epidemiologic and animal studies (U.S. EPA,
1992a). Carbophenothion is also toxic to freshwater and marine/estuarine
organisms and is highly toxic to birds (U.S. EPA, 1984c).
Carbophenothion has not been included in any national fish contaminant
monitoring program evaluated by the EPA Workgroup. Experimental evidence
indicates this compound accumulates in spot and sheepshead minnows (BCF
from 620 to 1,200) (U.S. EPA, 1993a); however; no consumption advisories are
currently in effect for carbophenothion (RTI, 1993).
Carbophenothion should be considered for inclusion in State fish and shellfish
contaminant monitoring programs in areas where its use is or has been
extensive. States should contact their appropriate State agencies to obtain
information on the historic and current uses of this pesticide.
4.3.3.2 Chlorpyrifos—
This organophosphate pesticide was first introduced in 1965 to replace the more
persistent organochlorine pesticides (e.g., DDT) (U.S. EPA, 1986e) and has
been used for a broad range of insecticide applications. Chlorpyrifos is used
primarily to control soil and foliar insects on cotton, peanuts, and sorghum
(Worthing, 1991; U.S. EPA, 1986e). Chlorpyrifos is also used to control root-
infesting and boring insects on a variety of fruits (e.g., apples, bananas, citrus,
grapes), nuts (e.g., almonds, walnuts), vegetables (e.g., beans, broccoli, brussel
sprouts, cabbage, cauliflower, peas, and soybeans), and field crops (e.g., alfalfa
_
-------
4. TARGET ANALYTES
and corn) (U.S. EPA, 1984d). As a household insecticide, chlorpyrifos has been
used to control ants, cockroaches, fleas, and mosquitoes (Worthing, 1991) and
is registered for use in controlling subsurface termites in California (U.S. EPA,
1983a). Based on use application, 57 percent of all chlorpyrifos manufactured
in the United States is used on corn, while 22 percent is used for pest control
and lawn and garden services (U.S. EPA, 1993a).
Chlorpyrifos has a moderate mammalian toxicity and has been assigned to
Toxicity Class II based on oral feeding studies (Farm Chemicals Handbook,
1989). No teratogenic or fetotoxic effects were found in mice or rats (IRIS,
1989). No carcinogenicity was found in chronic feeding studies with rats, mice
and dogs (U.S. EPA, 1983a). EPA has assigned chlorpyrifos a carcinogenicity
classification of D—not classifiable based on inadequate evidence of
carcinogenicity or lack of data in at least two animal studies or in both
epidemiologic and animal studies (U.S. EPA, 1992a).
Chlorpyrifos was recommended for monitoring by the U.S. FDA and has been
included in one national monitoring program, the National Study of Chemical
Residues in Fish (see Appendix B). In this latter study, chlorpyrifos was
detected at 26 percent of sites sampled nationally (U.S. EPA, 1992c, 1992d).
Eighteen percent of the sites with relatively high concentrations (2.5 to 344 ppb)
were scattered throughout the East, Midwest, and in California; the highest
concentrations detected (60 to 344 ppb) were found either in agricultural areas
or in urban areas with a variety of nearby industrial sources. It should be noted
that this national study did not specifically target agricultural sites where this
pesticide historically had been used or is currently used. Chlorpyrifos residues
in fish could be much higher if sampling were targeted for pesticide runoff.
Experimental evidence indicates this compound bioaccumulates in rainbow trout
(BCF from 1,280 to 3,903) (U.S. EPA, 1993a); however, no consumption
advisories are currently in effect for chlorpyrifos (RTI, 1993).
Chlorpyrifos should be considered for inclusion in State fish and shellfish
contaminant monitoring programs in areas where its use is or has been
extensive. States should contact their appropriate State agencies to obtain
information on the historic and current uses of this pesticide.
4.3.3.3 Diazinon—
Diazinon is a phosphorothiate insecticide and nematicide that was first registered
in 1952 for control of soil insects and pests of fruits, vegetables, tobacco, forage,
field crops, range, pasture, grasslands, and ornamentals; for control of
cockroaches and other household insects; for control of grubs and nematodes
in turf; as a seed treatment; and for fly control (U.S. EPA, 1986f).
Diazinon is moderately toxic to mammals and has been assigned to Toxicity
Class II based on oral toxicity tests (Farm Chemicals Handbook, 1989). EPA
has assigned diazinon to carcinogenicity classification D—not classifiable based
on a lack of data or inadequate evidence of carcinogenicity in at least two animal
_
-------
4. TARGET ANALYTES
tests or in both epidemiologic and animal studies (U.S. EPA, 1992a). This
compound is also highly toxic to birds, fish, and other aquatic invertebrates (U.S.
EPA, 1986f).
Diazinon has not been included in any national fish contaminant monitoring
program evaluated by the EPA Workgroup. Experimental evidence indicates this
compound accumulates in trout (BCF of 542) (U.S. EPA, 1993a); however, no
consumption advisories are currently in effect for diazinon (RTI, 1993).
Diazinon should be considered for inclusion in State fish and shellfish
contaminant monitoring programs in areas where its use is or has been
extensive. States should contact their appropriate State agencies to obtain
information on the historic and current uses of this pesticide.
4.3.3.4 Disulfoton—
Disulfoton is a multipurpose systemic insecticide and acaricide first registered in
1958 for use as a side dressing, broadcast, or foliar spray in the seed furrow to
control many insect and mite species and as a seed treatment for sucking
insects (Farm Chemicals Handbook, 1989).
Disulfoton is highly toxic to all mammalian systems and has been assigned to
Toxicity Class I on the basis of all routes of exposure (Farm Chemicals
Handbook, 1989). All labeling precautions and use restrictions are based on
human health risk. Disulfoton and its major metabolites are potent
cholinesterase inhibitors primarily attacking acetylcholinesterase. Contradictory
evidence is available on the mutagenicity of this compound and the EPA has
concluded that the mutagenic potential is not adequately defined (U.S. EPA,
1984e). EPA has assigned disulfoton to carcinogenicity classification D—not
classifiable based on a lack of data or inadequate evidence of carcinogenicity in
at least two animal tests or in both epidemiologic and animal studies (U.S. EPA,
1992a).
Disulfoton has not been included in any national fish contaminant monitoring
program evaluated by the EPA Workgroup. Experimental evidence indicates this
compound accumulates in fish (BCF from 460 to 700) (U.S. EPA, 1993a);
however, no consumption advisories are currently in effect for disulfoton (RTI,
1993).
Disulfoton should be considered for inclusion in State fish and shellfish
contaminant monitoring programs in areas where its use is or has been
extensive. States should contact their appropriate State agencies to obtain
information on the historic and current uses of this pesticide.
4.3.3.5 Ethlon—
Ethion is a multipurpose insecticide and acaricide that has been registered since
1965 for use on a wide variety of nonfood crops (turf, evergreen plantings, and
_
-------
4. TARGET ANALYTES
ornamentals), food crops (seed, fruit, nut, fiber, grain, forage, and vegetables),
and for domestic outdoor uses around dwellings and for lawns. Application to
citrus crops accounts for 86 to 89 percent of the ethion used in the United
States. The remaining 11 to 14 percent is applied to cotton and a variety of fruit
and nut trees and vegetables. Approximately 55 to 70 percent of all domestically
produced citrus fruits are treated with ethion (U.S. EPA, 1989e).
Acute oral toxicity studies have shown that technical-grade ethion is moderately
toxic to mammals (Toxicity Class II) (Farm Chemicals Handbook, 1989). In a
chronic rat toxicity study, a decrease in serum cholinesterase was observed in
both males and females. Ethion was not found to be carcinogenic in rats and
mice (U.S. EPA, 1989e). EPA has assigned ethion to carcinogenicity
classification D—not classifiable based on a lack of data or inadequate evidence
of carcinogenicity in at least two animal tests or in both epidemiologic and animal
studies (U.S. EPA, 1992a).
Ethion has not been included in any national fish contaminant monitoring
program evaluated by the EPA Workgroup. Experimental evidence indicates this
compound accumulates in Bluegill sunfish (BCF from 880 to 2,400) (U.S. EPA,
1993a); however, no consumption advisories are currently in effect for ethion
(RTI, 1993).
Ethion should be considered for inclusion in State fish and shellfish contaminant
monitoring programs in areas where its use is or has been extensive. States
should contact their appropriate State agencies to obtain information on the
historic and current uses of this pesticide.
4.3.3.6 Terbufos-—
Terbufos is a systemic organophosphate insecticide and nematicide registered
in 1974 principally for use on corn, sugar beets, and grain sorghum. The
primary method of application involves direct soil incorporation of a granular
formulation (Farm Chemicals Handbook, 1989).
Terbufos is highly toxic to humans and has been assigned to Toxicity Class I
(Farm Chemicals Handbook, 1989). Symptoms of acute cholinesterase inhibition
have been reported in all acute studies, and cholinesterase inhibition was
reported in several chronic mammalian feeding studies (U.S. EPA, 1985d). EPA
has assigned terbufos to carcinogenicity classification D—not classifiable based
on a lack of data or inadequate evidence of carcinogenicity in at least two animal
tests or in both epidemiologic and animal studies (U.S. EPA, 1992a). Terbufos
is also highly toxic to birds, fish, and other aquatic invertebrates (U.S. EPA,
1985d).
Terbufos has not been included in any national fish contaminant monitoring
program evaluated by the EPA Workgroup. Experimental evidence indicates this
compound accumulates in fish (BCF from 320 to 1,400) (U.S. EPA, 1993a);
4-22
-------
4. TARGET ANALYTES
however no consumption advisories are currently in effect for terbufos (RTI,
1993).
Terbufos should be considered for inclusion in State fish and shellfish
contaminant monitoring programs in areas where its use is or has been
extensive. States should contact their appropriate State agencies to obtain
information on the historic and current uses of this pesticide.
4.3.4 Chlorophenoxy Herbicides
Chlorophenoxy herbicides, which include oxyfluorfen, are nonselective foliar
herbicides that are most effective in hot weather (Ware, 1978).
4.3.4.1 Oxyfluorfen—
Oxyfluorfen is a pre- and postemergence herbicide that has been registered
since 1979 for use to control a wide spectrum of annual broadleaf weeds and
grasses in apples, artichokes, corn, cotton, jojoba, tree fruits, grapes, nuts,
soybeans, spearmint, peppermint, and certain tropical plantation and ornamental
crops (Farm Chemicals Handbook, 1989).
Evidence suggests that oxyfluorfen is moderately toxic to mammals and has
been assigned to Toxicity Class II based on a chronic mouse feeding study
(Farm Chemicals Handbook, 1989; IRIS, 1993). There is also evidence of
carcinogenicity (liver tumors) in mice (U.S. EPA, 1993a) and therefore
oxyfluorfen has been classified by EPA as a possible human carcinogen (C)
(U.S. EPA, 1992c).
Although oxyfluorfen has an active registration, it has not been included in any
national fish contaminant monitoring program evaluated by the EPA Workgroup.
Experimental evidence indicates this herbicide accumulates in Bluegill sunfish
(BCF from 640 to 1,800) (U.S. EPA, 1993a); however, no consumption
advisories are currently in effect for oxyfluorfen (RTI, 1993).
Oxyfluorfen should be considered for inclusion in State fish and shellfish
contaminant monitoring programs in areas where its use is or has been
extensive. States should contact their appropriate State agencies to obtain
information on the historic and current uses of this pesticide.
4.3.5 Polychlorlnated Biphenyls (Total)
PCBs are base/neutral compounds that are formed by the direct chlorination of
biphenyl. PCBs are closely related to many chlorinated hydrocarbon pesticides
(e.g., DDT, dieldrin, and aldrin) in their chemical, physical, and toxicologic
properties and in their widespread occurrence in the aquatic environment
(Nimmo, 1985). There are 209 different PCS compounds, termed congeners,
based on the possible chlorine substitution patterns. In the United States,
mixtures of various PCB congeners were formulated for commercial use under
4^23
-------
4. TARGET ANALYTES
the trade name Aroclor on the basis of their percent chlorine content. For
example, a common PCB mixture, Aroclor 1254, has an average chlorine content
of 54 percent by weight (Nimmo, 1985).
Unlike the organochlorine pesticides, PCBs were never intended to be released
directly into the environment; most uses were in industrial systems. Important
properties of PCBs for industrial applications include thermal stability, fire and
oxidation resistance, and solubility in organic compounds (Hodges, 1977). PCBs
were used as insulating fluids in electrical transformers and capacitors, as
plasticizers, as lubricants, as fluids in vacuum pumps and compressors, and as
heat transfer and hydraulic fluids (Hodges, 1977; Nimmo, 1985). Although use
of PCBs as a dielectric fluid in transformers and capacitors was generally
considered a closed-system application, the uses of PCBs, especially during the
1960s, were broadly expanded to many open systems where losses to the
environment were likely. Heat transfer systems, hydraulic fluids in die cast
machines, and uses in specialty inks are examples of more open-ended
applications that resulted in serious contamination in fish near industrial
discharge points (Hesse, 1976).
Although PCBs were once used extensively by industry, their production and use
in the United States were banned by the EPA in July 1979 (Miller, 1979). Prior
to 1979, the disposal of PCBs and PCB-containing equipment was not subject
to Federal regulation. Prior to regulation, of the approximately 1.25 billion
pounds purchased by U.S. industry, 750 million pounds (60 percent) were still
in use in capacitors and transformers, 55 million pounds (4 percent) had been
destroyed by incineration or degraded in the environment, and over 450 million
pounds (36 percent) were either in landfills or dumps or were available to biota
via air, water, soil, and sediments (Durfee et al., 1976).
PCBs are extremely persistent in the environment and are bioaccumulated
throughout the food chain (Eisler, 1986; Worthing, 1991). There is evidence that
PCB health risks increase with increased chlorination because more highly
chlorinated PCBs are retained more efficiently in fatty tissues (IRIS, 1992).
However, individual PCB congeners have widely varying potencies for producing
a variety of adverse biological effects including hepatotoxicity, developmental
toxicity, immunotoxicity, neurotoxicity, and carcinogenicity. The non-ortho-
substituted coplanar PCB congeners, and some of the mono-ortho-substituted
congeners, have been shown to exhibit "dioxin-like" effects (Golub et al., 1991;
Kimbrough and Jensen, 1989; McConnell, 1980; Poland and Knutson, 1982;
Safe, 1985, 1990; Tilson et al., 1990; U.S. EPA 1993c). The neurotoxic effects
of PCBs appear to be associated with some degree of ortho-chlorine substitution.
There is increasing evidence that many of the toxic effects of PCBs result from
alterations in hormonal function. However, because PCBs can act directly as
hormonal agonists or antagonists, PCB mixtures may have complex interactive
effects in biological systems (Korach et al., 1988; Safe et al., 1991; Shain et al.,
1991; U.S. EPA, 1993c). Because of the lack of sufficient toxicologic data, EPA
has hot developed quantitative estimates of health risk for specific congeners.
4-24
-------
4. TARGET ANALYTES
PCB mixtures have been classified as probable human carcinogens (IRIS, 1992;
U.S. EPA, 1988a).
Of particular concern are several studies that have suggested that exposure to
PCBs may be damaging to the health of fetuses and children (Fein et al., 1984;
Jacobson et al., 1985, 1990). However, these studies are inconclusive due to
a failure to assess confounding variables (J. Hesse, Michigan Department of
Public Health, personal communication, 1992). In a more recent study of
prenatal exposure to PCBs and reproductive outcome, birth size was found to
be associated positively with PCB exposure, contrary to expectations (Dar et al.,
1992). The results of these investigations clearly indicate the need for further
study. Nevertheless, it may be appropriate for States in which PCBs are found
to be a problem contaminant in fish or shellfish tissue to assess the need to
issue consumption advisories, particularly for pregnant women, nursing mothers,
and children.
PCBs have been included in eight major fish contaminant monitoring programs
(Appendix B). A recent summary of the National Contaminant Biomonitoring
Program data from 1976 through 1984 indicated a significant downward trend in
total PCBs, although PCB residues in fish tissue remained widespread (Schmitt
et al., 1990). Total PCBs were detected at 91 percent of 374 sites surveyed in
the National Study of Chemical Residues in Fish (U.S. EPA, 1992c, 1992d).
Currently, PCB contamination in fish and shellfish has resulted in the issuance
of consumption advisories in 31 States (Figure 4-3) (RTI, 1993).
PCBs may be analyzed quantitatively as Aroclor equivalents or as individual
congeners. Historically, Aroclor analysis has been performed by most
laboratories. This procedure can, however, result in significant error in
determining total PCB concentrations (Schwartz et al., 1987) and in assessing
the toxicologic significance of PCBs, because it is based on the assumption that
distribution of PCB congeners in environmental samples and parent Aroclors is
similar.
The distribution of PCB congeners in Aroclors is, in fact, altered considerably by
physical, chemical, and biological processes after release into the environment,
particularly when the process of biomagnification is involved (Norstrom, 1988;
Oliver and Niimi, 1988; Smith et al., 1990). Recent aquatic environmental
studies indicate that many of the most potent, dioxin-like PCB congeners are
preferentially accumulated in higher organisms (Bryan et al., 1987; Kubiak et al.,
1989; Oliver and Niimi, 1988). This preferential accumulation probably results
in a significant increase in the total toxic potency of PCB residues as they move
up the food chain. Consequently, the congener-specific analysis of PCBs is
required for more accurate determination of total PCB concentrations and for
more rigorous assessment of the toxicologic effects of PCBs.
Even though the large number of congeners of PCBs and their similar chemical
and physical properties present serious analytical difficulties, analytical methods
for the determination of PCB congeners have been improved in recent years so
4^25
-------
M American Samoa
D Guam
Source: RTI, 1993.
States issuing advisory (31)
Virgin Islands
D Puerto Rico
Current 7/15/93
3D
O
ro
o>
Figure 4-3. States issuing fish and shellfish advisories for PCBs.
-------
4. TARGET ANALYTES
that it is now possible to determine essentially all PCB congeners in mixtures
(Huckins et al., 1988; Kannan et al., 1989; MacLeod et al., 1985; Maack and
Sonzogni, 1988; Mes and Weber, 1989; NOAA, 1989b; Smith et al., 1990;
Tanabe et al., 1987). Both NOAA (MacLeod et al., 1985; NOAA, 1989b) and the
EPA Narragansett Research Laboratory conduct PCB congener analyses and
have adopted the same 18 PCB congeners for monitoring fish contamination.
However, quantitation of individual PCB congeners is relatively time-consuming
and expensive and many laboratories do not have the capability or expertise to
perform such analyses. Some States currently conduct both congener and
Aroclor analysis; however, most States routinely perform only Aroclor analysis.
For the purposes of screening tissue residues against potential levels of public
health concern in fish and shellfish contaminant monitoring programs, the issue
of whether to determine PCB concentrations as Aroclor equivalents or as
individual congeners cannot be resolved entirely satisfactorily at this time,
primarily because of a lack of toxicologic data for individual congeners.
Ideally, congener analysis should be conducted. However, at present, only an
Aroclor-based quantitative risk estimate of carcinogenicity is available (IRIS,
1993) for developing SVs and risk assessment. Consequently, until adequate
congener-specific toxicologic data are available to develop quantitative risk
estimates for a variety of toxicologic endpoints, the EPA Office of Water
recommends, as an interim measure, that PCBs be analyzed as Aroclor
equivalents, with total PCB concentrations reported as the sum of Aroclors.
States are encouraged to develop the capability to perform PCB congener
analysis. When congener analysis is conducted, the 18 congeners
recommended by NOAA (shown in Table 4-3) should be analyzed and summed
to determine a total PCB concentration according to the approach used by
NOAA (1989b). States may wish to consider including additional congeners
based on site-specific considerations. PCB congeners of potential environmental
importance identified by McFarland and Clarke (1989) are listed in Table 4-3.
This interim recommendation is intended to (1) allow States flexibility in PCB
analysis until reliable congener-specific quantitative risk estimates are available,
and (2) encourage the continued development of a reliable database of PCB
congener concentrations in fish and shellfish tissue in order to increase our
understanding of the mechanisms of action and toxicities of these chemicals.
The rationale for, and the uncertainties of, this recommended approach are
discussed further in Section 5.3.2.3.
4.3.6 Dioxlns and Dibenzofurans
Note: At this time, the EPA Office of Research and Development is reevaluating
the potency of dioxins and dibenzofurans. Information provided below as well
as information in Section 5.3.2.4 related to calculating toxicity equivalent
concentrations (TECs) and SVs for dioxins/dibenzofurans is subject to change
pending the results of this reevaluation.
_
-------
4. TARGET ANALYTES
Table 4-3. Polychlorlnated Blphenyl (PCB) Congeners Recommended for
Quantltatlon as Potential Target Analytes
Recommended by
PCB Congener8-" NOAAC
2,4' diCB 8
2,2',5 triCB 18
2,4,4' triCB 28
3,4,4' triCB
2,2'3,5' tetraCB 44
2,2'4,5' tetraCB
2,2',5,5' tetraCB 52
2,3',4,4' tetraCB 66
2,3',4',5 tetraCB
2,4,4',5 tetraCB
3,3',4,4' tetraCB 77
3,4,4',5 tetraCB
2,2',3,4,5' pentaCB
2,2',3,4',5 pentaCB
2,21,4,5,5' pentaCB 101
2I3,3',4,4' pentaCB 105
2,3,4,4',5 pentaCB
2,3',4,4'15 pentaCB 118
2,31,4,4',6 pentaCB
2',3,4,4',5 pentaCB
3,31,4,41,5 pentaCB 126
2',3,3',4.4' hexaCB 128
2,2',3,4I4',5' hexaCB 138
2,2',3,5,5',6 hexaCB
2,2',4,4',5,5' hexaCB 153
2,3,3',4,4>,5 hexaCB
2,3,3',4,4',5 hexaCB
2,3,3',414',6 hexaCB
2,3',414',5,5' hexaCB
2,3',4,4115'16 hexaCB
S.S'A^.S.S1 hexaCB 169
Recommended by McFarland
and Clarke (1989)
Highest Second
Priority" Priority*
18
37
44
99
52
70
74
77
81
87
49
101
105
114
116
119
123
126
128
138
151
153
156
157
158
167
168
169
(continued)
4-28
-------
4. TARGET ANALYTES
Table 4-3 (continued)
Recommended by
PCB Congener*1" NOAAC
2,2',3,3>,414',5 heptaCB 170
2121,3,4,41,5,5I heptaCB 180
2,2'13,4141,5',6 heptaCB
2,2',3,4,41I6,6' heptaCB
2,2I(3,4I,5,51,6 heptaCB 187
2,3I3II4,41,5,5I heptaCB
2.2',3131,4141,5,6 octaCB
2,21,3,3',4,5,5',6' octaCB
2.2',313',4I4',5,51,6 nonaCB
2>2>,3>3'14,4'15,51,616' decaCB
Recommended by McFarland
and Clarke (1989)
Highest
Priorltyd
170
180
183
184
195
206
209
Second
Priority"
187
189
201
a PCB congeners recommended for quantftation, from dichlorobiphenyl (diCB) through
decachlorobiphenyl (decaCB).
b Congeners are identified in each column by their International Union of Pure and Applied
Chemistry (IUPAC) number, as referenced in Ballschmitter and Zell (1980) and Mullin et al.
(1984).
0 EPA recommends that these 18 congeners be summed to determine total PCB concentration
(NOAA, 1989b).
d PCB congeners having highest priority for potential environmental importance based on potential
for toxicity, frequency of occurrence in environmental samples, and relative abundance in animal
tissues (McFarland and Clarke, 1989).
6 PCB congeners having second priority for potential environmental importance based on potential
for toxicity, frequency of occurrence in environmental samples, and relative abundance in animal
tissues (McFarland and Clarke, 1989).
4-29
-------
4. TARGET ANALYTES
The polychlorinated dibenzo-p-dioxins (PCDDs) and dibenzofurans (PCDFs) are
included as target analytes primarily because of the extreme potency of 2,3,7,8-
tetrachlorodibenzo-p-dioxin (2,3,7,8-TCDD). Extremely low doses of this isomer
have been found to elicit a wide range of toxic responses in animals, including
carcinogenicity, teratogenicity, fetotoxicity, reproductive dysfunction, and
immunotoxicity (U.S. EPA, 1987d). This compound is the most potent animal
carcinogen evaluated by EPA, and EPA has determined that there is sufficient
evidence to conclude that 2,3,7,8-TCDD is a probable human carcinogen (B2)
(IRIS, 1992). Concern over the health effects of 2,3,7,8-TCDD is increased
because of its persistence in the environment and its high potential to
bioaccumulate (U.S. EPA, 1987d).
Because dioxin/dibenzofuran contamination is found almost exclusively in
proximity to industrial sites (e.g., bleached kraft paper mills or facilities handling
2,4,5-trichlorophenoxyacetic acid [2,4,5-T], 2,4,5-trichlorophenol [2,4,5-TCP],
and/or silvex) (U.S. EPA, 1987d), it is recommended that each State agency
responsible for monitoring include these compounds as target analytes on a site-
specific basis based on the presence of industrial sites and results of any
environmental (water, sediment, soil, air) monitoring performed in areas adjacent
to these sites. All States should maintain a current awareness of potential
dioxin/dibenzofuran contamination.
Fifteen dioxin and dibenzofuran congeners have been included in two major fish
contaminant monitoring programs; however, one congener, 2,3,7,8-TCDD, has
been included in seven national monitoring programs (Appendix B). Six dioxin
congeners and nine dibenzofuran congeners were measured in fish tissue and
shellfish samples in the National Study of Chemical Residues in Fish. The
various dioxin congeners were detected at from 32 to 89 percent of the 388 sites
surveyed, while the furan congeners were detected at from 1 to 89 percent of the
388 sites surveyed (U.S. EPA, 1992c, 1992d). The dioxin/dibenzofuran
congeners detected at more than 50 percent of the sites are listed below:
• 1,2,3,4,6,7,8 HpCDD (89 percent)
• 2,3,7,8 TCDF (89 percent)
• 2,3,7,8 TCDD (70 percent)
• 1,2,3,6,7,8 HxCDD (69 percent)
• 2,3,4,7,8 PeCDF (64 percent)
• 1,2,3,4,6,7,8 HpCDF (54 percent)
• 1,2,3,7,8 PeCDD (54 percent).
Currently, 22 States have issued fish consumption advisories for
dioxins/dibenzofurans (Figure 4-4) (RTI, 1993).
4-30
-------
D American Samoa
D Guam
Source: RTI, 1993.
States issuing advisory (22)
D Virgin Islands
D Puerto Rico
Current 7/15/93
3J
O
5
>
>
|-
m
CO
Figure 4-4. States issuing fish and shellfish advisories for dioxin/dibenzofurans.
-------
4. TARGET ANALYTES
Table 4-4. Dibenzo-p-DioxIns and Dlbenzofurans Recommended
as Target Analytes
2,3,7,8-TCDD 1,2,3,7,8-PeCDF
2,3,4,7,8-PeCDF
1,2,3,7,8-PeCDD
1,2,3,4,7,8-HxCDF
1,2,3,4,7,8-HxCDD 1,2,3,6,7,8-HxCDF
1,2,3,6,7,8-HxCDD 1,2,3,7,8,9-HxCDF
1,2,3,7,8,9-HxCDD 2,3,4,6,7,8-HxCDF
1,2,3,4,6,7,8-HpCDD 1,2,3,4,6,7,8-HpCDF
1,2,3,4,7,8,9-HpCDF
OCDD
OCDF
2,3,7,8-TCDF
Source: Barnes and Bellin, 1989.
Dioxins/dibenzofurans should be considered for analysis primarily at sites of pulp
and paper mills using a chlorine bleaching process and at industrial sites where
the following organic compounds have been or are currently formulated:
herbicides (containing 2,4,5-trichlorophenoxy acids and 2,4,5-trichlorophenol),
hexachlorophene, pentachlorophenol, and PCBs (U.S. EPA, 1987d). If
resources permit, it is recommended that the 17 2,3,7,8-substituted tetra- through
octa-chlorinated dioxin and dibenzofuran congeners shown in Table 4-4 be
included as target analytes. At a minimum, 2,3,7,8-TCDD and 2,3,7,8-
tetrachlorodibenzofuran (2,3,7,8-TCDF) should be determined.
4.4 TARGET ANALYTES UNDER EVALUATION
At present, the EPA Office of Water is evaluating two metals (arsenic and lead)
and polycylic aromatic hydrocarbons for possible inclusion as recommended
target analytes in State fish and shellfish contaminant monitoring programs.
Toxicologic profiles for these compounds and the status of the evaluations are
provided in this section. Other contaminants will be evaluated and may be
recommended as target analytes as additional toxicologic data become available.
Note: Any time a State independently deems that any of the analytes currently
under evaluation and/or other contaminants are of public health concern within
its jurisdiction, the State should include these contaminants in its fish and
shellfish contaminant monitoring program.
4-32
-------
4. TARGET ANALYTES
4.4.1 MetalS
4.4.1.1 Arsenic-
Arsenic is not produced commercially within the United States in any significant
quantities, but it is a byproduct of nonferrous-metal (lead, zinc, and copper)
mining and smelting operations (NAS, 1977). Smelter solid waste is an arsenic
source because it is not commonly removed from waste streams (May and
McKinney, 1981). Arsenic compounds are imported to the United States
primarily for use in rodenticide and other pesticide formulations.
Seafood is a major source of trace amounts of arsenic in the human diet.
However, arsenic is generally present in the edible parts of fish as arsenic-
containing organic compounds (either arsenobetaine or arsenocholine) (NAS,
1991). These organic arsenic compounds are much less toxic than inorganic
forms and are not generally considered a threat to human health (ATSDR,
1989a). Inorganic forms of arsenic (e.g., arsenite and pentavalent arsenic) are
established carcinogens in humans (ATSDR, 1989a) and long-term effects
include dermal hyperkeratosis, dermal melanosis and carcinoma, hepatomegaly,
and peripheral neuropathy (NAS, 1991). Arsenic levels in seafood can be
extremely high (over 10 ppm) (NAS, 1991; G. Pollock, California EPA, personal
communication, 1993). To the degree that inorganic forms of arsenic are either
present in seafood or, upon consumption, may be produced as metabolites of
organic arsenic in seafood, some carcinogenic risk, while small, would be
expected (NAS, 1991).
Arsenic has been included in six national monitoring programs (Appendix B). A
relatively high incidence and magnitude of arsenic contamination has been
reported for bivalves in the southeastern States (Florida, Georgia, North
Carolina, South Carolina), California, and States bordering the Chesapeake Bay
(Virginia and Maryland), whereas Rhode Island, the Great Lakes States, and the
Pacific Coast States, including Alaska, have reported significant arsenic
contamination in their finfish populations (May and McKinney, 1981; NAS, 1991;
Schmitt and Brumbaugh, 1990). One State (Oregon) has a shellfish advisory
currently in effect for arsenic contamination (RTI, 1993).
Because of insufficient evidence for the carcinogenicity of the organic arsenic
compounds, which are predominant in edible fish and shellfish tissue (NAS,
1991), the EPA Office of Water has not included arsenic as a recommended
target analyte in fish and shellfish contaminant monitoring programs at this time.
4.4.1.2 Lead-
Lead is derived primarily from the mining and processing of limestone and
dolomite deposits, which are often sources of lead, zinc, and copper (May and
McKinney, 1981). It is also found as a minor component of coal. Historically,
lead has had a number of industrial uses, including use in paints, in solder used
in plumbing and food cans, and as a gasoline additive. As recently as the mid-
_
-------
4. TARGET ANALYTES
1980s, the primary source of lead in the environment was the combustion of
gasoline; however, use of lead in U.S. gasoline has fallen sharply in recent
years. At present, lead is used primarily in batteries, electric cable coverings,
some exterior paints, ammunition, and sound barriers. Currently, the major
points of entry of lead into the environment are from mining and smelting
operations, from fly ash resulting from coal combustion, and from the leachates
of landfills (May and McKinney, 1981).
Lead has been included in six national monitoring programs (Appendix B). Lead
has been shown to bioaccumulate, with the organic forms, such as tetraethyl
lead, appearing to have the greatest potential for bioaccumulation in fish tissues.
High concentrations of lead have been found in marine bivalves and finfish from
both estuarine and marine waters (NOAA, 1987, 1989a). Lead concentrations
in freshwater fish declined significantly from a geometric mean concentration of
0.28 ppm in 1976 to 0.11 ppm in 1984. This trend has been attributed primarily
to reductions in the lead content of U.S. gasoline (Schmitt and Brumbaugh,
1990). Currently three States (Massachusetts, Missouri, and Tennessee) and
American Samoa have issued fish advisories for lead contamination (RTI, 1993).
Lead is particularly toxic to children and fetuses. Subtle neurobehavioral effects
(e.g., fine motor dysfunction, impaired concept formation, and altered behavior
profile) occur in children exposed to lead at concentrations that do not result in
clinical encephalopathy (ATSDR, 1988). A great deal of information on the
health effects of lead has been obtained through decades of medical observation
and scientific research. This information has been assessed in the development
of air and water quality criteria by the Agency's Office of Health and
Environmental Assessment (OHEA) in support of regulatory decisionmaking by
the Office of Air Quality Planning and Standards (OAQPS) and by the Office of
Drinking Water (ODW). By comparison to most other environmental toxicants,
the degree of uncertainty about the health effects of lead is quite low. It appears
that some of these effects, particularly changes in the levels of certain blood
enzymes and in aspects of children's neurobehavioral development, may occur
at blood lead levels so low as to be essentially without a threshold. The
Agency's Reference Dose (RfD) Work Group discussed inorganic lead (and lead
compounds) in 1985 and considered it inappropriate to develop an RfD for
inorganic lead (IRIS, 1993). Lead and its inorganic compounds have been
classified as probable human carcinogens (B2) by EPA (IRIS, 1992). However,
at this time, a quantitative estimate of carcinogenic risk from oral exposure is not
available (IRIS, 1993).
Because of the lack of quantitative health risk assessment information for oral
exposure to inorganic lead, the EPA Office of Water has not included lead as a
recommended target analyte in fish and shellfish contaminant monitoring
programs at this time. Note: Because of the observation of virtually no-
threshold neurobehavioral developmental effects of lead in children, States
should include lead as a target analyte in fish and shellfish contaminant
programs if there is any evidence that this metal may be present at detectable
4-34
-------
4. TARGET ANALYTES
levels in fish or shellfish tissue. Additional information will be provided on this
issue in Volume 2—Risk Assessment—in this guidance series.
4.4.2 Organlcs
4.4.2.1 Polycycllc Aromatic Hydrocarbons—
Polycyclic aromatic hydrocarbons are base/neutral organic compounds that have
a fused ring structure of two or more benzene rings. PAHs are also commonly
referred to as polynuclear aromatic hydrocarbons (PNAs). PAHs with two to five
benzene rings (i.e., 10 to 24 skeletal carbons) are generally of greatest concern
for environmental and human health effects (Benkert, 1992). These PAHs
include
Acenaphthene • 2-6-Dimethylnaphthalene
Acenaphthylene • Fluoranthene
Anthracene • Fluorene
Benz[a]anthracene • lndeno[ 7,2,3-ccdpyrene
Benzo[a]pyrene • 1 -Methylnaphthalene
Benzo[t»jfluoranthene • 2-Methylnaphthalene
Benzo[/c]fluoranthene • 1 -Methylphenanthrene
Benzo[e]pyrene • Naphthalene
Benzo[g,/j,/]perylene • Perylene
2-Chloronaphthalene • Phenanthrene
Chrysene • Pyrene.
Dibenz[a,/?]anthracene
The metabolites of many of the high-molecular-weight PAHs (e.g., benz[a]an-
thracene, benzo[a]pyrene, benzo[b]fluoranthene, benzo[A]fluoranthene, chrysene,
dibenz[a,/j]anthracene, indeno[ 7,2,3-ccfjpyrene) have been shown in laboratory
test systems to be carcinogens, cocarcinogens, teratogens, and/or mutagens
(Moore and Ramamoorthy, 1984; U.S. DHHS, 1990). Benzo[a]pyrene, one of
the most widely occurring and potent PAHs, and several other PAHs have been
classified by EPA as probable human carcinogens (B2) (IRIS, 1992). Evidence
for the carcinogenicity of PAHs in humans comes primarily from epidemiologic
studies that have shown an increased mortality due to lung cancer in humans
exposed to PAH-containing coke oven emissions, roof-tar emissions, and
cigarette smoke (U.S. DHHS, 1990).
PAHs are ubiquitous in the environment and usually occur as complex mixtures
with other toxic chemicals. They are components of crude and refined petroleum
products and of coal. They are also produced by the incomplete combustion of
organic materials. Many domestic and industrial activities involve pyrosynthesis
of PAHs, which may be released into the environment in airborne particulates or
in solid (ash) or liquid byproducts of the pyrolytic process. Domestic activities
that produce PAHs include cigarette smoking, home heating with wood or fossil
4-35
-------
4. TARGET ANALYTES
fuels, waste incineration, broiling and smoking foods, and use of internal
combustion engines. Industrial activities that produce PAHs include coal coking;
production of carbon blacks, creosote, and coal tar; petroleum refining; synfuel
production from coal; and use of Soderberg electrodes in aluminum smelters and
ferrosilicum and iron works (Neff, 1985). Historic coal gasification sites have
also been identified as significant sources of PAH contamination (J. Hesse,
Michigan Department of Public Health, personal communication, March 1991).
Major sources of PAHs found in marine and fresh waters include biosynthesis
(restricted to anoxic sediments), spillage and seepage of fossil fuels, discharge
of domestic and industrial wastes, atmospheric deposition, and runoff (Neff,
1985). Urban stormwater runoff contains PAHs from leaching of asphalt roads,
wearing of tires, deposition from automobile exhaust, and oiling of roadsides and
unpaved roadways with crankcase oil (MacKenzie and Hunter, 1979). Solid
PAH-containing residues from activated sludge treatment facilities have been
disposed of in landfills or in the ocean (ocean dumping was banned in 1989).
Although liquid domestic sewage contains <1 \ig/L total PAH, the total PAH
content of industrial sewage is 5 to 15 pig/L (Borneff and Kunte, 1965) and that
of sewage sludge is 1 to 30 mg/kg (Grimmer et al., 1978; Nicholls et al., 1979).
In most cases, there is a direct relationship between PAH concentrations in river
water and the degree of industrialization and human activity in the surrounding
watersheds. Rivers flowing through heavily industrialized areas may contain 1
to 5 ppb total PAH, compared to unpolluted river water, ground water, or
seawater that usually contains less than 0.1 ppb PAH (Neff, 1979).
PAHs can accumulate in aquatic organisms from water, sediments, and food.
BCFs of PAHs in fish and crustaceans have frequently been reported to be in
the range of 100 to 2,000 (Eisler, 1987). In general, bioconcentration was
greater for the higher molecular weight PAHs than for the lower molecular weight
PAHs. Biotransformation by the mixed function oxidase system in the fish liver
can result in the formation of carcinogenic and mutagenic intermediates, and
exposure to PAHs has been linked to the development of tumors in fish (Eisler,
1987).
Sediment-associated PAHs can be accumulated by bottom-dwelling invertebrates
and fish (Eisler, 1987). For example, Great Lakes sediments containing elevated
levels of PAHs were reported by Eadie et al. (1983) to be the source of the body
burdens of the compounds in bottom-dwelling invertebrates. Varanasi et al.
(1985) found that benzo[a]pyrene was accumulated in fish, amphipod
crustaceans, shrimp, and clams when estuarine sediment was the source of the
compound. Approximate tissue-to-sediment ratios were 0.6 to 1.2 for
amphipods, 0.1 for clams, and 0.05 for fish and shrimp. NAS (1991) reported
that PAH contamination in bivalves has been found in all areas of the United
States. Bivalves are good bioaccumulators of some PAHs because they do not
metabolize these compounds as rapidly as fish. Varanasi et al. (1985) ranked
benzo[a]pyrene metabolism by aquatic organisms as follows: fish > shrimp >
amphipod crustaceans > clams. Half-lives for elimination of PAHs in fish ranged
_
-------
4. TARGET ANALYTES
from less than 2 days to 9 days (Niimi, 1987). If PAHs are included as target
analytes, bivalves should be selected as target species if available at a site.
Three States (Massachusetts, Michigan, and Ohio) have issued advisories for
PAH contamination in finfish (RTI, 1993).
The EPA Office of Water has not included PAHs on the recommended target
analyte list at this time, primarily because of the lack of quantitative estimates of
carcinogenic risk for all of the individual PAH compounds except benzo[a]pyrene
(IRIS, 1993). Although benzo[a]pyrene is one of the most lexicologically potent
PAHs, it may represent only a small fraction of the total PAH concentration in
fish or shellfish tissue. The carcinogenic potencies of other commonly occurring
PAHs vary widely (U.S. DHHS, 1990). Until reliable quantitative risk estimates
are available for other PAH compounds, States that choose to include PAHs as
target analytes in their fish and shellfish contaminant monitoring programs should
adopt the conservative approach of using the human health screening value
calculated for benzo[a]pyrene to determine the need for additional monitoring or
risk assessment.
4-37
-------
5. SCREENING VALUES FOR TARGET ANALYTES
SECTION 5
SCREENING VALUES FOR TARGET ANALYTES
For the purpose of this guidance document, screening values are defined as
concentrations of target analytes in fish or shellfish tissue that are of potential
public health concern and that are used as standards against which levels of
contamination in similar tissue collected from the ambient environment can be
compared. Exceedance of these SVs should be taken as an indication that more
intensive site-specific monitoring and/or evaluation of human health risk should
be conducted.
The EPA-recommended risk-based method for developing SVs (U.S. EPA,
1989d) is described in this section. This method is considered to be appropriate
for protecting the health of fish and shellfish consumers for the following reasons
(Reinert et al., 1991):
It gives full priority to protection of public health.
It provides a direct link between fish consumption rate and risk levels (i.e.,
between dose and response).
• It generally leads to conservative estimates of increased risk.
It is designed for protection of consumers of locally caught fish and shellfish,
including susceptible subpopulations such as sport and subsistence
fishermen who are at potentially greater risk than the general adult
population because they tend to consume greater quantities of fish and
because they frequently fish the same sites repeatedly.
At this time, the EPA Office of Water is recommending use of this method
because it is the basis for developing current water quality criteria and was the
approach used in the National Study of Chemical Residues in Fish (U.S. EPA,
1992c, 1992d). EPA recognizes that there are many other approaches and
models currently in use. Further discussion of the EPA Office of Water risk-
based approach, including a detailed description of the four steps involved in risk
assessment (hazard identification, dose-response assessment, exposure
assessment, and risk characterization) will be discussed in greater detail in the
second guidance document in this series.
5-1
-------
5. SCREENING VALUES FOR TARGET ANALYTES
5.1 GENERAL EQUATIONS FOR CALCULATING SCREENING VALUES
Risk-based SVs are derived from the general model for calculating the effective
ingested dose of a chemical m (Em) (U.S. EPA, 1989d):
where
Em = Effective ingested dose of chemical m in the population of concern
averaged over a 70-yr lifetime (mg/kg/d)
Cm = Concentration of chemical m in the edible portion of the species of
interest (mg/kg; ppm)
CR = Mean daily consumption rate of the species of interest by the general
population or subpopulation of concern averaged over a 70-yr lifetime
(kg/d)
Xm = Relative absorption coefficient, or the ratio of human absorption
efficiency to test animal absorption efficiency for chemical m
(dimensionless)
BW = Mean body weight of the general population or subpopulation of
concern (kg).
Using this model, the SV for the chemical m (SVm) is equal to Cm when the
appropriate measure of toxicologic potency of the chemical m (Pm) is substituted
for Em. Rearrangement of Equation (5-1), with these substitutions, gives
SVm = (Pm • BW) / (CR - XJ (5-2)
where
Pm = Toxicologic potency for chemical m; the effective ingested dose of
chemical m associated with a specified level of health risk as
estimated from dose-response studies; dose-response variable.
In most instances, relative absorption coefficients (Xm) are assumed to be 1 .0
(i.e., human absorption efficiency is assumed to be equal to that of the test
animal), so that
SVm = (Pm - BW) / CR . (5-3)
However, if Xm is known, Equation (5-2) should be used to calculate SVm.
Dose-response variables for noncarcinogens and carcinogens are defined in
Sections 5.1 .1 and 5.1 .2, respectively. These variables are based on an assess-
ment of the occurrence of a critical toxic or carcinogenic effect via a specific
route of exposure (i.e., ingestion, inhalation, dermal contact). Oral dose-
._
-------
5. SCREENING VALUES FOR TARGET ANALYTES
response variables for the recommended target analytes are given in Appendix
D. Because of the fundamental differences between the noncarcinogenic and
carcinogenic dose-response variables used in the EPA risk-based method, SVs
must be calculated separately for noncarcinogens and potential carcinogens as
shown in the following subsections.
5.1.1 Noncarcinogens
The dose-response variable for noncarcinogens is the reference dose (RfD).
The RfD is an estimate of a daily exposure to the human population (including
sensitive subpopulations) that is likely to be without appreciable risk of
deleterious effects during a lifetime. The RfD is derived by applying uncertainty
or modifying factors to a subthreshold dose (i.e., no observed adverse effect
level [NOAEL] or lowest observed adverse effect level [LOAEL] if the NOAEL is
indeterminate) observed in chronic animal bioassays. These uncertainty or
modifying factors range from 1 to 10,000 and are used to account for
uncertainties in sensitivity differences among human subpopulations; interspecies
extrapolation; short-term to lifetime exposure extrapolation; incomplete or
inadequate toxicity or pharmacokinetic databases; and, where applicable, the
use of a LOAEL instead of a NOAEL (U.S. EPA, 1989d).
The following equation should be used to calculate SVs for noncarcinogens:
SVn = (RfD • BW)/CR (5-4)
where
SVn = Screening value for a noncarcinogen (mg/kg; ppm)
RfD = Oral reference dose (mg/kg/d)
and BW and CR are defined as in Equation (5-1).
5.1.2 Carcinogens
According to The Risk Assessment Guidelines of 1986 (U.S. EPA, 1987f), the
default model for low-dose extrapolation of carcinogens is a version (GLOBAL
86) of the linearized multistage no-threshold model developed by Crump et al.
(1976). This extrapolation procedure provides an upper 95 percent bound risk
estimate (referred to as a q1*), which is considered by some to be a
conservative estimate of cancer risk. Other extrapolation procedures may be
used when justified by the data.
Screening values for carcinogens are derived from: (1) a carcinogenicity potency
factor or slope factor (SF), which is generally an upper bound risk estimate; and
(2) a risk level (RL), an assigned level of maximum acceptable individual
lifetime risk (e.g., RL = 10~5 for a level of risk not to exceed one excess case of
cancer per 100,000 individuals exposed over a 70-yr lifetime) (U.S. EPA, 1989d).
5-3
-------
5. SCREENING VALUES FOR TARGET ANALYTES
The following equation should be used to calculate SVs for carcinogens:
SVC = [(RL / SF) • BW] / CR (5-5)
where
SVC = Screening value for a carcinogen (mg/kg; ppm)
RL = Maximum acceptable risk level (dimensionless)
SF = Oral slope factor (mg/kg/d)'1
and BW and CR are defined as in Equation (5-1).
5.1.3 Recommended Values for Variables In Screening Value Equations
The recommended values in this section for variables used in Equations (5-4)
and (5-5) to calculate SVs are based upon assumptions for the general adult
population. For risk management purposes (e.g., to direct limited resources
toward protection of sensitive subpopulations), States may choose to use values
for consumption rate (CR), body weight (BW), and risk level (RL) different from
those recommended in this section.
5.1.3.1 Dose-Response Variables—
EPA has developed oral RfDs and/or SFs for all of the recommended target
analytes in Section 4 (see Appendix D). These are maintained in the EPA
Integrated Risk Information System (IRIS, 1992), an electronic database
containing health risk and EPA regulatory information on approximately 400
different chemicals. The IRIS RfDs and SFs are reviewed regularly and updated
as necessary when new or more reliable information on the toxic or carcinogenic
potency of chemicals becomes available.
When IRIS values for oral RFDs and SFs are available, they should be used to
calculate SVs for target analytes from Equations (5-4) and (5-5), respectively.
It is important that the most current IRIS values for oral RfDs and SFs be used
to calculate SVs for target analytes, unless otherwise recommended.
A summary description of IRIS and instructions for accessing information in IRIS
are found in U.S. EPA (1989d). Additional information can be obtained from
IRIS User Support (Tel: 513-569-7254). IRIS is also available on the National
Institutes of Health (NIH) National Library of Medicine TOXNET system (Tel:
301-496-6531).
In cases where IRIS values for oral RFDs or SFs are not available for calculating
SVs for target analytes, estimates of these variables should be derived from the
most recent water quality criteria (U.S. EPA, 1992e) according to procedures
described in U.S. EPA (1991 a, p. IV-12), or from the most current Reference
5-4
-------
5. SCREENING VALUES FOR TARGET ANALYTES
Dose List (U.S. EPA, 1993b) and the Classification List of Chemicals Evaluated
for Carcinogenicity Potential (U.S. EPA 1992a) from the Office of Pesticide
Programs Health Effects Division.
5.1.3.2 Body Weight (BW) and Consumption Rate (CR)—
Values for the variables BW and CR in Equations (5-4) and (5-5) are given in
Table 5-1 for the general adult population and various subpopulations. In this
document, the EPA Office of Water used a BW = 70 kg and a CR = 6.5 g/d to
calculate SVs for the general adult population. Note: The 6.5-g/d CR value that
is used to establish water quality criteria is currently under review by the EPA
Office of Water. This CR, which represents a consumption rate for the average
fish consumer in the general adult population (45 FR 231, Part V), may not be
appropriate for sport and subsistence fishermen who generally consume larger
quantities of fish and shellfish (U.S. EPA, 1990a).
With respect to consumption rates, EPA recommends that States always
evaluate any type of consumption pattern they believe could reasonably be
occurring at a site. Evaluating additional consumption rates only involves
calculating additional SVs and does not add to sampling or analytical costs.
The EPA has published detailed guidance on exposure factors (U.S. EPA,
1990a). In addition, EPA has published a review and analysis of survey
methods that can be used by States to determine fish and shellfish consumption
rates of local populations (U.S. EPA, 1992b). States should consult these
documents to ensure that appropriate values are selected to calculate SVs for
site-specific exposure scenarios.
5.1.3.3 Risk Level (RL)—
The EPA Office of Water recommends that an RL of 10~5 be used to calculate
screening values for the general adult population. However, States may choose
to use an appropriate RL value typically ranging from 10~4 to 10~7. This is the
range of risk levels employed in various U.S. EPA programs. Selection of the
appropriate RL is a risk management decision that is made by the State.
5.2 RECOMMENDED SCREENING VALUES FOR TARGET ANALYTES
Recommended target analyte SVs, and the dose-response variables used to
calculate them, are given in Table 5-2. These SVs were calculated from
Equations (5-4) or (5-5) using the following values for BW, CR, and RL and the
most current IRIS values for oral RfDs and SFs (IRIS, 1992) unless otherwise
noted:
5-5
-------
5. SCREENING VALUES FOR TARGET ANALYTES
Table 5-1. Recommended Values for Mean Body Weights (BWs)
and Fish Consumption Rates (CRs) for Selected Subpopulatlons
Variable Recommended value Subpopulation
BW
CR"
70kg
78kg
65kg
12kg
17kg
25kg
36kg
51 kg
61 kg
6.5 g/d (0.0065 kg/d)
14g/d(0.014kg/d)
15 g/d (0.015 kg/d)
30 g/d (0.030 kg/d)
140 g/d (0.140 kg/d)
All adults (U.S. EPA, 1990a)
Adult males (U.S. EPA, 1985b, 1990a)
Adult females (U.S. EPA, 1985b, 1990a)
Children <3 yr (U.S. EPA, 1985b, 1990a)
Children 3 to <6 yr (U.S. EPA, 1985b, 1990a)
Children 6 to <9 yr (U.S. EPA, 1985b, 1990a)
Children 9 to <12 yr (U.S. EPA, 1985b, 1990a)
Children 12 to <15 yr (U.S. EPA, 1985b, 1990a)
Children 15 to <18 yr (U.S. EPA, 1985b, 1990a)
Estimate of the average consumption of fish and
shellfish from estuarine and fresh waters by the
general U.S. population (45 PR 231, Part V)
Estimate of the average consumption of fish and
shellfish from marine, estuarine, and fresh waters by
the general U.S. population (45 FR 231, Part V)
Estimate of the average consumption of fish from the
Great Lakes by the 95th percentile of the regional
population (fishermen and nonfishermen) (U.S. EPA,
1992G)
Estimate of the average consumption of fish and
shellfish from marine, estuarine, and fresh waters by
the 50th percentile of recreational fishermen (U.S.
EPA, 1990a)
Estimate of the average consumption of fish and
shellfish from marine, estuarine, and fresh waters by
the 90th percentile of recreational fishermen (i.e.,
subsistence fishermen) (U.S. EPA, 1990a)
These are recommended consumption rates only. Note: EPA is currently evaluating the use of
6.5 g/d, 30 g/d, and 140 g/d as estimates of consumption rates for the general population, the 50th
percentile of recreational fishermen, and subsistence fishermen, respectively. When local
consumption rate data are available for these populations, they should be used to calculate SVs for
noncarcinogens and carcinogens, as described in Sections 5.1.1 and 5.1.2, respectively.
5-6
-------
5. SCREENING VALUES FOR TARGET ANALYTES
• For noncarclnogens:
BW = 70 kg, average adult body weight
CR = 6.5 g/d (0.0065 kg/d), estimate of average consumption of fish and
shellfish from estuarine and fresh waters by the general adult
population (45 FR 231, Part V).
• For carcinogens:
BW and CR, as above
RL = 10"5, a risk level corresponding to one excess case of cancer per
100,000 individuals exposed over a 70-yr lifetime.
Where both oral RfD and SF values are available for a given target analyte, both
noncarcinogenic and carcinogenic SVs are listed in Table 5-2. Unless otherwise
indicated, the lower of the two SVs should be used. EPA recommends that the
SVs in the shaded boxes (Table 5-2) be used by States when making the
decision to implement Tier 2 intensive monitoring. However, States may choose
to adjust these SVs for specific target analytes for the protection of sensitive
subpopulations (e.g., pregnant women, children, and recreational or subsistence
fishermen). EPA recognizes that States may use higher CRs that are more
appropriate for recreational and subsistence fishermen in calculating SVs for use
in their jurisdictions rather than the 6.5-g/d CR for the general adult population
used to calculate the SVs shown in Table 5-2.
Note: States should use the same SV (i.e., either for the general adult
population or adjusted for other subpopulations) for a given target analyte for
both screening and intensive studies. Therefore, it is critical that States clearly
define their program objectives and accurately characterize the population or
subpopulation(s) of concern in order to ensure that appropriate SVs are selected.
For noncarcinogens, adjusted SVs should be calculated from Equation (5-4)
using appropriate alternative values of BW and/or CR. For carcinogens,
adjusted SVs should be calculated from Equation (5-5) using an RL ranging from
10~4 to 10"7 and/or sufficiently protective alternative values of BW and CR.
Examples of SVs calculated for selected subpopulations of concern and for RL
values ranging from 10~4 to 10"7 are given in Table 5-3.
The need to accurately characterize the subpopulation of interest in order to
establish sufficiently protective SVs cannot be overemphasized. For example,
the recommended consumption rate of 140 g/d for subsistence fishermen may
be an underestimate of consumption rate for some subsistence populations. In
a recent study of Alaskan subsistence fishing economies (Wolf and Walker,
1987), daily consumption rates for subsistence fishermen were found to range
from 6 to 1,536 g/d, with an average daily consumption rate of 304 g/d. Using
5-7
-------
Table 5-2. Dose-Response
Target analyte
Metals
Cadmium
Mercury0
Selenium8
Organochlorine Pesticides
Total chlordane (sum of cis- and trans-
chlordane, cis- and trans-nonachlor,
and oxychlordane)'
Total DDT (sum of 4,4'- and 2,4'-
isomers of DDT, DDE, and ODD)8
Dicofol
Dieldrin
Endosulfan (1 and II)
Endrin
Heptachlor epoxide
Hexachlorobenzene
Lindane (y-hexachlorocyclohexane; y-HCH)
Mirex
Toxaphene
See notes at end of table
Variables and Recommended
Noncarcinogens
RfDb
(mg/kg/d)
1 x 10'3
6x 10'5d
5 x 10'3
6x 10'5
5 x 10~4
1x10'3h
5x 1Q-5
1.5x10'3h
3x 10'4
1.3x10-5
8x 10'4
3x10'4
2x10'4
2.5x10-4h'k
Screening Values
Carcinogens
SFb
(mg/kg/d)0
NA
NA
NA
1.3
0.34
NA
16
NA
NA
9.1
1.6
1.31
NAj
1.1
(SVs) for Target Analytes
SV» (ppm)
Carcinogens
Noncarcinogens (RL=10"5)
10 —
0.6*
50 —
0.6 0,08
5 0.3
10 —
0.6 7 X 10**
20 —
S —
0.1 0,0t
9 0,07
3 0.08
2 —
3 0*1
(continued)
Ul
i
m
m
z
z
o
m
31
iRGET ANALYTES
-------
V
-------
Table 5-2 (continued)
to one significant figure. EPA believes that using more than one significant figure would imply a degree of precision that is not warranted given the
large uncertainty factors generally used in deriving SVs. For target analytes with both carcinogenic and noncarcinogenic effects, the lower (more
conservative) of the calculated SVs should be used. Note: Values in the shaded boxes are SVs recommended for use in State fish/shellfish
consumption advisory programs for the general adult population. States may choose to use other SVs based on different CRs, BWs, and/or an RL
ranging from 10~4 to 10"7.
Unless otherwise noted, values listed are the most current oral RfDs and SFs in EPA's IRIS (IRIS, 1992).
Because most mercury in fish and shellfish tissue is present as methylmercury (MAS, 1991; Tollefson, 1989) and because of the relatively high cost
of analyzing for methylmercury, it is recommended that total mercury be analyzed and the conservative assumption be made that all mercury is
present as methylmercury. This approach is deemed to be most protective of human health and most cost-effective.
For the purpose of calculating an SV, the RfD for methylmercury currently available in the EPA IRIS database (3 x 10"4 mg/kg/d) has been lowered
by a factor of 5 to a value of 6 x 10's mg/kg/d. The EPA is reevaluating the RfO for methylmercury and is especially concerned about evidence that
the fetus, and possibly pregnant women, are at increased risk of adverse neurological effects from exposure to methylmercury (WHO, 1976, 1990;
Piotrowski and Inskip, 1981; Marsh et al., 1987). In the general adult population, blood methylmercury concentrations of 200 |ig/L (corresponding to
approximately 50 jig/g in hair) have been associated with a 5 percent risk of parasthesia; whereas for the fetus, a 5 percent risk of neurological and (/>
developmental abnormalities is associated with peak mercury concentrations of 10-20 jtg/g in the maternal hair (WHO, 1990). These findings suggest
a possible fivefold increase in fetal sensitivity to methylmercury exposure. Consequently, the EPA has chosen to apply an uncertainty factor of 5 to
the current IRIS RfD for methylmercury. This approach was deemed to be the most prudent as an interim measure until the current reevaluation of ~
the methylmercury RfD is completed.
The RfD for selenium is the IRIS (1992) value for selenious acid. The evidence of carcinogenicity for various selenium compounds in animal and >
O
mutagenicity studies is conflicting and difficult to interpret. However, evidence for selenium sulfide is sufficient for a B2 classification (IRIS, 1992).
m
to
The RfD and SF values listed are derived from studies using technical-grade chlordane (purity -95%) or a 90:10 mixture of chlordane:heptachlor or
analytical-grade chlordane (IRIS, 1992). No RfD or SF values are given in IRIS (1992) for the cis- and trans-chlordane isomers or the major O
chlordane metabolite, oxychlordane, or for the chlordane impurities cis- and trans-nonachlor. It is recommended that the total concentration of cis-
and trans-chlordane, cis- and trans-nonachlor, and oxychlordane be determined for comparison with the recommended SV. >
3}
O
(continued)
m
to
-------
Table 5-2 (continued)
9 The RfD value fisted is for DDT. The SF value is for DDT or DDE; the SF value for DDD is 0.24. The U.S. EPA Carcinogenicity Assessment Group
recommended the use of SF = 0.34 for any combination of DDT, DDE, DDD, and dicofol (Holder, 1986). It is recommended that the total
concentration of the 2,4'- and 4,4'-isomers of DDT and its metabolites, DDE and DDD, be determined for comparison with the recommended SV.
h The RfD value listed is from U.S. EPA (1993b).
1 IRIS (1992) has not provided an SF for lindane. The SF value listed for lindane was calculated from the water quality criteria (0.063 ng/L) (U.S. EPA,
1992e).
j The National Study of Chemical Residues in Fish (U.S. EPA, 1992c, 1992d) used a value of SF = 1.8 for mirex from HEAST (1989).
k The RfD value is the Office of Pesticide Programs value; this value was never submitted for verification.
' The National Study of Chemical Residues in Fish (U.S. EPA, 1992c, 1992d) used a value of RfD = 1x10'4 for Aroclor 1016 from ATSDR (1987c).
The Great Lakes Initiative uses an RfD = 8 x 10~6 for total PCBs (i.e., all PCB isomers and Aroclor mixtures) (U.S. EPA, 1992e). The EPA
Environmental Criteria and Assessment Office, Cincinnati, OH, is also currently developing RfDs for the noncancer toxicity of various commercial
mixtures of PCBs (Michael Doursan, Chief of Systemic Toxicants Assessment Branch, EPA Office of Research and Development, Cincinnati, OH,
personal communication, April 21, 1992). w
O
3J
The SF is based on a carcinogenicity assessment of Aroclor 1260. The SF of Aroclor 1260 is intended to represent the upper bound risk for all PCB rn
mixtures (IRIS, 1992).
The SF value listed is for 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)(U.S. EPA, 1986c). The National Study of Chemical Residues in Fish used a
value of RfD = 1x10"9 for 2,3,7,8-TCDD from ATSDR (1987d). It is recommended that, in both screening and intensive studies, the 17 2,3,7,8- >
substituted tetra- through octa-chlorinated dibenzo-p-dioxins and dibenzofurans be determined and a toxicity-weighted total concentration be _
calculated for each sample for comparison with the recommended SV, using the revised interim method for estimating Toxicity Equivalency nj
Concentrations (TECs) (Barnes and Bellin, 1989; U.S. EPA, 1991h). If resources are limited, the 2,3,7,8-TCDD and 2,3,7,8-TCDF congeners should
be determined at a minimum. 6
3)
3J
O
m
-------
5. SCREENING VALUES FOR TARGET ANALYTES
Table 5-3. Example Screening Values (SVs) for Various
Subpopulatlons and Risk Levels (RLs)a
Chemical
Subpopulatiorr
CRe BW
RfD
SF RL
SV (ppm)
Noncarclnogens
Chlorpyrifos
Cadmium
Standard adults
Children
Subsistence
fishermen
6.5 70 3 x 10'4
6.5 36d 3x10'3
140 70 3x10'3
Standard adults
Children
Subsistence
fishermen
6.5 70 1 x 10'J
6.5 36d 1 x 10'3
140 70 1 x 10'3
30
20
2
10
6
0.5
Carcinogens
Lindane
Toxaphene
Standard adults
6.5 70
1.3
1.3
1.3
1.3
io
-5
"2
8x
8x 10
8x 10'3
8 x 1Q-4
Children 6.5 36°
Subsistence 140 70
fishermen
Standard adults 6.5 70
Children 6.5 36d
Subsistence 140 70
fishermen
— 1.3
1.3
1.3
1.3
— 1.3
1.3
1.3
1.3
— 1.1
1.1
1.1
1.1
— 1.1
1.1
1.1
1.1
— 1.1
1.1
1.1
1.1
10'4
10'5
10'6
10'7
10'4
10'5
io-6
io-7
10'4
10'5
10'6
io-7
10'4
10'5
10"6
io-7
10'4
10'5
10'6
10"7
4x 10'1
4x 10'2
4x 10'3
4x 10'4
4x 10'2
4x 10'3
4x 10'4
4x 10'5
10X10'1
10 x 10'2
10 x 1Q-3
10 x 10'4
5 x 10-1
5 x 10'2
5x 1Q-3
5 x 10'4
5x 10'2
5 x 10'3
5 x 10'4
5 x 10'5
CR
BW
RfD
SF
RL
* See Equations (5-4) and (5-5).
See Table 5-2 for definitions of subpopulations.
c To calculate SVs, the CRs given in this table must be divided by 1,000 to convert g/d to kg/d.
° BW used is for children 9 to <12 yr (see Table 5-2).
Mean daily fish or shellfish consumption rate, averaged over a 70-yr lifetime for the population of concern (g/d).
Mean body weight, estimated for the population of concern (kg).
Oral reference dose for noncarcinogens (mq/kg/d).
Oral slope factor for carcinogens (mg/kg/d) .
Maximum acceptable risk level for carcinogens (dimensionless).
5-12
-------
5. SCREENING VALUES FOR TARGET ANALYTES
this average consumption rate and an estimated average body weight of 70 kg,
the SV for cadmium (RfD = 1 x 10"3 mg/kg/d) is, from Equation (5-4),
SV = (0.001 mg/kg/d • 70 kg) / (0.304 kg/d) = 0.2 mg/kg (ppm) . (5-7)
This value is significantly lower than the SV of 0.5 ppm for cadmium based on
the recommended consumption rate of 140 g/d for subsistence fishermen, as
shown in Table 5-3.
5.3 COMPARISON OF TARGET ANALYTE CONCENTRATIONS WITH
SCREENING VALUES
As noted previously, the same SV for a specific target analyte should be used
in both the screening and intensive studies. The measured concentrations of
target analytes in fish or shellfish tissue should be compared with their
respective SVs in both screening and intensive studies to determine the need for
additional monitoring and risk assessment.
Recommended procedures for comparing target analyte concentrations with SVs
are provided below. Related guidance on data analysis is given in Section 9.1.
5.3.1 Metals
For each of the metals recommended as target analytes (i.e., cadmium, mercury,
and selenium), the total metal tissue concentration should be determined for
comparison with the appropriate SV.
The determination of methylmercury is not recommended even though methyl-
mercury is the compound of greatest concern for human health (NAS, 1991;
Tollefson, 1989) and the recommended SV is for methylmercury (see Table 5-2).
Because most mercury in fish and shellfish tissue is present as methylmercury
(NAS, 1991; Tollefson, 1989), and because of the relatively high analytical cost
for methylmercury, it is recommended that total mercury be determined and the
conservative assumption be made that all mercury is present as methylmercury.
This approach is deemed to be most protective of human health and most cost-
effective.
Note: For the purposes of calculating an SV, the RfD for methylmercury
currently available in the EPA IRIS database (3 x 10~4 mg/kg/d) has been
lowered by a factor of 5 to a value of 6 x 10~5 mg/kg/d. The EPA is reevaluating
the RfD for methylmercury and is especially concerned about evidence that the
fetus, and possibly pregnant women, are at increased risk of adverse
neurological effects from exposure to methylmercury (Marsh et al., 1987;
Piotrowski and Inskip, 1981; WHO, 1976,1990). In the general adult population,
blood methylmercury concentrations of 200 \ig/L (corresponding to approximately
50 ng/g in hair) have been associated with a 5 percent risk of parasthesia;
whereas for the fetus, a 5 percent risk of neurological and developmental
abnormalities is associated with peak mercury concentrations of 10 to 20 n.g/g
-—
-------
5. SCREENING VALUES FOR TARGET ANALYTES
in the maternal hair (WHO, 1990). These findings suggest a possible fivefold
increase in fetal sensitivity to methylmercury exposure. Consequently, the EPA
has chosen to apply an uncertainty factor of 5 to the current IRIS RfD value for
methylmercury. This approach was deemed to be the most prudent as an
interim measure until the current reevaluation of the methylmercury RfD is
completed.
5.3.2 Organlcs
For each of the recommended organic target analytes that are single
compounds, the determination of tissue concentration and comparison with the
appropriate SV is straightforward. However, for those organic target analytes
that include a parent compound and structurally similar compounds or metabo-
lites (i.e., total chlordane, total DDT), or that represent classes of compounds
(i.e., PCBs, dioxins/dibenzofurans), additional guidance is necessary to ensure
that a consistent approach is used to determine appropriate target analyte
concentrations for comparison with recommended SVs.
5.3.2.1 Chlordane—
The SV for total chlordane is derived from technical-grade chlordane. Oral slope
factors are not available in IRIS (1992) for cis- and trans-chlordane, cis- and
trans-nonachlor, and oxychlordane. At this time, as a conservative approach,
EPA recommends that, in both screening and intensive studies, the
concentrations of cis- and trans-chlordane, cis- and trans-nonachlor, and
oxychlordane be determined and summed to give a total chlordane concentration
for comparison with the recommended SV for total chlordane (see Table 5-2).
5.3.2.2 DDT—
DDT and its metabolites (i.e., the 4,4'- and 2,4'-isomers of DDE and DDD) are
all potent toxicants, DDE isomers being the most prevalent in the environment.
As a conservative approach, EPA recommends that, in both screening and
intensive studies, the concentrations of 4,4'- and 2,4'-DDT and their DDE and
DDD metabolites be determined and a total DDT concentration be calculated for
comparison with the recommended SV for total DDT (see Table 5-2, footnote g).
5.3.2.3 PCBs—
Using the interim approach for PCB analysis recommended by the EPA Office
of Water (see Section 4.3.5), total PCB concentrations should be determined, in
both screening and intensive studies, as the sum of Aroclor equivalents. The
total PCB concentration should be compared with the recommended SV for
PCBs (see Table 5-2). Because this SV is based on the SF for Aroclor 1260,
the recommendation to use this SV for comparison with total Aroclor
concentration requires the assumption that Aroclor 1260 is representative of
other PCB mixtures, i.e., that the SF for Aroclor 1260 is an upper limit risk
estimate for all other PCB mixtures as well (IRIS, 1992; U.S. EPA, 1988a). The
_
-------
5. SCREENING VALUES FOR TARGET ANALYTES
Table 5-4. Toxlclty Equivalency Factors (TEFs) for Tetra-
through Octa-Chlorinated Dlbenzo-p-DloxIns and Dlbenzofurans
Analvte TEF8
2,3,7,8-TCDD 1.00
1,2,3,7,8-PeCDD 0.50
1,2,3,4,7,8-HxCDD 0.10
1,2,3,6,7,8-HxCDD 0.10
1,2,3,7,8,9-HxCDD 0.10
1,2,3,4,6,7,8-HpCDD 0.01
OCDD 0.001
2,3,7,8-TCDF 0.10
1,2,3,7,8-PeCDF 0.05
2,3,4,7,8-PeCDF 0.50
1,2,3,4,7,8-HxCDF 0.10
1,2,3,6,7,8-HxCDF 0.10
1,2,3,7,8,9-HxCDF 0.10
2,3,4,6,7,8-HxCDF 0.10
1,2,3,4,6,7,8-HpCDF 0.01
1,2,3,4,7,8,9-HpCDF 0.01
OCDF 0.001
Source: Barnes and Bellin, 1989.
aTEFs for all non-2,3,7,8-substituted congeners are zero.
5-15
-------
5. SCREENING VALUES FOR TARGET ANALYTES
EPA Office of Water recognizes that this assumption has significant
uncertainties.
The comparison of total PCB concentrations (determined as the sum of Aroclor
equivalents) with the Aroclor 1260-based SV may be overly conservative. The
EPA Carcinogen Assessment Group has reported a much lower SF for Aroclor
1254 (SF = 2.6) and data from studies of Aroclor 1242 (Schaeffer et al., 1984)
indicate that there are no statistically significant increases in liver tumors
compared to controls. A recent reassessment of the results of five PCB studies
in rats found significant differences between Aroclor 1260 and other Aroclors in
the types and incidence of pathological effects on rats (IEHR, 1991). On the
other hand, Aroclor 1260 may not represent an upper bound risk estimate
because the PCB congener distribution in fish and shellfish tissue is usually
markedly altered from, and may be more potent than, the parent Aroclor mixture
(Bryan et al., 1987; Kubiak et al., 1989; Norstrom, 1988; Oliver and Niimi, 1988;
Smith et al., 1990). This underscores the need to move toward congener-
specific analysis based on (1) pharmacokinetics and (2) relative potency at
specific site(s) of action (NAS, 1991).
EPA also recognizes that the current recommended SV of 10 ppb for total PCBs
will result in widespread exceedance in waterbodies throughout the country and
will drive virtually all fish and shellfish contaminant monitoring programs into the
risk assessment phase for PCBs. The decision on whether to issue a
consumption advisory for PCBs at this level is one that must be made by risk
managers in each State.
EPA is currently giving high priority to addressing the unresolved issues related
to PCB analysis and risk assessment. A work group has been convened to
examine the feasibility of TEFs for PCB congeners similar to those developed
for PCDDs and PCDFs (U.S. EPA, 1991J) and two EPA-sponsored national
workshops have been held recently to identify problematic issues and areas for
future research (U.S. EPA, 1993c; U.S. EPA, In preparation). Additional
guidance on PCB analyses will be provided in addenda to this document and in
subsequent documents in this series.
5.3.2.4 Dloxlns/Dibenzofurans—
Note: At this time, the EPA Office of Research and Development is reevaluating
the potency of dioxins/dibenzofurans. Consequently, the following recommen-
dation is subject to change pending the results of this reevaluation.
It is recommended in both screening and intensive studies that the 17 2,3,7,8-
substituted tetra- through octa-chlorinated PCDDs and PCDFs be determined
and that a toxicity-weighted total concentration be calculated for each sample for
comparison with the recommended SV for 2,3,7,8-TCDD (see Table 5-2).
5-16
-------
5. SCREENING VALUES FOR TARGET ANALYTES
The revised interim method for estimating toxicity equivalency concentrations
(Barnes and Bellin, 1989) should be used to estimate TCDD equivalent
concentrations according to the following equation:
TEC = I (TEFj - Cj) (5-8)
i
where
TEF, = Toxicity equivalency factor for the ith congener (relative to
2,3,7,8-TCDD)
Cj = Concentration of the ith congener.
TEFs for the 2,3,7,8-substituted tetra- through octa- PCDDs and PCDFs are
shown in Table 5-4.
If resources are limited, the 2,3,7.8-TCDD and 2,3,7,8-TCDF congeners should
be determined and the calculated TEC compared with the recommended SV for
2,3,7,8-TCDD (see Table 5-2).
5-17
-------
6. FIELD PROCEDURES
SECTION 6
FIELD PROCEDURES
This section provides guidance on sampling design of screening and intensive
studies and recommends field procedures for collecting, preserving, and shipping
samples to a processing laboratory for target analyte analysis. Planning and
documentation of all field procedures are emphasized to ensure that collection
activities are cost-effective and that sample integrity is preserved during all field
activities.
6.1 SAMPLING DESIGN
Prior to initiating a screening or intensive study, the program manager and field
sampling staff should develop a detailed sampling plan. As described in Section
2, there are seven major parameters that must be specified prior to the initiation
of any field collection activities:
• Site selection
Target species (and size class)
• Target analytes
• Target analyte screening values
• Sampling times
Sample type
Replicate samples.
In addition, personnel roles and responsibilities in all phases of the fish and
shellfish sampling effort should be defined clearly. All aspects of the final
sampling design for a State's fish and shellfish contaminant monitoring program
should be documented clearly by the program manager in a Work/QA Project
Plan (see Appendix E). Routine sample collection procedures should be
prepared as standard operating procedures (U.S. EPA, 1984b) to document the
specific methods used by the State and to facilitate assessment of final data
quality and comparability.
The seven major parameters of the sampling plan should be documented on a
sample request form prepared by the program manager for each sampling site.
The sample request form should provide the field collection team with readily
available information on the study objective, site location, site name/number,
target species and alternate species to be collected, target analytes to be
evaluated, anticipated sampling dates, sample type to be collected, number and
size range of individuals to be collected for each composite sample, sampling
6-1
-------
6. FIELD PROCEDURES
method to be used, and number of replicates to be collected. An example of a
sample request form is shown in Figure 6-1. The original sample request form
should be filed with the program manager and a copy kept with the field logbook.
The seven major parameters that must be specified in the sampling plan for
screening and intensive studies are discussed in Sections 6.1.1 and 6.1.2,
respectively.
6.1.1 Screening Studies (Tier 1)
The primary aim of screening studies is to identify frequently fished sites where
commonly consumed fish and shellfish species are contaminated and may pose
a risk to human health. Ideally, screening studies should include all waterbodies
where commercial, recreational, or subsistence fishing and shellfish harvesting
are practiced.
6.1.1.1 Site Selection-
Sampling sites should be selected to identify extremes of the bioaccumulation
spectrum, ranging from presumed undisturbed reference sites to sites where
existing data (or the presence of potential pollutant sources) suggest significant
contamination. Where resources are limited, States initially should target those
harvest sites suspected of having the highest levels of contamination and of
posing the greatest potential health risk to local fish and shellfish consumers.
Screening study sites should be located in frequently fished areas near
Point source discharges such as
— Industrial or municipal dischargers
— Combined sewer overflows (CSOs)
— Urban storm drains
Nonpoint source inputs such as
— Landfills, Resource Conservation and Recovery Act (RCRA) sites, or
Superfund Comprehensive Environmental Response, Compensation,
and Liability Act (CERCLA) sites
— Areas of intensive agricultural, silvicultural, or resource extraction
activities or urban land development
— Areas receiving inputs through multimedia mechanisms such as
hydrogeologic connections or atmospheric deposition (e.g., areas
affected by acid rain impacts, particularly lakes with pH <6.0 since
elevated mercury concentrations in fish have been reported for such
sites)
6-2
-------
6. FIELD PROCEDURES
Sample Request Form
Project
Objective
Sample
Type
Target
Contaminants
CD Screening Study
D Fish fillets only
d Shellfish (edible portions)
(Specify portions if other than
whole )
Whole fish or portions other
than fillet (Specify tissues used
if other than whole
CD All target contaminants
CD Additional contaminants
(Specify
D Intensive Study
D Fish fillets only
D Shellfish (edible portions)
(Specify portions if other than whole
CD Whole fish or portions other than fillet (Specify
tissues used if other than whole
J
O Contaminants exceeding screening study SVs
(Specify
J
INSTRUCTIONS TO SAMPLE COLLECTION TEAM
Project Number:.
County/Parish:
Target Species:
CD Freshwater
CD Estuarine
Site (Name/Number):
LatVLong.:
Alternate Species: (in order of preference)
Proposed Sampling Dates:.
Proposed Sampling Method:
CD Electrofishing
CD Seining
CD Trawling
CD Other (Specify,
G Mechanical grab or tongs
D Biological dredge
CD Hand collection
Number of Sample Replicates: CD No field replicates (1 composite sample only)
CD field replicates
(Specify number for each target species)
Number of Individuals
per Composite:
_ Fish per composite
.Shellfish per composite (specify number to obtain 200 grams of tissue)
Figure 6-1. Example of a sample request form.
6-3
-------
6. FIELD PROCEDURES
Areas acting as potential pollutant sinks where contaminated sediments
accumulate and bioaccumulation potential might be enhanced (i.e., areas
where water velocity slows and organic-rich sediments are deposited)
Areas where sediments are disturbed by dredging activities
• Unpolluted areas that can serve as reference sites for subsequent intensive
studies. For example, Michigan sampled lakes that were in presumed
unpolluted areas but discovered mercury contamination in fish from many of
these areas and subsequently issued a fish consumption advisory for all of
its inland lakes.
The procedures required to identify candidate screening sites near significant
point source discharges are usually straightforward. It is often more difficult,
however, to identify clearly defined candidate sites in areas affected by pollutants
from nonpoint sources. For these sites, assessment information summarized in
State Section 305(b) reports should be reviewed before locations are selected.
State 305(b) reports are submitted to the EPA Assessment and Watershed
Protection Division biennially and provide an inventory of the water quality in
each State. The 305(b) reports often contain Section 319 nonpoint source
assessment information that may be useful in identifying major sources of
nonpoint source pollution to State waters. States may also use a method for
targeting pesticide hotspots in estuarine watersheds that employs pesticide use
estimates from NOAA's National Coastal Pollutant Discharge Inventory (Farrow
etal., 1989).
It is important for States to identify and document at least a few unpolluted sites,
particularly for use as reference sites in subsequent monitoring studies.
Verification that targeted reference sites show acceptably low concentrations of
contaminants in fish or shellfish tissues also provides at least partial validation
of the methods used to select potentially contaminated sites. Clear differences
between the two types of sites support the site-selection methodology and the
assumptions about primary sources of pollution.
In addition to the intensity of subsistence, sport, or commercial fishing, factors
that should be evaluated (Versar, 1982) when selecting fish and shellfish
sampling sites include
Proximity to water and sediment sampling sites
Availability of data on fish or shellfish community structure
Bottom condition
Type of sampling equipment
Accessibility of the site.
The most important benefit of locating fish or shellfish sampling sites near sites
selected for water and sediment sampling is the possibility of correlating
contaminant concentrations in different environmental compartments (water,
sediment, and fish). Selecting sampling sites in proximity to one another is also
_
-------
6. FIELD PROCEDURES
more cost-effective in that it provides opportunities to combine sampling trips for
different matrices.
Availability of data on the indigenous fish and shellfish communities should be
considered in final site selection. Information on preferred feeding areas and
migration patterns is valuable in locating populations of the target species
(Versar, 1982). Knowledge of habitat preference provided by fisheries biologists
or commercial fishermen may significantly reduce the time required to locate a
suitable population of the target species at a given site.
Bottom condition is another site-specific factor that is closely related to the
ecology of a target fish or shellfish population (Versar, 1982). For example, if
only soft-bottom areas are available at an estuarine site, neither oysters
(Crassostrea virginica) nor mussels (Mytilus edulis and M. californianus) would
likely be present because these species prefer hard substrates. Bottom
condition also must be considered in the selection and deployment of sampling
equipment. Navigation charts provide depth contours and the locations of large
underwater obstacles in coastal areas and larger navigable rivers. Sampling
staff might also consult commercial fishermen familiar with the candidate site to
identify areas where the target species congregates and the appropriate
sampling equipment to use.
Another factor closely linked to equipment selection is the accessibility of the
sampling site. For some small streams or land-locked lakes (particularly in
mountainous areas), it is often impractical to use a boat (Versar, 1982). In such
cases the sampling site should have good land access. If access to the site is
by land, consideration should be given to the type of vegetation and local
topography that could make transport of collection equipment difficult. If access
to the sampling site is by water, consideration should be given to the location of
boat ramps and marinas and the depth of water required to deploy the selected
sampling gear efficiently and to operate the boat safely. Sampling equipment
and use are discussed in detail in Section 6.2.1.
The selection of each sampling site must be based on the best professional
judgment of the field sampling staff. Once the site has been selected, it should
be plotted and numbered on the most accurate, up-to-date map available.
Recent 7.5-minute (1:24,000 scale) maps from the U.S. Geologic Survey or blue
line maps produced by the U.S. Army Corps of Engineers are of sufficient detail
and accuracy for sample site mapping. The type of sampling to be conducted,
water depth, and estimated time to the sampling site from an access point
should be noted. The availability of landmarks for visual or range fixes should
be determined for each site, and biological trawl paths (or other sampling gear
transects) and navigational hazards should be indicated. Additional information
on site-positioning methods, including Loran-C, VIEWNAV, TRANSIT (NAVSAT),
GEOSTAR, and the NAVSTAR Global Positioning System (GPS), is provided in
Battelle (1986), Tetra Tech (1986), and Puget Sound Estuary Program (1990a).
6-5
-------
6. FIELD PROCEDURES
Each sampling site must be described accurately because State fish and
shellfish contaminant monitoring data may be stored in a database available to
users nationwide (see Section 9.2). For example, a sampling site may be
defined as a 2-mile section of river (e.g., 1 mile upstream and 1 mile down-
stream of a reference point) or a 2-mile stretch of lake or estuarine/marine
shoreline (U.S. EPA, 1990d). Each sampler should provide a detailed descrip-
tion of each site using a 7.5-minute USGS map to determine the exact latitude
and longitude coordinates for the reference point of the site. This information
should be documented on the sample request form and field record sheets (see
Section 6.2.3).
6.1.1.2 Target Species and Size Class Selection-
After reviewing information on each sampling site, the field collection staff should
identify the target species that are likely to be found at the site. Target species
recommended for screening studies in freshwater systems are shown in Tables
3-1 and 3-2, and Tables 3-7 through 3-13 list recommended species for
estuarine/marine areas. In freshwater ecosystems, one bottom-feeding and one
predator fish species should be collected. In estuarine/marine ecosystems,
either one bivalve species and one finfish species or two finfish species should
be collected. Second and third choice target species should be selected in the
event that the recommended target species are not collected at the site. The
same criteria used to select the recommended target species (Section 3.2)
should be used to select alternate target species. In all cases, the primary
selection criterion should be that the target species is commonly consumed
locally and is of harvestable size.
EPA recognizes that resource limitations may influence the sampling strategy
selected by a State. If monitoring resources are severely limited, precluding
performance of any Tier 2 intensive studies (Phase I and Phase II), EPA
recommends three sampling options to States for collecting additional samples
during the screening studies. These options are:
1. Collecting one composite sample for each of three size (age) classes of
each target species
2. Collecting replicate composite samples for each target species
3. Collecting replicate composite samples for each of three size (age) classes
of each target species.
Option 1 (single composite analysis for each of three size classes) provides
additional information on size-specific levels of contamination that may allow
States to issue an advisory for only the most contaminated size classes while
allowing other size classes of the target species to remain open to fishing. The
State could analyze the composite sample from the largest size class first. If
any SVs are exceeded, analysis of the smaller size class composite samples
could be conducted. This option, however, does not provide any additional
_
-------
6. FIELD PROCEDURES
information for estimating the variability of the contamination level in any specific
size class. To obtain information for estimating the variability of the contamina-
tion level in the target species, States could separately analyze each individual
fish specimen in any composite that exceeded the SVs. Note: This option of
analyzing individual fish within a composite sample is more resource-intensive
with respect to analytical costs but is currently used by some Great Lakes
States.
Option 2 (replicate analyses of one size class) provides additional statistical
power that would allow States to estimate the variability of contamination levels
within the one size class sampled; however, it does not provide information on
size-specific contamination levels.
Option 3 (replicate analyses of three size classes) provides both additional
information on size-specific contamination levels and additional statistical power
to estimate the variability of the contaminant concentrations in each of three size
classes of the target species. If resources are limited, the State could analyze
the replicate samples for the largest size class first; if the SVs are exceeded,
analysis of the smaller size class composite samples could then be conducted.
Note: The correlation between increasing size (age) and contaminant tissue
concentration observed for some freshwater finfish species (Voiland et al., 1991)
may be much less evident in estuarine/marine finfish species (G. Pollock,
California Environmental Protection Agency, personal communication, 1993).
The movement of estuarine and marine species from one niche to another as
they mature may change their exposure at a contaminated site. Thus, size-
based sampling in estuarine/marine systems should be conducted only when it
is likely to serve a potential risk management outcome.
6.1.1.3 Target Analyte Selection-
All recommended target analytes listed in Table 4-1 should be included in
screening studies unless reliable historic tissue, sediment, or pollutant source
data indicate that an analyte is not present at a level of concern for human
health. Additional regional or site-specific target analytes should be included in
screening studies when there is indication or concern that such contaminants are
a potential health risk to local fish or shellfish consumers. Historic data on water,
sediment, and tissue contamination and priority pollutant scans from known point
source discharges or nonpoint source monitoring should be reviewed to
determine whether analysis of additional analytes is warranted.
6.1.1.4 Target Analyte Screening Values—
To enhance national consistency in screening study data, States should use the
target analyte screening values listed in Table 5-3 to evaluate tissue contaminant
data. Specific methods used to calculate SVs for noncarcinogenic and
carcinogenic target analytes, including examples of SVs calculated for selected
subpopulations, are given in Sections 5.1 and 5.2. If target analytes in addition
__
-------
6. FIELD PROCEDURES
to those recommended in Table 5-3 are included in a screening study, these
calculation procedures should be used to estimate SVs based on typical
exposure assumptions for the general population for the additional compounds.
Note: If the State chooses to use a different risk level or consumption rate to
address site-specific considerations, the corresponding SVs should be calculated
prior to initiation of chemical analyses to ensure that the detection limits of the
analytical procedures are sufficiently low to allow reliable quantitation at or below
the chosen SV.
6.1.1.5 Sampling Times—
If program resources are sufficient, biennial screening of waterbodies is
recommended where commercial, recreational, or subsistence harvesting is
commonly practiced (as identified by the State). Data from these screenings can
then be used in the biennial State 305(b) reports to document the extent of
support of Clean Water Act goals. If biennial screening is not possible, then
waterbodies should be screened at least once every 5 years.
Selection of the most appropriate sampling period is very important, particularly
when screening studies may be conducted only once every 2 to 5 years. Note:
For screening studies, sampling should be conducted during the period when the
target species is most frequently harvested (U.S. EPA, 1989d; Versar, 1982).
In fresh waters, as a general rule, the most desirable sampling period is from
late summer to early fall (i.e., August to October) (Phillips, 1980; Versar, 1982).
The lipid content of many species (which represents an important reservoir for
organic pollutants) is generally highest at this time. Also, water levels are
typically lower during this time, thus simplifying collection procedures. This late
summer to early fall sampling period should not be used, however, if (1) it does
not coincide with the legal harvest season of the target species or (2) the target
species spawns during this period. Note: If the target species can be legally
harvested during its spawning period, however, then sampling to determine
contaminant concentrations should be conducted during this time.
A third exception to the late summer to early fall sampling recommendation
concerns monitoring for the organophosphate pesticides. Sampling for these
compounds should be conducted during late spring or early summer within 1 to
2 months following pesticide application because these compounds are
degraded and metabolized relatively rapidly compared to organochlorine
pesticides.
In estuarine and coastal waters, the most appropriate sampling time is during the
period when most fish are caught and consumed (usually summer for
recreational and subsistence fishermen). For estuarine/marine shellfish (bivalve
molluscs and crustaceans), two situations may exist. The legal harvesting
season may be strictly controlled for fisheries resource management purposes
or harvesting may be open year round. In the first situation, shellfish
contaminant monitoring should be conducted during the legal harvest period. In
__
-------
6. FIELD PROCEDURES
the second situation, monitoring should be conducted to correspond to the period
when the majority of harvesting is conducted during the legal season. State staff
may have to consider different sampling times for target shellfish species if
differences in the commercial and recreational harvesting period exist.
Ideally, the sampling period selected should avoid the spawning period of the
target species, including the period 1 month before and 1 month after spawning,
because many aquatic species are subject to stress during spawning. Tissue
samples collected during this period may not always be representative of the
normal population. For example, feeding habits, body fat (lipid) content, and
respiration rates may change during spawning and may influence pollutant
uptake and clearance. Collecting may also adversely affect some species, such
as trout or bass, by damaging the spawning grounds. Most fishing regulations
protect spawning periods to enhance propagation of important fishery species.
Species-specific information on spawning periods and other life history factors
is available in numerous sources (e.g., Carlander, 1969; Emmett et al., 1991;
Pflieger, 1975; Phillips, 1980). In addition, digitized life history information is
available in many States through the Multistate Fish and Wildlife Information
System (1990).
Exceptions to the recommended sampling periods for freshwater and
estuarine/marine habitats will be determined by important climatic, regional, or
site-specific factors that favor alternative sampling periods. For many States,
budgetary constraints may require that most sampling be conducted during June,
July, and August when temporary help or student interns are available for hire.
The actual sampling period and the rationale for its selection should be
documented fully and the final data report should include an assessment of
sampling period effects on the results.
6.1.1.6 Sample Type—
Composite samples of fish fillets or of the edible portions of shellfish are
recommended for analysis of target analytes in screening studies (U.S. EPA,
1987b; 1989d). For health risk assessments, a composite sample should consist
of that portion of the individual organism that is commonly consumed by the
population at risk. Skin-on fillets (with the belly flap included) are recommended
for most scaled finfish (see Sections 7.2.2.6 and 7.2.2.7). Other sample types
(e.g., skinless fillets) may be more appropriate for some target species (e.g.,
catfish and other scaleless finfish species). For shellfish, the tissue considered
to be edible will vary by target species (see Section 7.2.3.4) based on local food
preferences. A precise description of the sample type (including the number and
size of the individuals in the composite) should be documented in the program
records for each target species.
Note: Composite samples are homogeneous mixtures of samples from two or
more individual organisms of the same species collected at a particular site and
analyzed as a single sample. Because the costs of performing individual
chemical analyses are usually higher than the costs of sample collection and
_
-------
6. FIELD PROCEDURES
preparation, composite samples are most cost-effective for estimating average
tissue concentrations of target analytes in target species populations. Besides
being cost-effective, composite samples also ensure adequate sample mass to
allow analyses for all recommended target analytes. A disadvantage of using
composite samples, however, is that extreme contaminant concentration values
for individual organisms are lost.
In screening studies, EPA recommends that States analyze one composite
sample for each of two target species at each screening site. Organisms used
in a composite sample
Must all be of the same species
Should satisfy any legal requirements of harvestable size or weight, or at
least be of consumable size if no legal harvest requirements are in effect
Should be of similar size so that the smallest individual in a composite is no
less than 75 percent of the total length (size) of the largest individual
• Should be collected at the same time (i.e., collected as close to the same
time as possible but no more than 1 week apart) [Note: This assumes that
a sampling crew was unable to collect all fish needed to prepare the
composite sample on the same day. If organisms used in the same
composite are collected on different days (no more than 1 week apart), they
should be processed within 24 hours as described in Section 7.2 except that
individual fish may have to be filleted and frozen until all the fish to be
included in the composite are delivered to the laboratory. At that time, the
composite homogenate sample may be prepared.]
Should be collected in sufficient numbers to provide a 200-g composite
homogenate sample of edible tissue for analysis of recommended target
analytes.
Individual organisms used in composite samples must be of the same species
because of the significant species-specific bioaccumulation potential. Accurate
taxonomic identification is essential in preventing the mixing of closely related
species with the target species. Note: Under no circumstance should
individuals from different species be used in a composite sample (U.S. EPA,
1989d, 1990d).
For cost-effectiveness, EPA recommends that States collect only one size class
for each target species and focus on the larger individuals commonly harvested
by the local population. Ideally, the individuals within each target species
composite should be of similar size within a target size range. For persistent
chlorinated organic compounds (e.g., DDT, PCBs, and toxaphene) and organic
mercury compounds, the larger (older) individuals within a population are
generally the most contaminated (Phillips, 1980; Voiland et al., 1991). As noted
earlier, this correlation between increasing size and increasing contaminant
6-10
-------
6. FIELD PROCEDURES
concentration is most striking in freshwater finfish species but is less evident in
estuarine and marine species. Size is used as a surrogate for age, which
provides some estimate of the total time the individual organism has been at risk
of exposure. Therefore, the primary target size range ideally should include the
larger individuals harvested at each sampling site. In this way, the States will
maximize their chances of detecting high levels of contamination in the single
composite sample collected for each target species. If this ideal condition
cannot be met, the field sampling team should retain individuals of similar length
that fall within a secondary target size range.
Individual organisms used in composite samples should be of similar size
(WDNR, 1988). Note: Ideally, for fish or shellfish, the total length (or size) of
the smallest individual in any composite sample should be no less than 75
percent of the total length (or size) of the largest individual in the composite
sample (U.S. EPA, 1990d). For example, if the largest fish is 200 mm, then the
smallest individual included in the composite sample should be at least 150 mm.
In the California Mussel Watch Program, a predetermined size range (55 to 65
mm) for the target bivalves (Mytilus californianus and M. edulis) is used as a
sample selection criterion at all sampling sites to reduce size-related variability
(Phillips, 1988). Similarly, the Texas Water Commission (1990) specifies the
target size range for each of the recommended target fish species collected in
the State's fish contaminant monitoring program.
Individual organisms used in a composite sample ideally should be collected at
the same time so that temporal changes in contaminant concentrations
associated with the reproduction cycle of the target species are minimized.
Each composite sample should contain 200 g of tissue so that sufficient material
will be available for the analysis of recommended target analytes. A larger
composite sample mass may be required when the number of target analytes is
increased to address regional or site-specific concerns. However, the tissue
mass may be reduced in the Tier 2 intensive studies (Phase I and II) when a
limited number of specific analytes of concern have been identified (see Section
7.2.2.9). Given the variability in size among target species, only approximate
ranges can be suggested for the number of individual organisms to collect to
achieve adequate mass in screening studies (U.S. EPA, 1989d; Versar, 1982).
For fish, 3 to 10 individuals should be collected for a composite sample for each
target species; for shellfish, 3 to 50 individuals should be collected for a
composite sample. In some cases, however, more than 50 small shellfish (e.g.,
mussels, shrimp, crayfish) may be needed to obtain the recommended 200-g
sample mass. Note: The same number of individuals should be used in each
composite sample for a given target species at each sampling site.
As alluded to above, a serious limitation of using composite samples is that
information on extreme levels of contamination in individual organisms is lost.
Therefore, EPA recommends that the residual individual homogenates be saved
to allow for analyses of individual specimens if resources permit (Versar, 1982).
Analysis of individual homogenates allows States to estimate the underlying
__
-------
6. FIELD PROCEDURES
population variance which, as described in Section 6.1.2.6, facilitates sample
size determination for the intensive studies. Furthermore, individual
homogenates may also be used to provide materials for split and spike samples
for routine QC procedures either for composites or individual organisms (see
Section 8.3).
Recommended sample preparation procedures are discussed in Section 7.2.
6.1.1.7 Replicate Samples—
The collection of sufficient numbers of individual organisms from a target species
at a site to allow for the independent preparation of more than one composite
sample (i.e., sample replicates) is strongly encouraged but is optional in
screening studies. If resources and storage are available, single replicate (i.e.,
duplicate) composite samples should be collected at a minimum of 10 percent
of the screening sites (U.S. EPA, 1990d). The collection and storage of replicate
samples, even if not analyzed at the time due to inadequate resources, allow for
followup QC checks. These sites should be identified during the planning phase
and sample replication specifications noted on the sample request form. If
replicate field samples are to be collected, States should follow the guidance
provided in Section 6.1.2.7. Note: Additional replicates must be collected at
each site for each target species if statistical comparisons with the target analyte
SVs are required in the State monitoring programs. The statistical advantages
of replicate sampling are discussed in detail in Section 6.1.2.7.
6.1.2 Intensive Studies (Tier 2)
The primary aim of intensive studies is to characterize the magnitude and
geographic extent of contamination in harvestable fish and shellfish species at
those screening sites where concentrations of target analytes in tissues were
found to be above selected SVs. Intensive studies should be designed to verify
results of the screening study, to identify specific fish and shellfish species and
size classes for which advisories should be issued, and to determine the
geographic extent of the fish contamination. In addition, intensive studies should
be designed to provide data for States to tailor their advisories based on the
consumption habits or sensitivities of specific local human subpopulations.
State staff should plan the specific aspects of field collection activities for each
intensive study site after a thorough review of the aims of intensive studies
(Section 2.2) and the fish contaminant data obtained in the screening study. All
the factors that influence sample collection activities should be considered and
specific aspects of each should be documented clearly by the program manager
on the sample request form for each site.
6.1.2.1 Site Selection-
Intensive studies should be conducted at all screening sites where the selected
SV for one or more target analytes was exceeded. The field collection staff
_
-------
6. FIELD PROCEDURES
should review a 7.5-minute (1:24,000 scale) USGS hydrologic map of the study
site and all relevant water, sediment, and tissue contaminant data. The site
selection factors evaluated in the screening study (Section 6.1.1.1) must be
reevaluated before initiating intensive study sampling.
States should conduct Tier 2 intensive studies in two phases if program
resources allow. Phase I Intensive studies should be more extensive
investigations of the magnitude of tissue contamination at suspect screening
sites. Phase II Intensive studies should define the geographic extent of the
contamination around these suspect screening sites in a variety of size (age)
classes for each target species. The field collection staff must evaluate the
accessibility of these additional sites and develop a sampling strategy that is
scientifically sound and practicable.
Selection of Phase II sites may be quite straightforward where the source of
pollutant introduction is highly localized or if site-specific hydrologic features
create a significant pollutant sink where contaminated sediments accumulate and
the bioaccumulation potential might be enhanced (U.S. EPA, 1986f). For
example, upstream and downstream water quality and sediment monitoring to
bracket point source discharges, outfalls, and regulated disposal sites showing
contaminants from surface runoff or leachate can often be used to characterize
the geographic extent of the contaminated area. Within coves or small
embayments where streams enter large lakes or estuaries, the geographic extent
of contamination may also be characterized via multilocational sampling to
bracket the areas of concern. Such sampling designs are clearly most effective
where the target species are sedentary or of limited mobility (Gilbert, 1987). In
addition, the existence of barriers to migration, such as dams, should be taken
into consideration.
6.1.2.2 Target Species and Size Class Selection—
Whenever possible, the target species found in the screening study to have
elevated tissue concentrations of one or more of the target analytes should be
resampled in the intensive study. Recommended target species for freshwater
sites are listed in Tables 3-1 and 3-2; target species for estuarine/marine waters
are listed in Tables 3-7 through 3-9 for Atlantic Coast estuaries, in Table 3-10
for Gulf Coast estuaries, and in Tables 3-11 through 3-13 for Pacific Coast
estuaries. If the target species used in the screening study are not collected in
sufficient numbers, alternative target species should be selected using criteria
provided in Section 3.2. The alternative target species should be specified on
the sample request form.
For Phase I intensive studies, States should collect replicate composite samples
of one size class for each target species and focus sampling on larger
individuals commonly harvested by the local population (as appropriate). If
contamination of this target size class is high, Phase II studies should include
collection of replicate composite samples of three size classes within each target
species.
6-13
-------
6. FIELD PROCEDURES
EPA recognizes that resource limitations may influence the sampling strategy
selected by a State. If monitoring resources are limited for intensive studies,
States may determine that it is more resource-efficient to collect replicate
composite samples of three size classes (as required for Phase II studies) during
Phase I sampling rather than revisit the site at a later time to conduct Phase II
intensive studies. In this way, the State may save resources by reducing field
sampling costs associated with Phase II intensive studies.
By sampling three size (age) classes, States collect data on the target species
that may provide them with additional risk management options. If contaminant
concentrations are positively correlated with fish and shellfish size, frequent
consumption of smaller (less contaminated) individuals may be acceptable even
though consumption of larger individuals may be restricted by a consumption
advisory. In this way, States can tailor an advisory to protect human health and
still allow restricted use of the fishery resource. Many Great Lakes States have
used size (age) class data to allow smaller individuals within a given target
species to remain fishable while larger individuals are placed under an advisory.
6.1.2.3 Target Analyte Selection—
Phase I intensive studies should include only those target analytes found in the
screening study to be present in fish and shellfish tissue at concentrations
exceeding selected SVs (Section 5.2). Phase II studies should include only
those target analytes found in Phase I intensive studies to be present at
concentrations exceeding SVs. In most cases, the number of target analytes
evaluated in Phase I and II intensive studies will be significantly smaller than the
number evaluated in screening studies.
6.1.2.4 Target Analyte Screening Values-
Target analyte SVs used in screening studies should also be used in Phase I
and II intensive studies. Specific methods used to calculate SVs for
noncarcinogenic and carcinogenic target analytes, including examples of SVs
calculated for various exposure scenarios, are given in Section 5.1.
6.1.2.5 Sampling Times—
To the extent that program resources allow, sampling in intensive studies should
be conducted during the same period or periods during which screening studies
were conducted (i.e., when the target species are most frequently harvested for
consumption) and should be conducted preferably within 1 year of the screening
studies. In some cases, it may be best to combine Phase I and Phase II
sampling to decrease both the time required to obtain adequate data for
issuance of specific advice relative to species, size classes, and geographic
extent and/or the monitoring costs entailed in revisiting the site (see Section
6.1.2.2).
6-14
-------
6. FIELD PROCEDURES
States should follow the general guidance provided in Section 6.1.1.5 for
recommended sampling times. The actual sampling period and rationale for its
selection should be documented fully for Phase I and II studies.
6.1.2.6 Sample Type—
Composite samples of fish fillets or the edible portions of shellfish are
recommended for analysis of target analytes in intensive studies. The general
guidance in Section 6.1.1.6 should be followed to prepare composite samples
for each target species. In addition, separate composite samples may be
prepared for selected size (age) classes within each target species, particularly
in Phase II studies after tissue contamination has been verified in Phase I
studies. Because the number of replicate composite samples and the number
of fish and shellfish per composite required to test whether the site-specific mean
contaminant concentration exceeds an SV are intimately related, both will be
discussed in the next section.
Note: The same number of individual organisms should be used to prepare all
replicate composite samples for a given target species at a given site. If this
number is outside the recommended range, documentation should be provided.
Recommended sample preparation procedures are discussed in Section 7.2.
6.1.2.7 Replicate Samples—
In intensive studies (Phases I and II), EPA recommends that States analyze
replicate composite samples of each target species at each sampling site.
Replicate composite samples should be as similar to each other as possible. In
addition to being members of the same species, individuals within each
composite should be of similar length (size) (see Section 6.1.1.6). The relative
difference between the average length (size) of individuals within any composite
sample from a given site and the average of the average lengths (sizes) of
individuals in all composite samples from that site should not exceed 10 percent
(U.S. EPA, 1990d). In order to determine this, States should first calculate the
average length of the target species fish constituting each composite replicate
sample from a site. Then, States should take the average of these averages for
the site. In the following example, the average of the average lengths of
individuals (±10 percent) in five replicate composite samples is calculated to be
310 (±31) mm.
6-15
-------
6. FIELD PROCEDURES
Replicate
1
2
3
4
5
Average of the
Average Length of Individual
Fish In Composite Sample (mm)
300
320
330
280
320
average length (±10%) = 310 (±31) mm.
Therefore, the acceptable range for the average length of individual composite
samples is 279 to 341 mm, and the average length of individual fish in each of
the five replicate composites shown above falls within the acceptable average
size range.
All replicate composite samples for a given sampling site should be collected
within no more than 1 week of each other so that temporal changes in target
analyte concentrations associated with the reproductive cycle of the target
species are minimized.
The remainder of this section provides general guidelines for estimating the
number of replicate composite samples per site (n) and the number of individuals
per composite (m) required to test the null hypothesis that the mean target
analyte concentration of replicate composite samples at a site is equal to the SV
versus the alternative hypothesis that the mean target analyte concentration is
greater than the SV. These guidelines are applicable to any target species and
any target analyte.
Note: It is not possible to recommend a single set of sample size requirements
(e.g., number of replicate composite samples per site and the number of
individuals per composite sample) for all fish and shellfish contaminant
monitoring studies. Rather, EPA presents'a more general approach to sample
size determination that is both scientifically defensible and cost-effective. At
each site, States must determine the appropriate number of replicate composite
samples and of individuals per composite sample based on
• Site-specific estimations of the population variance of the target analyte
concentration
• Fisheries management considerations
• Statistical power consideration.
If the population variance of the target analyte concentrations at a site is small,
fewer replicate composite samples and/or fewer individuals per composite
sample may be required to test the null hypothesis of interest with the desired
6-16
-------
6. FIELD PROCEDURES
statistical power. In this case, using sample sizes that are larger than required
to achieve the desired statistical power would not be cost-effective.
Alternatively, suppose EPA recommended sample sizes based on an analyte
concentration with a population variance that is smaller than that of the target
analyte. In this case, the EPA-recommended sample size requirements may be
inadequate to test the null hypothesis of interest at the statistical power level
selected by the State. Therefore, EPA recommends an approach that provides
the flexibility to sample less in those waters where the target analyte
concentrations are less variable, thereby reserving sampling resources for those
site-specific situations where the population variance of the target analyte tissue
concentration is greater.
The EPA recommends the following statistical model, which assumes that z( is
the contaminant concentration of the ith replicate composite sample at the site
of interest where i=1,2,3,...,n and, furthermore, that each replicate composite
sample is comprised of m individual fish fillets of equal mass. Let z be the mean
target analyte concentration of observed replicate composite samples at a site.
Ignoring measurement error, the variance of z is
Var(z) = ^/(nm) (6-1)
where
o2 = Population variance
n = Number of replicate composite samples
m = Number of individual samples in each composite sample.
To test the null hypothesis that the mean target analyte concentration across the
n replicate composite samples is equal to the SV versus the alternative
hypothesis that the mean target analyte concentration is greater than the SV, the
estimate of the Var(z), s2, is
s2 = [Z(zi-z)2]/[n(n-1)] (6-2)
where the summation occurs over the n composite samples. Under the null
hypothesis, the following statistic
(z - SV) / s (6-3)
has a Student-t distribution with (n - 1) degrees of freedom (Cochran, 1977;
Kish, 1965). The degrees of freedom are one less than the number of
composite samples.
An optimal sampling design would specify the minimum number of replicate
composite samples (n) and of individuals per composite (m) required to detect
a minimum difference between the SV and the mean target analyte concentration
6-17
-------
6. FIELD PROCEDURES
of replicate composite samples at a site. Design characteristics necessary to
estimate the optimal sampling design include
Minimum detectable difference between the site-specific mean target analyte
concentration and the SV
• Power of the hypothesis test (i.e., the probability of detecting a true
difference when one exists)
Level of significance (i.e., the probability of rejecting the null hypothesis of
no difference between the site-specific mean target analyte concentration
and the SV when a difference does exist)
Population variance, o2 (i.e., the variance in target analyte concentrations
among individuals from the same species, which the statistician often must
estimate from prior information)
Cost components (including fixed costs and variable sample collection,
preparation, and analysis costs).
In the absence of such design specifications, guidance for selecting the number
of replicate composite samples at each site and the number of fish per
composite sample is provided. This guidance is based on an investigation of the
precision of the estimate of o2/nm and of statistical power.
Note: Under optimal field and laboratory conditions, at least two replicate
composite samples are required at each site for variance estimation. To
minimize the risk of a destroyed or contaminated composite sample precluding
the site-specific statistical analysis, a minimum of three replicate composite
samples should be collected at each site if possible. Because three replicate
composite samples provide only two degrees of freedom for hypothesis testing,
additional replicate composite samples are recommended.
The stability of the estimated standard error of z must also be considered
because this estimated standard error is the denominator of the statistic for
testing the null hypothesis of interest. A measure of the stability of an estimate
is its statistical precision. The assumption is made that the Zj's come from a
normal distribution, and then the standard error of a2/nm is defined as a product
of o2 and a function of n (the number of replicate composite samples) and m
(the number of fish per composite). A fortunate aspect of composite sampling
is that the composite target analyte concentrations tend to be normally
distributed via the Central Limit Theorem. This formulation is used to determine
which combinations of n and m are associated with a more precise estimate of
a2/nm.
6-18
-------
6. FIELD PROCEDURES
Modifying Cochran (1963) to reflect the normality assumption and the sampling
design of n replicate composite samples and m fish per composite sample, the
function of n and m of interest is shown in square brackets:
se
.21
nm
1/2
(6-4)
Table 6-1 provides values of this function for various combinations of m and n.
The data presented in Table 6-1 suggest that, as either n or m increases, the
standard error of a*/nm decreases. The advantage of increasing the number of
replicate composite samples can be described in terms of this standard error.
For example, the standard error of o^/nm from a sample design of five replicate
composite samples and six fish per composite (0.024) will be more than 50
percent smaller than that from a sample design of three replicate composite
samples and six fish per composite (0.056). In general, holding the number of
fish per composite fixed, the standard error of o2/nm estimated from five
replicate samples will be about 50 percent smaller than that estimated from three
replicate samples.
The data in Table 6-1 also suggest that greater precision in the estimated
standard error of z is gained by increasing the number of replicate samples (n)
than by increasing the number of fish per composite (m). If the total number of
individual fish caught at a site, for example, is fixed at 50 fish, then, with a
design of 10 replicate samples of 5 fish each, the value of the function of n and
m in Table 6-1 is 0.009; with 5 replicate samples of 10 fish each, the value is
Table 6-1. Values of
n2m
(n-1)J
for Various Combinations of n and m
No. of
replicate
composite
samples (n)
3
4
5
6
7
10
15
Number of fish per composite sample
3
0.111
0.068
0.047
0.035
0.027
0.016
0.008
4
0.083
0.051
0.035
0.026
0.021
0.012
0.006
5
0.067
0.041
0.028
0.021
0.016
0.009
0.005
6
0.056
0.034
0.024
0.018
0.014
0.008
0.004
7
0.048
0.029
0.020
0.015
0.012
0.007
0.004
8
0.042
0.026
0.018
0.013
0.010
0.006
0.003
9
0.037
0.023
0.016
0.012
0.009
0.005
0.003
(m)
10
0.033
0.020
0.014
0.011
0.008
0.005
0.003
12
0.028
0.017
0.012
0.009
0.007
0.004
0.002
15
0.022
0.014
0.009
0.007
0.005
0.003
0.002
6-19
-------
6. FIELD PROCEDURES
0.014. Thus, there is greater precision in the estimated standard error of z
associated with the first design as compared with the second design.
Two assumptions are made to examine the statistical power of the test of the
null hypothesis of interest. First, it is assumed that the true mean of the site-
specific composite target analyte concentrations (n) is either 10 percent or 50
percent higher than the screening value. Second, it is presumed that a factor
similar to a coefficient of variation, the ratio of the estimated population standard
deviation to the screening value (i.e., o/SV), is 50 to 100 percent. Four
scenarios result from joint consideration of these two assumptions. The power
of the test of the null hypothesis that the mean composite target analyte
concentration at a site is equal to the SV versus the alternative hypothesis that
the mean target analyte concentration is greater than the SV is estimated under
each set of assumptions. Estimates of the statistical power for two of the four
scenarios are shown in Table 6-2.
Power estimates for the two scenarios where the true mean of the site-specific
composite target analyte concentration was assumed to be only 10 percent
higher than the screening value are not presented. The power to detect this
small difference was very poor: for 125 of the resulting 140 combinations of n
and m, the power was less than 50 percent.
Several observations can be made concerning the data in Table 6-2. Note: The
statistical power increases as either n (number of replicate composite samples)
or m (number of fish per composite) increases. However, greater power is
achieved by increasing the number of replicate composite samples as opposed
to increasing the number of fish per composite. Furthermore, if the number of
replicate composite samples per site and the number of fish per composite are
held constant, then, as the ratio of the estimated population variance to the SV
increases (i.e., o/SV), the statistical power decreases.
States may use these tables as a starting point for setting the number of
replicate composite samples per site and the number of fish per composite in
their fish and shellfish contaminant monitoring studies. The assumption
regarding the ratio of the estimated population variance to the SV presented in
Section A of Table 6-2 is probably unrealistic. Data in Section B, which reflect
more realistic assumptions concerning the estimated population variance, show
that States will be able to detect only large differences between the site-specific
mean target analyte concentrations and the SV. Specifically, using five replicate
composite samples and six to seven fish per composite sample, the power to
detect a 50 percent increase over the SV is between 70 and 80 percent.
However, when the number of fish per composite increases to 8 to 10, the power
increases by about 10 percentage points.
One final note on determining the number of replicate composite samples per
site and the number of fish per composite should be emphasized. According to
Section 6.1.2.3, Phase I intensive studies will focus on those target analytes that
exceeded the selected SV used in the screening study. Thus, multiple target
6-20
-------
6. FIELD PROCEDURES
Table 6-2. Estimates of Statistical Power of
Hypothesis of Interest Under Specified Assumptions
No. of
replicate
composite
samples
(n)
• Number of fish per composite (m)
3 4 5 6 7 8 9 10 12 15
A. Ratio of o/SV = 0.5 and n = 1.5 x SV:
3
4
5
6
7
10
15
6
8
9
9
9
9
9
6
9
9
9
9
9
9
7
9
9
9
9
9
9
8
9
9
9
9
9
9
9
9
9
9
9
9
9
9
9
9
9
9
9
9
9
9
9
9
9
9
9
9
9
9
9
9
9
9
9
9
9
9
9
9
9
9
9
9
9
9
9
9
B. Ratio of o/SV = 1.0 and n = 1.5 x SV:
3
4
5
6
7
10
15
-
-
-
5
6
8
9
-
-
5
6
7
8
9
-
-
6
7
8
9
9
-
5
7
8
8
9
9
-
6
8
8
9
9
9
-
6
8
8
9
9
9
-
7
8
9
9
9
9
-
7
8
9
9
9
9
5
8
9
9
9
9
9
6
8
9
9
9
9
9
-: Power less than 50 percent.
5: Power between 50 and 60 percent.
6: Power between 60 and 70 percent.
7: Power between 70 and 80 percent.
8: Power between 80 and 90 percent
9: Power above 90 percent.
analytes may be under investigation during Phase I intensive studies, and the
population variances of these analytes are likely to differ. Note: States should
use the target analyte that exhibits the largest population variance when
selecting the number of replicate composite samples per site and the number of
fish per composite. This conservative approach supports use of the data in
Section B of Table 6-2 where the ratio of o/SV is twice that of the data in Section
A. States may estimate population variances from historic fish contaminant data
or from composite data as described by EPA (1989d). This estimate of a2 can
be used to determine whether the sampling design (i.e., number of replicate
composite samples [n] and number of individuals per composite [m]) should be
modified to achieve a desired statistical power.
6-21
-------
6. FIELD PROCEDURES
After States have implemented their fish and shellfish contaminant monitoring
program, collected data on cost and variance components, and addressed other
design considerations, they may want to consider using an optimal composite
sampling protocol as described in Rohlf et al. (1991) for refining their sampling
design. An optimal sampling design is desirable because it detects a specified
minimum difference between the site-specific mean contaminant concentration
and the SV at minimum cost.
6.2 SAMPLE COLLECTION
Sample collection activities should be initiated in the field only after an approved
sampling plan has been developed. This section discusses recommended
sampling equipment and its use, considerations for ensuring preservation of
sample integrity, and field recordkeeping and chain-of-custody procedures
associated with sample processing, preservation, and shipping.
6.2.1 Sampling Equipment and Use
In response to the variations in environmental conditions and target species of
interest, fisheries biologists have had to devise sampling methods that are
intrinsically selective for certain species and sizes of fish and shellfish (Versar,
1982). Although this selectivity can be a hindrance in an investigation of
community structure, it is not a problem where tissue contaminant analysis is of
concern because tissue contaminant data can best be compared only if factors
such as differences in taxa and size are minimized.
Collection methods can be divided into two major categories, active and passive.
Each collection method has advantages and disadvantages. Various types of
sampling equipment, their use, and their advantages and disadvantages are
summarized in Table 6-3 for fish and in Table 6-4 for shellfish. Note: Either
active or passive collection methods may be used as long as the methods
selected result in collection of a representative fish sample of the type consumed
by local sport and subsistence fishermen.
A basic checklist of field sampling equipment and supplies is shown in Table 6-5.
Safety considerations associated with the use of a boat in sample collection
activities are summarized in Table 6-6.
6.2.1.1 Active Collection-
Active collection methods employ a wide variety of sampling devices. Devices
for fish sampling include electroshocking units, seines, trawls, and angling
equipment (hook and line). Devices for shellfish sampling include seines, trawls,
mechanical grabs (e.g., pole- or cable-operated grab buckets and tongs),
biological and hydraulic dredges, scoops and shovels, rakes, and dip nets.
Shellfish can also be collected manually by SCUBA divers. Although active
collection requires greater fishing effort, it is usually more efficient than passive
collection for covering a large number of sites and catching the relatively small
6^22
-------
6. FIELD PROCEDURES
number of individuals needed from each site for tissue analysis (Versar, 1982).
Active collection methods are particularly useful in shallow waters (e.g., streams,
lake shorelines, and shallow coastal areas of estuaries).
Active collection methods have distinct disadvantages for deep water sampling.
They require more field personnel and more expensive equipment than passive
collection methods. This disadvantage may be offset by coordinating sampling
efforts with commercial fishing efforts. Purchasing fish and shellfish from
commercial fishermen using active collection devices is acceptable; however,
field sampling staff should accompany the commercial fishermen during the
collection operation to ensure that samples are collected and handled properly
and to verify the sampling site location. The field sampling staff then remove the
target species directly from the sampling device and ensure that sample
collection, processing, and preservation are conducted as prescribed in sample
collection protocols, with minimal chance of contamination. This is an excellent
method of obtaining specimens of commercially important target species,
particularly from the Great Lakes and coastal estuarine areas (Versar, 1982).
The EPA advises against the use of chemical poisons as a collection technique
for fish and shellfish contaminant monitoring programs because these toxicants
may induce physiological changes that could alter contaminant concentrations
in the tissues (Versar, 1982).
More detailed descriptions of active sampling devices and their use are provided
in Battelle (1975); Bennett (1970); Gunderson and Ellis (1986); Hayes (1983);
Mearns and Allen (1978); Pitt, Wells, and McKone (1981); Puget Sound Estuary
Program (1990b); Versar (1982); and Weber (1973).
6.2.1.2 Passive Collection-
Passive collection methods employ a wide array of sampling devices for fish and
shellfish, including gill nets, fyke nets, trammel nets, hoop nets, pound nets, and
d-traps. Passive collection methods generally require less fishing effort than
active methods but are usually less desirable for shallow water sample collection
because of the ability of many species to evade these entanglement and
entrapment devices. These methods normally yield a much greater catch than
would be required for a contaminant monitoring program and are time consuming
to deploy. In deep water, however, passive collection methods are generally
more efficient than active methods. Crawford and Luoma (1993) caution that
passive collection devices (e.g., gill nets) should be checked frequently to ensure
that captured fish do not deteriorate prior to removal from the sampling device.
Versar (1982, 1984) and Hubert (1983) describe passive sampling devices and
their use in more detail.
Purchasing fish and shellfish from commercial fishermen using passive collection
methods is acceptable; however, field sampling staff should accompany the
fishermen during both the deployment and collection operations to ensure that
samples are collected and handled properly and to verify the sampling site
_
-------
Table 6-3. Summary of Fish Sampling Equipment
Device
Use
Advantages
Disadvantages
ACTIVE METHODS!
Electrofishing
Seines
Trawls
Angling
Purchasing specimens
from commercial
fishermen
Shallow rivers, lakes, and streams.
Shallow rivers, lakes, and streams.
Shoreline areas of estuaries.
Various sizes can be used from boats
in moderate to deep open bodies
of water (10 to >70 m depths).
Generally species selective involving
use of hook and line.
Only in areas where target
species are commercially harvested.
Most efficient nonselective method. Minimal
damage to fish. Adaptable to a number of
sampling conditions (e.g., boat, wading, shore-
lines). Particularly useful at sites where other
active methods cannot be used (e.g., around
snags and irregular bottom contours).
Relatively inexpensive and easily operated.
Mesh size selection available for target species.
Effective in deep waters not accessible by
other methods. Allows collection of a large
number of samples.
Most selective method. Does not require use
of large number of personnel or expensive
equipment
Most cost-effective and efficient means of
obtaining commercially valuable species
from harvested waters.
Nonselective — stuns or kills most fish. Cannot
be used in brackish, salt, or extremely soft
water. Requires extensive operator training.
DANGEROUS when not used properly.
Cannot be used in deep water or over substrates
with an irregular contour. Not completely efficient
as fish can evade the net during seining operation
Requires boat and trained operators.
Inefficient and not dependable.
Limited use; commercially harvested areas may
not include sampling sites chosen for fish
contaminant monitoring. The field collection staff
should accompany the commercial fishermen and
should remove the required samples from the
collection device. This will ensure the proper
handling of the specimens and accurate recording
of the collection time and sampling location.
PASSIVE METHODS I""".
Gill nets
Trammel nets
Lakes, rivers, and estuaries. Where
fish movement can be expected or
anticipated.
Lakes, rivers, and estuaries. Where
fish movement can be expected or
anticipated. Frequently used
where fish may be scared into the net
Effective for collecting pelagic fish species.
Relatively easy to operate. Requires
less fishing effort than active methods. Selec-
tivity can be controlled by varying mesh size.
Slightly more efficient than a straight gill net.
Not effective for bottom-dwelling fish or popula-
tions that do not exhibit movement patterns. Nets
prone to tangling or damage by large and sharp
spirted fish. Gill nets will kill captured specimens,
which, when left for extended periods, may
undergo physiological changes.
(Same as for gill nets.) Tangling problems may
be more severe. Method of scaring fish into net
requires more personnel or possibly boats in
deep water areas.
(continued)
o>
10
en
m
5
•o
31
O
O
m
o
3J
m
-------
TABLE 6-3. (continued)
Device
Use
Advantages
Disadvantages
PASSIVE METHODS 1
Hoop, Fyke and
Pound Nets
D-Traps
Shallow rivers, lakes, and estuaries
where currents are present or when
movements of fish are predictable.
Frequently used in commercial
operations.
Used for long-term capture of slow-
moving fish, particularly bottom
species. Can be used in all environ-
ments.
Unattended operation. Very efficient in regard
to long-term return and expended effort.
Particularly useful in areas where active
methods are impractical.
Easy to operate and set. Unattended operation.
Particularly useful for capturing bottom dwelling
organisms in deep waters or other types of
inaccessible areas. Relatively inexpensive—
often can be hand made.
Inefficient for short term. Difficult to set up and
maintain.
Efficiency is highly variable. Not effective for
pelagic fish or fish that are visually oriented.
Less efficient for all species when water is clear
rather than turbid. Not a good choice for a
primary sampling technique, but valuable as
backup for other methods.
Source: Versar, 1982.
O>
Tl
m
o
•o
30
o
O
m
o
ro
Ol
m
-------
Table 6-4. Summary of Shellfish Sampling Equipment
Device
ACTIVE METHODS |
Seines
Trawls
Mechanical grabs
Double-pole-
operated grab
buckets
Tongs or double-
handled grab
sampler
Line or Cable-Operated
Grab Buckets:
Ekman grab
Petersen grab
Ponar grab
Orange peel grab
Use
Shallow shoreline areas of
estuaries.
Various sizes can be used from boats
in moderate to deep open bodies
of water (10 to >70 m depths).
Used from boat or pier. Most useful
in shallow water areas less than
6 m deep including lakes, rivers,
and estuaries.
Most useful in shallow water, lakes,
rivers, and estuaries. Generally used
from a boat.
Used from boat or pier to sample soft
to semisoft substrates.
Deep lakes, rivers, and estuaries for
sampling most substrates.
Deep lakes, rivers, and estuaries for
sampling sand, silt or clay substrates.
Deep lakes, rivers, and estuaries for
sampling most substrates.
Advantages
Relatively inexpensive and easily operated.
Mesh size selection available for target crusta-
cean species (e.g., shrimp and crabs).
Effective in deeper waters not accessible by
other methods. Allows collection of a large
number of samples.
Very efficient means of sampling bivalves
(e.g., clams and oysters) that are located on
or buried in bottom sediments.
Very efficient means of sampling oysters, clams,
and scallops. Collection of surrounding or
overlying sediments is not required and the
jaws are generally open baskets. This reduces
the weight of the device and allows the washing
of collected specimens to remove sediments.
Can be used in water of varying depths in
lakes, rivers, and estuaries.
Large sample is obtained; grab can penetrate
most substrates.
Most universal grab sampler. Adequate on
most substrates. Large sample is obtained
intact.
Designed for sampling hard substrates.
Disadvantages
Cannot be used in deep water or over substrates
with an irregular contour. Not completely efficient
as crustaceans can evade the net during seining
operation.
Requires boat and trained operators.
At depths greater than 6 m, the pole-operated
devices become difficult to operate manually.
At depths greater than 6 m, the pole-operated
devices become difficult to operate manually.
Possible incomplete closure of jaws can result in
sample loss. Must be repeatedly retrieved and
deployed. Grab is small and is not particularly
effective in collecting large bivalves (clams and
oysters).
Grab is heavy, may require winch for deploy-
ment. Possible incomplete closure of jaws can
result in sample loss. Must be repeatedly retrieved
and deployed.
Possible incomplete closure of jaws can result in
sample loss. Must be repeatedly retrieved and
deployed.
Grab is heavy, may require winch for deployment.
Possible incomplete closure of jaws can result in
sample loss. Must be repeatedly retrieved and
deployed. Grab is small and not particularly
effective in collecting large bivalves (clams
and oysters).
en
m
5
•o
31
O
O
m
o
c
31
m
to
(continued)
-------
TABLE 6-4. (continued)
Device
Use
Advantages
Disadvantages
Biological or
hydraulic dredges
Dragged along the bottom of deep
waterbodies to collect large stationary
invertebrates.
Qualitative sampling of large area of bottom
substrate and benthic community. Length of
tows can be relatively short if high density
of shellfish exists in sampling area.
If the length of the tow is long, it is difficult to
pinpoint the exact location of the sample collec-
tion area. Because of the scouring operation of
the dredge, bivalve shells may be damaged. All
bivalve specimens should be inspected and
individuals with cracked or damaged shells
should be discarded.
Scoops, shovels Used in shallow waters accessible by
wading or SCUBA equipment for
collection of hard clams (Mercenaria
mercenaries or soft-shell clam (Mya
arenaria).
Does not require a boat; sampling can be
done from shore.
Care must be taken not to damage the shells of
bivalves while digging in substrate.
Scrapers
Used in shallow waters accessible by
wading or SCUBA equipment for
collection of oysters. (Crassostrea
virginica) or mussels (Mytilus sp).
Does not require a boat; sampling can be
done from shore.
Care must be taken not to damage shells of
bivalves while removing them from hard
substrate.
Rakes
Used in shallow waters accessible by
wading or can be used from a boat.
Does not require a boat; sampling can be done
close to shore. Can be used in soft sediments
to collect clams or scallops, and can also be
used to dislodge oysters or mussels that are
attached to submerged objects such as rocks
and pier pilings.
Care must be taken not to damage the shells of
the bivalves while raking or dislodging them from
the substrate.
Purchasing specimens Only in areas where target species
from commercial are commercially harvested.
fishermen
Most cost-effective and efficient means of
obtaining bivalves for pollutant analysis from
commercially harvested waters.
Limited use; commercially harvested areas may
not include sampling sites chosen for shellfish
contaminant monitoring. The field collection staff
should accompany the commercial fishermen and
should remove the required samples from the
collection device. This will ensure the proper
handling of the specimens and accurate recording
of the exact collection time and sampling location.
PASSIVE METHODS L
D-traps
Used for capture of slow-moving
crustaceans (crabs and lobsters)
that move about on or just above
the substrate.
Can be used in a variety of environments.
Particularly useful for capturing bottom
dwelling organisms in deep water or other
inaccessible areas. Relatively inexpensive,
can be hand made.
Catch efficiency is highly variable. Not a good
choice for a primary sampling technique, but
valuable as a backup for other methods.
Source: Versar. 1982.
O>
rb
o>
m
5
•o
a
o
o
m
o
c
3)
m
-------
6. FIELD PROCEDURES
Table 6-5. Checklist of Field Sampling Equipment and Supplies
for Fish and Shellfish Contaminant Monitoring Programs
Boat supplies
- Fuel supply (primary and auxiliary supply)
- Spare parts repair kit
- Life preservers
- First aid kit (including emergency phone numbers of local hospitals, family contacts for each
member of the sampling team)
- Spare oars
- Nautical charts of sampling site locations
Collection equipment (e.g., nets, traps, electroshocking device)
Recordkeeping/documentation supplies
- Field logbook
- Sample request forms
- Specimen identification labels
- Chain-of-Custody (COC) Forms and COC tags or labels
- Indelible pens
Sample processing equipment and supplies
- Holding trays
- Fish measuring board (metric units)
- Calipers (metric units)
- Shucking knife
- Balance to weigh representative specimens for estimating tissue weight (metric units)
- Aluminum foil (extra heavy duty)
- Freezer tape
- String
- Several sizes of plastic bags for holding individual or composite samples
- Resealable watertight plastic bags for storage of Field Records, COC Forms, and Sample
Request Forms
Sample preservation and shipping supplies
- Ice (wet ice, blue ice packets, or dry ice)
- Ice chests
- Filament-reinforced tape to seal ice chests for transport to the central processing laboratory
6-28
-------
6. FIELD PROCEDURES
Table 6-6. Safety Considerations for Field Sampling Using a Boat
Field collection personnel should not be assigned to duty alone in boats.
Life preservers should be worn at all times by field collection personnel near the water or
on board boats.
If electrofishing is the sampling method used, there must be two shutoff switches-one at
the generator and a second on the bow of the boat.
All deep water sampling should be performed with the aid of an experienced, licensed
boat captain.
All sampling during nondaylight hours, during severe weather conditions, or during
periods of high water should be avoided or minimized to ensure the safety of field
collection personnel.
All field collection personnel should be trained in CPR, water safety, boating safety, and
first aid procedures for proper response in the event of an accident. Personnel should
have local emergency numbers readily available for each sampling trip and know the
location of the hospitals or other medical facilities nearest each sampling site.
location. The field sampling staff can then ensure that sample collection,
processing, and preservation are conducted as prescribed in sample collection
protocols, with minimal chance of contamination.
6.2.2 Preservation of Sample Integrity
The primary QA consideration in sample collection, processing, preservation, and
shipping procedures is the preservation of sample integrity to ensure the
accuracy of target analyte analyses. Sample integrity is preserved by prevention
of loss of contaminants already present in the tissues and prevention of
extraneous tissue contamination (Smith, 1985).
Loss of contaminants already present in fish or shellfish tissues can be
prevented in the field by ensuring that the skin on fish specimens has not been
lacerated by the sampling gear or that the carapace of crustaceans or shells of
bivalves have not been cracked during sample collection resulting in loss of
tissues and/or fluids that may contain contaminants. Once the samples have
reached the laboratory, further care must be taken during thawing (if specimens
are frozen) to ensure that all liquids from the thawed specimens are retained with
the tissue sample as appropriate (see Section 7.2.2).
Sources of extraneous tissue contamination include contamination from sampling
gear, grease from ship winches or cables, spilled engine fuel (gasoline or diesel),
_
-------
6. FIELD PROCEDURES
engine exhaust, dust, ice chests, and ice used for cooling. All potential sources
of contamination in the field should be identified and appropriate steps taken to
minimize or eliminate them. For example, during sampling, the boat should be
positioned so that engine exhausts do not fall on the deck. Ice chests should be
scrubbed clean with detergent and rinsed with distilled water after each use to
prevent contamination. To avoid contamination from melting ice, samples should
be placed in waterproof plastic bags (Stober, 1991). Sampling equipment that
has been obviously contaminated by oils, grease, diesel fuel, or gasoline should
not be used. All utensils or equipment that will be used directly in handling fish
or shellfish (e.g., fish measuring board or calipers) should be cleaned in the
laboratory prior to each sampling trip, rinsed in acetone and pesticide-grade
hexane, and stored in aluminum foil until use (Versar, 1982). Between sampling
sites, the field collection team should clean each measurement device by rinsing
it with ambient water and rewrapping it in aluminum foil to prevent contamination.
Sources of contamination in the laboratory should be minimized by resecting
(i.e., surgically removing) tissues and preparing composite homogenate samples
for analysis in a controlled environment. All resection and sample preparation
should be conducted in a processing laboratory under cleanroom conditions to
reduce contamination of specimens (Schmitt and Finger, 1987; Stober, 1991).
Procedures for laboratory processing and resection are described in Section 7.2.
Procedures for assessing sources of sample contamination through the analyses
of field and processing blanks are described in Section 8.3.3.6.
6.2.3 Field Recordkeeplng
Thorough documentation of all field sample collection and processing activities
is necessary for proper interpretation of field survey results. For fish and
shellfish contaminant studies, it is advisable to use preprinted waterproof data
forms, indelible ink, and writing implements that can function when wet (Puget
Sound Estuary Program, 1990b). When multicopy forms are required, no-
carbon-required (NCR) paper is recommended because it allows information to
be forwarded on the desired schedule and retained for the project file at the
same time.
Four separate preprinted sample tracking forms should be used for each
sampling site to document field activities from the time the sample is collected
through processing and preservation until the sample is delivered to the
processing laboratory. These are
Field record form
Sample identification label
Chain-of-custody (COC) label or tag
COC form.
6-30
-------
6. FIELD PROCEDURES
6.2.3.1 Field Record Form—
The following information should be included on the field record for each
sampling site in both Tier 1 screening (Figures 6-2 and 6-3) and Tier 2 intensive
studies as appropriate (Figures 6-4 and 6-5):
Project number
Sampling date and time (specify convention used, e.g., day/month/year and
24-h clock)
Sampling site location (including site name and number, county/parish,
latitude/longitude, waterbody name/segment number, waterbody type, and
site description)
Sampling depth
Collection method
Collectors' names and signatures
Agency (including telephone number and address)
Species collected (including species scientific name, composite sample
number, individual specimen number, number of individuals per composite
sample, number of replicate samples, total length/size [mm], sex [male,
female, indeterminate])
Note: States should specify a unique numbering system to track samples for
their own fish and shellfish contaminant monitoring programs.
Percent difference in size between the smallest and largest specimens to be
composited (smallest individual length [or size] divided by the largest
individual length [or size] x 100; should be >75 percent) and mean
composite length or size (mm)
Notes (including visible morphological abnormalities, e.g., fin erosion, skin
ulcers, cataracts, skeletal and exoskeletal anomalies, neoplasms, or
parasites).
6.2.3.2 Sample Identification Label—
A sample identification label should be completed in indelible ink for each
individual fish or shellfish specimen after it is processed to identify each sample
uniquely (Figure 6-6). The following information should be included on the
sample identification label:
6-31
-------
6. FIELD PROCEDURES
Field Record for Fish Contaminant Monitoring Program — Screening Study
Project Number:.
Sampling Date and Time:
SITE LOCATION £
Site Name/Number:
County/Parish:
. LatAong.:
Waterbody Name/Segment Number:.
Waterbody Type: D RIVER
Site Description:
D LAKE D ESTUARY
Collection Method:
Collector Name: _
(print and sign)
Agency: _
Address:
Phone:
FISH COLLECTED
Bottom Feeder—Species Name: _
Composite Sample #:
Fish # Length (mm) Sex
_ Number of Individuals:
Fish # Length (mm)
Sex
001
002
003
004
005
Minimum size
Maximum size
Notes (e.g., morphological anomalies):
006
007
008
009
010
x100 =
. >75% Composite mean length.
mm
Predator—Species Name:
Composite Sample #:
Fish # Length (mm) Sex
_ Number of Individuals:
Fish # Length (mm)
Sex
001
002
003
004
005
Minimum size
006
007
008
009
010
x100 =
Z 75%
Maximum size
Notes (e.g., morphological anomalies):
Composite mean length.
mm
Figure 6-2. Example of a field record for fish contaminant monitoring
program—screening study.
6-32
-------
6. FIELD PROCEDURES
Field Record for Shellfish Contaminant Monitoring Program — Screening Study
Project Number:
Sampling Date and Time:
SITE LOCATION
Site Name/Number:
County/Parish:
Waterbody Name/Segment Numt
Waterbody Type: D RIVER
Site Description:
jer:
D
LatAong.:
LAKE D ESTUARY
Collection Method:
Collector Name:
(print and sign)
Agency:
Address:
Phone: ( )
SHELLFISH COLLECTED 1 < - ' , - O ,\ < * ^ *> — >>* >, , , — 1
Bivalve Species Name:
Composite Sample #:
Bivalve # Size (mm)
001
002
003
004
005
006
007
008
009
010
011
012
013
014
015
016
017
Minimum size
x100 =
Maximum size
Notes (e.g., morphological anoms
Bivalve #
018
019
020
021
022
023
024
025
026
027
028
029
030
031
032
033
034
2 75%
lies):
Number of Individuals:
Size (mm) Bivalve # Size (mm)
035
036
037
038
039
040
041
042
043
044
045
046
047
048
049
050
Composite mean size mm
Figure 6-3. Example of a field record for shellfish contaminant monitoring
program—screening study.
6-33
-------
6. FIELD PROCEDURES
Reid Record for Fish
Project Number:
Contaminant Monitoring Program — Intensive Study
Samolina Date and Time:
SITE LOCATION
Site Name/Number:
County/Parish:
Waterbody Name/Segment Number:
Waterbody Type: D RIVER
Site Description:
LatAona:
D LAKE D ESTUARY
Collection Method:
Collector Name:
(print and sign)
Aaency:
Address:
Phone: (
>
FISH COLLECTED F - 1
Soecies Name:
Composite Sample #:
Fish # Length (mm) Sex (M
001
002
003
004
005
Minimum length
= — x100 =
Maximum length
Notes (e.g., morphological anomalies)
Species Name:
Composite Samole #:
Fish # Length (mm) Sex (M
001
002
003
004
005
Replicate
Number of Individuals:
, F, or I) Fish* Length (mm) Sex(M,
006
007
008
009
010
% Composite mean lenqth
Replicate
Number of Individuals:
, F, or I) Fish* Length (mm) Sex(M,
006
007
008
009
010
Minimum length „„ -,,„„..._
Maximum length
Notes (e.g., morphological anomalies)
Number
F, or 1)
mm
Number:
F, or 1)
mm
page 1 of 2
Figure 6-4. Example of a field record for fish contaminant monitoring
program—intensive study.
6-34
-------
6. FIELD PROCEDURES
Field Record for Fish Contaminant
Project Number:
SITE LOCATION:
Monitoring Program — Intensive Study (con.)
Sampling Date and Time:
Site Name/Number:
County/Parish:
LatAong.:
FISH COLLECTED .
Species Name:
Composite Sample #:
Fish # Length (mm) Sex (M, F, or 1)
001
002
003
004
005
Minimum length x1QO
Maximum length
Notes (e.g.. morphological anomalies):
Species Name:
Composite Sample #:
Fish # Length (mm) Sex (M, F, or I)
001
002
003
004
005
Minimum length
x 1 00 = %
Maximum length
Notes (e.g.. morpholoaical anomalies):
Species Name:
Composite Sample #:
Fish # Length (mm) Sex (M, F, or I)
001
002
003
004
005
Minimum length
— x 1 00 - > 75%
Maximum length
Notes (e.g., morphological anomalies):
Replicate Number:
Number of Individuals:
Fish # Length (mm) Sex (M, F, or I)
006
007
008
009
010
Composite mean length mm
Replicate Number:
Number of Individuals:
Fish # Length (mm) Sex (M, F, or I)
006
007
008
009
010
Composite mean length mm
Replicate Number:
Number of Individuals:
Fish # Length (mm) Sex (M, F, or I)
006
007
008
009
010
Composite mean length mm
page 2 of 2
Figure 6-4 (continued)
6-35
-------
6. FIELD PROCEDURES
Field Record for Shellfish Contaminant Monitoring Program — Intensive Study
Project Number:
Samolina Date and Time:
SITE LOCATION
Site Name/Number:
County/Parish:
Waterbody Name/Segment Number:
Waterbody Type: D RIVER
Site Description:
LatAona.:
D LAKE D ESTUARY
Collection Method:
Collector Name:
(print and sign)
Aaencv:
Address:
Phone: ( )
SHELLFISH COLLECTED •" : : "["n""'' ""' '"':.':: ""'":'."::" '•::": ...:::." ". . . : •::. • . ..::.: : ":. ':.,: :.: . ::.:":•. :::.: r:::..11.:::;!
Species Name:
Composite Sample #:
Shellfish # Size (mm) Sex Shellfish *
001
002
003
004
005
006
007
008
009
01-0
011
012
013
.014
015
016
017
Minimum size
x100= >
Maximum size
Notes (e.g., morphological anomalies):
018
019
020
021
022
023
024
025
026
027
028
029
030
031
032
033
034
75%
Replicate Number:
Number of Individuals:
Size (mm) Sex Shellfish # Size (mm) Sex
035
036
037
038
039
040
041
042
043
044
045
046
047
048
049
050
Composite mean size mm
Figure 6-5. Example of a field record for shellfish contaminant monitoring
program—Intensive study.
6-3G
-------
6. FIELD PROCEDURES
Species Name or Code
Total Length or Size (cm)
Sample Type
Sampling Site (name/number)
Specimen Number
Sampling Date (d/mo/yr)
Time (24-h clock)
Figure 6-6. Example of a sample Identification label.
Project Number
Sampling Site (name and/or ID number)
Collecting Agency (name, address, phone)
Composite Number/Specimen Number(s)
Sampling Date (d/m/yr)/Time (24-hr clock)
Species Name or Code
Chem
DAII
Dot
Sampler (name and signature)
ical Analyses
target analytes
hers (specify)
Processing
Whole Body
Comments
Resection
Study Type
Screening
Intensive
Phase! D
Phase II D
Type of Ice
Wet
Dry
Figure 6-7. Example of a chaln-of-custody tag or label.
6-37
-------
6. FIELD PROCEDURES
• Species scientific name or code number
Total length/size of specimen (mm)
Specimen number
• Sample type: F (fish fillet analysis only)
S (shellfish edible portion analysis only)
W (whole fish analysis)
0 (other fish tissue analysis)
Sampling site—waterbody name and/or identification number
Sampling date/time (specify convention, e.g., day/month/year and 24-h
clock).
A completed sample identification label should be taped to each
aluminum-foil-wrapped specimen and the specimen should be placed in a
waterproof plastic bag.
6.2.3.3 Chain-of-Custody Label or Tag—
A COC label or tag should be completed in indelible ink for each individual fish
specimen. The information to be completed for each fish is shown in Figure 6-7.
After all information has been completed, the COC label or tag should be taped
or attached with string to the outside of the waterproof plastic bag containing the
individual fish sample. Information on the COC label/tag should also be
recorded on the COC form (Figure 6-8).
Because of the generally smaller size of shellfish, several individual aluminum-
foil-wrapped shellfish specimens (within the same composite sample) may be
placed in the same waterproof plastic bag. A COC label or tag should be
completed in indelible ink for each shellfish composite sample. If more than 10
individual shellfish are to be composited, several waterproof plastic bags may
have to be used for the same composite. It is important not to place too many
individual specimens in the same plastic bag to ensure proper preservation
during shipping, particularly during summer months. Information on the COC
label/tag should also be recorded on the COC form.
6.2.3.4 Chaln-of-Custody Form—
A COC form should be completed in indelible ink for each shipping container
(e.g., ice chest) used. Information recommended for documentation on the COC
form (Figure 6-8) is necessary to track all samples from field collection to receipt
at the processing laboratory. In addition, this form can be used for tracking
samples through initial laboratory processing (e.g., resection) as described in
Section 7.2.
6-38
-------
6. FIELD PROCEDURES
Chain-of-Custody Record
Proiect Number
Collecting Agency (name, address, phone)
Samplers (print and sign)
Composite
Number
Specimer
Nos.
Sampling
Time
Study Type
Scr
Int
Sampling Date
Container
ol
Sampling Site (name/number)
Delivery Shipment Record
Delivery Method
Q Hand cany
n Shipped
Relinquished by: (signature)
Relinquished by
(signature)
J/ /
/£> /
'///
'//
w
y • / -' / Comments
Deliver/Ship to: (name, address and phone)
Date /Time
Date 11
Ime
Received by: (signature)
Relinquished by:
(signature)
Received lor Central Processing
Laboratory by: (signature)
Date
Time
Date
Date/Time Shipped:
/Time
Received by: (signature)
Remarks:
Laboratory Custody:
Released
Name/Date
Received
Name/Date
Purpose
Location
Figure 6-8. Example of a chaln-of-custody record form.
6-39
-------
6. FIELD PROCEDURES
Prior to sealing the ice chest, one copy of the COC form and a copy of the field
record sheet should be sealed in a resealable waterproof plastic bag. This
plastic bag should be taped to the inside cover of the ice chest so that it is
maintained with the samples being tracked. Ice chests should be sealed with
reinforced tape for shipment.
6.2.3.5 Field Logbook—
In addition to the four sample tracking forms discussed above, the field collection
team should document in a field logbook any additional information on sample
collection activities, hydrologic conditions (e.g., tidal stage), weather conditions,
boat or equipment operations, or any other unusual activities observed (e.g.,
dredging) or problems encountered that would be useful to the program manager
in evaluating the quality of the fish and shellfish contaminant monitoring data.
6.3 SAMPLE HANDLING
6.3.1 Sample Selection
6.3.1.1 Species Identification—
As soon as fish and shellfish are removed from the collection device, they should
be identified by species. Nontarget species or specimens of target species that
do not meet size requirements (e.g., juveniles) should be returned to the water.
Species identification should be conducted only by experienced personnel
knowledgeable of the taxonomy of species in the waterbodies included in the fish
and shellfish contaminant monitoring program. Taxonomic keys, appropriate for
the waters being sampled, should be consulted for species identification.
Because the objective of both the screening and intensive monitoring studies is
to determine the magnitude of contamination in specific fish and shellfish
species, it is necessary that all individuals used in a composite sample be of a
single species. Note: Correct species identification is important and different
species should never be combined in a single composite sample.
When sufficient numbers of the target species have been identified to make up
a composite sample, the species name and all other appropriate information
should be recorded on the field record forms (Figures 6-2 through 6-5).
6.3.1.2 Initial Inspection and Sorting-
Individuals of the selected target species should be rinsed in ambient water to
remove any foreign material from the external surface. Individual fish of the
selected target species then should be placed in clean holding trays to prevent
contamination. Fish and shellfish may be placed on ice immediately after
capture to stun them, thereby facilitating processing and packaging procedures.
Bivalves (oysters, clams, scallops, and mussels) adhering to one another should
be separated and scrubbed with a nylon or natural fiber brush to remove any
6^40
-------
6. FIELD PROCEDURES
adhering detritus or fouling organisms from the exterior shell surfaces (NOAA,
1987). All bivalves should be inspected carefully to ensure that the shells have
not been cracked or damaged by the sampling equipment and damaged
specimens should be discarded (Versar, 1982). Crustaceans, including shrimp,
crabs, crayfish, and lobsters, should be inspected to ensure that their
exoskeletons have not been cracked or damaged during the sampling process,
and damaged specimens should be discarded (Versar, 1982). After shellfish
have been rinsed, individual specimens should be grouped by target species and
placed in clean holding trays to prevent contamination.
A few shellfish specimens may be resected (edible portions removed) to
determine wet weight of the edible portions. This will provide an estimate of the
number of individuals required to ensure that the recommended sample weight
(200 g) is attained. Note: Individuals used to determine the wet weight of the
edible portion should not be used for target analyte analyses.
6.3.1.3 Length or Size Measurements-
Each fish within the selected target species should be measured to determine
total body length (mm). To be consistent with the convention used by most
fisheries biologists in the United States, maximum body length should be
measured as shown in Figure 6-9. The maximum body length is defined as the
length from the anterior-most part of the fish to the tip of the longest caudal fin
ray (when the lobes of the caudal fin are compressed dorsoventrally) (Anderson
and Gutreuter, 1983).
For shellfish, each individual specimen should be measured to determine the
appropriate body size (mm). As shown in Figure 6-9, the recommended body
measurements differ depending on the type of shellfish being collected. Height
is a standard measurement of size for oysters, mussels, clams, scallops, and
other bivalve molluscs (Abbott, 1974; Galtsoff, 1964). The height is the distance
from the umbo to the anterior (ventral) shell margin. For crabs, the lateral width
of the carapace is a standard size measurement (U.S. EPA, 1990c); for shrimp
and crayfish, the standard measurement of body size is the length from the
rostrum to the tip of the telson (Texas Water Commission, 1990); and for
lobsters, two standard measurements of body size are commonly used. For
clawed and spiny lobsters, the standard size is the length of the carapace. For
spiny lobsters, the length of the tail is also used as a standard size
measurement.
6.3.1.4 Sex Determination (Optional)—
An experienced fisheries biologist can often make a preliminary sex
determination for fish by visual inspection. The body of the fish should not be
dissected in the field to determine sex; sex can be determined through internal
examination of the gonads during laboratory processing (Section 7.2.2.4).
6-41
-------
6. FIELD PROCEDURES
Maximum body length3
Fish
Carapace widthb
Crab
Height0
Bivalve
Rostrum
Length
Shrimp, Crayfish
Telson
a Maximum body length is the length from the anterior-most part of the fish to the tip of the
longest caudal fin ray (when the lobes of the caudal fin are compressed dorso ventrally
(Anderson and Gutreuter, 1983).
b Carapace width is the lateral distance across the carapace (from tip of spine to tip of spine)
(U.S. EPA, 1990c).
c Height is the distance from the umbo to the anterior (ventral) shell margin (Galtsoff, 1964).
d Length is the distance from the tip of the rostrum to the tip of the telson (Texas Water
Commission, 1990).
Figure 6-9. Recommended measurements of body length and
size for fish and shellfish.
6-42
-------
6. FIELD PROCEDURES
Spiny Lobster
Clawed Lobster
Carapace
length9
8 Carapace length is the distance from the anterior-most edge of the groove between the horns
directly above the eyes, to the rear edge of the top part of the carapace as measured along the
middorsal line of the back (Laws of Florida Chapter 46-24.003).
f Tail length is the distance measured lengthwise along the top middorsal line of the entire tail
to the rear-most extremity (this measurement shall be conducted with the tail in a flat straight
position with the tip of the tail closed (Laws of Florida Chapter 46-24.003).
9 Carapace length is the distance from the rear of the eye socket to the posterior margin of the
carapace (New York Environmental Conservation Law 13-0329.5.a and Massachusetts General
Laws Chapter 130).
Figure 6-9 (continued)
6-43
-------
6. FIELD PROCEDURES
For shellfish, a preliminary sex determination can be made by visual inspection
only for crustaceans. Sex cannot be determined in bivalve molluscs without
shucking the bivalves and microscopically examining gonadal material. Bivalves
should not be shucked in the field to determine sex; sex determination through
examination of the gonads can be performed during laboratory processing if
desired (Section 7.2.3.2).
6.3.1.5 Morphological Abnormalities (Optional)—
If resources allow, States may wish to consider documenting external gross
morphological conditions in fish from contaminated waters. Severely polluted
aquatic habitats have been shown to produce a higher frequency of gross
pathological disorders than similar, less polluted habitats (Krahn et al., 1986;
Malins et al., 1984, 1985; Mix, 1986; Sinderman, 1983; and Sinderman et al.,
1980).
Sinderman et al. (1980) reviewed the literature on the relationship of fish
pathology to pollution in marine and estuarine environments and identified four
gross morphological conditions acceptable for use in monitoring programs:
• Fin erosion
Skin ulcers
Skeletal anomalies
• Neoplasms (i.e., tumors).
Fin erosion is the most frequently observed gross morphological abnormality in
polluted areas and is found in a variety of fishes (Sinderman, 1983). In demersal
fishes, the dorsal and anal fins are most frequently affected; in pelagic fishes,
the caudal fin is primarily affected.
Skin ulcers have been found in a variety of fishes from polluted waters and are
the second most frequently reported gross abnormality. Prevalence of ulcers
generally varies with season and is often associated with organic enrichment
(Sinderman, 1983).
Skeletal anomalies include abnormalities of the head, fins, gills, and spinal
column (Sinderman, 1983). Skeletal anomalies of the spinal column include
fusions, flexures, and vertebral compressions.
Neoplasms or tumors have been found at a higher frequency in a variety of
polluted areas throughout the world. The most frequently reported visible tumors
are liver tumors, skin tumors (i.e., epidermal papillomas and/or carcinomas), and
neurilemmomas (Sinderman, 1983).
The occurrence of fish parasites and other gross morphological abnormalities
that are found at a specific site should be noted on the field record form. States
interested in documenting morphological abnormalities in fish should review the
6-44
-------
6. FIELD PROCEDURES
protocols for fish pathology studies recommended in the Puget Sound Estuary
Program (1990c) and those described by Goede and Barton (1990).
6.3.2 Sample Packaging
6.3.2.1 Fish-
After initial processing to determine species, size, sex, and morphological
abnormalities, each fish should be individually wrapped in extra heavy duty
aluminum foil. Spines on fish should be sheared to minimize punctures in the
aluminum foil packaging (Stober, 1991). The sample identification label shown
in Figure 6-6 should be taped to the outside of each aluminum foil package,
each individual fish should be placed into a waterproof plastic bag and sealed,
and the COC tag or label should be attached to the outside of the plastic bag
with string or tape. All of the packaged specimens in a composite sample should
be kept together (if possible) in the same shipping container (ice chest) for
transport. Once packaged, samples should be cooled on ice immediately.
6.3.2.2 Shellfish-
After initial processing to determine species, size, sex, and morphological
abnormalities, each shellfish specimen should be wrapped individually in extra
heavy duty aluminum foil. A completed sample identification label (Figure 6-6)
should be taped to the outside of each aluminum foil package. Note: Some
crustacean species (e.g., blue crabs and spiny lobsters) have sharp spines on
their carapace that might puncture the aluminum foil wrapping. Carapace spines
should never be sheared off because this would destroy the integrity of the
carapace. For such species, one of the following procedures should be used to
reduce punctures to the outer foil wrapping:
• Double-wrap the entire specimen in extra heavy duty aluminum foil.
Place clean cork stoppers over the protruding spines prior to wrapping the
specimen in aluminum foil.
Wrap the spines with multiple layers of foil before wrapping the entire
specimen in aluminum foil.
For shellfish, several individual aluminum-foil-wrapped specimens (in the same
composite sample) may be placed in the same waterproof plastic bag. In this
case, a COC tag or label should be completed for the composite sample and
appropriate information recorded on the field record sheet and COC form. The
COC label or tag should then be attached to the outside of the plastic bag with
string or tape. For composite samples containing more than 10 shellfish
specimens or especially large individuals, additional waterproof plastic bags
should be used to ensure proper preservation. Note: It is important not to place
too many individual specimens in the same waterproof plastic bag to ensure
proper preservation during shipping. This is especially important when samples
6^45
-------
6. FIELD PROCEDURES
Table 6-7. Recommendations for Preservation of Fish and Shellfish Samples from
Time of Collection to Delivery at the Processing Laboratory
Sample
type
Number
per
composite
Container
Preservation
Maximum
shipping
time
Fish'
Whole fish
(to be filleted)
3-10 Extra heavy duty
aluminum foil wrap
of each fish.b
Each fish is placed
in a waterproof
plastic bag.
Cool on wet ice or
blue ice packets
(preferred method)
or
Freeze on dry ice
only if shipping
time will exceed 24
hours
24 hours
48 hours
Whole fish
3-10
Same as above.
Cool on wet ice or
blue ice packets
or
Freeze on dry ice
24 hours
48 hours
Shellfish8
L
Whole shellfish
(to be resected for
edible tissue)
3-50c Extra heavy duty
aluminum foil wrap
of each specimen."
Shellfish in the
same composite
sample may be
placed in the same
waterproof plastic
bag.
Cool on wet ice or
blue ice packets
(preferred method)
or
Freeze on dry ice
if shipping time
will exceed 24
hours
24 hours
48 hours
Whole shellfish 3-50c Same as above. Cool on wet ice or
blue ice packets
or
Freeze on dry ice
24 hours
48 hours
a Use only individuals that have attained at least legal harvestable or consumable size.
b Aluminum foil should not be used for long-term storage of any sample (i.e., whole organisms,
fillets, or homogenates) that will be analyzed for metals.
0 Species and size dependent. For very small shellfish species, more than 50 individuals may be
required to achieve the 200-g composite sample mass recommended for screening studies.
6-46
-------
6. FIELD PROCEDURES
are collected during hot weather and/or when the time between field collection
and delivery to the processing laboratory approaches the maximum holding time
(Table 6-7). Once packaged, composite samples should be cooled on ice
immediately.
6.3.3 Sample Preservation
The type of ice to be used for shipping should be determined by the length of
time the samples will be in transit to the processing laboratory and the sample
type to be analyzed (Table 6-7).
6.3.3.1 Fish or Shellfish To Be Resected—
Note: Fish and shellfish specimens should not be frozen prior to resection if
analyses will include edible tissue only because freezing may cause some
internal organs to rupture and contaminate fillets or other edible tissues (Stober,
1991; U.S. EPA, 1986b). Wet ice or blue ice (sealed prefrozen ice packets) is
recommended as the preservative of choice when the fish fillet or shellfish edible
portions are the primary tissues to be analyzed. Samples shipped on wet or blue
ice should be delivered to the processing laboratory within 24 hours (Smith,
1985; U.S. EPA, 1990d). If the shipping time to the processing laboratory will
exceed 24 hours, dry ice should be used.
Note: One exception to the use of dry ice for long-term storage is if fish are
collected as part of extended offshore fish surveys. States involved in these
types of field surveys may employ shipboard freezers to preserve samples for
extended periods rather than using dry ice. Ideally, all fish should be resected
in cleanrooms aboard ship prior to freezing.
6.3.3.2 Fish or Shellfish for Whole-Body Analysis—
At some sites, States may deem it necessary to collect fish for whole-body
analysis if a local subpopulation of concern typically consumes whole fish or
shellfish. If whole fish or shellfish samples are to be analyzed, either wet ice,
blue ice, or dry ice may be used; however, if the shipping time to the processing
laboratory will exceed 24 hours, dry ice should be used.
Dry ice requires special packaging precautions before shipping by aircraft to
comply with U.S. Department of Transportation (DOT) regulations. The Code of
Federal Regulations (49 CFR 173.217) classifies dry ice as Hazard Class 9
UN1845 (Hazardous Material). These regulations specify the amount of dry ice
that may be shipped by air transport and the type of packaging required. For
each shipment by air exceeding 5 pounds of dry ice per package, advance
arrangements must be made with the carrier. Not more than 441 pounds of dry
ice may be transported in any one cargo compartment on any aircraft unless the
shipper has made special written arrangements with the aircraft operator.
6-47
-------
6. FIELD PROCEDURES
The regulations further specify that the packaging must be designed and
constructed to permit the release of carbon dioxide gas to prevent a buildup of
pressure that could rupture the package. If samples are transported in a cooler,
several vent holes should be drilled to allow carbon dioxide gas to escape. The
vents should be near the top of the vertical sides of the cooler, rather than in the
cover, to prevent debris from falling into the cooler. Wire screen or cheesecloth
should be installed in the vents to keep foreign materials from contaminating the
cooler. When the samples are packaged, care should be taken to keep these
vents open to prevent the buildup of pressure.
Dry ice is exempted from shipping certification requirements if the amount is less
than 441 pounds and the package meets design requirements. The package
must be marked "Carbon Dioxide, Solid" or "Dry Ice" with a statement indicating
that the material being refrigerated is to be used for diagnostic or treatment
purposes (e.g., frozen tissue samples).
6.3.4 Sample Shipping
The fish and shellfish samples should be hand-delivered or shipped to the
processing laboratory as soon as possible after collection. The time the samples
were collected and time of their arrival at the processing laboratory should be
recorded on the COC form (Figure 6-8).
If the sample is to be shipped rather than hand-delivered to the processing
laboratory, field collection staff must ensure the samples are packed properly
with adequate ice layered between samples so that sample degradation does not
occur. In addition, a member of the field collection staff should telephone ahead
to the processing laboratory to alert them to the anticipated delivery time of the
samples and the name and address of the carrier to be used. Field collection
staff should avoid shipping samples for weekend delivery to the processing
laboratory unless prior plans for such a delivery have been agreed upon with the
processing laboratory staff.
6-48
-------
7. LABORATORY PROCEDURES I — SAMPLE HANDLING
SECTION 7
LABORATORY PROCEDURES I — SAMPLE HANDLING
This section provides guidance on laboratory procedures for sample receipt,
chain-of-custody, processing, distribution, analysis, and archiving. Planning,
documentation, and quality assurance and quality control of all laboratory
activities are emphasized to ensure that (1) sample integrity is preserved during
all phases of sample handling and analysis, (2) chemical analyses are performed
cost-effectively and meet program data quality objectives, and (3) data produced
by different States and Regions are comparable.
Laboratory procedures should be documented in a Work/QA Project Plan (U.S.
EPA, 1980b) as described in Appendix E. Routine sample processing and
analysis procedures should be prepared as standard operating procedures
(SOPs) (U.S. EPA, 1984b).
7.1 SAMPLE RECEIPT AND CHAIN-OF-CUSTODY
Fish and shellfish samples may be shipped or hand-carried from the field
according to one or more of the following pathways:
From the field to a State laboratory for sample processing and analysis
• From the field to a State laboratory for sample processing and shipment of
composite sample aliquots to a contract laboratory for analysis
• From the field to a contract laboratory for sample processing and analysis.
Sample processing and distribution for analysis ideally should be performed by
one processing laboratory. Transportation of samples from the field should be
coordinated by the sampling team supervisor and the laboratory supervisor
responsible for sample processing and distribution (see Section 6.3.4). An
accurate written custody record must be maintained so that possession and
treatment of each sample can be traced from the time of collection through
analysis and final disposition.
Fish and shellfish samples should be brought or shipped to the sample
processing laboratory in sealed containers accompanied by a copy of the sample
request form (Figure 6-1), a chain-of-custody form (Figure 6-10), and the field
records (Figures 6-4 through 6-7). Each time custody of a sample or set of
samples is transferred, the Personnel Custody Record of the COC form must be
__
-------
7. LABORATORY PROCEDURES I — SAMPLE HANDLING
completed and signed by both parties. Corrections to the COC form should be
made in indelible ink by drawing a single line through the original entry, entering
the correct information and the reason for the change, and initialing and dating
the correction. The original entry should never be obscured.
When custody is transferred from the field to the sample processing laboratory,
the following procedure should be used:
• Note the shipping time. If samples have been shipped on wet or blue ice,
check that the shipping time has not exceeded 24 hours.
Check that each shipping container has arrived undamaged and that the
seal is intact.
Open each shipping container and remove the copy of the sample request
form, the COC form, and the field records.
Note the general condition of the shipping container (samples iced properly
with no leaks, etc.) and the accompanying documentation (dry, legible, etc.).
Locate individuals in each composite sample listed on the COC form and
note the condition of their packaging. Individual specimens should be
properly wrapped and labeled. Note any problems (container punctured,
illegible labels, etc.) on the COC form.
If individuals in a composite are packaged together, check the contents of
each composite sample container against the field record for that sample to
ensure that the individual specimens are properly wrapped and labeled.
Note any discrepancies or missing information on the COC form.
Initial the COC form and record the date and time of sample receipt.
Enter the following information for each composite sample into a permanent
laboratory record book and, if applicable, a computer database:
— Sample identification number (specify conventions for the composite
sample number and the specimen number)
— Receipt date (specify convention, e.g., day/month/year)
— Sampling date (specify convention, e.g., day/month/year)
— Sampling site (name and/or identification number)
— Fish species (scientific name or code number)
— Total length of each fish or size of each shellfish (mm)
7-2
-------
7. LABORATORY PROCEDURES I — SAMPLE HANDLING
Table 7-1. Recommendations for Container Materials, Preservation, and Holding
Times for Fish and Shellfish Tissues from Receipt at Sample Processing
Laboratory to Analysis
Storage
Analyte
Matrix
Sample
container
Preservation
Holding time
Mercury Tissue (fillets and edible
portions, homogenates)
Other Tissue (fillets and edible
metals portions, homogenates)
Organics Tissue (fillets and edible
portions, homogenates)
Metals and Tissue (fillets and edible
organics portions, homogenates)
Plastic, borosilicate Freeze at £-20 °C
glass, quartz,
PTFE
Plastic, borosilicate Freeze at <-2Q °C
glass, quartz,
PTFE
Borosilicate glass, Freeze at _<-20 °C
PTFE, quartz,
aluminum foil
Borosilicate glass, Freeze at £-20 °C
quartz, PTFE
28 days
1 year
1 year
28 days
(for mercury)
and 1 year
(for other
metals arid
organics)
PTFE = Polytetrafluoroethylene; Teflon.
If samples have been shipped on wet or blue ice, distribute them
immediately to the technician responsible for resection (see Section 7.2.2).
If samples have been shipped on dry ice, they may be distributed
immediately to the technician for processing or stored in a freezer at <-20 °C
for later processing. Once processed, fillets or edible portions of fish or
shellfish, or tissue homogenates, should be stored according to the
procedures described in Section 7.2 and in Table 7-1. Note: Holding times
in Table 7-1 are maximum times recommended for holding samples from the
time they are received at the laboratory until they are analyzed. To ensure
sample integrity and analytical data quality, these times should not be
exceeded.
7.2 SAMPLE PROCESSING
This section includes recommended procedures for preparing composite
homogenate samples of fish fillets and edible portions of shellfish as required in
screening and intensive studies. Recommended procedures for preparing whole
fish composite homogenates are included in Appendix F for use by States in
assessing the potential risk to local subpopulations known to consume whole fish
or shellfish.
7-3
-------
7. LABORATORY PROCEDURES I — SAMPLE HANDLING
7.2.1 General Considerations
All laboratory personnel performing sample processing procedures (see Sections
7.2.2 and 7.2.3) should be trained or supervised by an experienced fisheries
biologist. Care must be taken during sample processing to avoid contaminating
samples. Schmitt and Finger (1987) have demonstrated that contamination of
fish flesh samples is likely unless the most exacting clean dissection procedures
are used. Potential sources of contamination include dust, instruments, utensils,
work surfaces, and containers that may contact the samples. All sample
processing (i.e., filleting, removal of other edible tissue, homogenizing,
compositing) should be done in an appropriate laboratory facility under
cleanroom conditions (Stober, 1991). Cleanrooms or work areas should be free
of metals and organic contaminants. Ideally, these areas should be under
positive pressure with filtered air (HEPA filter class 100) (California Department
of Fish and Game, 1990). Periodic wipe tests should be conducted in clean
areas to verify the absence of significant levels of metal and organic
contaminants. All instruments, work surfaces, and containers used to process
samples must be of materials that can be cleaned easily and that are not
themselves potential sources of contamination.
To avoid cross-contamination, all equipment used in sample processing (i.e.,
resecting, homogenizing, and compositing) should be cleaned thoroughly before
each composite sample is prepared. Verification of the efficacy of cleaning
procedures should be documented through the analysis of processing blanks or
rinsates (see Section 8.3.3.6).
Because sources of organic and metal contaminants differ, it is recommended
that duplicate samples be collected, if time and funding permit, when analyses
of both organics and metals are required (e.g., for screening studies). One
sample can then be processed and analyzed for organics and the other can be
processed independently and analyzed for metals (Batelle, 1989; California
Department of Fish and Game, 1990; Puget Sound Estuary Program, 1990c,
1990d). If fish are of adequate size, separate composites of individual fillets may
be prepared and analyzed independently for metals and organics. If only one
composite sample is prepared for the analyses of metals and organics, the
processing equipment must be chosen and cleaned carefully to avoid
contamination by both organics and metals.
Suggested sample processing equipment and cleaning procedures by analysis
type are discussed in Sections 7.2.1.1 through 7.2.1.3. Other procedures may
be used if it can be demonstrated, through the analysis of appropriate blanks,
that no contamination is introduced (see Section 8.3.3.6).
7.2.1.1 Samples for Organics Analysis—
Equipment used in processing samples for organics analysis should be of
stainless steel, anodized aluminum, borosilicate glass, polytetrafluoroethylene
(PTFE), ceramic, or quartz. Polypropylene and polyethylene (plastic) surfaces,
_
-------
7. LABORATORY PROCEDURES I — SAMPLE HANDLING
implements, gloves, and containers are a potential source of contamination by
organics and should not be used. If a laboratory chooses to use these materials,
there should be clear documentation that they are not a source of contamination.
Filleting should be done on glass or PIPE cutting boards that are cleaned
properly between fish or on cutting boards covered with heavy duty aluminum
foil that is changed after each filleting. Tissue should be removed with clean,
high-quality, corrosion-resistant stainless steel or quartz instruments or with
knives with titanium blades and PTFE handles (Lowenstein and Young, 1986).
Fillets or tissue homogenates may be stored in borosilicate glass, quartz, or
PTFE containers with PTFE-lined lids or in heavy duty aluminum foil (see Table
7-1).
Prior to preparing each composite sample, utensils and containers should be
washed with detergent solution, rinsed with tap water, soaked in pesticide-grade
isopropanol or acetone, and rinsed with organic-free, distilled, deionized water.
Work surfaces should be cleaned with pesticide-grade isopropanol or acetone,
washed with distilled water, and allowed to dry completely. Knives, fish sealers,
measurement boards, etc., should be cleaned with pesticide-grade isopropanol
or acetone followed by a rinse with contaminant-free distilled water between
each fish sample (Stober, 1991).
7.2.1.2 Samples for Metals Analysis—
Equipment used in processing samples for metals analyses should be of quartz,
PTFE, ceramic, polypropylene, or polyethylene. The predominant metal
contaminants from stainless steel are chromium and nickel. If these metals are
not of concern, the use of high-quality, corrosion-resistant stainless steel for
sample processing equipment is acceptable. Quartz utensils are ideal but
expensive. For bench liners and bottles, borosilicate glass is preferred over
plastic (Stober, 1991). Knives with titanium blades and PTFE handles are
recommended for performing tissue resections (Lowenstein and Young, 1986).
Borosilicate glass bench liners are recommended. Filleting may be done on
glass or PTFE cutting boards that are cleaned properly between fish or on
cutting boards covered with heavy duty aluminum foil that is changed after each
fish. Fillets or tissue homogenates may be stored in plastic, borosilicate glass,
quartz, or PTFE containers (see Table 7-1).
Prior to preparing each composite sample, utensils and containers should be
cleaned thoroughly with a detergent solution, rinsed with tap water, soaked in
acid, and then rinsed with metal-free water. Quartz, PTFE, glass, or plastic
containers should be soaked in 50% HN03, for 12 to 24 hours at room
temperature. Note: Chromic acid should not be used for cleaning any
materials. Acids used should be at least reagent grade. Stainless steel parts
may be cleaned as stated for glass or plastic, omitting the acid soaking step
(Stober, 1991).
7-5
-------
7. LABORATORY PROCEDURES I — SAMPLE HANDLING
7.2.1.3 Samples for Both Organlcs and Metals Analyses—-
As noted above, several established monitoring programs, including the Puget
Sound Estuary Program (1990c, 1990d), the NOAA Mussel Watch Program
(Battelle, 1989), and the California Mussel Watch Program (California
Department of Fish and Game, 1990), recommend different procedures for
processing samples for organics and metals analyses. However, this may not
be feasible if fish are too small to allow for preparing separate composites from
individual fillets or if resources are limited. If a single composite sample is
prepared for the analyses of both organics and metals, precautions must be
taken to use materials and cleaning procedures that are noncontaminating for
both organics and metals.
Quartz, ceramic, borosilicate glass, and PTFE are recommended materials for
sample processing equipment. If chromium and nickel are not of concern, high-
quality, corrosion-resistant stainless steel utensils may be used. Knives with
titanium blades and PTFE handles are recommended for performing tissue
resections (Lowenstein and Young, 1986). Borosilicate glass bench liners are
recommended. Filleting should be done on glass or PTFE cutting boards that
are cleaned properly between fish or on cutting boards covered with heavy duty
aluminum foil that is changed after each filleting. Fillets or tissue homogenates
should be stored in clean borosilicate glass, quartz, or PTFE containers with
PTFE-lined lids.
Prior to preparing each composite sample, utensils and containers should be
cleaned thoroughly with a detergent solution, rinsed with tap water, soaked in
50% HNO3, for 12 to 24 hours at room temperature, and then rinsed with
organics- and metal-free water. Note: Chromic acid should not be used for
cleaning any materials. Acids used should be at least reagent grade. Stainless
steel parts may be cleaned using this recommended procedure with the acid
soaking step method omitted (Stober, 1991).
Aliquots of composite homogenates taken for metals analysis (see Section 7.3.1)
may be stored in plastic containers that have been cleaned according to the
procedure outlined above, with the exception that aqua regia must not be used
for the acid soaking step.
7.2.2 Processing Fish Samples
Processing in the laboratory to prepare fish fillet composite homogenate samples
for analysis (diagrammed in Figure 7-1) involves
Inspecting individual fish
Weighing individual fish
Removing scales and/or otoliths for age determination (optional)
7-6
-------
7. LABORATORY PROCEDURES I — SAMPLE HANDLING
Log in fish samples using COC procedures
Unwrap and inspect individual fish
Weigh individual fish
Remove and archive scales and/or otoliths for age determination (optional)
Determine sex (optional); note morphological abnormalities (optional)
Remove scales from all scaled fish
Remove skin from scaleless fish (e.g., catfish)
Fillet fish
Weigh fillets (g)
Homogenize fillets
Divide homogenized sample into quarters, mix opposite
quarters, and then mix halves (3 times)
Optional
Composite equal weights (g) of w*J^s! Save remainder of fillet
homogenized fillet tissues from the
selected number of fish (200-g)
Seal and label (200-g) composite
homogenate in appropriate container(s)
and store at -20 °C until analysis (see
Table 7-1 for recommended container
materials and holding times).
homogenate from each fish
Seal and label individual fillet
homogenates in appropriate
container(s) and archive at
-20 °C (see Table 7-1 for
recommended container
materials and holding times).
COC = Chain of custody.
Figure 7-1. Preparation of fish fillet composite homogenate samples.
7-7
-------
7. LABORATORY PROCEDURES I — SAMPLE HANDLING
• Determining the sex of each fish (optional)
• Examining each fish for morphological abnormalities (optional)
Scaling all fish with scales (leaving belly flap on); removing skin of scaleless
fish (e.g., catfish)
• Filleting (resection)
Weighing fillets
Homogenizing fillets
Preparing a composite homogenate
Preparing aliquots of the composite homogenate for analysis
Distributing frozen aliquots to one or more analytical laboratories.
Whole fish should be shipped or brought to the sample processing laboratory
from the field on wet or blue ice within 24 hours of sample collection. Fillets
should be resected within 48 hours of sample collection. Ideally, fish should not
be frozen prior to resection because freezing may cause internal organs to
rupture and contaminate edible tissue (Stober, 1991; U.S. EPA, 1986b).
However, if resection cannot be performed within 48 hours, the whole fish should
be frozen at the sampling site and shipped to the sample processing laboratory
on dry ice. Fish samples that arrive frozen (i.e., on dry ice) at the sample
processing laboratory should be placed in a <-20 °C freezer for storage until
filleting can be performed. The fish should then be partially thawed prior to
resection. If rupture of organs is noted for an individual fish, the specimen
should be eliminated from the composite sample.
Sample processing procedures are discussed in the following sections. Data
from each procedure should be recorded directly in a bound laboratory notebook
or on forms that can be secured in the laboratory notebook. An example sample
processing record for fish fillet composites is shown in Figure 7-2.
7.2.2.1 Sample Inspection-
Individual fish received for filleting should be unwrapped and inspected carefully
to ensure that they have not been compromised in any way (i.e., not properly
preserved during shipment). Any specimen deemed unsuitable for further
processing and analysis should be discarded and identified on the sample
processing record.
7-8
-------
Sample Processing Record for Fish Contaminant Monitoring Program — Fish Fillet Composites
Project Number Sampling Date and Time:
STUDY PHASE:
SITE LOCATION
Site Name/Number:
County/Parish:
Screening Study | |;
Intensive Study: Phase I
Phase II
Waterbody Name/Segment Number:
Sample Type (bottom feeder, predator, etc.)_
Composite Sample #:
LatVLong.:
Waterbody Type:.
Species Name:
Replicate Number:
Number of Individuals:
First Fillet (F1)
or Combined Fillets (C)
Second Fillet (F2)
Fish 8
Weight
(9)
Scales/Otollths Sex Resection Weight Homogenate Wt. of Homog. Weight Homogenate Wt. of Homog.
Removed (/) (M,F) Performed (S) (g) Prepared (/) for Composite (g) (g) Prepared (/) for Composite (g)
001
002
003
004
005
006
007
008
009
010
Analyst
Date
Notes: .
Total Composite Weight (g)
(F1orC).
-------
7. LABORATORY PROCEDURES I — SAMPLE HANDLING
7.2.2.2 Sample Weighing—
A wet weight should be determined for each fish. All samples should be
weighed on balances that are properly calibrated and of adequate accuracy and
precision to meet program data quality objectives. Balance calibration should be
checked at the beginning and end of each weighing session and after every 20
weighings in a weighing session.
Fish shipped on wet or blue ice should be weighed directly on a foil-lined
balance tray. To prevent cross contamination between individual fish, the foil
lining should be replaced after each weighing. Frozen fish (i.e., those shipped
on dry ice) should be weighed in clean, tared, noncontaminating containers if
they will thaw before the weighing can be completed. Note: Liquid from the
thawed whole fish sample will come not only from the fillet tissue but from the
gut and body cavity, which are not part of the final fillet sample. Consequently,
inclusion of this liquid with the sample may result in an overestimate of target
analyte and lipid concentrations in the fillet hpmogenate. Nevertheless, it is
recommended, as a conservative approach, that all liquid from the thawed whole
fish sample be kept in the container as part of the sample.
AH weights should be recorded to the nearest gram on the sample processing
record and/or in the laboratory notebook.
7.2.2.3 Age Determination (Optional)—
Age provides a good indication of the duration of exposure to pollutants (Versar,
1982). A few scales or otoliths (Jearld, 1983) should be removed from each fish
and delivered to a fisheries biologist for age determination. For most warm
water inland gamefjsh, 5 to 10 scales should be removed from below the lateral
line and behind the pectoral fin. On soft-rayed fish such as trout and salmon,
the scales should be taken just above the lateral line (WDNR, 1988). For catfish
and other scaleless fish, the pectoral fin spines should be clipped and saved
(Versar, 1982). The scales, spines, or otoliths may be stored by sealing them
in small envelopes (such as coin envelopes) or plastic bags labeled with, and
cross-referenced by, the identification number assigned to the tissue specimen
(Versar, 1982). Removal of scales, spines, or otoliths from each fish should be
noted (by a check mark) on the sample processing record.
7.2.2.4 Sex Determination (Optional)—
Fish sex should be determined before filleting. To determine the sex of a fish,
an incision should be made on the ventral surface of the body from a point
immediately anterior to the anus toward the head to a point immediately posterior
to the pelvic fins. If necessary, a second incision should be made on the left
side of the fish from the initial point of the first incision toward the dorsal fin. The
resulting flap should be folded back to observe the gonads. Ovaries appear
whitish to greenish to golden brown and have a granular texture. Testes appear
7-10
-------
7. LABORATORY PROCEDURES I — SAMPLE HANDLING
creamy white and have a smooth texture (Texas Water Commission, 1990). The
sex of each fish should be recorded on the sample processing form.
7.2.2.5 Assessment of Morphological Abnormalities (Optional)—
Assessment of gross morphological abnormalities in finfish is optional. This
assessment may be conducted in the field (see Section 6.3.1.5) or during initial
inspection at the processing laboratory prior to filleting. States interested in
documenting morphological abnormalities should consult Sinderman (1983) and
review recommended protocols for fish pathology studies used in the Puget
Sound Estuary Program (1990c) and those described by Goede and Barton
(1990).
7.2.2.6 Scaling or Skinning—
To control contamination, separate sets of utensils and cutting boards should be
used for skinning or scaling fish and for filleting fish. Fish with scales should be
scaled and any adhering slime removed prior to filleting. Fish without scales
(e.g., catfish) should be skinned prior to filleting. These fillet types are
recommended because it is believed that they are most representative of the
edible portions of fish prepared and consumed by sport anglers. However, it is
the responsibility of each program manager, in consultation with State fisheries
experts, to select the fillet or sample type most appropriate for each target
species based on the dietary customs of local populations of concern.
A fish is scaled by laying it flat on a clean glass or PTFE cutting board or on one
that has been covered with heavy duty aluminum foil and removing the scales
and adhering slime by scraping from the tail to the head using the blade edge
of a clean stainless steel, ceramic, or titanium knife. Cross-contamination is
controlled by rinsing the cutting board and knife with contaminant-free distilled
water between fish. If an aluminum foil covered cutting board is used, the foil
should be changed between fish. The skin should be removed from fish without
scales by loosening the skin just behind the gills and pulling it off between knife
blade and thumb.
Once the scales and slime have been scraped off or the skin removed, the
outside of the fish should be washed with contaminant-free distilled water and
it should be placed on a second clean cutting board for filleting.
7.2.2.7 Filleting—
Filleting should be conducted only by or under the supervision of an experienced
fisheries biologist. If gloves are worn, they should be talc- or dust-free, and of
non- contaminating materials. Prior to filleting, hands should be washed with
Ivory soap and rinsed thoroughly in tap water, followed by distilled water (U.S.
EPA, 1991d). Specimens should come into contact with noncontaminating
surfaces only. Fish should be filleted on glass or PTFE cutting boards that are
cleaned properly between fish or on cutting boards covered with heavy duty
_
-------
7. LABORATORY PROCEDURES I — SAMPLE HANDLING
aluminum foil that is changed between fish (Puget Sound Estuary Program,
1990d, 1990e). Care must be taken to avoid contaminating fillet tissues with
material released from inadvertent puncture of internal organs.
Ideally, fish should be filleted while ice crystals are still present in the muscle
tissue. Therefore, if fish have been frozen, they should not be allowed to thaw
completely prior to filleting. Fish should be thawed only to the point where it
becomes possible to make an incision into the flesh (U.S. EPA, 1991d).
Clean, high-quality stainless steel, ceramic, or titanium utensils should be used
to remove one or both fillets from each fish, as necessary. The general
procedure recommended for filleting fish is illustrated in Figure 7-3 (U.S. EPA,
1991d).
The belly flap should be included in each fillet. Any dark muscle tissue in the
vicinity of the lateral line should not be separated from the light muscle tissue
that constitutes the rest of the muscle tissue mass. Bones still present in the
tissue after filleting should be removed carefully (U.S. EPA, 1991d).
If both fillets are removed from a fish, they can be combined or kept separate for
duplicate QC analysis, analysis of different analytes, or archival of one fillet.
Fillets should be weighed (either individually or combined, depending on the
analytical requirements) and the weight(s) recorded to the nearest gram on the
sample processing record.
If fillets are to be homogenized immediately, they should be placed in a properly
cleaned glass or PTFE homogenization container. If samples are to be analyzed
for metals only, plastic homogenization containers may be used. To facilitate
homogenization it may be necessary or desirable to chop each fillet into smaller
pieces using a titanium or stainless steel knife prior to placement in the
homogenization container.
If fillets are to be homogenized later, they should be wrapped in heavy duty
aluminum foil and labeled with the sample identification number, the sample type
(e.g., "F" for fillet), the weight (g), and the date of resection. If composite
homogenates are to be prepared from only a single fillet from each fish, fillets
should be wrapped separately and the designation "F1" and "F2" should be
added to the sample identification number for each fillet. The individual fillets
from each fish should be kept together. All fillets from a composite sample
should be placed in a plastic bag labeled with the composite identification
number, the individual sample identification numbers, and the date of resection
and stored at £-20 °C until homogenization.
7.2.2.8 Preparation of Individual Homogenates—
To ensure even distribution of contaminants throughout tissue samples and to
facilitate extraction and digestion of samples, the fillets from individual fish must
be ground and homogenized prior to analysis. The fillets from an individual fish
_
-------
7. LABORATORY PROCEDURES I — SAMPLE HANDLING
Scaled Fish
Scaleless Fish
After removing the scales (by
scraping with the edge of a
knife) and rinsing the fish:
Grasp the skin at the base of the head
(preferably with pliers) and pull toward
the tail.
Note: This step
applies only for
catfish and
other scaleless
species.
Make a shallow cut through the
skin (on either side of the dorsal
fin) from the top of the head to
the base of the tail.
Make a cut behind the entire
length of the gill cover, cutting
through the skin and flesh to the
bone.
Make a shallow cut along the belly
from the base of the pectoral fin to
the tail. A single cut is made from
behind the gill cover to the anus
and then a cut is made on both
sides of the anal fin. Do not cut into
the gut cavity as this may
contaminate fillet tissues.
Remove the fillet.
Source: U.S. EPA, 1991d.
Figure 7-3. Illustration of basic fish filleting procedure.
7-13
-------
7. LABORATORY PROCEDURES I — SAMPLE HANDLING
may be ground and homogenized separately, or combined, depending on the
analytical requirements and the sample size.
Fish fillets should be ground and homogenized using an automatic grinder
(Hobart model 84186 or equivalent) or high-speed blender or homogenizer
(Tekmar Tissumizer, 1/4-hp Hobart Model 4616, 1-hp Hobart Model 4822, or
equivalent). Large fillets may be cut into 2.5-cm cubes with high-quality stainless
steel or titanium knives or with a food service band saw (Hobart Model 5212 or
equivalent) prior to homogenization. Parts of the blender or homogenizer used
to grind the tissue (i.e., blades, probes) should be made of tantalum or titanium
rather than stainless steel. Stainless steel blades and/or probes have been
found to be a potential source of nickel and chromium contamination (due to
abrasion at high speeds) and should be avoided.
Grinding and homogenization of tissue is easier when it is partially frozen
(Stober, 1991). Chilling the grinder/blender briefly with a few chips of dry ice will
also help keep the tissue from sticking to it (Smith, 1985).
The fillet sample should be ground until it appears to be homogeneous. The
ground sample should then be divided into quarters, opposite quarters mixed
together by hand, and the two halves mixed together. The grinding, quartering,
and hand-mixing steps should be repeated at least two more times. If chunks
of tissue are present at this point, the grinding and homogenization should be
repeated. No chunks of tissue should remain because these may not be
extracted or digested efficiently. If the sample is to be analyzed for metals only,
the ground tissue may be mixed by hand in a polyethylene bag (Stober, 1991).
The preparation of each individual homogenate should be noted (marked with
a check) on the sample processing record. At this time, individual homogenates
may be either processed further to prepare composite homogenates or frozen
separately and stored at <-20 °C (see Table 7-1).
7.2.2.9 Preparation of Composite Homogenates—
Composite homogenates should be prepared from equal weights of individual
homogenates. The same type of individual homogenate (i.e., either single fillet
or combined fillet) should always be used in a given composite sample.
If individual homogenates have been frozen, they should be thawed partially and
rehomogenized prior to weighing and compositing. Any associated liquid should
be kept as a part of the sample. The weight of each individual homogenate
used in the composite homogenate should be recorded, to the nearest gram, on
the sample processing record.
Each composite homogenate should be blended as described for individual
homogenates in Section 7.2.2.8. The composite homogenate may be processed
immediately for analysis or frozen and stored at <-20 °C (see Table 7-1).
7-14
-------
7. LABORATORY PROCEDURES I — SAMPLE HANDLING
The remainder of each individual homogenate should be archived at <-20 °C with
the designation "Archive" and the expiration date recorded on the sample label.
The location of the archived samples should be indicated on the sample
processing record under "Notes."
It is essential that the weights of individual homogenates yield a composite
homogenate of adequate size to perform all necessary analyses. Weights of
individual homogenates required for a composite homogenate, based on the
number of fish per composite and the weight of composite homogenate
recommended for analyses of all screening study target analytes (see Table 4-1),
are given in Table 7-2. The total composite weight required for intensive studies
may be less than that for screening studies if the number of target analytes is
reduced significantly.
The recommended sample size of 200 g for screening studies is intended to
provide sufficient sample material to (1) analyze for all recommended target
analytes (see Table 4-1) at appropriate detection limits; (2) meet minimum QC
requirements for the analyses of laboratory duplicate, matrix spike, and matrix
spike duplicate samples (see Sections 8.3.3.4 and 8.3.3.5); and (3) allow for
Table 7-2. Weights (g) of Individual Homogenates
Required for Screening Study Composite Homogenate Sample"'"
Number of
fish per
sample
3
4
5
6
7
8
9
10
Total composite weight
100 g
(minimum)
33
25
20
17
14
13
11
10
200 g
(recommended)
67
50
40
33
29
25
22
20
500 g
(maximum)
167
125
100
84
72
63
56
50
aBased on total number of fish per composite and the total composite weight required for
analysis in screening studies. The total composite weight required in intensive studies may
be less if the number of target analytes is reduced significantly.
blndividual homogenates may be prepared from one or both fillets from a fish. A composite
homogenate should be prepared only from individual homogenates of the same type (i.e.,
either from individual homogenates each prepared from a single fillet or from individual
homogenates each prepared from both fillets).
7-15
-------
7. LABORATORY PROCEDURES I — SAMPLE HANDLING
reanalysis if the QC control limits are not met or if the sample is lost. However,
sample size requirements may vary among laboratories and the analytical
methods used. Each program manager must consult with the analytical
laboratory supervisor to determine the actual weights of composite homogenates
required to analyze for all selected target analytes at appropriate detection limits.
7.2.3 Processing Shellfish Samples
Laboratory processing of shellfish to prepare edible tissue composite
homogenates for analysis (diagrammed in Figure 7-4) involves
• Inspecting individual shellfish
Determining the sex of each shellfish (optional)
Examining each shellfish for morphological abnormalities (optional)
Removing the edible parts from each shellfish in the composite sample (3
to 50 individuals, depending upon the species)
• Combining the edible parts in an appropriate noncontaminating container
Weighing the composite sample
Homogenizing the composite sample
Preparing aliquots of the composite homogenate for analysis
- Distributing frozen aiiquots to one or more analytical laboratories.
Sample aliquotting and shipping are discussed in Section 7.3; all other
processing steps are discussed in this section. Shellfish samples should be
processed following the general guidelines in Section 7.2.1 to avoid
contamination. In particular, it is recommended that separate composite
homogenates be prepared for the analysis of metals and organics if resources
allow. An example sample processing record for shellfish edible tissue
composite samples is shown in Figure 7-5.
Shellfish samples should be shipped or brought to the sample processing
laboratory either on wet or blue ice (if next-day delivery is assured) or on dry ice
(see Section 6.3.3). Shellfish samples arriving on wet ice or blue ice should
have edible tissue removed and should be frozen to <-20 °C within 48 hours
after collection. Shellfish samples that arrive frozen (i.e., on dry ice) at the
processing laboratory should be placed in a <-20 °C freezer for storage until
edible tissue is removed.
7-16
-------
7. LABORATORY PROCEDURES I — SAMPLE HANDLING
Log in shellfish samples using COC procedures
Unwrap and inspect individual shellfish
Determine sex (optional); note morphological
abnormalities (optional)
Remove edible tissue from each shellfish in composite
Combine edible tissue from individual shellfish in
composite in a tared container (g)
Weigh the filled container (g)
Homogenize the composite sample
Divide homogenized sample into quarters, mix opposite
quarters and then mix halves (3 times)
Seal and label (200-g) composite
homogenate in appropriate
container(s) and store at -20 °C until
analysis (see Table 7-1 for
recommended container materials
and holding times).
Seal and label remaining
composite homogenate in
appropriate container(s) and
archive at -20 °C (see Table 7-1 for
recommended container materials
and holding times).
COC = Chain of custody.
Figure 7-4. Preparation of shellfish edible tissue composite homogenate samples.
7-17
-------
7. LABORATORY PROCEDURES I — SAMPLE HANDLING
Sample Processing Record for Shellfish Contaminant Monitoring Program — Edible Tissue Composites
Project Number
STUDY PHASE: Screening Study
SITE LOCATION
Site Name/Number
County/Parish:
Waterbody Name/Segment Number
SHELLFISH COLLECTED
Species Name:
Description of Edible Tissue
Composite Sample #:
Q
Shellfish Included in
# Composite (/) Shellfish #
001
002
003
004
005
006
007
008
009
010
011
012
013
014
015
016
017
Preparation of Composite:
Weight of container + shellfish
Weight of container (tare weight)
Total weight of composite
Notes:
018
019
020
021
022
023
024
025
026
027
028
029
030
031
032
033
034
Sampling Date and Time:
Intensive Study: Phase I I I Phase II I
Lat/Lona.:
Waterbody Type:
Number of Individuals:
Included in Included in
Composite (/) Shellfish # Composite (/)
035
036
037
038
039
040
041
042
043
044
045
046
047
048
049
050
g
g
a + =
# of specimens Average weight
of specimen
Analvst
Date
Figure 7-5. Example of a sample processing record for shellfish contaminant
monitoring program—edible tissue composites.
7-18
-------
7. LABORATORY PROCEDURES I — SAMPLE HANDLING
7.2.3.1 Sample Inspection—
Individual shellfish should be unwrapped and inspected carefully to ensure that
they have not been compromised in any way (i.e., not properly preserved during
shipment). Any specimen deemed unsuitable for further processing and analysis
should be discarded and identified on the sample processing record.
7.2.3.2 Sex Determination (Optional)—
The determination of sex in shellfish species is impractical if large numbers of
individuals of the target species are required for each composite sample.
For bivalves, determination of sex is a time-consuming procedure that must be
performed after shucking but prior to removal of the edible tissues. Once the
bivalve is shucked, a small amount of gonadal material can be removed using
a Pasteur pipette. The gonadal tissue must then be examined under a
microscope to identify egg or sperm cells.
For crustaceans, sex also should be determined before removal of the edible
tissues. For many species, sex determination can be accomplished by visual
inspection. Sexual dimorphism is particularly striking in many species of
decapods. In the blue crab, Callinectes sapidus, the female possesses a broad
abdomen suited for retaining the maturing egg mass or sponge, while the
abdomen of the male is greatly reduced in width. For shrimp, lobsters, and
crayfish, sexual variations in the structure of one or more pair of pleopods are
common.
States interested in determining the sex of shellfish should consult taxonomic
keys for specific information on each target species.
7.2.3.3 Assessment of Morphological Abnormalities (Optional)—
Assessment of gross morphological abnormalities in shellfish is optional. This
assessment may be conducted in the field (see Section 6.3.1.5) or during initial
inspection at the processing laboratory prior to removal of the edible tissues.
States interested in documenting morphological abnormalities should consult
Sinderman and Rosenfield (1967), Rosen (1970), and Murchelano (1982) for
detailed information on various pathological conditions in shellfish and review
recommended protocols for pathology studies used in the Puget Sound Estuary
Program (1990c).
7.2.3.4 Removal of Edible Tissue—
Edible portions of shellfish should consist only of those tissues that the
population of concern might reasonably be expected to eat. Edible tissues
should be clearly defined in site-specific sample processing protocols. A brief
description of the edible portions used should also be provided on the sample
7-19
-------
7. LABORATORY PROCEDURES I — SAMPLE HANDLING
processing record. General procedures for removing edible tissues from a
variety of shellfish are illustrated in Appendix G.
Thawing of frozen shellfish samples should be kept to a minimum during tissue
removal to avoid loss of liquids. Shellfish should be rinsed well with organics-
and metal-free water prior to tissue removal to remove any loose external debris.
Bivalve molluscs (oysters, clams, mussels, and scallops) typically are prepared
by severing the adductor muscle, prying open the shell, and removing the soft
tissue. The soft tissue includes viscera, meat, and body fluids (Smith, 1985).
Byssal threads from mussels should be removed with a knife before shucking
and should not be included in the composite sample.
Edible tissue for crabs typically includes all leg and claw meat, back shell meat,
and body cavity meat. Internal organs generally are removed. Inclusion of the
hepatopancreas should be determined by the eating habits of the local
population or subpopulations of concern. If the crab is soft-shelled, the entire
crab should be used in the sample. Hard- and soft-shelled crabs must not be
combined in the same composite (Smith, 1985).
Typically, shrimp and crayfish are prepared by removing the cephalothorax and
then removing the tail meat from the shell. Only the tail meat with the section
of intestine passing through the tail muscle is retained for analysis (Smith, 1985).
Edible tissue for lobsters typically includes the tail and claw meat. If the
tomalley (hepatopancreas) and gonads or ovaries are consumed by local
populations of concern, these parts should also be removed and analyzed
separately (Duston et al., 1990).
7.2.3.5 Sample Weighing-
Edible tissue from all shellfish in a composite sample (3 to 50 individuals) should
be placed in an appropriate preweighed and labeled noncontaminating container.
The weight of the empty container (tare weight) should be recorded to the
nearest gram on the sample processing record. All fluids accumulated during
removal of edible tissue should be retained as part of the sample. As the edible
portion of each shellfish is placed in the container, it should be noted on the
sample processing record. When the edible tissue has been removed from all
shellfish in the composite, the container should be reweighed and the weight
recorded to the nearest gram on the sample processing record. The total
composite weight should be approximately 200 g for screening studies. If the
number of target analytes is significantly reduced in intensive studies, a smaller
composite homogenate sample may suffice (see Section 7.2.2.9). At this point,
the composite sample may be processed for analysis or frozen and stored at
<-20°C (see Table 7-1).
7-20
-------
7. LABORATORY PROCEDURES I — SAMPLE HANDLING
7.2.3.6 Preparation of Composite Homogenates—
Composite samples of the edible portions of shellfish should be homogenized in
a grinder, blender, or homogenizer that has been cooled briefly with dry ice
(Smith, 1985). For metals analysis, tissue may be homogenized in 4-oz
polyethylene jars (California Department of Fish and Game, 1990) using a
Polytron (e.g., Brinkman Model PT10-35) equipped with a titanium generator
(e.g., Brinkman Model PTA 20). If the tissue is to be analyzed for organics only,
or if chromium and nickel contamination are not of concern, a commercial food
processor with stainless steel blades and glass container may be used. The
composite should be homogenized to a paste-like consistency. Larger samples
may be cut into 2.5-cm cubes with high-quality stainless steel or titanium knives
before grinding. If samples were frozen after dissection, they can be cut without
thawing with either a knife-and-mallet or a clean bandsaw. The ground samples
should be divided into quarters, opposite quarters mixed together by hand, and
the two halves mixed together. The quartering and mixing should be repeated
at least two more times until a homogeneous sample is obtained. No chunks
should remain in the sample because these may not be extracted or digested
efficiently. At this point, the composite homogenates may be processed for
analysis or frozen and stored at £-20 °C (see Table 7-1).
7.3 SAMPLE DISTRIBUTION
The sample processing laboratory should prepare aliquots of the composite
homogenates for analysis, distribute the aliquots to the appropriate laboratory (or
laboratories), and archive the remainder of each composite homogenate.
7.3.1 Preparing Sample Aliquots
Note: Because lipid material tends to migrate during freezing, frozen composite
homogenates must be thawed and rehomogenized before aliquots are prepared
(U.S. EPA, 1991d). Samples may be thawed overnight in an insulated cooler or
refrigerator and then homogenized. Recommended aliquot weights and
appropriate containers for different types of analyses are shown in Table 7-3.
The actual sample size required will depend on the analytical method used and
the laboratory performing the analysis. Therefore, the exact sample size
required for each type of analysis should be determined in consultation with the
analytical laboratory supervisor.
The exact quantity of tissue required for each digestion or extraction and
analysis should be weighed and placed in an appropriate container that has
been labeled with the aliquot identification number, sample weight (to the nearest
0.1 g), and the date aliquots were prepared (Stober, 1991). The analytical
laboratory can then recover the entire sample, including any liquid from thawing,
by rinsing the container directly into the digestion or extraction vessel with the
appropriate solvent. It is also the responsibility of the processing laboratory to
provide a sufficient number of aliquots for laboratory duplicates, matrix spikes,
and matrix spike duplicates so that the QC requirements of the program can be
_
-------
7. LABORATORY PROCEDURES I — SAMPLE HANDLING
Table 7-3. Recommended Sample Aliquot Weights and Containers
for Various Analyses
Analysis
Metals
Organics
Dioxins/dibenzofurans
Aliquot weight (g)
1-5
20-50
20-50
Shipping/storage container
Polystyrene, borosilicate glass, or PTFE
jar with PTFE-lined lid
Glass or PTFE jar with PTFE-lined lid
Glass or PTFE jar with PTFE-lined lid
PTFE = Polytetrafluoroethylene (Teflon).
met (see Sections 8.3.3.4 and 8.3.3.5), and to provide extra aliquots to allow for
reanalysis if the sample is lost or if QC control limits are not met.
It is essential that accurate records be maintained when aliquots are prepared
for analysis. Use of a carefully designed form is recommended to ensure that
all the necessary information is recorded. An example of a sample aliquot
record is shown in Figure 7-6. The composite sample identification number
should be assigned to the composite sample at the time of collection (see
Section 6.2.3.1) and carried through sample processing (plus "F1," "F2," or "C"
if the composite homogenate is comprised of individual or combined fillets). The
aliquot identification number should indicate the analyte class (e.g., MT for
metals, OR for organics, DX for dioxins) and the sample type (e.g., R for routine
sample; RS for a routine sample that is split for analysis by a second laboratory;
MS1 and MS2 for sample pairs, one of which will be prepared as a matrix spike).
For example, the aliquot identification number may be of the form WWWWW-XX-
YY-ZZZ, where WWWWW is a 5-digit sample composite identification number;
XX indicates individual (F1 or F2), or combined (C) fillets; YY is the analyte
code; and 777 is the sample type.
Blind laboratory duplicates should be introduced by preparing two separate
aliquots of the same composite homogenate and labeling one aliquot with a
"dummy" composite sample identification. However, the analyst who prepares
the laboratory duplicates must be careful to assign a "dummy" identification
number that has not been used for an actual sample and to indicate clearly on
the processing records that the samples are blind laboratory duplicates. The
analytical laboratory should not receive this information.
When the appropriate number of aliquots of a composite sample have been
prepared for all analyses to be performed on that sample, the remainder of the
composite sample should be labeled with "ARCHIVE" and the expiration date
and placed in a secure location at <20 °C in the sample processing laboratory.
The location of the archived samples should be indicated on the sample aliquot
record. Unless analyses are to be performed immediately by the sample
_
-------
Fish and Shellfish Monitoring Program
Sample Aliquot Record
Aliquot prepared bv Date Time
(name)
Comments
Samples from:
Project No. Site # D Sc
Composite Sample ID
Archive Location:
Analyte Code
Aliquot ID
Aliquot Weight (g)
Analyze ton
Ship to:
sreening study Intensive study D Phase 1 D Phase II
Analyte Code
Aliquot ID
Aliquot Weight (g)
Analyze for:
Ship to:
Analyte Code
Aliquot ID
Analyze for:
Ship to:
Aliquot Weight (g)
Figure 7-6. Example of a fish and shellfish monitoring program sample aliquot record.
Page.
of
ro
CO
CD
O
3J
§
30
-<
•o
30
O
O
m
o
c
33
m
en
CO
>
TJ
m
o
o
-------
7. LABORATORY PROCEDURES I — SAMPLE HANDLING
processing laboratory, aliquots for sample analysis should be frozen at <-20 °C
before they are transferred or shipped to the appropriate analytical laboratory.
7.3.2 Sample Transfer
The frozen aliquots should be transferred on dry ice to the analytical laboratory
(or laboratories) accompanied by a sample transfer record such as the one
shown in Figure 7-7. Further details on Federal regulations for shipping
biological specimens in dry ice are given in Section 6.3.3.2. The sample transfer
record may include a section that serves as the analytical laboratory COC
record. The COC record must be signed each time the samples change hands
for preparation and analysis.
7-24
-------
7. LABORATORY PROCEDURES I — SAMPLE HANDLING
Fish and Shellfish Monitoring Program
Sample Transfer Record
Date
DO MM YY
Released by:
Time : (24-h clock)
HH MM
(name)
At:
(location)
Shipment Method.
Shipment Destination
Date
Time
(24-h clock)
DD MM
Received by:
YY
HH MM
(name)
At:
Comments
(location)
Study Type: D Screening—Analyze for: D Trace metals D Organics D Lipid
Intensive Phase 1 D Phase II D — Analyze for (specify)
Sample IDs:
Laboratory Chain of Custody
Relinquished by
Received by
Purpose
Location
Figure 7-7. Example of a fish and shellfish monitoring program
sample transfer record.
7-25
-------
8. LABORATORY PROCEDURES II — SAMPLE ANALYSES
SECTION 8
LABORATORY PROCEDURES II — SAMPLE ANALYSES
Sample analyses may be conducted by one or more State or private contract
laboratories. Because of the toxicity of dioxins/dibenzofurans and the difficulty
and cost of these analyses, relatively few laboratories currently have the
capability of performing them. Table 8-1 lists contract laboratories experienced
in dioxin/dibenzofuran analyses. This list is provided for information purposes
only and is not an endorsement of specific laboratories.
8.1 RECOMMENDED ANALYTES
8.1.1 Target Analytes
All recommended target analytes listed in Table 4-1 should be included in
screening studies unless reliable historic tissue, sediment, or pollutant source
data indicate that an analyte is not present at a level of concern for human
health. Additional target analytes should be included in screening studies if
States have site-specific information (e.g., historic tissue or sediment data,
discharge monitoring reports from municipal and industrial sources) that these
contaminants may be present at levels of concern for human health.
8.1.2 Llpid
Intensive studies should include only those target analytes found to exceed
screening values in screening studies (see Section 5.2).
A lipid analysis should also be performed and reported (as percent lipid by wet
weight) for each composite tissue sample in both screening and intensive
studies. This measurement is necessary to ensure that gel permeation
chromatography columns are not overloaded when used to clean up tissue
extracts prior to analysis of organic target analytes. In addition, because
bioconcentration of nonpolar organic compounds is dependent upon lipid content
(i.e., the higher the lipid content of the individual organism, the higher the residue
in the organism), lipid analysis is often considered essential by users of fish and
shellfish monitoring data. Consequently, it is important that lipid data are
obtained for eventual inclusion in a national database of fish and shellfish
contaminant data.
8-1
-------
8. LABORATORY PROCEDURES II — SAMPLE ANALYSES
Table 8-1. Contract Laboratories Conducting Dioxln/Dlbenzofuran
Analyses In Fish and Shellfish Tissues8
Alta Analytical Laboratory13
5070 Robert J. Matthews Parkway, Suite 2
Eldorado Hills, CA 95630
916/933-1640
FAX: 916/933-0940
Bill Luksemburg
Battelle-Columbus Laboratories0
505 King Avenue
Columbus, OH 43201
614/424-7379
! Karen Riggs/Gerry Pitts
Enseco-Califomia Analytical Labs'3
2544 Industrial Blvd.
West Sacramento, CA 95691
916/372-1393
916/372-1059
Kathy Gill/Michael Filigenzi/Mike Millie
IT Corporation
Technology Development Laboratory0
304 Directors Drive
Knoxville, TN 37923
615/690-3211
Duane Root/Nancy Conrad/Bruce Wagner
Midwest Research Institute0
425 Volker Boulevard
Kansas City, MO 64110
816/753-7600 ext. 190/ext. 160
Paul Kramer/John Stanley
New York State Department of Health0
Wadsworth Laboratories
Empire State Plaza
P.O. Box 509
Albany, NY 12201-0509
518/474-4151
Arthur Richards/Kenneth Aldous
Pacific Analytical Inc.0
1989-B Palomar Oaks Way
Carlsbad, CA 92009
619/931-1766
Phil Ryan/Bruce Colby
Seakem Analytical Services0
P.O. Box2219
2045 Mills Road
Sidney, BC V8L 351
Canada
604/656-0881
Valerie Scott/Allison Peacock/Coreen Hamilton
TMS Analytical Services0
7726 Moiler Road
Indianapolis, IN 46268
317/875-5894
FAX: 317/872-6189
Dan Denlinger/Don Eickhoff/
Kelly Mills/Janet Sachs
Triangle Laboratories0
Alston Technical Park
801 Capitola Drive, Suite 10
Research Triangle Park, NC 27713
919/544-5729
Laurie White
Twin City Testing Corporation0
662 Cromwell Avenue
St. Paul, MN 55114
612/649-5502
Chuck Sueper/Fred DeRoos
University of Nebraska
Mid-West Center for Mass Spectrometry
12th and T Street
Lincoln, NE 68588
402/472-3507
Michael Gross
Wellington Environmental Consultants0
395 Laird Road
Guelph, Ontario N1G 3X7
Canada
519/822-2436
Judy Sparling/Brock Chittin
Wright State University0
175Brehm Laboratory
3640 Colonel Glen Road
Dayton, OH 45435
513/873-2202
Thomas Tiernan/Garrett Van Ness
aThis list should not be construed as an endorsement of these laboratories, but is provided for information
purposes only.
°Laboratory participating in Method 1613 interlaboratory (round-robin) dioxin study (May 1991).
8-2
-------
8. LABORATORY PROCEDURES II — SAMPLE ANALYSES
8.2 ANALYTICAL METHODS
This section provides guidance on selecting methods for analysis of
recommended target analytes. Analytical methods should include appropriate
procedures for sample preparation (i.e., for digestion of samples to be analyzed
for metals and for extraction and extract cleanup of samples to be analyzed for
organics).
8.2.1 Llpld Method
It is recommended that a gravimetric method be used for lipid analysis (e.g.,
Bligh and Dyer, 1959; California Department of Fish and Game, 1990; U.S. FDA,
1990). This method is in common use by numerous laboratories and is easy to
perform. Because there can be substantial differences (factors of 2 or 3) in lipid
measurements depending on the solvent system used in extraction (D. Swack-
hamer, University of Minnesota, personal communication, 1993; D. Murphy,
Maryland Department of the Environment, Water Quality Toxics Division,
personal communication, 1993), it is recommended that petroleum ether be used
as the extraction solvent in all lipid analyses.
8.2.2 Target Analyte Methods
EPA has published interim procedures for sampling and analysis of priority
pollutants in fish tissue (U.S. EPA, 1981b); however, at present, official EPA-
approved methods are available only for the analysis of low parts-per-billion
concentrations of metals in fish and shellfish tissues (U.S. EPA, 1991g).
Because of the lack of official EPA-approved methods for all recommended
target analytes, and to allow States and Regions flexibility in developing their
analytical programs, specific analytical methods for recommended target
analytes in fish and shellfish monitoring programs are not included in this
guidance document.
Note: A performance-based analytical program is recommended for the analysis
of target analytes. This recommendation is based on the assumption that the
analytical results produced by different laboratories and/or different methods will
be comparable if appropriate QC procedures are implemented within each
laboratory and if comparable analytical performance on round-robin comparative
analyses of standard reference materials or split sample analyses of field
samples can be demonstrated. This approach is intended to allow States to use
cost-effective procedures and to encourage the use of new or improved
analytical methods without compromising data quality. Performance-based
analytical programs currently are used in several fish and shellfish monitoring
programs, including the NOAA Status and Trends Program (Battelle, 1989b;
Cantillo, 1991; NOAA, 1987), the EPA Environmental Monitoring and
Assessment Program (EMAP) (U.S. EPA, 1991e), and the Puget Sound Estuary
Program (1990d, 1990e).
8-3
-------
8. LABORATORY PROCEDURES II — SAMPLE ANALYSES
Analytical methods used in fish and shellfish contaminant monitoring programs
should be selected using the following criteria:
Technical merit—Methods should be technically sound; they should be
specific for the target analytes of concern and based on current, validated
analytical techniques that are widely accepted by the scientific community.
Sensitivity—Method detection and quantitation limits should be sufficiently
low to allow reliable quantitation of the target analytes of concern at or below
selected SVs. Ideally, the method detection limit (in tissue) should be at
least five times lower than the selected SV for a given target analyte (Puget
Sound Estuary Program, 1990e).
Data quality—The accuracy and precision should be adequate to ensure that
analytical data are of acceptable quality for program objectives.
Cost-efficiency—Resource requirements should not be unreasonably high.
A review of current EPA guidance for chemical contaminant monitoring programs
and of analytical methods currently used or recommended in several of these
programs (as shown in Table 8-2) indicates that a limited number of analytical
techniques are most commonly used for the determination of the recommended
target analytes. These techniques are listed in Table 8-3. As shown in Table
8-4 and Appendix H, analytical methods employing these techniques have
typically achievable detection and/or quantitation limits that are well below the
recommended SVs for most target analytes, with the possible exception of
dieldrin, heptachlor epoxide, toxaphene, PCBs, and dioxins/dibenzofurans.
Recommended procedures for determining method detection and quantitation
limits are given in Section 8.3.3.3.
If lower SVs are used in a study (e.g., for susceptible populations), it is the
responsibility of program managers to ensure that the detection and quantitation
limits of the analytical methods are sufficiently low to allow reliable quantitation
of target analytes at or below these SVs. If analytical methodology is not
sensitive enough to reliably quantitate target analytes at or below selected SVs
(e.g., dieldrin, heptachlor epoxide, toxaphene, PCBs, dioxins/dibenzofurans),
program managers must determine appropriate fish consumption guidance based
on lowest detectable concentrations or provide justification for adjusting SVs to
values at or above achievable method detection limits.
The analytical techniques identified in Table 8-3 are recommended for use in
State fish and shellfish contaminant monitoring programs. However, alternative
techniques may be used if acceptable detection limits, accuracy, and precision
can be demonstrated.
Laboratories should select analytical methods for routine analyses of target
analytes that are most appropriate for their programs based on available
resources, experience, program objectives, and data quality requirements. A
-------
8. LABORATORY PROCEDURES II — SAMPLE ANALYSES
Table 8-2. Current References for Analytical Methods for
Contaminants In Fish and Shellfish Tissues
Bioaccumulation Monitoring Guidance: 4. Analytical Methods for U.S. EPA Priority Pollutants and
301 (h) Pesticides in Tissues from Marine and Estuarine Organisms (U.S. EPA, 1986b)
Quality Assurance/Quality Control (QA/QC) for 301 (h) Monitoring Programs: Guidance on Field and
Laboratory Methods (U.S. EPA, 1987e)
Pesticide Analytical Manual (PAM Vols. I and II) (U.S. FDA, 1990)
Standard Analytical Procedures of the NOAA National Analytical Facility (Krahn et al., 1988; MacLeod
et al., 1985)
Official Methods of Analysis of the Association of Official Analytical Chemists (Williams, 1984)
Analytical Procedures and Quality Assurance Plan for the Determination of Mercury in Fish (U.S.
EPA, 1989a)
Analytical Procedures and Quality Assurance Plan for the Determination of PCDD/PCDF in Fish (U.S.
EPA, 1989b)
Analytical Procedures and Quality Assurance Plan for the Determination of Xenobiotic Chemical
Contaminants in Fish (U.S.EPA, 1989c)
Interim Methods for the Sampling and Analysis of Priority Pollutants in Sediments and Fish Tissue
(U.S. EPA, 1981b)
Methods for the Determination of Metals in Environmental Samples (U.S. EPA, 1991g)
Puget Sound Estuary Program Plan (1990d, 1990e)
Environmental Monitoring and Assessment Program (EMAP) Near Coastal Virginian Province Quality
Assurance Project Plan (Draft) (U.S. EPA, 1991e)
U.S. EPA Contract Laboratory Program Statement of Work for Inorganic Analysis (U.S. EPA, 1991b)
U.S. EPA Contract Laboratory Program Statement of Work for Organic Analysis (U.S. EPA, 1991c)
U.S. EPA Method 625: Base/Neutrals and Acids by GC/MS (40 CFR 136, Appendix A).
U.S. EPA Method 1625: Semivolatile Organic Compounds by Isotope Dilution GC/MS (40 CFR 136,
Appendix A)
U.S. EPA Method 8290: Polychlorinated Dibenzodioxins (PCDDs) and Polychlorinated Dibenzofurans
(PCDFs) by High Resolution Gas Chromatography/High Resolution Mass Spectrometry
(HRGC/HRMS) (U.S. EPA, 1990b)
Test Methods for the Evaluation of Solid Waste, Physical/Chemical Methods (SW-846) (U.S. EPA,
1986f)
Standard Methods for the Examination of Water and Wastewater (Greenburg et al., 1992)
Test Methods for the Chemical Analysis of Municipal and Industrial Wastewater (U.S. EPA, 1982)
Methods for Organic Analysis of Municipal and Industrial Wastewater (40 CFR 136, Appendix A).
Methods for the Chemical Analysis of Water and Wastes (U.S. EPA, 1979b)
Laboratory Quality Assurance Program Plan (California Department of Fish and Game, 1990)
Assessment and Control of Bioconcentratable Contaminants in Surface Water (U.S. EPA, 1991 a).
Guidelines for Studies of Contaminants in Biological Tissues for the National Water-Quality
Assessment Program (Crawford and Luoma, 1993)
Analytical Chemistry of PCBs (Erickson, 1991)
Analytical Methods for Pesticides and Plant Growth Regulators, Vol. 11 (Zweig and Sherma, 1980)
8-5
-------
8. LABORATORY PROCEDURES II — SAMPLE ANALYSES
Table 8-3. Recommended Analytical Techniques for Target Analytes
Target analyte Analytical technique
Metals
Cadmium GFAA or ICPa
Mercury CVAA
Selenium GFAA, ICP, or HAAa-b
Organlcs
PCBs (total Arochlors)6 GC/ECDd'e'f
Organochlorine pesticides GC/ECDd'e
Organophosphate pesticides GC/MS, GC/FPD, or GC/NPD9
Chlorophenoxy herbicides GC/ECDd'e
Dioxins/dibenzofurans HRGC/HRMSh-'
CVAA - Cold vapor atomic absorption spectrophotometry.
GC/ECD o Gas chromatography/electron capture detection.
GC/FPD = Gas chromatography/flame photometric detection.
GC/MS - Gas chromatography/mass spectrometry.
GC/NPD o Gas chromatography/nitrogen-phosphorus detection.
GFAA - Graphite furnace atomic absorption spectrophotometry.
HAA o Hydride generation atomic absorption spectrophotometry.
HRGC/HRMS - High-resolution gas chrpmatography/high-resolution mass spectrometry.
ICP o Inductively coupled plasma emission spectrometry.
PCBs = Polychlorinated biphenyls.
a Atomic absorption methods require a separate determination for each element, which increases the time
and cost relative to the broad-scan ICP method. However, GFAA detection limits are typically more than an
order of magnitude lower than those achieved with ICP.
b Use of HAA can lower detection limits for selenium by a factor of 10-100 (Crecelius, 1978; Skoog, 1985).
c Analysis of total PCBs, as the sum of Aroclor equivalents, is recommended in both screening and intensive
studies because of the lack of adequate toxicologic data to develop screening values (SVs) for individual
PCB congeners (see Section 4.3.5). However, because of the wide range of toxicities among different PCB
congeners and the effects of metabolism and degradation on Aroclor composition in the environment,
congener analysis is deemed to be a more scientifically sound and accurate method for determining total
PCB concentrations. Consequently, States are encouraged to develop the capability to conduct PCB
congener analysis.
d GC/ECD does not provide definitive compound identification, and false positives due to interferences are
commonly reported. Confirmation by an alternative GC column phase (with ECD), or by GC/MS with
selected ion monitoring, is required for positive identification of PCBs, organochlorine pesticides, and
Chlorophenoxy herbicides.
8 GC/MS with selected ion monitoring may be used for quantitative analyses of these compounds if
acceptable detection limits can be achieved.
' If PCB congener analysis is conducted, capillary GC columns are recommended (NOAA, 1989b; Dunn et
al, 1984; Schwartz et al., 1984; Mullin et al., 1984; Stalling et al., 1987). An enrichment step, employing an
activated carbon column, may also be required to separate and quantify coeluting congeners or congeners
present at very low concentrations (Smith, 1981; Schwartz et al., 1993).
9 Some of the chlorinated organophosphate pesticides (i.e., chlorpyrifos, diazinon, ethion) may be analyzed
by GC/ECD (USGS, 1987).
h The analysis of the 17 2,3,7,8-substrtuted congeners of tetra- through octa-chlorinated dibenzo-p-dioxins
(PCDDs) and dibenzofurans (PCDFs) using isotope dilution is recommended. Note: If resources are
limited, at a minimum, 2,3,7,8-TCDD and 2,3,7,8-TCDF should be analyzed.
1 Because of the toxic'rty of dioxins/dibenzofurans and the difficulty and cost of these analyses, relatively few
laboratories currently have the capability of performing these analyses. Contract laboratories experienced
in conducting dioxin/dibenzofuran analyses are listed in Table 8-1.
8-6
-------
CO
Table 8-4. Range of Detection
Target analyte SV*
Metals
Cadmium 1 0 ppm
Mercury 0.6 ppm
Selenium 50 ppm
Organochlorine
Pesticides9
Chlordane (total) 80 ppb
cis-Chlordane
trans-Chlordane
cis-Nonachlor
trans-Nonachlor
Oxychlordane
DDT (total) 300 ppb
4.4'-DDT
2,4'-DDT
4,4'-DDD
2,4'-DDD
4.4'-DDE
2,4'-DDE
Dicofol 10, 000 ppb
Dieldrin 7 ppb
Endosulfan (total) 20,000 ppb
Endosulfan I
Endosulfan II
PCBs = Polychlorinated biphenyls. SV =
and Quantltatlon
Range of
detection limits
0.005-0.046 ppmc;
0.4 ppmd
0.0013-0.1 ppm6
0.017-0.15 ppmf;
0.02 ppm4;
0.6 ppma
< 1.5-5 ppb
<1. 5-5 ppb
<1.5-5 ppb
<1.5-7 ppb
<1.5-5 ppb
0.1 -13 ppb
0.1 -10 ppb
0.1 -10 ppb
0.1 -10 ppb
0.1-38 ppb
0.1-10 ppb
100 ppb
0.1 -5 ppb
5 ppb
5-70 ppb
Screening value (wet
Limits of Current Analytical Methods for Recommended Target
Range of
quantftation
limits
—
—
—
2-20 ppb
2-1 5 ppb
2-1 5 ppb
2-1 5 ppb
2-1 5 ppb
2-1 5 ppb
2-15 ppb
2-1 5 ppb
2-15 ppb
2-1 5 ppb
2-1 5 ppb
2.5 ppb
2-15 ppb
—
~
Target analyte
Organochlorine
Pesticides9
(continued)
Endrin
Heptachlor epoxide
Hexachlorobenzene
Lindane
Mirex
Toxaphene
Organophosphate
Pesticides
Carbophenothion
Chlorpyrifos
Diazinon
Disulfoton
Ethion
Turbufos
Chlorophenoxy
Herbicides
Oxyfluorfen
PCBs9
(total Aroclors)
Dioxins/dibenzo-
furansh (total)
TCDD/TCDF
PeCDD/PeCDF
HxCDD/HxCDF
HpCDD/HpCDF
weight).
SV*
3,000 ppb
10 ppb
70 ppb
80 ppb
2,000 ppb
1 00 ppb
1 ,000 ppb
30,000 ppb
900 ppb
500 ppb
5,000 ppb
10,000 ppb
800 ppb
10 ppb
0.7 ppt
Range of
detection limits
<1-15 ppb
0.1-5 ppb
0.1 -2 ppb
0.1 -5 ppb
0.1 -5 ppb
3-100 ppb
—
10 ppb9
50 ppb9
—
20 ppb9
—
—
20-62 ppb
1 ppt
2 ppt
4 ppt
10 ppt
Analytes"
Range of
quant Station
limits
2-15 ppb
2-15 ppb
2-15 ppb
2-1 5 ppb
2-1 5 ppb
60-153 ppb
—
2.5 ppb9
—
—
—
—
—
11 0-1 70 ppb
—
—
—
~
(continued)
00
r~
>
CD
31
>
3
3)
-<
•a
o
O
m
c
3)
0)
1
1
>
TJ
r™
m
i-
<
m
CO
-------
Table 8-4 (continued)
00
00
8 Wet weight. Summarized from Appendix H.
b From Table 5-2. Except for mercury, SVs are for general adult population using RfDs or oral slope factors available in the EPA IRIS database and
assuming a consumption rate (CR) - 6.5 g/d, average body weight (BW) = 70 kg, lifetime (70-yr) exposure, and, for carcinogens, a risk level (RL) - 10 .
The IRIS RfD for methylmercury was towered by a factor of 5 to calculate the recommended SV = 0.6 ppm in order to account for a possible fivefold
increase in fetal sensitivity to methylmercury exposure (WHO, 1990). This approach is deemed to be most prudent as an interim measure until the
current revaluation of the methylmercury RfD is completed (IRIS, 1993). NOTE: Increasing CR, decreasing BW, and/or using an RL <10'5 will decrease
the SV. Program managers must ensure that detection and quantitation limits of analytical methods are sufficient to allow reliable quantitation of target
analytes at or below selected SVs. If analytical methodology is not sensitive enough to reliably quantitate target analytes at or below selected SVs (e.g.,
dieldrin, heptachlor epoxide, toxaphene, PCBs, dioxins/dibenzofurans), the program managers must determine appropriate fish consumption guidance
based on lowest detectable concentrations, or provide justification for adjusting SVs to values at or above achievable method detection or quantitation
limits.
c Analysis by graphite furnace atomic absorption spectrophotometry (GFAA). co
d Analysis by inductively coupled plasma atomic emission spectrophotometry (ICP).
OJ
Analysis by cold vapor atomic absorption spectrophotometry (CVAA). O
' Analysis by hydride generation atomic absorption spectrophotometry (HAA). H
8 Analysis by gas chromatography/electron capture detection (GC/ECD). <
•o
h Analysis by high-resolution GC/high-resolution mass spectrometry (HRGC/HRMS). 3J
O
m
o
3J
m
T
W
TJ
m
!
W
-------
8. LABORATORY PROCEDURES II — SAMPLE ANALYSES
recent evaluation of current methods for the analyses of organic and trace metal
target analytes in fish tissue provides useful guidance on method selection,
validation, and data reporting procedures (Capuzzo et al., 1990).
The references in Table 8-2 should be consulted in selecting appropriate
analytical methods. An additional resource for method selection is the EPA
Environmental Monitoring Methods Index System (EMMI), an automated
inventory of information on environmentally significant analytes and methods for
their analysis (U.S. EPA, 1991f). At present, the EMMI database includes
information on more than 2,600 analytes from over 80 regulatory and
nonregulatory lists and more than 900 analytical methods in a variety of
matrices, including tissue. When fully implemented, this database will provide
a comprehensive cross-reference between analytes and analytical methods with
detailed information on each analytical method, including sponsoring
organization, sample matrix, and estimates of detection limits, accuracy, and
precision.
EMMI is available from the EPA Sample Control Center for all EPA personnel
and from National Technical Information Service (NTIS) for all other parties.
Please contact the EMMI Coordinator at the EPA Sample Control Center, (703)
557-5040, for further information. To order EMMI from NTIS, call (703) 487-4650
and request reference number PB92-503093. Additional information on EMMI
may be obtained through EPA's ALL-IN-1 electronic mail system (for assistance
call 1-800-334-2405) with inquiries directed to mailbox EPA 4258,
EMMI.SUPPORT.
A future source of information on analytical methods will be the NOAA National
Status and Trends Methods document currently in preparation (G. Lowenstein,
NOAA National Status and Trends Program, personal communication, 1992).
Because chemical analysis is frequently one of the most expensive components
of a sampling and analysis program, the selection of an analytical method often
will be influenced by its cost. In general, analytical costs may be expected to
increase with increased sensitivity (i.e., lower detection limits) and reliability (i.e.,
accuracy and precision). Analytical costs will also be dependent on the number
of samples to be analyzed, the requested turnaround time, the number and type
of analytes requested, the level of QC effort, and the amount of support
documentation requested (Puget Sound Estuary Program, 1990d). However,
differences in protocols, laboratory experience, and pricing policies of
laboratories often introduce large variation into analytical costs. Approximate
costs per sample for the analysis of target analytes by the recommended
analytical techniques are provided in Table 8-5.
8.3 QUALITY ASSURANCE AND QUALITY CONTROL CONSIDERATIONS
Quality assurance and quality control must be integral parts of each chemical
analysis program. The QA process consists of management review and
oversight at the planning, implementation, and completion stages of the
8-9
-------
8. LABORATORY PROCEDURES II — SAMPLE ANALYSES
Table 8-5. Approximate Range of Costs per Sample for
Analysis of Recommended Target Analytes3
Target analyte Approximate cost range (1992 $)
Metalsb
Cadmium 25 - 50
Mercury 35-50
Selenium 25 - 50
Organochlorlne pestlcldesc>d 285 - 500
Organophosphate pesticides" 250 - 500
Chlorophenoxy herbicides' 250 - 500
PCBsc
Total Aroclors 210-500
Dloxlns/dlbenzofurans9
TCDD/TCDF only 200 - 1,000
TCDD/TCDF through
OCDD/OCDF isomers 450 - 1,600
Llpld 30-40
OCDD = Octachlorodibenzo-p-dioxin. TCDD = 2,3,7,8-Tetrachlorodibenzo-p-dioxin.
OCDF = Octachlorodibenzofuran. TCDF = 2,3,7,8-Tetrachlorodibenzofuran.
PCBs = Polychlorinated biphenyls.
a These costs include sample digestion or extraction and cleanup, but not sample preparation
(i.e., resection, grinding, homogenization, compositing). Estimated cost of sample preparation
for a composite homogenate of five fish is $200 to $500.
b Analysis of cadmium by graphite furnace atomic absorption spectrophotometry (GFAA).
Analysis of selenium by GFAA or hydride generation atomic absorption spectrophotometry
(HAA). Analysis of mercury by cold vapor atomic absorption spectrophotometry (CVAA).
0 Analysis by gas chromatography/electron capture detection (GC/ECD).
d Estimated costs are for analysis of all recommended target analyte organochlorine pesticides
(see Table 4-1).
6 Analysis by gas chromatography/flame photometric detection (GC/FPD) or gas
chromatography/nitrogen-phosphorus detection (GC/NPD). Some of the chlorinated
organophosphate pesticides (i.e., chlorpyrifos, diazinon, ethion) may be analyzed as
organochlorine pesticides by GC/ECD (USGS, 1987).
1 Analysis by GC/ECD.
9 Analysis by HRGC/HRMS.
8-10
-------
8. LABORATORY PROCEDURES II — SAMPLE ANALYSES
analytical data collection activity to ensure that data provided are of the quality
required. The QC process includes those activities required during data
collection to produce the data quality desired and to document the quality of the
collected data.
During the planning of a chemical analysis program, QA activities focus on
defining data quality criteria and designing a QC system to measure the quality
of data being generated. During the implementation of the data collection effort,
QA activities ensure that the QC system is functioning effectively and that the
deficiencies uncovered by the QC system are corrected. After the analytical data
are collected, QA activities focus on assessing the quality of data obtained to
determine its suitability to support decisions for further monitoring, risk
assessments, or issuance of advisories.
The purpose of this section is to describe the general QA and QC requirements
for chemical analysis programs.
8.3.1 QA Plans
Each laboratory performing chemical analyses in fish and shellfish contaminant
monitoring programs must have an adequate QA program (U.S. EPA, 1984c).
The QA program should be documented fully in a QA plan or in a combined
Work/QA Project Plan (U.S. EPA, 1980b). (See Appendix E.) Each QA and QC
requirement or procedure should be described clearly. Documentation should
clearly demonstrate that the QA program meets overall program objectives and
data quality requirements. The QA guidelines in the Puget Sound Estuary
Program (1990d, 1990e), the NOAA Status and Trends Program (Battelle,
1989b; Cantillo, 1991; NOAA, 1987), the EPA 301 (h) Monitoring Programs (U.S.
EPA, 1987e), the EPA EMAP Near Coastal (EMAP-NC) Program (U.S. EPA,
1991e), and the EPA Contract Laboratory (CLP) Program (U.S. EPA, 1991b,
1991c) are recommended as a basis for developing program-specific QA
programs. Additional method-specific QC guidance is given in references In
Table 8-2.
8.3.2 Method Documentation
Methods used routinely for the analyses of contaminants in fish and shellfish
tissues must be documented thoroughly, preferably as formal standard operating
procedures (U.S. EPA, 1984b). Recommended contents of an analytical SOP
are shown in Figure 8-1. Analytical SOPs must be followed exactly as written.
A published method may serve as an analytical SOP only if the analysis is
performed exactly as described. Any significant deviations from analytical SOPs
must be documented in the laboratory records (signed and dated by the
responsible person) and noted in the final data report. Adequate evidence must
be provided to demonstrate that an SOP deviation did not adversely affect
method performance (i.e., detection or quantitation limits, accuracy, precision).
8-11
-------
8. LABORATORY PROCEDURES II — SAMPLE ANALYSES
Scope and application
Method performance characteristics (accuracy, precision,
method detection and quantitation limits) for each analyte
Interferences
Equipment, supplies, and materials
Sample preservation and handling procedures
Instrument calibration procedures
Sample preparation (i.e., extraction, digestion, cleanup)
procedures
Sample analysis procedures
Quality control procedures
Corrective action procedures
Data reduction and analysis procedures (with example
calculations)
Recordkeeping procedures (with standard data forms, if
applicable)
Safety procedures and/or cautionary notes
Disposal procedures
References
Figure 8-1. Recommended contents of analytical
standard operating procedures (SOPs).
Otherwise, the effect of the deviation on data quality must be assessed and
documented and all suspect data must be identified.
8.3.3 Minimum QA and QC Requirements for Sample Analyses
The guidance provided in this section is derived primarily from the protocols
developed for the Puget Sound Estuary Program (1990d, 1990e). These
protocols have also provided the basis for the EPA EMAP-NC QA and QC
8-12
-------
8. LABORATORY PROCEDURES II — SAMPLE ANALYSES
requirements (U.S. EPA, 1991e). QA and QC recommendations specified in this
document are intended to provide a uniform performance standard for all
analytical protocols used in State fish and shellfish contaminant monitoring
programs and to enable an assessment of the comparability of results generated
by different laboratories and different analytical procedures. These recommen-
dations are intended to represent minimum QA and QC procedures for any given
analytical method. Additional method-specific QC procedures should always be
followed to ensure overall data quality.
For sample analyses, minimum QA and QC requirements consist of (1) initial
demonstration of laboratory capability and (2) routine analyses of appropriate QA
and QC samples to demonstrate continued acceptable performance and to
document data quality.
Initial demonstration of laboratory capability (prior to analysis of field samples)
should include
Instrument calibration
Documentation of detection and quantitation limits
Documentation of accuracy and precision
Analysis of an accuracy-based performance evaluation sample provided by
an external QA program.
Ongoing demonstration of acceptable laboratory performance and documentation
of data quality should include
• Routine calibration and calibration checks
Routine assessment of accuracy and precision
Routine monitoring of interferences and contamination
• Regular assessment of performance through participation in external QA
interlaboratory comparison exercises, when available.
The QA and QC requirements for the analyses of target analytes in tissues
should be based on specific performance criteria (i.e., warning or control limits)
for data quality indicators such as accuracy and precision. Warning limits are
numerical criteria that serve to alert data reviewers and data users that data
quality may be questionable. A laboratory is not required to terminate analyses
when a warning limit is exceeded, but the reported data may be qualified during
subsequent QA review. Control limits are numerical data criteria that, when
exceeded, require suspension of analyses and specific corrective action by the
laboratory before the analyses may resume.
8-13
-------
8. LABORATORY PROCEDURES II — SAMPLE ANALYSES
Typically, warning and control limits for accuracy are based on the historical
mean recovery plus or minus two or three standard deviation units, respectively.
Warning and control limits for precision are typically based on the historical
standard deviation or coefficient of variation (or mean relative percent difference
for duplicate samples) plus two or three standard deviation units, respectively.
Procedures incorporating control charts (ASTM, 1976; Taylor, 1985) and/or
tabular presentations of historical data should be in place for routine monitoring
of analytical performance. Procedures for corrective action in the event of
excursion outside warning and control limits should also be in place.
The results for the various QC samples analyzed with each batch of samples
should be reviewed by qualified laboratory personnel immediately following the
analysis of each sample batch to determine when warning or control limits have
been exceeded. When established control limits are exceeded, appropriate
corrective action should be taken and, if possible, all suspect samples
reanalyzed before resuming routine analyses. If reanalyses cannot be
performed, all suspect data should be identified clearly. Note: For the purposes
of this guidance manual, a batch is defined as any group of samples from the
same source that is processed at the same time and analyzed during the same
analytical run.
Recommended QA and QC samples (with definitions and specifications),
frequencies of analyses, control limits, and corrective actions are summarized
in Table 8-6.
Note: EPA recognizes that resource limitations may prevent some States from
fully implementing all recommended QA and QC procedures. Therefore, as
additional guidance, the minimum numbers of QA and QC samples
recommended for routine analyses of target analytes are summarized in Table
8-7. It is the responsibility of each program manager to ensure that the
analytical QC program is adequate to meet program data quality objectives for
method detection limits, accuracy, precision, and comparability.
Recommended QA and QC procedures and the use of appropriate QA and QC
samples are discussed in Sections 8.3.3.2 through 8.3.3.8. Recommended
procedures for documenting and reporting analytical and QA and QC data are
given in Section 8.4. Because of their importance in assessing data quality and
interlaboratory comparability, reference materials are discussed separately in the
following section.
8.3.3.1 Reference Materials—
The appropriate use of reference materials is an essential part of good QA and
QC practices for analytical chemistry. The following definitions of reference
materials (Puget Sound Estuary Program, 1990d) are used in this guidance
document:
8-14
-------
Table 8-6. Recommended Quality Assurance and Quality Control Samples
Sample type
(definition;
specifications)
Recommended frequency
Objective of analysis"
Recommended
control limits'"
Recommended
corrective action
Calibration standards
(3-5 standards over the
expected range of
sample target analyte
concentrations, with the
lowest concentration
standard at or near the
MDL; see Section
8.3.3.2.1)
Full calibration:
Establish relationship
between instrument
response and target
analyte concentration.
Used for organics
analysis by GC/ECD
and for metals
analysis.
Instrument/method dependent;
follow manufacturer's
recommendations or procedures
in specific analytical protocols. At
a minimum, perform a 3-point
calibration each time an
instrument is set up for analysis,
after each major equipment
change or disruption, and when
routine calibration check exceeds
specific control limits.
Organics: RSD of RFs of
calibration standards <20%.
Metals: %R of all calibration
standards = 95-105.
Determine cause of problem
(e.g., instrument instability or
malfunction, contamination,
inaccurate preparation of
calibration standards) and
take appropriate corrective
action. Recalibrate and
reanalyze all suspect
samples or flag all suspect
data.
internal Standard Calibration
Instrument internal
standards (e.g., 2,2'-
difluorobiphenyl)
(see Section 8.3.3.2.1 for
definition)
Full calibration:
Determine RRFs of
organic target analytes
for quantitative
analysis. Required for
internal calibration of
GC/MS systems.
Optional calibration
technique for GC/ECD.
In every calibration standard,
sample, and blank analyzed;
added to final sample extract.
Internal standard calibration
performed at same frequency
recommended for external
calibration.
RSD of RRFs of calibration
standards <30%.
Determine cause of problem
(e.g., instrument instability or
malfunction, contamination,
inaccurate preparation of
internal standards or calibra-
tion standards) and take
appropriate corrective action.
Recalibrate and reanalyze all
suspect samples or flag all
suspect data.
£
03
O
3J
o
33
m
o
c
3)
m
CO
m
>
>
m
v>
CO
I
en
(continued)
-------
Table 8-6 (continued)
Sample type
(definition;
specifications) Objective
Recommended frequency Recommended
of analysis" control limits'*
Recommended
corrective action
| .Calibration Verification
Calibration check Verify calibration.
standards
(minimum of one mid-
range standard prepared
independently from initial
calibration standards; an
instrument internal
standard must be added
to each calibration check
standard when internal
standard calibration is
being used; see Section
8.3.3.2.1)
Organics (GC/MS): After initial Organics: Percent difference
calibration or recalibration. At between the average RF (or
beginning and end of each RRF) from initial calibration
work shift, and once every 12 h and the RF (or RRF) from
(or every 10-12 analyses, the calibration check £25%.
whichever is more frequent). Mercury. %R = 80-120.
Organics (GC/ECD): After initial Other Metals: %R = 90-1 1 0.
calibration or recalibration. At
beginning and end of each
work shift, and once every 6 h
(or every 6 samples, whichever
is less frequent).
Metals: After initial calibration or
recalibration. Every 10
samples or every 2 h,
whichever is more frequent.
I
Determine cause of problem
(e.g., instrument instability or
malfunction, contamination,
inaccurate preparation of
calibration standards) and
take appropriate corrective
action. Recalibrate and
reanalyze all suspect
samples or flag all suspect
data.
[ Method Detection Limit Determination |
Spiked matrix samples Establish or confirm
(analyte-free tissue MDL for analyte of
samples to which known interest (Keith, 1991 a;
amounts of target Keith et al., 1983).
analytes have been
added; one spike for
each target analyte at 3-
5 times the estimated
MDL; see Section
8.3.3.3.1)
Seven replicate analyses prior to Determined by program
use of method for routine manager.
analyses, and after any significant
change to a method currently in
use. Reevaluation of MDL
annually.
Redetermine MDL.
(continued)
00
r~
03
o
3)
H
O
31
•o
31
8
m
a
c
31
to
^2
|
to
1-
m
z
to
m
to
-------
Table 8-6 (continued)
Sample type
(definition;
specifications)
Objective
Recommended frequency
of analysis*
Recommended
control limits'*
Recommended
corrective action
Accuracy «rwt Precision Assessment
Reference materials6
(see Section 8.3.3.1 for
definitions)
(SRMs or CRMs,
prepared from actual
contaminated fish or
shellfish tissue if
possible, covering the
range of expected target
analyte concentrations.
Assess method
performance (initial
method validation and
routine accuracy
assessment).
Method validation: As many as
required to assess accuracy (and
precision) of method before
routine analysis of samples (i.e.,
when using a method for the first
time or after any method
modification).
Routine accuracy assessment
one (preferably blind) per 20
samples or one per batch,
whichever is more frequent.
Organics: Measured value
<95% confidence intervals, if
certified. Otherwise,
%R = 70-130.d
Metals: %R-85-115.d
Organics: Measured value
<95% confidence intervals, if
certified. Otherwise,
%R - 70-130.d
Metals: %R-85-115.d
NA
Determine cause of problem
(e.g., inaccurate calibration,
contamination), take
appropriate corrective action,
and reanalyze all suspect
samples or flag all suspect
data.
oo
09
CD
O
31
I
•o
3J
O
O
m
a
3)
m
w
1
S
I
CO
m
(continued)
-------
Table 8-6 (continued)
Sample type
(definition;
specifications)
Objective
Recommended frequency
of analysis"
Recommended
control limits'1
Recommended
corrective action
Laboratory control
samples
(Accuracy-based
samples consisting of
fish or shellfish tissue
homogenates spiked with
target analytes of
interest; may be SRMs
or CRMs; sometimes
referred to as QC
samples. When
available, EPA-CRMs
are recommended for
routine use as laboratory
control samples; see
Appendix I)
Matrix spikes
(composite tissue
homogenates of field
samples to which known
amounts of target
analytes have been
added; 0.5 to 5 times the
concentration of the
analyte of interest or 5
times the MQL)
Assess method
performance (initial
method validation and
routine accuracy
assessment). Used
for initial accuracy
assessment only if
reference materials
prepared from actual
contaminated fish or
shellfish are not
available.
Assess matrix effects
and accuracy (%R)
routinely.
Method validation: As many as
required to assess accuracy (and
precision) of method before
routine analysis of samples (i.e.,
when using a method for the first
time or after any method
modification).
Routine accuracy assessment
One per 20 samples or one per
batch, whichever is more
frequent.
Determined by program
manager.
NA
Organics: %R = 70-130.d
Afefafe:%R = 85-115.d
One per 20 samples or one per
batch, whichever is more
frequent.
Organics: %R £50 with good
precision.
Metals: %R = 75-125.
Determine cause of problem
(e.g., inaccurate calibration,
inaccurate preparation of
control samples), take
appropriate corrective action,
and reanalyze all suspect
samples or flag all suspect
data. Zero percent recovery
requires rejection of all
suspect data
Determine cause of problem
(e.g., incomplete extraction
or digestion, contamination),
take appropriate corrective
action, and reanalyze all
suspect samples or flag all
suspect data. Zero percent
recovery requires rejection of
all suspect data.
J>
CD
O
3)
I
3)
TJ
31
O
O
m
o
c
31
m
I
>
•o
m
>
CO
I
CO
CO
(continued)
-------
Table 8-6 (continued)
Sample type
(definition;
specifications)
Objective
Recommended frequency
of analysis*
Recommended
control limits'"
Recommended
corrective action
Matrix spike replicates
(replicate aliquots of
matrix spike samples;
0.5 to 5 times the
concentration of the
analyte of interest or 5
times the MQL)
Assess method
precision routinely.
One duplicate per 20 samples or
one per batch, whichever is more
frequent.
Laboratory replicates"
(replicate aliquots of
composite tissue
homogenates of field
samples)
Assess method
precision routinely.
One blind duplicate sample per
20 samples or one per batch,
whichever is more frequent.
Organics: A difference of no
more than a factor of 2
among replicates (i.e.,
approximately 50%
coefficient of variation).
Note: Pooling of variances
in duplicate analyses from
different sample batches is
recommended for estimating
the standard deviation or
coefficient of variation of
replicate analyses.
Metals: |RPD| <20 for
duplicates.
Organics: A difference of no
more than a factor of 2
among replicates (i.e.,
approximately 50%
coefficient of variation).
Note: Pooling of variances
in duplicate analyses from
different sample batches is
recommended for estimating
the standard deviation or
coefficient of variation of
replicate analyses.
Metals: |RPD| <20 for
duplicates.
Determine cause of problem
(e.g., incomplete extraction
or digestion, contamination,
instrument instability or
malfunction), take
appropriate corrective action,
and reanalyze all suspect
samples or flag all suspect
data.
Determine cause of problem
(e.g., composite sample not
homogeneous, instrument
instability or malfunction),
take appropriate corrective
action, and reanalyze all
suspect samples or flag all
suspect data.
CO
£
00
O
3J
o
33
-<
•o
33
O
O
m
o
30
m
OT
TJ
r~
m
00
i
CO
(continued)
m
-------
Table 8-6 (continued)
Sample type
(definition;
specifications)
Objective
Recommended frequency
of analysis*
Recommended
control limits'1
Recommended
corrective action
Analytical Replicates
(replicate aliquots of final
sample extract or
digestate)
Field replicates
(replicate composite
tissue samples)
Assess analytical
precision.
Assess total variability
(i.e., population
variability, field or
sampling variability,
and analytical method
variability).
Duplicate injections for all metal Determined by program
• I
analyses.
Screening studies: OPTIONAL; if
program resources allow, a
minimum of one blind replicate
(i.e., duplicate) for each primary
target species at 10 percent of
screening sites.8
Intensive studies: Blind replicate
samples for each target species
(and size, age or sex class, if
appropriate) at each sampling
site. Number of replicates
determined by program manager
(see Section 6.1.2.7).
manager,
g
Determined by program
manager.
g
Determined by program
manager.9
Determine cause of problem
(e.g., instrument instability or
malfunction), take
appropriate corrective action,
and reanalyze sample.
Determined by program
manager.
Determined by program
manager.
£
CD
O
31
o
31
<
•o
31
O
O
m
o
31
m
CO
co
>
m
>
00
&
CO
m
CO
(continued)
-------
Table 8-6 (continued)
Sample type
(definition;
specifications)
Objective
Recommended frequency
of analysis'
Recommended
control limits6
Recommended
corrective action
| Contamination Assessment
Blanks (field, method,
processing, bottle,
reagent)
(see Section 8.3.3.6 for
definitions)
Assess contamination
from equipment,
reagents, etc.
One field blank per sampling site.
One method blank per 20
samples or one per batch,
whichever is more frequent. At
least one processing blank per
study. At least one bottle blank
per lot or per batch of samples,
whichever is more frequent. One
reagent blank prior to use of a
new batch of reagent and
whenever method blank exceeds
control limits.
Concentration of any anatyte
•o
m
>
>
W
m
[ Routine Monitoring
-------
Table 8-6 (continued)
Sample type
(definition;
specifications)
Objective
Recommended frequency
, of analysis*
Recommended
control limits'*
Recommended
corrective action
Prepared from other
surrogate
compounds
Assess method
performance and
estimate the recovery
of organic target
analytes analyzed by
GC/MS or GC/ECD.
In every calibration standard,
sample, and blank analyzed for
organics, unless isotope dilution
technique is used:
Semivolatiles:
3 for neutral fraction
2 for acid fraction
Volatile* 3
Pesticides/PCBs: 1
Added to samples prior to
extraction.
Determined by program
manager according to most
recent EPA CLP guidelines.*1
Determine cause of problem
(e.g., incomplete extraction
or digestion, contamination,
inaccurate preparation of
surrogates), take appropriate
corrective action, and
reanalyze all suspect
samples or flag all suspect
data.
i External QA Assessment
E
03
O
33
O
3J
TJ
3)
O
O
m
o
c
3J
m
I
>
TJ
m
>
>
<
m
OT
Accuracy-based per-
formance evaluation
samples
(QA samples from NOAA
interlaboratory
comparison program;
see Section 8.3.3.8.1}
Initial demonstration of
laboratory capability.
Once prior to routine analysis of
field samples (blind).
Organics: %R=70-130.d
Metals: %R=85-115.d
Ongoing
demonstration of
laboratory capability.
One exercise (four to six
samples) per year (blind).
Determined by NOAA. Based
on consensus value of all
participating laboratories.
Determine cause of problem
and reanalyze sample. Do
not begin analysis of field
samples until performance
evaluation sample results
are acceptable.
Determine cause of problem.
Do not continue analysis of
field samples until laboratory
capability is clearly
demonstrated.
CO
rb
ro
(continued)
-------
Table 8-6 (continued)
Sample type
(definition;
specifications)
Objective
Recommended frequency
of analysis*
Recommended
control limits'*
Recommended
corrective action
Split samples
(laboratory replicates
analyzed by different
laboratories; see Section
8.3.3.8.2)
Assess interlaboratory
comparability.
5-10% of composite homogenates Determined by program
split between States and/or managers.
Regions that routinely share
monitoring results, or as
determined by program
managers.9
Review sampling and
analytical methods. Identify
sources of noncomparability.
Standardize and validate
methods to document
comparability.
CLP = Contract laboratory program.
CRM = Certified reference material (see Section 8.3.3.1).
GC/ECD = Gas chromatography/electron capture detection.
GC/MS = Gas chromatography/mass spectrometry.
MDL = Method detection limit (see Section 8.3.3.3.1).
MQL = Method quantrtatbn limit (see Section 8.3.3.3.2).
NA = Not applicable.
NOAA = National Oceanic and Atmospheric Administration.
PCBs = Polychlorinated biphenyls.
QA = Quality assurance.
%R = Percent recovery (see Sections 8.3.3.4 and 8.3.3.7.1).
RF = Response factor (see Section 8.3.3.2.1).
RPD = Relative percent difference (see Section 8.3.3.5).
RRF = Relative response factor (see Section 8.3.3.2.1).
RSD = Relative standard deviation (see Section 8.3.3.5).
SRM = Standard reference material (see Section 8.3.3.1).
a Recommended frequencies are based primarily on recommendations in U.S. EPA (1986f, 1987e, 1989c, 1991b, 1991c), Puget Sound Estuary Program (1990d,
1990e), and Battelle (1989b).
b From Puget Sound Estuary Program (1990d, 1990e) action limits, except where otherwise noted. Note: Individual programs may require more stringent control
limits. It is the responsibility of each program manager to set control limits that will ensure that the measurement data meet program data quality objectives.
c As available (see Table 8-8 and Appendix I).
d FromU.S.EPA(1991e).
8 Sometimes referred to as analytical replicates (e.g., in Puget Sound Estuary Program, 1990d).
f From U.S. EPA (1987e).
9 Recommended by EPA for this guidance document.
h From U.S. EPA (1991b, 1991c).
£
a
o
31
o
3J
m
o
a
m
I
>
•o
m
!
u>
s
-------
8. LABORATORY PROCEDURES II — SAMPLE ANALYSES
Table 8-7. Minimum Recommended QA and QC Samples for
Routine Analysis of Target Analytes"
Sample Type
Target analyte
Metals
Organics
Accuracy-based performance
evaluation sample6
Method blank
Once prior to routine
analysis of field samples,
plus one exercise (four
to six samples) per year.
1
Once prior to routine
analysis of field samples,
plus one exercise (four
to six samples) per year.
1
Laboratory duplicate
Matrix spike
Laboratory control sample
(SRM or CRM, if available)
Calibration check standard
Surrogate spike (isotopically labeled
target analyte or other surrogate
compound added prior to extraction)
NA
Each sample
Instrument (injection) internal standard;
added prior to injection
NA
Each calibration or
calibration check
standard and each
sample or blank
analyzed by GC/MSd
CRM = Certified reference material (see Section 8.3.3.1).
GC/MS = Gas chromatography/mass spectroscopy.
NA = Not applicable.
QA = Quality assurance.
QC = Quality control.
SRM = Standard reference material (see Section 8.3.3.1).
a Unless otherwise specified, the number given is the recommended number of QC samples per
20 samples or per batch, whichever is more frequent. Additional method-specific QC
requirements should always be followed provided these minimum requirements have been met.
b QA samples from National Oceanic and Atmospheric Administration interlaboratory comparison
program (see Section 8.3.3.8.1).
c One every 10 samples (plus one at beginning and end of each analytical run).
d Optional for analyses by GC/electron capture detection (ECD), GC/flame ionization detection
(FID), or GC with other nonspecific detectors.
8-24
-------
8. LABORATORY PROCEDURES II — SAMPLE ANALYSES
A reference material is any material or substance of which one or more
properties have been sufficiently well established to allow its use for
instrument calibration, method evaluation, or characterization of other
materials.
A certified reference material (CRM) is a reference material of which the
value(s) of one or more properties has (have) been certified by a variety of
technically valid procedures. CRMs are accompanied by or traceable to a
certificate or other documentation that is issued by the certifying organization
(e.g., U.S. EPA, NIST, National Research Council of Canada [NRCC]).
A standard reference material (SRM) is a CRM issued by the NIST.
Reference materials may be used to (1) provide information on method accuracy
and, when analyzed in replicate, on precision, and (2) obtain estimates of
intermethod and/or interlaboratory comparability. An excellent discussion of the
use of reference materials in QA and QC procedures is given in Taylor (1985).
The following general guidelines should be followed to ensure proper use of
reference materials (NOAA, 1992):
When used to assess the accuracy of an analytical method, the matrix of the
reference material should be as similar as possible to that of the samples of
interest. If reference materials in matrices other than fish or shellfish tissue
are used, possible matrix effects should be addressed in the final data
analysis or interpretation.
Concentrations of reference materials should cover the range of possible
concentrations in the samples of interest. Note: Because of a lack of low-
and high-concentration reference materials for most analytes in fish and
shellfish tissue matrices, potential problems at low or high concentrations
often cannot be documented.
Reference materials should be analyzed prior to beginning the analyses of
field samples to assess laboratory capability and regularly thereafter to
detect and document any changes in laboratory performance over time.
Appropriate corrective action should be taken whenever changes are
observed outside specified performance limits (e.g., accuracy, precision).
If possible, reference material samples should be introduced into the sample
stream as double blinds, that is, with identity and concentration unknown to
the analyst. However, because of the limited number of certified fish and
shellfish tissue reference materials available, the results of analyses of these
materials may be biased by an analyst's increasing ability to recognize these
materials with increased use.
• Results of reference material analyses are essential to assess interlaboratory
or intermethod comparability. However, the results of sample analyses
should not be corrected based on percent recoveries of reference materials.
8^25
-------
8. LABORATORY PROCEDURES II — SAMPLE ANALYSES
Final reported results should include both uncorrected sample results and
percent recoveries of reference materials.
Sources of EPA-certified and other recommended reference materials for the
analysis of priority pollutants and selected related compounds in fish and
shellfish tissues are given in Appendix I. Currently available marine or estuarine
tissue reference materials that may be appropriate for use by analytical
laboratories in fish and shellfish contaminant monitoring programs are given in
Table 8-8.
8.3.3.2 Calibration and Calibration Checks-
General guidelines for initial calibration and routine calibration checks are
provided in this section. Method-specific calibration procedures are included in
the references in Table 8-2. It is the responsibility of each program manager to
ensure that proper calibration procedures are developed and followed for each
analytical method to ensure the accuracy of the measurement data.
All analytical instruments and equipment should be maintained and calibrated
properly to ensure optimum operating conditions throughout a measurement
program. Calibration and maintenance procedures should be performed
according to SOPs based on the manufacturers' specifications and the
requirements of specific analytical procedures. Calibration procedures must
include provisions for documenting calibration frequencies, conditions, standards,
and results to describe adequately the calibration history of each measurement
system. Calibration records should be inspected regularly to ensure that these
procedures are being performed at the required frequency and according to
established SOPs. Any deficiencies in the records or deviations from
established procedures should be documented and appropriate corrective action
taken.
Calibration standards of known and documented accuracy must be used to
ensure the accuracy of the analytical data. Each laboratory should have a
program for verifying the accuracy and traceability of calibration standards
against the highest quality standards available. If possible, NIST-SRMs or
EPA-certified standards should be used for calibration standards (see Section
8.3.3.4 and Appendix I). A log of all calibration materials and standard solutions
should be maintained. Appropriate storage conditions (i.e., container
specifications, shelf-life, temperature, humidity, light condition) should be
documented and maintained.
8.3.3.2.1 Initial and routine calibration
Prior to beginning routine analyses of samples, a minimum of three (and
preferably five) calibration standards should be used to construct a calibration
curve for each target analyte, covering the normal working range of the
instrument or the expected target analyte concentration range of the samples to
be analyzed. The lowest-concentration calibration standard should be at or near
8-26
-------
8. LABORATORY PROCEDURES II — SAMPLE ANALYSES
Table 8-8. Fish and Shellfish Tissue Reference Materials
Identification
code
DOLT-1
DORM-1
LUTS-1
TORT-1
GBW-08571
GBW-08572
MA-A-1/OC
MA-A-3/OC
MA-B-3/OC
MA-M-2/OC
MA-A-1/TM
MA-A-2/TM
MA-B-3/TM
MA-B-3/RN
IAEA-350
IAEA-351
IAEA-352
CRM-278
CRM-422
EPA-FISH
EPA-SRS903
EPA-0952
EPA-2165
RM-50
SRM-1566a
SRM-1974
NIES-6
Sources:
BCR - C
Analytes
Elements
Elements
Elements
Elements
Elements
Elements
Organic compounds
Organic compounds
Organic compounds
Organic compounds
Elements
Elements
Elements
Isotopes
Elements
Organic compounds
Isotopes
Elements
Elements
Pesticides
Chlordane
Mercury
Mercury
Elements
Elements
Organics
Elements
Source
NRCC
NRCC
NRCC
NRCC
NRCCRM
NRCCRM
IAEA
IAEA
IAEA
IAEA
IAEA
IAEA
IAEA
IAEA
IAEA
IAEA
IAEA
BCR
BCR
EPA1
EPA2
EPA1
EPA1
NIST
NIST
NIST
NIES
Matrix
Dogfish liver (freeze-dried)
Dogfish muscle (freeze-dried)
Non-defatted lobster hepatopancreas
Lobster hepatopancreas
Mussel tissue (freeze-dried)
Prawn tissue
Copepod homogenate (freeze-dried)
Shrimp homogenate (freeze-dried)
Fish tissue (freeze-dried)
Mussel tissue
Copepod homogenate (freeze-dried)
Fish flesh homogenate
Fish tissue (freeze-dried)
Fish tissue (freeze-dried)
Tuna homogenate (freeze-dried)
Tuna homogenate (freeze-dried)
Tuna homogenate (freeze-dried)
Mussel tissue (freeze-dried)
Cod muscle (freeze-dried)
Fish tissue
Fish tissue
Fish tissue
Fish tissue
Albacore tuna (freeze-dried)
Oyster tissue (freeze-dried)
Mussel tissue
Mussel tissue
ommunity Bureau of Reference, Commission of the European Communities,
Directorate General for Science, Research and Development, 200 rue de la Loi, B-
1049 Brussels, Belgium.
EPA - U.S. Environmental Protection Agency, Quality Assurance Branch, EMSL-Cincinnati,
Cincinnati, OH, 45268, USA. (EPA1: Material available from Supelco, Inc., Supelco
Park, Bellefonte, PA, 16823-0048, USA. EPA2: Material available from Fisher
Scientific, 711 Forbes Ave., Pittsburgh, PA 15219.)
IAEA o International Atomic Energy Agency, Analytical Quality Control Service, Laboratory
Seibersdorf, P. O. Box 100, A-1400 Vienna, Austria.
NRCCRM = National Research Center for CRMs, Office of CRMs, No. 7, District 11, Hepingjie,
Chaoyangqu, Beijing, 100013, China.
NRCC = National Research Council of Canada, Institute for Environmental Chemistry, Marine
Analytical Chemistry Standards Program, Division of Chemistry, Montreal Road,
Ottawa, Ontario K1A OR9, Canada.
NIST - National Institute of Standards and Technology, Office of Standard Reference
Materials, Gaithersburg, MD, 20899, USA.
NIES «• National Institute for Environmental Studies, Yatabe-machi, Tsukuba, Ibaraki, 305,
Japan.
8-27
-------
8. LABORATORY PROCEDURES II — SAMPLE ANALYSES
the estimated method detection limit (see Section 8.3.3.3.1). Calibration
standards should be prepared in the same matrix (i.e., solvent) as the final
sample extract or digestate. Criteria for acceptable calibration (e.g., acceptable
limits for r2, slope, intercept, percent recovery, response factors) should be
established for each analytical method. If these control limits are exceeded, the
source of the problem (e.g., inaccurate standards, instrument instability or
malfunction) should be identified and appropriate corrective action taken. No
analyses should be performed until acceptable calibration has been achieved
and documented.
In addition to the initial calibration, an established schedule for the routine
calibration and maintenance of analytical instruments should be followed, based
on manufacturers' specifications, historical data, and specific procedural
requirements. At a minimum, calibration should be performed each time an
instrument is set up for analysis, after any major disruption or failure, after any
major maintenance, and whenever a calibration check exceeds the
recommended control limits (see Table 8-6).
Two types of calibration procedures are used in the analytical methods
recommended for the quantitation of target analytes: external calibration and
internal standard calibration.
External calibration
In external calibration, calibration standards with known concentrations of target
analytes are analyzed, independent of samples, to establish the relationship
between instrument response and target analyte concentration. External
calibration is used for the analyses of metals and, at the option of the program
manager, for the analyses of organics by gas chromatography/electron capture
detection (GC/ECD), gas chromatography/flame ionization detection (GC/FID),
or GC methods using other nonspecific detectors.
External calibration for metals analysis is considered acceptable if the percent
recovery of all calibration standards is between 95 and 105 percent; external
calibration for organic analyses is considered acceptable if the relative standard
deviation (RSD) of the response factors (RFs) is <20 percent (see Table 8-6).
If these limits are exceeded, the initial calibration should be repeated.
Internal standard calibration
Calibration of GC/mass spectrometry (MS) systems used for the analysis of
organic target analytes requires the addition of an Internal standard to each
calibration standard and determination of the response of the target analyte of
interest relative to that of the internal standard. Internal standard calibration may
also be used with nonspecific detector GC methods such as GC/ECD and
GC/FID. Internal standards used to determine the relative response factors
(RRFs) are termed instrument or injection internal standards (Puget Sound
Estuary Program, 1990d; U.S. EPA, 1991e). The addition of instrument internal
8-28
-------
8. LABORATORY PROCEDURES II — SAMPLE ANALYSES
standards to both calibration standards and sample extracts ensures rigorous
quantitation, particularly accounting for shifts in retention times of target analytes
in complex sample extracts relative to calibration standards. Recommended
instrument internal standards for semivolatile organic compounds are included
in analytical methods for these compounds (see references in Table 8-2).
The RRF for each target analyte is calculated for each calibration standard as
follows:
RRFt = (A,) (Cis) / (/y (Ct) (8-1)
where
A, = Measured response (integrated peak area) for the target analyte
Cis = Concentration of the instrument internal standard in the calibration
standard
Ais = Measured response (integrated peak area) for the instrument internal
standard
C, = Concentration of the target analyte in the calibration standard.
If the relative standard deviation (RSD) of the average RRF, for all calibration
standards (RRF,) is <30 percent, RRFt can be assumed to be constant across
the working calibration range and RRF, can be used to quantitate target analyte
concentrations in the samples as follows:
C, (ppm or ppb, wet weight) = (At) (Cjs) (Ve) / (AJ (RRF,) (W) (8-2)
where
C, = Concentration of the target analyte in the sample
Cis = Concentration of the instrument internal standard in the sample
extract
Ve = Volume of the final sample extract (ml)
W = Weight of sample extracted (g)
and At, AJS, and RRFt are defined as in Equation (8-1).
If the RSD of RRF, for all calibration standards is >30 percent, the initial
calibration should be repeated (see Table 8-6).
8-29
-------
8. LABORATORY PROCEDURES II — SAMPLE ANALYSES
8.3.3.2.2 Routine calibration checks
After initial calibration has been achieved and prior to the routine analyses of
samples, the accuracy of the calibration should be verified by the analysis of a
calibration check standard. A calibration check standard is a mid-range
calibration standard that has been prepared independently (i.e., using a different
stock) from the initial calibration standards. When internal standard calibration
is being used, an instrument internal standard must be added to each calibration
check standard.
Routine calibration checks should be conducted often enough throughout each
analysis run to ensure adequate maintenance of instrument calibration (see
Table 8-6). A calibration check should always be performed after analyzing the
last sample in a batch and at the end of each analysis run.
If a calibration check does not fall within specified calibration control limits, the
source of the problem should be determined and appropriate corrective action
taken (see Table 8-6). After acceptable calibration has been reestablished, all
suspect analyses should be repeated. If resources permit, it is recommended
that all samples after the last acceptable calibration check be reanalyzed.
Otherwise, the last sample analyzed before the unacceptable calibration check
should be reanalyzed first and reanalysis of samples should continue in reverse
order until the difference between the reanalysis and initial results is within the
specified control limits. If reanalysis is not possible, all suspect data (i.e., since
the last acceptable calibration check) should be identified clearly in the
laboratory records and the data report.
8.3.3.2.3 Calibration range and data reporting
As noted in Section 8.3.2.1, the lowest-concentration calibration standard should
be at or near the method detection limit. The highest-concentration calibration
standard should be selected to cover the full range of expected concentrations
of the target analyte in fish and shellfish tissue samples. If a sample
concentration occurs outside the calibration range, the sample should be diluted
or concentrated as appropriate and reanalyzed or the calibration range should
be extended. Extremely high concentrations of organic compounds may indicate
that the extraction capabilities of the method have been saturated and extraction
of a smaller sample or modification of the extraction procedure may be required.
All reported concentrations must be within the upper limit of the demonstrated
working calibration range. Procedures for reporting data, with appropriate
qualifications for data below method detection and quantitation limits, are given
in Section 8.3.3.3.3.
8.3.3.3 Assessment of Detection and Quantitation Limits-
It is the responsibility of each laboratory to determine appropriate detection and
quantitation limits for each analytical method for each target analyte in a fish or
8-30
-------
8. LABORATORY PROCEDURES II — SAMPLE ANALYSES
shellfish tissue matrix. When available scientific literature demonstrates that the
selected SVs are analytically attainable, the laboratory is responsible for
ensuring that these limits are sufficiently low to allow reliable quantitation of the
analyte at or below the selected SVs (see Section 5.2). Detection and
quantitation limits must be determined prior to the use of any method for routine
analyses and after any significant changes are made to a method during routine
analyses. Several factors influence achievable detection and quantitation limits
regardless of the specific analytical procedure. These include amount of sample
available, matrix interferences, and stability of the instrumentation. The limits of
detection given in Table 8-4 and Appendix H are considered to be representative
of typically attainable values. Depending upon individual laboratory capabilities
and fish tissue matrix properties, it should be noted that SVs for some
recommended target analytes (e.g., dieldrin, heptachlor epoxide, toxaphene,
PCBs, and dioxins/dibenzofurans) may not always be analytically attainable
quantitation limits. In these instances, all historic and current data on
contaminant sources and on water, sediment, and fish and shellfish contaminant
tissue data should be reviewed to provide additional information that could aid
in the risk assessment process and in making risk management decisions.
The EPA has previously issued guidance on detection limits for trace metal and
organic compounds for analytical methods used in chemical contaminant
monitoring programs (U.S. EPA, 1985a). However, at present there is no clear
consensus among analytical chemists on a standard procedure for determining
and reporting the limits of detection and quantitation of analytical procedures.
Furthermore, detection and quantitation limits reported in the literature are
seldom clearly defined. Appendix H clearly illustrates the widespread
inconsistency in defining and reporting limits of detection and quantitation.
Reported detection limits may be based on instrument sensitivity or determined
from the analyses of method blanks or low-level matrix spikes; quantitation limits
may be determined from the analyses of method blanks or low-level matrix
spikes (Puget Sound Estuary Program, 1990d).
8.3.3.3.1 Detection limits
The EPA recommends that the method detection limit (MDL) defined below and
determined according to 40 CFR 136, Appendix B, be used to establish the limits
of detection for the analytical methods used for analyses of all target analytes:
Method Detection Limit (MDL): The minimum concentration of an analyte
in a given matrix (i.e., fish or shellfish tissue homogenates for the purposes
of this guidance) that can be measured and reported with 99 percent
confidence that the concentration is greater than zero. The MDL is
determined by multiplying the appropriate (i.e., n-1 degrees of freedom)
one-sided 99 percent Student's t-statistic (t0 99) by the standard deviation (S)
obtained from a minimum of seven replicate analyses of a spiked matrix
sample containing the analyte of interest at a concentration three to five
times the estimated MDL (Glaser et al., 1981; 40 CFR 136, Appendix B):
8-31
-------
8. LABORATORY PROCEDURES II — SAMPLE ANALYSES
MDL = (to.99) (S). (8-3)
It is important to emphasize that all sample processing steps of the analytical
method (e.g., digestion, extraction, cleanup) must be included in the
determination of the MDL
In addition to the MDL, three other types of detection limits have been defined
by the American Chemical Society Committee on Environmental Improvement
(Keith, 1991 a):
• Instrument Detection Limit (IDL): The smallest signal above background
noise that an instrument can detect reliably.
Limit of Detection (LOD): The lowest concentration that can be determined
to be statistically different from a method blank at a specified level of
confidence. The recommended value for the LOD is three times the
standard deviation of the blank in replicate analyses, corresponding to a 99
percent confidence level.
Reliable Detection Limit (RDL): The concentration level of an analyte in
a given matrix at which a detection decision is extremely likely. The RDL is
generally set higher than the MDL. When RDL=MDL, the risk of a false
positive at 3o from zero is <1 percent, whereas the corresponding risk of a
false negative is 50 percent. When RDL=2MDL, the risk of either a false
positive or a false negative at 3o from zero is <1 percent.
Each of these estimates has its practical limitations. The IDL does not account
for possible blank contaminants or matrix interferences. The LOD accounts for
blank contaminants but not for matrix effects or interferences. In some
instances, the relatively high value of the MDL or RDL may be too stringent and
result in the rejection of valid data; however, these are the only detection limit
estimates that account for matrix effects and interferences and provide a high
level of statistical confidence in sample results. The MDL is the recommended
detection limit in the EPA EMAP-NC Program (U.S. EPA, 1991e).
The MDL, expressed as the concentration of target analyte in fish tissue, is
calculated from the measured MDL of the target analyte in the sample extract
or digestate according to the following equation:
(ppm or ppb) = (MDL^, • V) /W (8-4)
where
V = Final extract or digestate volume, after dilution or concentration (mL)
W = Weight of sample digested or extracted (g).
8-32
-------
8. LABORATORY PROCEDURES II — SAMPLE ANALYSES
Equation (8-4) clearly illustrates that the MDL in tissue may be improved
(reduced) by increasing the sample weight (W) and/or decreasing the final
extract or digestate volume (V).
The initial MDL is a statistically derived empirical value that may differ in actual
samples depending on several factors, including sample size, matrix effects, and
percent moisture. Therefore, it is recommended that each laboratory reevaluate
annually all MDLs for the analytical methods used for the sample matrices
typically encountered (U.S. EPA, 1991e).
Experienced analysts may use their best professional judgment to adjust the
measured MDL to a lower "typically achievable" detection limit (Puget Sound
Estuary Program, 1990e; U.S. EPA, 1985a) or to derive other estimates of
detection limits. For example, EPA recommends the use of lower limits of
detection (LLDs) for GG/MS methods used to analyze organic pollutants in
bioaccumulation monitoring programs (U.S. EPA, 1986b). Estimation of the LLD
for a given analyte involves determining the noise level in the retention window
for the quantitation mass of the analyte for at least three field samples in the
sample set being analyzed. The LLD is then estimated as the concentration
corresponding to the signal required to exceed the average noise level observed
by at least a factor of 2. Based on the best professional judgment of the analyst,
this LLD is applied to samples in the set with comparable or lower interference;
samples with significantly higher interferences (i.e., by at least a factor of 2) are
assigned correspondingly higher LLDs. LLDs are greater than IDLs but usually
are less than the more rigorously defined MDLs. Thus, data quantified between
the LLD and the MDL have a lower statistical confidence associated with them
than data quantified above the MDL. However, these data are considered valid
and useful in assessing low-level environmental contamination.
If estimates of detection limits other than the MDL are developed and used to
qualify reported data, they should be clearly defined in the analytical SOPs and
in all data reports, and their relationship to the MDL should be clearly described.
8.3.3.3.2 Quantitation limits
In addition to the MDL, a method quantitation limit (MQL), or minimum
concentration allowed to be reported at a specified level of confidence without
qualifications, should be derived for each analyte. Ideally, MQLs should account
for matrix effects and interferences. The MQL can be greater than or equal to
the MDL. At present, there is no consistent guidance in the scientific literature
for determining MQLs; therefore, it is not possible to provide specific
recommendations for determining these limits at this time.
The American Chemical Society Committee on Environmental Improvement
(Keith, 1991b; Keith et al., 1983) has defined one type of quantitation limit:
Limit of Quantitation (LOQ): The concentration above which quantitative
results may be obtained with a specified degree of confidence. The
-------
8. LABORATORY PROCEDURES II — SAMPLE ANALYSES
recommended value for the LOQ is 10 times the standard deviation of a
method blank in replicate analyses, corresponding to an uncertainty of ±30
percent in the measured value (10o± 3o) at the 99 percent confidence level.
The LOQ is the recommended quantitation limit in the EPA EMAP-NC Program
(U.S. EPA, 1991e). However, the LOQ does not account for matrix effects or
interferences.
The U.S. EPA (1986d) has defined another type of quantitation limit:
Practical Quantitation Limit (PQL): The lowest concentration that can be
reliably reported within specified limits of precision and accuracy under
routine laboratory operating conditions.
The Puget Sound Estuary Program (1990d) and the National Dioxin Study (U.S.
EPA, 1987d) used a PQL based on the lowest concentration of the initial
calibration curve (C, in ^ig/mL), the amount of sample typically analyzed (W, in
g), and the final extract volume (V, in mL) of that method:
PQL (ng/g;ppm) - CQig/mLWmL) . (8-5)
However, this PQL is also applicable only to samples without substantial matrix
effects or interferences.
A reliable detection limit (RDL) equal to 2 MDL may also be used as an estimate
of the MQL (see Section 8.3.3.3.1). The RDL accounts for matrix effects and
provides a high level of statistical confidence in analytical results.
Analysts must use their expertise and professional judgment to determine the
best estimate of the MQL for each target analyte. MQLs, including the estimated
degree of confidence in analyte concentrations above the quantitation limit,
should be clearly defined in the analytical SOPs and in all data reports.
8.3.3.3.3 Use of detection and quantitation limits In reporting data
The analytical laboratory does not have responsibility or authority to censor data.
Therefore, all data should be reported with complete documentation of limitations
and problems. Method detection and quantitation limits should be used to
qualify reported data for each composite sample as follows (Keith, 1991b):
"Zero" concentration (no observed response) should be reported as not
detected (ND) with the MDL noted, e.g., "ND(MDL=X)".
Concentrations below the MDL should be reported with the qualification that
they are below the MDL.
8-34
-------
8. LABORATORY PROCEDURES II — SAMPLE ANALYSES
Concentrations between the MDL and the MQL should be reported with the
qualification that they are below the quantitation limit.
• Concentrations at or above the MQL may be reported and used without
qualification.
The use of laboratory data for comparing target analyte concentrations to SVs
in screening and intensive studies is discussed in Sections 9.1.1 and 9.1.2.
8.3.3.4 Assessment of Method Accuracy—
The accuracy of each analytical method should be assessed and documented
for each target analyte of interest, in a fish or shellfish tissue matrix, prior to
beginning routine analyses and on a regular basis during routine analyses.
Method accuracy may be assessed by analysis of appropriate reference
materials (i.e., SRMs or CRMs prepared from actual contaminated fish or
shellfish tissue, see Table 8-8 and Appendix I), laboratory control samples
(i.e., accuracy-based samples consisting of fish and shellfish tissue
homogenates spiked with compounds representative of the target analytes of
interest), and/or matrix spikes. If possible, laboratory control samples should
be SRMs or CRMs. Note: Only the analysis of fish or shellfish tissue SRMs or
CRMS prepared from actual contaminated fish or shellfish tissue allows rigorous
assessment of total method accuracy, including the accuracy with which an
extraction or digestion procedure isolates the target analyte of interest from
actual contaminated fish or shellfish. The analysis of spiked laboratory control
samples or matrix spikes provides an assessment of method accuracy including
sample handling and analysis procedures, but does not allow rigorous
assessment of the accuracy or efficiency of extraction or digestion procedures
for actual contaminated fish or shellfish. Consequently, these samples should
not be used for the primary assessment of total method accuracy unless SRMs
or CRMs prepared from actual contaminated fish or shellfish tissue are not
available.
The concentrations of target analytes in samples used to assess accuracy
should fall within the range of concentrations found in the field samples;
however, this may not always be possible for reference materials or laboratory
control samples because of the limited number of these samples available in fish
and shellfish tissue matrices (see Table 8-8 and Appendix I). Matrix spike
samples should be prepared using spike concentrations approximately equal to
the concentrations found in the unspiked samples. An acceptable range of spike
concentrations is 0.5 to 5 times the expected sample concentrations (U.S. EPA,
1987e). Spikes should always be added to the sample homogenates prior to
digestion or extraction.
Accuracy is calculated as percent recovery from the analysis of reference
materials, or laboratory control samples, as follows:
8-35
-------
8. LABORATORY PROCEDURES II — SAMPLE ANALYSES
% Recovery = 100 (M/T) (8-6)
where
M = Measured value of the concentration of target analyte
T = "True" value of the concentration of target analyte.
Accuracy is calculated as percent recovery from the analysis of matrix spike
samples as follows:
% Recovery = [(Ms-Mu)/TJx 100 (8-7)
where
Ms = Measured concentration of target analyte in the spiked sample
Mu = Measured concentration of target analyte in the unspiked sample
Ts = "True" concentration of target analyte added to the spiked sample.
When sample concentrations are less than the MDL, the value of one-half the
MDL should be used as the concentration of the unspiked sample (Ma) in
calculating spike recoveries.
8.3.3.4.1 Initial assessment of method accuracy
As discussed above, method accuracy should be assessed initially by analyzing
appropriate SRMs or CRMs that are prepared from actual contaminated fish or
shellfish tissue. The number of reference samples required to be analyzed for
the initial assessment of method accuracy should be determined by each
laboratory for each analytical procedure with concurrence of the program
manager. If such SRMs or CRMs are not available, laboratory control samples
or matrix spikes may be used for initial assessment of method accuracy.
8.3.3.4.2 Routine assessment of method accuracy
Laboratory control samples and matrix spikes should be analyzed for continuous
assessment of accuracy during routine analyses. It is recommended that one
laboratory control sample and one matrix spike sample be analyzed with every
20 samples or with each sample batch, whichever is more frequent (Puget
Sound Estuary Program, 1990d, 1990e). Ideally, CRMs or SRMs should also
be analyzed at this recommended frequency; however, limited availability and
cost of these materials may make this impractical.
For organic compounds, isotopically labeled or surrogate recovery standards
which must be added to each sample to monitor overall method performance
also provide an assessment of method accuracy (see Section 8.3.3.7.1).
Percent recovery values for spiked samples must fall within established control
limits (see Table 8-6). If the percent recovery falls outside the control limit, the
8-36
-------
8. LABORATORY PROCEDURES II — SAMPLE ANALYSES
analyses should be discontinued, appropriate corrective action taken, and, if
possible, the samples associated with the spike reanalyzed. If reanalysis is not
possible, all suspect data should be clearly identified.
Note: Reported data should not be corrected for percent recoveries. Recovery
data should be reported for each sample to facilitate proper evaluation and use
of analytical results.
Poor performance on the analysis of reference materials or poor spike recovery
may be caused by inadequate mixing of the composite homogenate sample
before aliquotting, inconsistent digestion or extraction procedures, matrix
interferences, or instrumentation problems. If replicate analyses are acceptable
(see Section 8.3.3.5), matrix interferences or loss of target analytes during
sample preparation are indicated. To check for loss of target analytes during
sample preparation, a step-by-step examination of the procedure using spiked
blanks should be conducted. For example, to check for loss of metal target
analytes during digestion, a postdigestion spike should be prepared and
analyzed and the results compared with those from a predigestion spike. If the
results are significantly different, the digestion technique should be modified to
obtain acceptable recoveries. If there is no significant difference in the results
of pre- and postdigestion spikes, the sample should be diluted by at least a
factor of 5 and reanalyzed. If spike recovery is still poor, then the method of
standard additions or use of a matrix modifier is indicated (U.S. EPA, 1987e).
8.3.3.5 Assessment of Method Precision—
The precision of each analytical method should be assessed and documented
for each target analyte prior to the performance of routine analyses and on a
regular basis during routine analysis.
Precision is defined as the agreement among a set of replicate measurements
without assumption of knowledge of the true value. Method precision (i.e., total
variability due to sample preparation and analysis) is estimated by means of the
analyses of duplicate or replicate tissue homogenate samples containing
concentrations of the target analyte of interest above the MDL. All samples used
for assessment of total method precision must be carried through the complete
analytical procedure, including extraction or digestion.
The most commonly used estimates of precision are the relative standard
deviation (RSD) or coefficient of variation (CV) for multiple samples, and the
relative percent difference (RPD) when only two samples are available. These
are defined as follows:
RSD = CV = 100S/x (8-8)
8-37
-------
8. LABORATORY PROCEDURES II — SAMPLE ANALYSES
where
S = Standard deviation of the X| measurements
Xj = Arithmetic mean of the X| measurements, and
RPD = 100 {(x, - XgVRx, + x2)/2]} . (8-9)
8.3.3.5.1 Initial assessment of method precision
Method precision should be assessed prior to routine sample analyses by
analyzing replicate samples of the same reference materials, laboratory control
samples, and/or matrix spikes that are used for initial assessment of method
accuracy (see Section 8.3.3.4.1). The number of replicates required to be
analyzed for the initial assessment of method precision should be determined by
each Jaboratory for each analytical procedure with concurrence of the program
manager. Because precision may be concentration-dependent, initial assess-
ments of precision across the estimated working range should be obtained.
8.3.3.5.2 Routine assessment of method precision
Ongoing assessment of method precision during routine analysis should be
performed by analyzing replicate aliquots of tissue homogenate samples taken
prior to sample extraction or digestion (i.e., laboratory replicates) and matrix
spike replicates. Matrix spike concentrations should approximate unspiked
sample concentrations; an acceptable range for spike concentrations is 0.5 to
5 times the sample concentrations (U.S. EPA, 1987e).
For ongoing assessment of method precision, it is recommended that one
laboratory duplicate and one matrix spike duplicate be analyzed with every 20
samples or with each sample batch, whichever is more frequent. In addition, it
is recommended that a laboratory control sample be analyzed at the above
frequency to allow an ongoing assessment of method performance, including an
estimate of method precision over time. Specific procedures for estimating
method precision by laboratory and/or matrix spike duplicates and laboratory
control samples are given in ASTM (1983). This reference also includes
procedures for estimating method precision from spike recoveries and for testing
for significant change in method precision over time.
Precision estimates obtained from the analysis of laboratory duplicates, matrix
spike duplicates, and repeated laboratory control sample analyses must fall
within specified control limits (see Table 8-7). If these values fall outside the
control limits, the analyses should be discontinued, appropriate corrective action
taken, and, if possible, the samples associated with the duplicates reanalyzed.
If reanalysis is not possible, all suspect data should be clearly identified.
Unacceptable precision estimates derived from the analysis of duplicate or
replicate samples may be caused by inadequate mixing of the sample before
8-38
-------
8. LABORATORY PROCEDURES II — SAMPLE ANALYSES
aliquotting; inconsistent contamination; inconsistent digestion, extraction, or
cleanup procedures; or instrumentation problems (U.S. EPA, 1987e).
8.3.3.5.3 Routine assessment of analytical precision
The analysis of replicate aliquots of final sample extracts or digestates
(analytical replicates) provides an estimate of analytical precision only; it does
not provide an estimate of total method precision. For organic target analytes,
analytical replicates may be included at the discretion of the program manager
or laboratory supervisor. For the analysis of target metal analytes by graphite
furnace atomic absorption spectrophotometry (GFAA) and cold vapor atomic
absorption spectrophotometry (CVAA), it is recommended that duplicate
injections of each sample be analyzed and the mean concentration be reported.
The RPD should be within control limits established by the program manager or
laboratory supervisor, or the sample should be reanalyzed (U.S. EPA, 1987e).
8.3.3.5.4 Assessment of overall variability
Estimates of the overall variability of target analyte concentrations in a sample
fish or shellfish population and of the sampling and analysis procedures can be
obtained by collecting and analyzing field replicates. Replicate field samples
are optional in screening studies; however, if resources permit, it is
recommended that duplicate samples be collected at 10 percent of the screening
sites as a minimal QC check. In intensive studies, replicate samples should be
collected at each sampling site (see Section 6.1.2.7). Although the primary
purpose of replicate field samples in intensive studies is to allow more reliable
estimates of the magnitude of contamination, extreme variability in the results of
these samples may also indicate that sampling and/or analysis procedures are
not adequately controlled.
8.3.3.6 Routine Monitoring of Interferences and Contamination—
Because contamination can be a limiting factor in the reliable quantitation of
target contaminants in tissue samples, the recommendations for proper materials
and handling and cleaning procedures given in Sections 6.2.2 and 7.2 should be
followed carefully to avoid contamination of samples in the field and laboratory.
Many metal contamination problems are due to airborne dust. High zinc blanks
may result from airborne dust or galvanized iron, and high chromium arid nickel
blanks often indicate contamination from stainless steel. Mercury thermometers
should not be used in the field because broken thermometers can be a source
of significant mercury contamination. In the laboratory, samples to be analyzed
for mercury should be isolated from materials and equipment (e.g., polarographs)
that are potential sources of mercury contamination. Cigarette smoke is a
source of cadmium. Consequently, care should be taken to avoid the presence
of cigarette smoke during the collection, handling, processing, and analysis of
samples for cadmium. In organic analyses, phthalates, methylene chloride, and
8-39
-------
8. LABORATORY PROCEDURES II — SAMPLE ANALYSES
toluene are common laboratory contaminants that are often detected in blanks
at concentrations above the MDL (U.S. EPA, 1987e).
Cross-contamination between samples should be avoided during all steps of
analysis of organic contaminants by GC-based methods. Injection micro-
syringes must be cleaned thoroughly between uses. If separate syringes are
used for the injection of solutions, possible differences in syringe volumes should
be assessed and, if present, corrected for. Particular care should be taken to
avoid carryover when high- and low-level samples are analyzed sequentially.
Analysis of an appropriate method blank (see next page) may be required
following the analysis of a high-level sample to assess carryover (U.S. EPA,
1987e).
To monitor for interferences and contamination, the following blank samples
should be analyzed prior to beginning sample collection and analyses and on a
routine basis throughout each study (U.S. EPA, 1987e):
Field blanks are rinsates of empty field sample containers (i.e., aluminum
foil packets and plastic bags) that are prepared, shipped, and stored as
actual field samples. Field blanks should be analyzed to evaluate field
sample packaging materials as sources of contamination. Each rinsate
should be collected and the volume recorded. The rinsate should be
analyzed for target analytes of interest and the total amount of target analyte
in the rinsate recorded. It is recommended that one field blank be analyzed
with every 20 samples or with each batch of samples, whichever is more
frequent.
Processing blanks are rinsates of utensils and equipment used for
dissecting and homogenizing fish and shellfish. Processing blanks should
be analyzed, using the procedure described above for field blanks, to
evaluate the efficacy of the cleaning procedures used between samples. It
is recommended that processing blanks be analyzed at least once at the
beginning of a study and preferably once with each batch of 20 or fewer
samples.
• Bottle blanks are rinsates of empty bottles used to store and ship sample
homogenates. Bottle blanks should be collected after the bottles are
cleaned prior to use for storage or shipment of homogenates. They should
be analyzed, using the procedure described above for field blanks, to
evaluate their potential as sources of contamination. It is recommended that
one bottle blank be analyzed for each lot of bottles or with each batch of 20
or fewer samples, whichever is more frequent.
Method blanks are samples of extraction or digestion solvents that are
carried through the complete analytical procedure, including extraction or
digestion; they are also referred to as procedural blanks. Method blanks
should be analyzed to evaluate contaminants resulting from the total
analytical method (e.g., contaminated glassware, reagents, solvents, column
8-40
-------
8. LABORATORY PROCEDURES II — SAMPLE ANALYSES
packing materials, processing equipment). It is recommended that one
method blank be analyzed with every 20 samples or with each batch of
samples, whichever is more frequent.
Reagent blanks are samples of reagents used in the analytical procedure.
It is recommended that each lot of analytical reagents be analyzed for target
analytes of interest prior to use to prevent a potentially serious source of
contamination. For organic analyses, each lot of alumina, silica gel, sodium
sulfate, or Florasil used in extract drying and cleanup should also be
analyzed for target analyte contamination and cleaned as necessary.
Surrogate mixtures used in the analysis of organic target analytes have also
been found to contain contaminants and the absence of interfering impurities
should be verified prior to use (U.S. EPA, 1987e).
Because the contamination in a blank sample may not always translate into
contamination of the tissue samples, analysts and program managers must use
their best professional judgment when interpreting blank analysis data. Ideally,
there should be no detectable concentration of any target analyte in any blank
sample (i.e., the concentration of target analytes in all blanks should be less than
the MDL). However, program managers may set higher control limits (e.g.,
-------
8. LABORATORY PROCEDURES II — SAMPLE ANALYSES
technique, RRFs used for quantitation may be calculated from measured isotope
ratios in calibration standards and not from instrument internal standards.
However, an instrument internal standard still must be added to the final sample
extract prior to analysis to determine the percent recoveries of isotopically
labeled recovery standards added prior to extraction. Thus, in isotope dilution
methods, instrument internal standards may be used only for QC purposes (i.e.,
to assess the quality of data) and not to quantify analytes. Control limits for the
percent recovery of each isotopically labeled recovery standard should be
established by the program manager, consistent with program data quality
requirements. Control limits for percent recovery and recommended corrective
actions given in EPA Method 1625 (40 CFR 136, Appendix A) should be used
as guidance.
If isotopically labeled analogs of target analytes are not available or if the isotope
dilution technique cannot be used (e.g., for chlorinated pesticides and PCBs
analyzed by GC/ECD), other surrogate compounds should be added as recovery
standards to each sample prior to extraction and to each calibration standard.
These surrogate recovery standards should have chemical and physical
properties similar to the target analytes of interest and should not be expected
to be present in the original samples. Recommended surrogate recovery
standards are included in the methods referenced in Table 8-2 and in EMMI
(U.S. EPA, 1991f).
Samples to which surrogate recovery standards have been added are termed
surrogate spikes. The percent recovery of each surrogate spike (% Rs) should
be determined for all samples as follows:
%Rs=100(Cm/Ca) (8-10)
where
% Rs = Surrogate spike percent recovery
Cm = Measured concentration of surrogate recovery standard
Ca = Actual concentration of surrogate recovery standard added to the
sample.
Control limits for the percent recovery of each surrogate spike should be
established by the program manager consistent with program data quality
requirements. The control limits in the most recent EPA CLP methods (U.S.
EPA, 1991c) are recommended for evaluating surrogate recoveries.
Note: Reported data should not be corrected for percent recoveries of surrogate
recovery standards. Recovery data should be reported for each sample to
facilitate proper evaluation and use of the analytical results.
8-42
-------
8. LABORATORY PROCEDURES II — SAMPLE ANALYSES
8.3.3.7.2 Other performance evaluation procedures
The following additional procedures are required to evaluate the performance of
GC-based analytical systems prior to the routine analysis of field samples (U.S.
EPA, 1989c; U.S. EPA, 1991c). It is the responsibility of each program manager
to determine specific evaluation procedures and control limits appropriate for
their data quality requirements.
Evaluation of the GC System
GC system performance should be evaluated by determining the number of
theoretical plates of resolution and the relative retention times of the internal
standards.
Column Resolution: The number of theoretical plates of resolution, N,
should be determined at the time the calibration curve is generated (using
chrysene-d10) and monitored with each sample set. The value of N should
not decrease by more than 20 percent during an analysis session. The
equation for N is given as follows:
N = 16(RT/W)2 (8-11)
where
RT = Retention time of chrysene-d10 (s)
W = Peak width of chrysene-d10 (s).
Relative Retention Time: Relative retention times of the internal standards
should not deviate by more than ±3 percent from the values calculated at the
time the calibration curve was generated.
If the column resolution or relative retention times are not within the specified
control limits, appropriate corrective action (e.g., adjust GC parameters, flush GC
column, replace GC column) should be taken.
Evaluation of the MS System
The performance of the mass spectrometer should be evaluated for sensitivity
and spectral quality.
Sensitivity: The signal-to-noise value should be at least 3.0 or greater for
m/z 198 from an injection of 10 ng decafluorotriphenylphosphine (DFTPP).
Spectral Quality: The intensity of ions in the spectrum of a 50-ng injection
of DFTPP should meet the following criteria (U.S. EPA, 1991c):
8-43
-------
8. LABORATORY PROCEDURES II — SAMPLE ANALYSES
m/z Criteria
51 30-80% mass 198
68 <2% mass 69
69 present
70 <2% mass 69
127 25-75% mass 198
197 <1% mass 198
198 base peak, 100% relative abundance
199 5-9% mass 198
275 10-30% mass 198
365 >0.75% mass 198
441 present and
-------
8. LABORATORY PROCEDURES II — SAMPLE ANALYSES
8.3.3.8.1 Round-robin analysis Intel-laboratory comparison program
At present, the only external round-robin QA program available for analytical
laboratories conducting fish and shellfish tissue analyses for environmental
pollutants is administered by NOAA in conjunction with its National Status and
Trends (NS&T) Program (Cantillo, 1991). This QA program has been designed
to ensure proper documentation of sampling and analysis procedures and to
evaluate both the individual and collective performance of participating
laboratories. Recently, NOAA and the EPA have agreed to conduct the NS&T
Program and the EMAP-NC Program as a coordinated effort. As a result,
EMAP-NC now cosponsors and cooperatively funds the NS&T QA Program, and
the interlaboratory comparison exercises include all EMAP-NC laboratories (U.S.
EPA, 1991e).
Note: Participation in the NS&T QA program by all laboratories performing
chemical analyses for State fish and shellfish contaminant monitoring programs
is recommended to enhance the credibility and comparability of analytical data
among the various laboratories and programs.
Each laboratory participating in the NS&T QA program is required to
demonstrate its analytic capability prior to the analysis of field samples by the
blind analysis of a fish and shellfish tissue sample that is uncompromised,
homogeneous, and contains the target analytes of interest at concentrations of
interest. A laboratory's performance generally will be considered acceptable if
its reported results are within ±30 percent (for organics) and ±15 percent (for
metals) of the actual or certified concentration of each target analyte in the
sample (U.S. EPA, 1991e). If any of the results exceed these control limits, the
laboratory will be required to repeat the analysis until all reported results are
within the control limits. Routine analysis of field samples will not be allowed
until initial demonstration of laboratory capability is acceptable.
Following the initial demonstration of laboratory capability, each participating
laboratory is required to participate in one intercomparison exercise per year as
a continuing check on performance. This intercomparison exercise includes both
organic and inorganic (i.e., trace metals) environmental and standard reference
samples. The organic analytical intercomparison program is coordinated by
NIST, and the inorganic analytical intercomparison program is coordinated by the
NRCC. Sample types and matrices vary yearly. Performance evaluation
samples used in the past have included accuracy-based solutions, sample
extracts, and representative matrices (e.g., tissue or sediment samples).
Laboratories are required to analyze the performance evaluation samples blind
and to submit their results to NIST or NRCC, as instructed. Individual laboratory
performance is evaluated against the consensus values (i.e., grand means) of
the results reported by all participating laboratories. Laboratories that fail to
achieve acceptable performance must take appropriate corrective action. NIST
and NRCC will provide technical assistance to participating laboratories that have
problems with the intercomparison analyses. At the end of each calendar year,
the results of the intercomparison exercises are reviewed at a workshop
8-45
-------
8. LABORATORY PROCEDURES II — SAMPLE ANALYSES
sponsored by NIST and NRCC. Representatives from each laboratory are
encouraged to participate in these workshops, which provide an opportunity for
discussion of analytical problems encountered in the intercomparison exercises.
Note: Nonprofit laboratories (e.g., EPA and other Federal laboratories, State,
municipal, and nonprofit university laboratories) may participate in the NS&T QA
program at no cost on a space-available basis. In 1993, the estimated cost of
participation in the NIST Intercomparison Exercise Program for Organic Contami-
nants in the Marine Environment will be $2,000 and $2,300 for private labora-
tories within and outside the United States, respectively. This cost covers
samples for one exercise per year. Samples may be obtained directly from NIST
by contacting Ms. Reenie Parris, NIST, Chemistry B158, Gaithersburg, MD
20899; Tel:301-975-3103, FAX:301-926-8671. At present, the cost of
participation in trace inorganic exercises by private laboratories has not been
established. Once this cost has been set, trace inorganic samples will be
available directly from NRCC.
To obtain additional information about participation in the NS&T QA program,
contact Dr. Adriana Cantillo, QA Manager, NOAA/National Status and Trends
Program, N/ORCA21, Rockville, MD 20852, Tel: 301-443-8655.
8.3.3.8.2 Split sample analysis Interlaboratory comparison programs
Another useful external QA procedure for assessing interlaboratory comparability
of analytical data is a split-sample analysis program in which a percentage
(usually 5 to 10 percent) of all samples analyzed by each State or Region are
divided and distributed for analyses among laboratories from other States or
Regions. Because actual samples are used in a split-sample analysis program,
the results of the split-sample analyses provide a more direct assessment of the
comparability of the reported results from different States or Regions.
The NS&T QA program does not include an interlaboratory split-sample analysis
program. However, it is recommended that split-sample analysis programs be
established by States and/or Regions that routinely share results.
8.4 DOCUMENTATION AND REPORTING OF DATA
The results of all chemical analyses must be documented adequately and
reported properly to ensure the correct evaluation and interpretation of the data.
8.4.1 Analytical Data Reports
The documentation of analytical data for each sample should include, at a
minimum, the following information:
• Study identification (e.g., project number, title, phase)
8-46
-------
8. LABORATORY PROCEDURES II — SAMPLE ANALYSES
Description of the procedure used, including documentation and justification
of any deviations from the standard procedure
Method detection and quantitation limits for each target analyte
Method accuracy and precision for each target analyte
Discussion of any analytical problems and corrective action taken
Sample identification number
Sample weight (wet weight)
Final dilution volume
Date(s) of analysis
Identification of analyst
Identification of instrument used (manufacturer, model number, serial
number, location)
Summary calibration data, including identification of calibration materials,
dates of calibration and calibration checks, and calibration range(s); for
GC/MS analyses, include DFTPP and bromofluorobenzene (BFB) spectra
and quantitation report
Reconstructed ion chromatograms for each sample analyzed by GC/MS
Mass spectra of detected target compounds for each sample analyzed by
GC/MS
Chromatograms for each sample analyzed by GC/ECD and/or GC/FID
Raw data quantitation reports for each sample
Description of all QC samples associated with each sample (e.g., reference
materials, field blanks, rinsate blanks, method blanks, duplicate or replicate
samples, spiked samples, laboratory control samples) and results of all QC
analyses. QC reports should include quantitation of all target analytes in
each blank, recovery assessments for all spiked samples, and replicate
sample summaries. Laboratories should report all surrogate and matrix spike
recovery data for each sample; the range of recoveries should be included
in any reports using these data.
Analyte concentrations with reporting units identified (as ppm or ppb wet
weight, to two significant figures unless otherwise justified). Note: Reported
data should not be recovery- or blank-corrected.
8-47
-------
8. LABORATORY PROCEDURES II — SAMPLE ANALYSES
Lipid content (as percent wet weight)
Specification of all tentatively identified compounds (if requested) and any
quantitation data.
Data qualifications (including qualification codes and their definitions, if
applicable, and a summary of data limitations).
To ensure completeness and consistency of reported data, standard forms
should be developed and used by each laboratory for recording and reporting
data from each analytical method. Standard data forms used in the EPA
Contract Laboratory Program (U.S. EPA, 1991b, 1991 c) may serve as useful
examples for analytical laboratories.
All analytical data should be reviewed thoroughly by the analytical laboratory
supervisor and, ideally, by a qualified chemist who is independent of the
laboratory. In some cases, the analytical laboratory supervisor may conduct the
full data review, with a more limited QA review provided by an independent
chemist. The purpose of the data review is to evaluate the data relative to data
quality specifications (e.g., detection and quantitation limits, precision, accuracy)
and other performance criteria established in the Work/QA Project Plan. In many
instances, it may be necessary to qualify reported data values; qualifiers should
always be defined clearly in the data report.
8.4.2 Summary Reports
Summaries of study data should be prepared for each target species at each
sampling site. Specific recommendations for reporting data for screening and
intensive studies are given in Section 9.2.
8-48
-------
9. DATA ANALYSIS AND REPORTING
SECTION 9
DATA ANALYSIS AND REPORTING
This section provides guidance on (1) analysis of laboratory data for both
screening and intensive studies that should be included in State data reports and
(2) data reporting requirements for a national database (National Fish Tissue
Data Repository) for fish and shellfish contaminant monitoring programs.
All data analysis and reporting procedures should be documented fully as part
of the Work/QA Project Plan for each study, prior to initiating the study (see
Appendix E). All routine data analysis and reporting procedures should be
described in standard operating procedures. In particular, the procedures to be
used to determine if the concentration of a target analyte in fish or shellfish
tissue differs significantly from the selected SV must be clearly documented.
9.1 DATA ANALYSIS
9.1.1 Screening Studies
The primary objective of Tier 1 screening studies is to assist States in identifying
potentially contaminated harvest areas where further investigation of fish and
shellfish contamination may be warranted. The criteria used to determine
whether the measured target analyte concentration in a fish or shellfish tissue
composite sample is different from the SV (greater than or less than) should be
clearly documented. If a reported target analyte concentration exceeds the SV
in the screening study, a State should initiate a Tier 2, Phase I, intensive study
(see Section 6.1.2.1) to verify the level of contamination in the target species.
Because of resource limitations, some States may choose to conduct a risk
assessment using screening study data; however, this approach is not
recommended because a valid statistical analysis cannot be performed on a
single composite sample. If a reported analyte concentration is close to the SV
but does not exceed the SV, the State should reexamine historic data on water,
sediment, and fish tissue contamination at the site, and evaluate data on
laboratory performance. If these data indicate that further examination of the site
is warranted, the State should initiate a Tier 2, Phase I, intensive study to verify
the magnitude of the contamination.
Because replicate composite samples are not required as part of a screening
study, estimating the variability of the composite target analyte concentration at
any site is precluded. The following procedure is recommended for use by
9-1
-------
9. DATA ANALYSIS AND REPORTING
States for analysis of the individual target analyte concentration for each
composite sample from reported laboratory data (see Section 8.3.3.3)
• A datum reported below the method detection limit (MDL), including a datum
reported as not detected (i.e., ND, no observed response) should be
assigned a value of one-half the MDL.
A datum reported between the MDL and the method quantitation limit (MQL)
should be assigned a value of the MDL plus one-half the difference between
the MQL and the MDL
A datum reported at or above the MQL should be used as reported.
This approach is similar to that published in 40 CFR Parts 122, 123, 131, and
132—Proposed Water Quality Guidance for the Great Lakes System.
i
If resources permit and replicate composite samples are collected at a suspected
site of contamination, then a State may conduct a statistical analysis of
differences between the mean target analyte concentration and the SV, as
described in Section 9.1.2.
9.1.2 Intensive Studies
The primary objectives of Tier 2 intensive studies are to confirm the findings of
the screening study by assessing the magnitude and geographic extent of the
contamination in various size classes of selected target species. The EPA Office
of Water recommends that States collect replicate composite samples of three
size classes of each target species in the study area to verify whether the mean
target analyte concentration of replicate composite samples for any size class
exceeds the SV for any target analyte identified in the screening study.
The following procedure is recommended for use by States in calculating the
mean arithmetic target analyte concentration from reported laboratory data (see
Section 8.3.3.3.3).
• Data reported below the method detection limit (MDL), including data
reported as not detected (i.e., ND, no observed response) should be
assigned a value of one-half the MDL.
• Data reported between the MDL and the method quantitation limit (MQL)
should be assigned a value of the MDL plus one-half the difference between
the MQL and the MDL.
Data reported at or above the MQL should be used as reported.
This approach is similar to that published in 40 CFR Parts 122, 123, 131, and
132—Proposed Water Quality Guidance for the Great Lakes System.
9-2
-------
9. DATA ANALYSIS AND REPORTING
State staff should consult a statistician in interpreting intensive study tissue
residue results to determine the need for additional monitoring, risk assessment,
and issuance of a fish or shellfish consumption advisory. Additional information
on risk assessment, risk management, and risk communication procedures will
be provided in later volumes in this guidance series.
9.2 DATA REPORTING
9.2.1 State Data Reports
State data reports should be prepared by the fish contaminant monitoring
program manager responsible for designing the screening and intensive studies.
Summaries of Tier 1 screening study data should be prepared for each target
species sampled at each screening site. For Tier 2 intensive studies (Phase I
and Phase II), data reports should be prepared for each target species (by size
class, as appropriate) at each sampling site within the waterbody under
investigation (see Section 6.1.2). Screening and intensive study data reports
should include, at a minimum, the information shown in Figure 9-1.
9.2.2 Reports to the National Fish Tissue Data Repository
The Ocean Data Evaluation System (ODES) database, managed by the EPA
Office of Water, is a primary source for maintaining, retrieving, and analyzing
freshwater, estuarine, and marine data. The EPA Office of Water selected the
ODES database to serve as a national repository for fish and shellfish
contaminant monitoring data for both inland and coastal waters. The National
Fish Tissue Data Repository (NFTDR) is a collection of fish and shellfish
contaminant monitoring data gathered by various Federal, State, and local
agencies. The NFTDR was established to facilitate exchange of fish and
shellfish contaminant monitoring data nationally.
ODES is an integrated database management system that contains analytically
powerful, user-friendly software that allows users to store, access, and analyze
various types of environmental data. ODES resides on an IBM 9000 mainframe
computer at the EPA's National Computer Center in North Carolina. Users can
access ODES by telephone toll free from anywhere in the United States using
almost any type of personal computer or mainframe computer terminal.
ODES provides a variety of features to aid users in storing and analyzing data:
• The user interface makes ODES easy to access and operate. No knowl-
edge of computer programming is required to operate the menu-driven
system.
• The statistical, graphical, and modeling tools enable the user to perform a
wide variety of statistical tests, conduct model simulations, and produce
reports and graphics.
9-3
-------
9. DATA ANALYSIS AND REPORTING
• Study identification (e.g., project number, title, and study type)
• Program manager
• Sampling site name
• Latitude (in degrees, minutes, and seconds)
• Longitude (in degrees, minutes, and seconds)
• Type of waterbody (lake, river, estuary, etc.)
• Name of waterbody
• Sampling date (e.g., DD, MM, YY)
• Sampling time (e.g., HH, MM in a 24-h format)
• Sampling gear type used (e.g., dredge, seine, trawl)
• Sampling depth
• Scientific name of target species
• Common name of target species
• Composite sample numbers
• Number of individuals in each composite sample
• Number of replicate composite samples
• Predominant characteristics of specimens used in each composite sample
- Predominant life stage of individuals in composite
- Predominant sex of individuals in composite (if applicable)
- Average age of individuals in composite (if applicable)
- Average body length or size (mm)
- Description of edible portion (tissue type)
(continued)
Figure 9-1. Recommended data reporting requirements for screening
and Intensive studies.
9-4
-------
9. DATA ANALYSIS AND REPORTING
Analytical methods used (including method for lipid analysis)
Method detection and quantitation limits for each target analyte
Sample cleanup procedures (e.g., additional steps taken to further purify the
sample extracts or digestates)
Data qualifiers (e.g., additional qualifying information about the
measurement)
Percent lipid (wet weight basis) in each composite sample
For each target analyte in each composite sample:
- Total wet weight of composite sample (g) used in analysis
- Measured concentration (wet weight basis) as reported by the laboratory
(see Section 8.3.3.3.3)
- Units of measurement for target analyte concentration
- Evaluation of laboratory performance (i.e., description of all QA and QC
samples associated with the sample(s) and results of all QA and QC
analyses)
In screening studies with only one composite sample for each target
species, the State should provide for each target analyte a comparison of
reported concentration with selected SV and indication of whether SV was
exceeded (see Section 9.1.1).
In intensive studies, for each target analyte in each set of replicate
composite samples, the State should provide
- Range of target analyte concentrations for each set of replicate
composite samples
- Mean (arithmetic) target analyte concentration for each set of replicate
composite samples (see Section 9.1.2)
- Standard deviation of mean target analyte concentration
- Comparison of target analyte arithmetic mean concentration with selected
SV and indication of whether SV was exceeded.
Figure 9-1 (continued)
9-5
-------
9. DATA ANALYSIS AND REPORTING
• The data in ODES are subjected to extensive QA and QC checks.
A QA and QC report describing the data collection methods and procedures
is produced. This report may be accessed online for each data set.
• An ODES data set can be easily downloaded to a personal computer or
routed to a mainframe file in ASCII format for use with software packages
such as SAS, SPSS, LOTUS 1 -2-3, or ARC/INFO (a Geographic Information
System Software).
• A bridge exists between ODES and STORET, the EPA's Water Quality
Storage and Retrieval System, allowing the user to download or analyze
STORET water quality data using several of ODES statistical, graphical, and
modeling tools.
For reporting purposes, State fish and shellfish contaminant monitoring data from
screening and intensive monitoring studies are equivalent and, when submitted
to ODES, should not be separated.
Further information on the ODES database is contained in the following
documents:
1. ODES User Manual- This document provides background information on the
National Fish Tissue Data Repository and ODES database and provides
detailed examples of how to use the reporting and analysis software features
available in ODES.
2. ODES 123 Data Submissions Manual - This document explains how to
submit data, lists which data fields to include in the submission file, provides
specifications for each data field, and includes an easy-to-use Lotus 1-2-3
template for data entry. Two versions of this template are available—one for
Lotus 1-2-3 for Windows and one for Lotus 1-2-3 V.3 or higher.
State, regional, and local agency staff may obtain copies of these documents
from
NFTDR Data Manager
Fish Contamination Section
U.S. Environmental Protection Agency
401 M Street, SW (WH-585)
Washington, DC 20460
9-6
-------
10. LITERATURE CITED
SECTION 10
LITERATURE CITED
Abbott, R.T. 1974. American Seashells-The Marine Molluscs of the Atlantic
and Pacific Coasts of North America. 2nd Edition. Van Nostrand Reinhold
Company, New York, NY.
Agocs, M.M., R.A. Etzel, R.G. Parrish, D.C. Paschal, P.R. Campagna, D.S.
Cohon, E.M. Kilbourne, and J.L Hesse. 1990. Mercury exposure from
interior latex paint. New England Journal of Medicine 323(16):1096-1101.
Anderson, R.O., and S.J. Gutreuter. 1983. Length, weight, and associated
structural indices, pp. 283-300. In: Fisheries Techniques. L.A. Nielson and
D.L. Johnson (eds). American Fisheries Society, Bethesda, MD.
Armbruster, G., K. Gerow, W. Gutenmann, C. Littmann, and D. Lisk. 1987. The
effects of several methods of fish preparation on residues of polychlorinated
biphenyls and sensory characteristics in striped bass. J. Food Safety 8:235-
243.
Armbruster, G., K.L Gall, W.H. Gutenmann, and D.J. Lisk. 1989. Effects of
trimming and cooking by several methods on polychlorinated biphenyls
(PCBs) residues in bluefish. J. Food Safety 9:235-244.
ASTM (American Society for Testing and Materials). 1976. ASTM Manual on
Presentation of Data and Control Chart Analysis. ASTM STP-ILSD.
Committee E-11, Philadelphia, PA.
ASTM (American Society for Testing and Materials). 1983. Standard Practice
for Intralaboratory Quality Control Procedures and a Discussion on Reporting
Low-Level Data. D4210-83. Committee D-19, Philadelphia, PA.
ATSDR (Agency for Toxic Substances and Disease Registry). 1987a. Draft
Toxicological Profile for Aldrin/Dieldrin. U.S. Public Health Service,
Washington, DC.
ATSDR (Agency for Toxic Substances and Disease Registry). 1987b. Draft
Toxicological Profile for Heptachlor. U.S. Public Health Service, Washington,
DC.
10-1
-------
10. LITERATURE CITED
ATSDR (Agency for Toxic Substances and Disease Registry). 1987c.
Toxicological Profile for Selected PCBs. U.S. Public Health Service,
Washington, DC.
ATSDR (Agency for Toxic Substances and Disease Registry). 1987d.
Toxicological Profile for 2,3,7,8-TCDD (Dioxin). Draft. U.S. Public Health
Service in collaboration with the U.S. Environmental Protection Agency,
Washington, DC.
ATSDR (Agency for Toxic Substances and Disease Registry). 1988.
Toxicological Profile for Lead. Draft. Prepared by Technical Resources, Inc.
Oak Ridge National Laboratory, Oak Ridge, TN.
ATSDR (Agency for Toxic Substances and Disease Registry). 1989a.
Toxicological Profile for Arsenic. ATSDR/TP-88/02. Prepared by Life
Systems, Inc., for ATSDR. U.S. Public Health Service in collaboration with
U.S. Environmental Protection Agency, Washington, DC.
ATSDR (Agency for Toxic Substances and Disease Registry). 1989b.
Toxicological Profile for Hexachlorobenzene. Draft. Prepared by Life
Systems, Inc., for ATSDR. U.S. Public Health Service in collaboration with
U.S. Environmental Protection Agency, Washington, DC.
ATSDR (Agency for Toxic Substances and Disease Registry). 1990.
Toxicological Profile for Endosulfan. Draft. Prepared by Clement
Associates, Inc., for ATSDR, Atlanta, GA.
Bache, C.A., W.H. Gutenmann, and DJ. Lisk. 1971. Residues of total mercury
and methylmercuric salts in lake trout as a function of age. Science
172:951.
Ballschmitter, K., and M. Zell. 1980. Analysis of polychlorinated biphenyls
(PCBs) by glass capillary gas chromatography, composition of technical
aroclpr-and clophen-PCB mixtures. Fresenius Anal. Chem. 302:20-31.
Barnes, D.G., and J.S. Bellin. 1989. Interim Procedures for Estimating Risks
Associated with Exposures to Mixtures of Chlorinated Dibenzo-p-Dioxins and
-Dibenzofurans (CDDs and CDFs). Risk Assessment Forum, U.S.
Environmental Protection Agency, Washington, DC.
Battelle Memorial Institute. 1975. Environmental Impact Monitoring of Nuclear
Power Plants: Source Book of Monitoring Methods. Volume 2. Atomic
Industrial Forum, Inc., Washington, DC.
10-2
-------
10. LITERATURE CITED
Battelle Memorial Institute. 1989. Work/Quality Assurance Project Plan for the
National Status and Trends Mussel Watch Program. Collection of Bivalve
Molluscs and Superficial Sediments for Coastal U.S. Atlantic and Pacific
Locations and Analyses for Organic Chemicals and Toxic Trace Elements.
Prepared for U.S. Department of Commerce, National Oceanic and
Atmospheric Administration by Battelle Ocean Sciences, Duxbury, MA.
Battelle New England Marine Research Laboratory. 1986. Phase II Mussel
Watch Field Manual. NOAA Contract No. 50-DGNC-5-0263. Prepared for
U.S. Department of Commerce. National Oceanic and Atmospheric
Administration by Battelle Ocean Sciences. Duxbury, MA.
Benkert, K.A. 1992. Contaminant Assessment of Biota and Sediments in the
Albemarle-Pamlico Region. Contract 40190-0-6823. Prepared for U.S. Fish
and Wildlife Service, Raleigh, NC.
Bennett, G.W. 1970. Management of Lakes and Ponds. Van Nostrand
Reinhold Company, New York, NY.
Bligh, E.G., and W.J. Dyer. 1959. A rapid method of total lipid extraction and
purification. Can J. Biochem. Physiol. 37:911-917.
Borneff, J., and H. Kunte. 1965. Carcinogenic substances in water and soil.
Part XVII. Concerning the origin and estimation of the polycyclic aromatic
hydrocarbons in water. Arch: Hyg. (Berlin) 149:226-243.
Broutman, M.A., and D.L. Leonard. 1988. The Quality of Shellfish Growing
Waters in the Gulf of Mexico. Strategic Assessment Branch, National
Oceanic and Atmospheric Administration, U.S. Department of Commerce,
Rockville, MD.
Brown, D.W., A.J. Friedman, and W.D. MacLeod, Jr. 1985. Quality Assurance
Guidelines for Chemical Analysis of Aquatic Environmental Samples.
Prepared for Seattle District, U.S. Army Corps of Engineers, Seattle,
Washington. National Analytical Facility, National Oceanographic and
Atmospheric Administration, Seattle, WA.
Bryan, A.M., W.B. Stone, and P.G. Olafsson. 1987. Disposition of toxic PCB
congeners in snapping turtle eggs: expressed as toxic equivalents of TCDD.
Bull. Environ. Contam. Toxicol. 39(5):791-796.
California Department of Fish and Game. 1990. Laboratory Quality Assurance
Program Plan. Environmental Services Division, Sacramento, CA.
California Environmental Protection Agency. 1991. A Study of Chemical
Contamination of Marine Fish from Southern California. Office of
Environmental Health Hazard Assessment, Sacramento, CA.
10-3
-------
10. LITERATURE CITED
Cantillo, A.Y. 1991. Reference materials for marine science. Sea Technology
May:45-47.
Capuzzo, J.M., J.W. Farrington, G.T. Wallace, and A.E. McElroy. 1990.
Chemical Contaminants in Fish and Shellfish: Development of Uniform
Testing and Reporting Standards. Project No. NA89-EA-D-00014. Prepared
for National Oceanic and Atmospheric Administration. Woods Hole
Oceanographic Institution, Woods Hole, MA.
Carlander, K.D. 1969. Handbook of Freshwater Fishes of the United States and
Canada. 3rd Ed. Iowa State University Press, Ames, IA.
Cichy, R.F., M.E. Zabik, and C.M. Weaver. 1979. Polychlorinated biphenyl
reduction in lake trout by irradiation and broiling. Bull. Environ. Contam.
Tpxicol. 22:807-812.
Clarkson, T.W. 1990. Human health risks from methyl mercury in fish.
Environmental Toxicology and Chemistry 9:957-961.
Cochran, W.G. 1963. Sampling Techniques. John Wiley & Sons, New York,
NY.
Cochran, W.G. 1977. Sampling Techniques. John Wiley & Sons, New York,
NY.
Crawford, J.K., and S.N. Luoma. 1993. Guidelines for Studies of Contaminants
in Biological Tissues for the National Water-Quality Assessment Program.
USGS Open-File Report 92-494. U.S. Geological Survey, Lemoyne, PA.
Crecelius, E.D. 1978. Modification of the arsenic speciation technique using
hydride generation. Anal. Chem 50(6):826-827.
Crump, K.S., D.G. Hoel, C.H. Langley, and R. Peto. 1976. Fundamental
carcinogenic processes and their implications for low dose risk assessment.
Cancer Res. 36:2973-2979.
Cunningham, P.A. 1979. The use of bivalve molluscs in heavy metal pollution
research: \r\MarinePollution: Functional Responses. W.B. Vernberg, F.P.
Thurberg, A. Calabrese and F.J. Vernberg (eds.). Academic Press, New
York, NY.
Cunningham, P.A., J.M. McCarthy, and D. Zeitlin. 1990. Results of the 1989
Census of State Fish/Shellfish Consumption Advisory Programs. Prepared
for Assessment and Watershed Protection Division, Office of Water
Regulations and Standards, U.S. Environmental Protection Agency.
Research Triangle Institute, Research Triangle Park, NC.
10-4
-------
10. LITERATURE CITED
Cunningham, P.A., and C.O. Whitaker. 1989. A Survey of the Status of
Biomonitoring in State NPDES and Nonpoint Source Monitoring Programs.
Prepared for the Office of Policy, Planning, and Evaluation, U.S.
Environmental Protection Agency. Research Triangle Institute, Research
Triangle Park, NC.
Czapla, T.C., M.E. Pattillo, D.M. Nelson, and M.E. Monaco. 1991. Distribution
and Abundance of Fishes and Invertebrates in Central Gulf of Mexico
Estuaries. ELMR Report No. 7. Strategic Assessment Branch, National
Oceanic and Atmospheric Administration, U.S. Department of Commerce,
Rockville, MD.
Dar, E., M.S. Kanarek, H.A. Anderson, and W.C. Sonzogni. 1992. Fish
consumption and reproductive outcomes in Green Bay Wisconsin.
Environmental Research. 59(1 ):189-201.
Dourson, M.L., and J.M. Clark. 1990. Fish consumption advisories: Toward a
unified, scientifically credible approach. Regulatory Toxicology and
Pharmacology 12:161 -178.
Dunn, W.J., III, D.L Stallings, T.R. Schwartz, J.W. Hogan, J.D. Petty, E.
Johanson, and S. Wold. 1984. Pattern recognition for classification and
determination of polychlorinated biphenyls in environmental samples. Anal.
Chem. 56:1308-1313.
Durfee, R.L., G. Contos, F.C. Whitmore, J.D. Borden, E.E. Hackman, and R.A.
Westin. 1976. PCBs in the United States; Industrial Use and Environmental
Distributions. Office of Toxic Substances, U.S. Environmental Protection
Agency, Washington, DC.
Duston, N.M., C.A. Batdorf, and J.P. Schwartz. 1990. Progress Report: Metal
Concentrations in Marine Fish and Shellfish from Boston and Salem
Harbors, and Coastal Massachusetts. Executive Office of Environmental
Affairs, Department of Fisheries, Wildlife, and Environmental Law
Enforcement, Division of Marine Fisheries, Salem, MA.
Eadie, B.J., W.R. Faust, P.F. Landrum et al. 1983. Bioconcentrations of PAH
by some benthic organisms of the Great Lakes, pp. 437-449. In:
Polynuclear Aromatic Hydrocarbons: Formation, Metabolism and
Measurement. M. Cooke, A.J. Dennis, (eds.) Battelle Press, Columbus,
OH.
Eichers, T.R., P.A. Andrilenas, and T.W. Anderson. 1978. Farmers Use of
Pesticides in 1976. Agric. Econ. Rep. No. 41B. Economics Statistics and
Cooperative Service, U.S. Department of Agriculture, Washington, DC.
10-5
-------
10. LITERATURE CITED
Eisler, R. 1986. Polychlorinated Biphenyl Hazards to Fish, Wildlife and
Invertebrates: A Synoptic Review. U.S. Fish and Wildlife Services Biol.
Rep. 85(1.7). Patuxent Wildlife Research Center, Laurel, MD. 72 pp.
Eisler, R. 1987. Polycyclic Aromatic Hydrocarbon Hazards to Fish, Wildlife and
Invertebrates: A Synoptic Review. U.S. Fish and Wildlife Services Biol.
Rep. 85(1.14). Patuxent Wildlife Research Center, Laurel MD. 134 pp.
Emmett, R.L, S.A. Hinton, S.L. Stone, and M.E. Monaco. 1991. Distribution
and Abundance of Fishes and Invertebrates in West Coast Estuaries.
Volume II: Life History Summaries. ELMR Report No. 8. Strategic
Assessment Division, National Oceanic and Atmospheric Administration,
Rockville, MD.
Erickson, M.D. (ed.). 1991. Analytical Chemistry of PCBs. OCLC No.
24542805. Lewis Publishers, Boca Raton, FL
Farrow, D.R., A.S. Pait, and D.J. Basta. 1989. Targeting pesticide hotspots in
estuarine watersheds. In: Coastal Zone '89, Proceedings of the Sixth
Symposium on Coastal and Ocean Management, Charleston, SC.
Farm Chemicals Handbook. 1989. Meister Publishing Company, Willoughby,
OH.
Fein, G.C., J.L. Jacobson, P.M. Schwarz, and J.K. Dowler. 1984. Prenatal
exposure to polychlorinated biphenyls: Effects on birth size and gestation
age. Journal of Pediatrics 150(2):315-20.
Foran, J.A., and D. Vander-Ploeg. 1989. Consumption advisories for sport fish
in the Great Lakes Basin: Jurisdictional inconsistencies. J. Great Lakes
Res. 1513:476-485.
Foran, J.A., M. Cox, and D. Croxton. 1989. Sport fish consumption advisories
and projected cancer risks in the Great Lakes basin. Amer. J. Public Health
79(3):322-325.
Friberg, L., and D. Vostal (eds.). 1972. Mercury in the Environment. The
Chemical Rubber Company, CRC Press, Cleveland, OH.
Gall, K.L., and M. Voiland. 1990. Contaminants in Sportfish: Managing Risks.
Cornell Cooperative and Sea Grant Extension Fact Sheet. Cornell
University, Ithaca, NY.
Galtsoff, P.S. 1964. The American Oyster (Crassotrea virginica Gmelin).
Fishery Bulletin of the Fish and Wildlife Service, Volume 64. United States
Department of the Interior, Washington, DC.
10-6
-------
10. LITERATURE CITED
Gilbert, P.O. 1987. Statistical Methods for Environmental Pollution Monitoring.
Van Nostrand Reinhold Company, New York, NY.
Glaser, J.A., D.L Forest, G.D. McKee, S. Quave, and W.L. Budde. 1981. Trace
analyses for wastewaters. Environ. Sci. Technol. 15:1425-1450.
Glass, G.E., J.A. Sorenson, K.W. Schmidt, and G.R. Rapp. 1990. New source
identification of mercury contamination in the Great Lakes. Environ. Sci.
Technol. 24:1059-1069.
Goede, R.W., and B.A. Barton. 1990. Organismic indices and an autopsy-
based assessment as indicators of health and condition of fish, pp., 93-108.
In: American Fisheries Society Symposium, 8. Biological Indicators of
Stress in Fish. S.M. Adams (ed.) American Fisheries Society, Bethesda,
MD.
Golub, M.S., J.M. Donald, and J.A. Reyes. 1991. Reproductive toxicity of
commercial PCB Mixtures: LOAELS and NOAELS from animal studies.
Environ. Health Perspect. 94:245-253.
Greenburg, A.E., L.S. Clersceri, and A.D. Eaton (eds.). 1992. Standard
Methods for the Examination of Water and Wastewater. 18th edition.
American Public Health Association and American Water Works Association,
Washington, DC. 1268 pp.
Grimmer, G., H. Bohnke, and H. Borwitzky. 1978. Profile analysis of polycyclic
aromatic hydrocarbons in sewage sludge by gas chromatography. Fresenius
Z.Anal. Chem. 289:91-95.
Gunderson, D.R., and I.E. Ellis. 1986. Development of a plumb staff beam trawl
for sampling demersal fauna. Fisheries Review 4:35-41.
Hayes, M.L. 1983. Active capture techniques, pp. 123-146. In: Fisheries
Techniques. L.A. Nielsen and D.L Johnson (eds.). American Fisheries
Society, Bethesda, MD.
HEAST. 1989. Health Effects Summary Tables. Office of Emergency and
Remedial Response, U.S. Environmental Protection Agency, Washington,
DC.
Hesse, J.L. 1976. Polychlorinated biphenyl usage and sources of loss to the
environment. In: National Conference on Polychlorinated Biphenyls. J.L.
Buckly et al. (eds.). QV 633 N277c 1975. U.S. Environmental Protection
Agency, Washington, DC.
Hodges, L. 1977. Environmental Pollution. Holt, Rinehart and Winston, New
York, NY.
10-7
-------
10. LITERATURE CITED
Holder, J.W. 1986. The Assessment of the Carcinogenicity of Dicofol
(Kelthane), DDT, DDE, and ODD (TDE). EPA-600/6-86/001. Carcinogen
Assessment Group, U.S. Environmental Protection Agency, Washington, DC.
Hubert, W.A. 1983. Passive capture techniques, pp. 95-122. In: Fisheries
Techniques. L.A. Nielsen and D.L Johnson (eds.). American Fisheries
Society, Bethesda, MD.
Huckins, J.N., T.R. Schwartz, J.D. Petty, and LM. Smith. 1988. Determination,
fate, and potential significance of PCBs in fish and sediment with emphasis
on selected AHH-inducing congeners. Chemosphere 17(10):1995-2016.
IARC (International Agency for Research on Cancer). 1987. IARC Monographs
on the Evaluation of the Carcinogenic Risk of Chemicals to Humans.
Supplement 7. World Health Organization, Lyon, France.
IEHR. 1991. Reassessment of Liver Findings in Five PCB Studies in Rats.
Institute for Evaluating Health Risks. 1101 Vermont Ave., NW, Washington,
DC 20005.
IRIS (Integrated Risk Information System). 1989. U.S. Environmental Protection
Agency, Duluth, MN.
IRIS (Integrated Risk Information System). 1992. U.S. Environmental Protection
Agency, Duluth, MN.
IRIS (Integrated Risk Information System). 1993. U.S. Environmental Protection
Agency, Duluth, MN.
Jacobson, S.W., G.G. Fein, J.L. Jacobson, P.M. Schwartz, and J.K. Douter.
1985. The effects of intrauterine PCB exposure on visual recognition
memory. Child Development 56:853-60.
Jacobson, J.L., S.W. Jacobson, and H.E. Humphrey. 1990. Effects of in utero
exposure to polychlorinated biphenyls and related contaminants on cognitive
functioning in young children. J. Pediatrics 116(1):38-45.
Jearld, A. 1983. Age determination, pp. 301-324. In: Fisheries Techniques.
L.A. Nielsen and D. Johnson (eds.). American Fisheries Society, Bethesda,
MD.
Kaiser, K.L.E. 1978. The rise and fall of mirex. Environ. Sci. Technol.
12:520-528.
10-8
-------
10. LITERATURE CITED
Kannan, N., S. Tanabe, M. Ono, and R. Tatsukawa. 1989. Critical evaluation
of polychlorinated biphenyl toxicity in terrestrial and marine mammals:
increasing impact of non-ortho and mono-ortho coplanar polychlorinated
biphenyls from land to ocean. Arch. Environ. Contam. Toxicol. 18(6):850-
857.
Keith, LH. 1991 a. Report Results Right! Parti. Chemtech June:352-356.
Keith, LH. 1991b. Report Results Right! Part 2. Chemtech August:486-489.
Keith, LH., W. Crommett, J. Deegan, Jr., R.A. Libby, J.K. Taylor, and G.
Wentler. 1983. Principles of environmental analysis. Analyt. Chem.
55:1426-1435.
Kimbrough, R.D., and A.A. Jensen. 1989. Topics in Environmental Health:
Halogenated Biphenyls, Terphenyls, Naphthalenes, Dibenzodioxins and
Related Products. Vol. 4, 2nd Edition. Elsevier, North Holland, NY.
Kish, L. 1965. Survey Sampling. John Wiley & Sons, New York, NY.
Klaassen, C.D., M.D. Amdur, and J. Doull (eds.). 1986. Casarett and Doull's
Toxicology, The Basic Science of Poisons. 3rd edition. MacMillan Publishing
Co., New York, NY.
Korach, K.S. P. Sarver, K. Chae, J.A. McLachlan, and J.D. McKinney. 1988.
Estrogen receptor-binding activity of polychlorinated hydroxybiphenyls:
conformationally restricted structural probes. Molecular Pharmacology
33:120-126.
Krahn, M.M., LD. Rhodes, M.S. Myers, LK. Moore, W.D. MacLeod, and D.C.
Malins. 1986. Associations between metabolites of aromatic compounds
in bile and the occurrence of hepatic lesions in English sole (Parophyrs
vetulus) from Puget Sound, Washington. Arch. Environ. Contam. Toxicol.
15:61-67.
Krahn, M.M., C.A. Wigren, R.W. Pearce, L.K. Moore, R.G. Bogar, W.D.
MacLeod, Jr., S.L Chan, and D.W. Brown. 1988. Standard Analytical
Procedures for the NOAA National Analytical Facility. New HPLC Cleanup
and Revised Extraction Procedures for Organic Contaminants. NOAA Tech.
Memo NMFS F/NWC-153. National Oceanic and Atmospheric
Administration, U.S. Department of Commerce, Northwest and Alaska
Fisheries Center, Seattle, WA. 52 pp.
Kubiak, T.J., H.J. Harris, LM. Smith, T.R. Schwartz, D.L. Stalling, J.A. Trick, L.
Sileo, D.E. Docherty, and T.C. Erdman. 1989. Microcontaminants and
reproductive impairment of the Forster's tern on Green Bay, Lake
Michigan—1983. Arch. Environ. Contam. Toxicol. 18(5):706-727.
10-9
-------
10. LITERATURE CITED
Kurland, L.T., S.N. Faro, and H. Siedler. 1960. Minamata Disease: The
outbreak of a neurological disorder in Minamata, Japan, and its relationship
to the ingestion of seafood contaminated by mercuric compounds. World
A/euro/. 1:370-391.
Leonard, D.L., M.A. Broutman, and K. E. Harkness. 1989. The Quality of
Shellfish Growing Waters on the East Coast of the United States. Strategic
Assessment Branch, National Oceanic and Atmospheric Administration, U.S.
Department of Commerce, Rockville, MD.
Leonard, D.L., and E.A. Slaughter. 1990. The Quality of Shellfish Growing
Waters on the West Coast of the United States. Strategic Assessment
Branch, National Oceanic and Atmospheric Administration, U.S. Department
of Commerce, Rockville, MD.
Li, M. 1975. Pollution in nation's estuaries originating from the agricultural use
of pesticides, pp. 451-466. In: Estuarine Pollution Control and
Assessment. Office of Water Planning and Standards, U.S. Environmental
Protection Agency, Washington, DC.
Lowe, T.P., T.W. May, W.G. Brumbaugh, and D.A. Kane. 1985. National
Contaminant Biomonitoring Program: Concentrations of seven elements in
freshwater fish, 1978-1981. Arch. Environ. Contam. Toxicol. 14:363-388.
Lowenstein, G.G., and D.R. Young. 1986. National Status and Trends Program
for Marine Environmental Quality, Benthic Surveillance Project: Cycle III
Field Manual. NOAA Tech. Memorandum NOS OMA 28. National Oceanic
and Atmospheric Administration, U.S. Department of Commerce, Rockville,
MD.
Maack, L., and W.C. Sonzogni. 1988. Analysis of polychlorobiphenyl congeners
in Wisconsin fish. Arch. Environ. Toxicol. 17(6):711-719.
MacKenzie, M.J., and J.V. Hunter. 1979. Sources and fates of aromatic
compounds in urban stormwater runoff. Environ. Sci. Technol. 13(2):179-
183.
MacLeod W., Jr., D. Brown, A. Friedman, O. Maynes, and R. Pierce. 1985.
Standard Analytical Procedures of the NOAA National Analytical Facility,
1984-85, Extractable Toxic Organic Compounds. NOAA Technical
Memorandum NMFS F/NWC-64. Prepared for the National Status and
Trends Program, National Oceanic and Atmospheric Administration, U.S.
Department of Commerce, Rockville, MD.
10-10
-------
10. LITERATURE CITED
Malins, D.C., B.B. McCain, D.W. Brown, S.L. Chan, M.S. Myers, J.T. Landahl,
P.G. Prohaska, A.J. Friedman, LD. Rhodes, D.G. Burrows, W.D. Gronlund,
and H.O. Hodgins. 1984. Chemical pollutants in sediments and diseases of
bottom-dwelling fish in Puget Sound, WA. Environ. Sci. Technol.
18:705-713.
Malins, D.C., M.M. Krahn, M.S. Myers, LD. Rhodes, D.W. Brown, C.A. Krone,
B.B. McCain, and S.L. Chan. 1985. Toxic chemicals in sediments and biota
from a creosote-polluted harbor: Relationships with hepatic neoplasms and
other hepatic lesions in English sole (Parophrys vetulus). Carcinogenesis
6:1463-1469.
Marsh, D.O. 1987. Dose-response relationships in humans: Methyl mercury
epidemics in Japan and Iraq. In: The Toxicity of Methyl Mercury. C.U.
Eccles and Z. Annau (eds). Johns Hopkins University Press, Baltimore, MD.
Marsh, D.O., T.W. Clarkson, C. Cox, G.J. Meyers, L Amin-Zaki and S. AI-Tikriti.
1987. Fetal methylmercury poisoning: relationship between concentration
in single strands of maternal hair and child effects. Archives of Neurology
44:1017-1022.
May, T.W., and G.L. McKinney. 1981. Cadmium, lead, mercury, arsenic and
selenium concentrations in freshwater fish, 1976-1977--National Pesticide
Monitoring Program. Pesticides Monitoring Journal 15(1):14-38
McConnell, E.E. 1980. pp. 109-150. In: Topics in Environmental Health:
Halogenated Biphenyls, Terphenyls, Naphthalenes, Dibenzodioxins and
Related Products. R.D. Kimbrough (ed.). Elsevier, North Holland, New
York.
McFarland, V.A., and J.U. Clarke. 1989. Environmental occurrence, abundance,
and potential toxicity of polychlorinated biphenyl congeners: considerations
for a congener-specific analysis. Environ. Health Perspect. 81:225-239.
Mearns, A.J., and M.J. Allen. 1978. Use of Small Trawls in Coastal Biological
Surveys. Final Report. Prepared for Corvallis Environmental Research
Laboratory, Office of Research and Development, U.S. Environmental
Protection Agency, Corvallis, OR. Southern California Coastal Water
Research Project, El Segundo, CA.
Mes, J., and D. Weber. 1989. Non-orthochlorine substituted coplanar
polychlorinated biphenyl congeners in Canadian adipose tissue, breast milk,
and fatty foods. Chemosphere. 19(8-9):1357-1365.
Miller, G.T. 1979. Living in the Environment. Wadsworth Publishing Company,
Belmont, CA.
10-11
-------
10. LITERATURE CITED
Mix, M.C. 1986. Cancerous diseases in aquatic animals and their association
with environmental pollutants: A critical literature review. Mar. Environ. Res.
20:1-141.
Monaco, M.E., D.M. Nelson, T.C. Czapla, and M.E. Patillo. 1989. Distribution
and Abundance of Fishes and Invertebrates in Texas Estuaries. ELMR
Report No. 3. Strategic Assessment Branch, National Oceanic and
Atmospheric Administration, U.S. Department of Commerce, Rockville, MD.
Monaco, M.E., D.M. Nelson, R.L Emmett, and S.A. Hinton. 1990. Distribution
and Abundance of Fishes and Invertebrates in West Coast Estuaries.
Volume I: Data Summaries. ELMR Report No. 4. Strategic Assessment
Branch, National Oceanic and Atmospheric Administration, U.S. Department
of Commerce, Rockville, MD.
Moore, J.W., and Ramamoorthy, S. 1984. Organic Chemicals in Natural
Waters. Springer-Verlag, New York. 289 pp.
Morris, C.R., and J.R.P. Cabral (eds.). 1986. Hexachlorobenzene: Proceedings
of an International Symposium. IARC Scientific Publication No. 77. World
Health Organization, Lyon, France.
Mullin, M.D., C.M. Pochini, S. McCrindle, M. Romkes, S.H. Safe, and L.M. Safe.
1984. High-resolution PCB analysis: synthesis and chromatographic
properties of all 209 PCB congeners. Environ. Sci. Technol. 18:468-476.
Multistate Fish and Wildlife Information Systems. 1990. Second Annual
Progress Report (July 1-July 30, 1990). Department of Fisheries and
Wildlife, Virginia Polytechnical Institute, Blacksburg, VA.
Murchelano, R.A. 1982. Some pollution-associated diseases and abnormalities
of marine fish and shellfish: A perspective for the New York Bight, pp. 327-
346. In: Ecological Stress and the New York Bight: Science and
Management. G.F. Mayer (ed.). Estuarine Research Federation, Columbia,
SC.
NAS (National Academy of Sciences). 1976. Selenium. Committee on Medical
and Biologic Effects of Environmental Pollutants, National Research Council,
Washington, DC.
. 1977. Arsenic. Committee on Medical and Biologic Effects
of Environmental Pollutants, National Research Council, Washington, DC.
1991. Seafood Safety. Committee on Evaluation of the
Safety of Fishing Products, National Academy Press, Washington, DC.
10-12
-------
10. LITERATURE CITED
National Marine Fisheries Service. 1987a. Marine Recreational Fishery
Statistics Survey, Atlantic and Gulf Coasts, 1986. Current Fishery Statistics
Number 8392. National Oceanic and Atmospheric Administration, U.S.
Department of Commerce, Rockville, MD.
National Marine Fisheries Service. 1987b. Marine Recreational Fishery
Statistics Survey, Pacific Coast, 1986. Current Fishery Statistics Number
8393. National Oceanic and Atmospheric Administration, U.S. Department
of Commerce, Rockville, MD.
NCI (National Cancer Institute). 1978. Bioassay of Dicofol for Possible
Carcinogenicity. NCI Carcinogenesis Technical Report Series #90.
Rockville, MD.
Neff, J.M. 1979. Polycyclic Aromatic Hydrocarbons in the Aquatic Environment:
Sources, Fates and Biological Effects. Applied Science, London, England.
Neff, J.M. 1985. Polycyclic aromatic hydrocarbons. In: Fundamentals of
Aquatic Toxicology. G.M. Rand and S.R. Petrocelli (eds.). Hemisphere
Publishing Corporation, Washington, DC.
Nelson, D.M., M.E. Monaco, E.A. Irlandi, L.R. Settle, and L Coston-Clements.
1991. Distribution and Abundance of Fishes and Invertebrates in Southeast
Estuaries. ELMR Report No. 9. Strategic Assessment Division, National
Oceanic and Atmospheric Administration, U.S. Department of Commerce,
Rockville, MD.
Nicholls, T.P., R. Perry, and J.N. Lester. 1979. The influence of heat treatment
on the metallic and polycyclic aromatic hydrocarbon content of sewage
sludge. Sci. Total Environ. 12:137-150.
Niimi, A.J. 1987. Biological half-lives of chemicals in fishes. Rev. Environ.
Contam. Toxicol. 99:1-46. Springer-Verlag, New York, NY.
Nimmo D.R. 1985. Pesticides. In: Fundamentals of Aquatic Toxicology, G.M.
Rand and S.R. Petrocelli (eds.). Hemisphere Publishing Corporation,
Washington, DC.
NOAA (National Oceanic and Atmospheric Administration). 1987. National
Status and Trends Program for Marine Environmental Quality-Progress
Report: A Summary of Selected Data on Chemical Contaminants in Tissues
Collected During 1984, 1985 and 1986. NOAA Technical Memorandum
NOS OMA 38. U.S. Department of Commerce, Rockville, MD.
10-13
-------
10. LITERATURE CITED
NOAA (National Oceanic and Atmospheric Administration). 1989a. National
Status and Trends Program for Marine Environmental Quality - Progress
Report: A Summary of Selected Data on Tissue Contamination from the
First Three Years (1986-1988) of the Mussel Watch Project. NOAA
Technical Memorandum NOS OMA 49. U.S. Department of Commerce,
Rockville, MO.
NOAA (National Oceanic and Atmospheric Administration). 1989b. Standard
analytical procedures of the NOAA National Analytical Facility. 2nd ed.
NOAA Tech. Mem. NMFS F/NWC-92,1985-86. National Status and Trends
Program, U.S. Department of Commerce, Rockville, MD.
NOAA (National Oceanic and Atmospheric Administration). 1992. Standard and
Reference Materials for Marine Science. Third Edition. U.S. Department of
Commerce, Rockville, Maryland.
Norstrom, R.J. 1988. In: Hazards, Decontamination and Replacement of
Polychlorinated Biphenyls. J.P. Crine (ed.). Plenum Publishing Corporation,
New York.
NPCA (National Paint and Coatings Association). 1988. Annual Report.
Washington, DC.
Oliver, B.G., and A.J. Niimi. 1988. Trophodynamic analysis of polychlorinated
biphenyl congeners and other chlorinated hydrocarbons in the Lake Ontario
ecosystem. Environ. Sci. Technol. 22(4):388-397.
Oregon Department of Environmental Quality. 1990. Rationale for Aquatic
Tissue Monitoring. Water Quality Division, Portland, OR.
Pflieger, W.H. 1975. The Fishes of Missouri. Missouri Department of
Conservation, Jefferson City, MO.
Phillips, D.J.H. 1980. Quantitative Aquatic Biological Indicators. Pollution
Monitoring Series. Applied Science Publishers Ltd, London, England
Phillips, P.T. 1988. California State Mussel Watch-Ten Year Data Summary
(1977-1987) Water Quality Monitoring Report No. 87-3. State Water
Resources Control Board, Division of Water Quality, Sacramento, CA.
Pillay, K.K.S., C.C. Thomas, and J.W. Kaminski. 1969. Neutron activation
analysis of the selenium content of fossil fuels. Nucl. Appl. Technol.
7:478-483.
Piotrowski, J.K., and M.J. Inskip. 1981. Health Effects of Mercury; A Technical
Report (1981). MARC Report Number 24. Chelsea College, University of
London. 82 pp.
10-14
-------
10. LITERATURE CITED
Pitt, T.K., R. Wells and W.D. McKone. 1981. A critique of research otter trawl
surveys by the St. John's Research and Resource Services. Canadian
Spec. Publ. Fish Aquatic Sci. 58:42-61.
Poland, A., and J.C. Knutson. 1982. 2,3,7,8-Tetrachlorodibenzo-p-dioxin and
related halogenated aromatic hydrocarbons: examination of the mechanism
of toxicity. Annu. Rev. Pharmacol. Toxicol. 22:517-554.
Puget Sound Estuary Program. 1990a (revised). Recommended protocols for
station positioning in Puget Sound. In: Recommended Protocols and
Guidelines for Measuring Selected Environmental Variables in Puget Sound.
Prepared by PTI Environmental Services, Bellevue, WA. Region 10, U.S.
Environmental Protection Agency, Seattle, WA. (Looseleaf)
Puget Sound Estuary Program. 1990b (revised). Recommended protocols for
sampling soft-bottom demersal fishes by beach seine and trawl in Puget
Sound. In: Recommended Protocols and Guidelines for Measuring
Selected Environmental Variables in Puget Sound. Prepared by PTI
Environmental Services, Bellevue, WA. Region 10, U.S. Environmental
Protection Agency, Seattle, WA. (Looseleaf)
Puget Sound Estuary Program. 1990c (revised). Recommended protocols for
fish pathology studies in Puget Sound. In: Recommended Protocols and
Guidelines for Measuring Selected Environmental Variables in Puget Sound.
Prepared by PTI Environmental Services, Bellevue, WA. Region 10, U.S.
Environmental Protection Agency, Seattle, WA. (Looseleaf)
Puget Sound Estuary Program. 1990d (revised). Recommended guidelines for
measuring organic compounds in Puget Sound sediments and tissue
samples. In: Recommended Protocols and Guidelines for Measuring
Selected Environmental Variables in Puget Sound. Prepared by PTI
Environmental Services, Bellevue, WA. Region 10, U.S. Environmental
Protection Agency, Seattle, WA. (Looseleaf)
Puget Sound Estuary Program. 1990e (revised). Recommended protocols for
measuring metals in Puget Sound water, sediment, and tissue samples. In:
Recommended Protocols and Guidelines for Measuring Selected
Environmental Variables in Puget Sound. Prepared by PTI Environmental
Services, Bellevue, WA. Region 10, U.S. Environmental Protection Agency,
Seattle, WA. (Looseleaf)
Reinert, R.E., D. Stewart, and H.L. Seagram. 1972. Effects of dressing and
cooking on DDT concentrations in certain fish from Lake Michigan. J.
Fisheries Research Board Canada 29:525-529.
Reinert, R.E., B.A. Knuth, M.A. Kamrin, and Q.J. Stober. 1991. Risk
assessment, risk management, and fish consumption advisories in the
United States. Fisheries (6):5-12.
10-15
-------
10. LITERATURE CITED
Rohlf, F.J., H.R. Akcakaya, and S.P. Ferraro. 1991. Optimizing Composite
Sampling Protocols. Contract 68-CO-0051. Prepared for the U.S.
Environmental Protection Agency. Applied Biomathematics, Corvallis, OR.
Rosen, B. 1970. Shell Disease of Aquatic Crustaceans, pp. 409-415. In: A
Symposium of Diseases of Fish and Shellfishes. S.F. Sniezko (ed.). Amer.
Fish. Soc. Spec. Publ. No. 5. American Fisheries Society, Washington, DC.
RTI (Research Triangle Institute). 1993. National Listing of State Fish and
Shellfish Consumption Advisories and Bans. (Current as of July 22,1993.)
Prepared for Office of Science and Technology, U.S. Environmental
Protection Agency. Research Triangle Park, NC.
Safe, S. 1985. CRC Critical Reviews in Toxicology. Polychlorinated Biphenyls
(PCBs) and Polybrominated Biphenyls (PBBs): Biochemistry, Toxicology
and Mechanism of Action. CRC Press, Cleveland, OH.
Safe, S. 1990. Polychlorinated biphenyls (PCBs), dibenzo-p-dioxins (PCDDs),
dibenzofurans (PCDFs), and related compounds: environmental and
mechanistic considerations which support the development of toxic
equivalency factors (TEFs). Critical Reviews in Toxicology 21 (1 ):51 -88.
Safe, S., B. Astroff, M. Harris, T. Zacharewski, R. Dickerson, M. Romkes, and
L Biegel. 1991. 2,3,7,8-Tetrachlorodibenzo-p-dioxin and related
compounds as antiestrogens: characterization and mechanism of action.
Pharmacology and Toxicology 69:400-409.
Sanders, M., and B.L Haynes. 1988. Distribution pattern and reduction of
Polychlorinated biphenyls (PCBs) in bluefish, Pomatomus saltatrix fillets
through adipose tissue removal. Bulletin of Environmental Contamination
and Toxicology 41:670-677.
Sax, I.N. 1984. Dangerous Properties of Industrial Materials. 6th Edition. Van
Nostrand Reinhold Company, New York, NY.
Schaeffer, E.H., H. Greim, and W. Goessner. 1984. Pathology of chronic
Polychlorinated biphenyl (PCB) feeding in rats. Toxicol. Appl. Pharmacol.
75:278-288.
Schmitt, C.J., and S.E. Finger. 1987. The effects of sample preparation on
measured concentrations of eight elements in edible tissues of fish from
streams contaminated by lead mining: Archives of Environmental
Contamination and Toxicology 16:185-207.
Schmitt, C.J., M.A. Ribick, J.K. Ludke, and T.W. May. 1983. National Pesticide
Monitoring Program: Organochlorine Residues in Freshwater Fish, 1976-
1979. Resource Publication 152. Fish and Wildlife Service, U.S.
Department of the Interior, Washington, DC.
10-16
-------
10. LITERATURE: CITED
Schmitt, C.J., J.L. Zajicek, and M.A. Ribick. 1985. National Pesticide Monitoring
Program: Residues of organochlorine chemicals in freshwater fish,
1980-1981. Arch. Environ. Contam. Toxicol. 14:225-260.
Schmitt, C.J., and W.G. Brumbaugh. 1990. National Contaminant Biomonitoring
Program: Concentrations of arsenic, cadmium, copper, lead, mercury,
selenium, and zinc in U.S. freshwater fish, 1978-1984. Arch. Environ.
Contam. Toxicol. 19:731-747.
Schmitt, C.J., J.L. Zajicek, and P.M. Peterman. 1990. National Contaminant
Biomonitoring Program: Residues of organochlorine chemicals in U.S.
freshwater fish, 1976-1984. Arch. Environ. Contam. Toxicol. 19:748-781.
Schwartz, T.R., R.D. Campbell, D.L Stalling, R.L Little, J.D. Petty, J.W. Hogan,
and E.M. Kaiser. 1984. Laboratory data base for isomer-specific
determination of polychlorinated biphenyls. Anal. Chem. 56:1303-1308.
Schwartz, T.R., D.L. Stalling, and C.L. Rice. 1987. Are polychlorinated biphenyl
residues adequately described by Aroclor mixture equivalents? Isomer-
specific principal components analysis of such residues in fish and turtles.
Environ. Sci. Technol. 21(1)72-76.
Schwartz, T.R., D.E. Tillit, K.P. Feltz, and P.M. Peterman. In press.
Determination on non- and non-O.O'-chlorine substituted polychlorinated
biphenyls in Aroclors and environmental samples. Submitted to
Chemosphere.
Shain, W., B. Bush, and R. Seegal. 1991. Neurotoxicity of polychlorinated
biphenyls: structure-activity relationship of individual congeners. Toxicology
and Applied Pharmacology 111:33-42.
Sheffy, T.B. 1987. A Review of Mercury in Wisconsin's Environment.
Recommendations for Studying and Identifying the Cause of the Problem.
Bureau of Air Management, Wisconsin Department of Natural Resources,
Madison, Wl.
Sinderman, C.J. 1983. An examination of some relationships between pollution
and disease. Rapp. P. V. Reun. Cons. Int. Explor. Mer. 182:37-43.
Sinderman, C.J., F.B. Bang, N.O. Christensen, V. Dethlefsen, J.C. Harshbarger,
J.R. Mitchell, and M.F. Mulcahy. 1980. The role and value of pathology in
pollution effects monitoring programs. Rapp. P. V. Reun. Cons. Int. Explor.
Mer. 179:135-151.
Sinderman, C.J., and A. Rosenfield. 1967. Principal diseases of commercially
important marine bivalve mollusca and Crustacea. Fish. Bull. 66:335-385.
10-17
-------
10. LITERATURE CITED
Sittig, M. (ed.). 1980. Pesticide Manufacturing and Toxic Materials Control
Encyclopedia. Noyes Data Corporation, Park Ridge, NJ.
Skea, J.C., H.A. Simonin, E.J. Harris, S. Jackling, J.J. Spagnoli, J. Synula, and
J.R. Colghoun. 1979. Reducing levels of Mirex, Aroclor 1254, and DDE by
trimming and cooking Lake Ontario brown trout and smallmouth bass. J.
Great Lakes Research 5:153-150.
Skerfving, S. 1988. Mercury in women exposed to methylmercury through fish
consumption, and in their newborn babies and breast milk. Bull. Environ.
Contam. Toxicol. 41:475-482.
Skoog, D.A. 1985. Principles of Instrumental Analysis. 3rd ed. Saunders
Press, Philadelphia, PA. pp. 270-277, 282-284, 303-304.
Smith, L.M. 1981. Carbon dispersed on glass fibers as an adsorbent for
contaminant enrichment and fractionation. Anal. Chem. 53:2152-2154.
Smith, R.L. 1985. Guidance on Sampling Aquatic Organisms for Tissue
Analyses during FY1986. Environmental Services Division, Region 3, U.S.
Environmental Protection Agency, Philadelphia, PA.
Smith, R.A., R.B. Alexander, and M.G. Wolman. 1987. Water quality trends in
the nation's rivers. Science 235:1607-1615.
Smith, W.E., K. Funk, and M. Zabik. 1973. Effects of cooking on concentrations
of PCB and DDT compounds in Chinook and coho salmon from Lake
Michigan. J. Fish. Res. Board Can. 30:702-706.
Smith, L.M., T.R. Schwartz, and K. Feltz. 1990. Determination and occurrence
of AHH-active polychlorinated biphenyls, 2,3,7,8-tetrachloro-p-dioxion and
2,3,7,8-tetrachlorodibenzofuran in Lake Michigan sediment and biota, the
question of their relative toxicological significance. Chemosphere
21 (9):1063-1085.
Stalling, D.L, T.R. Schwartz, W.J. Dunn III, and S. Wold. 1987. Classification
of polychlorinated biphenyl residues. Anal. Chem. 59:1853-1859.
Stober, Q.J. 1991. Guidelines for Fish Sampling and Tissue Preparation for
Bioaccumulative Contaminants. Environmental Services Division, Region 4,
U.S. Environmental Protection Agency, Athens, GA.
Tanabe, S., N. Kannan, A. Subramanian, S. Watanabe, and R. Tatsukawa.
1987. Environ. Poll. 47:147-163.
10-18
-------
10. LITERATURE CITED
Taylor, J.K. 1985. Standard Reference Materials: Handbook for SRM Users.
Center for Analytical Chemistry, U.S. Department of Commerce, National
Bureau of Standards, Gaithersburg, MD. NBS Special Publication 260-100.
Tetra Tech. 1986. Evaluation of Survey Positioning Method for Near-Shore and
Estuarine Waters. Prepared for Office of Marine and Estuarine Protection,
U.S. Environmental Protection Agency. Bellevue, WA.
Texas Water Commission. 1990. Texas Tissue Sampling Guidelines. Texas
Water Commission, Austin, TX.
Tilson, H.A., J.L. Jacobson, and W.J. Rogan. 1990. Polychlorinated biphenyls
and the developing nervous systems: cross-species comparisons.
Neurotox. Teratol. 12:239-248.
Tollefson, Linda. 1989. Methylmercury in fish: Assessment of risk for U.S.
consumers. In: The Risk Assessment of Environmental and Human Health
Hazards: A Textbook of Case Studies. Dennis J. Paustenback (ed.). John
Wiley & Sons, New York, NY.
TVA (Tennessee Valley Authority). 1991. Fish Tissue Studies in the Tennessee
Valley in 1989. Draft. Chattanooga, TN.
U.S. DHHS (U.S. Department of Health and Human Services). 1990.
Toxicological Profile for Polycyclic Aromatic Hydrocarbons. TP-90-20.
Agency for Toxic Substances and Disease Registry, Public Health Service,
Atlanta, GA.
U.S. EPA (U.S. Environmental Protection Agency). 1978. Metal
Bioaccumulation in Fish and Aquatic Invertebrates. EPA-600/3-78-103.
Environmental Research Laboratory, Office of Research and Development,
Springfield, VA.
U.S. EPA (U.S. Environmental Protection Agency). 1979a. Health Assessment
Document for Cadmium. EPA-600/8-79-003. Environmental Standards and
Criteria, Office of Research and Development, Research Triangle Park, NC.
U.S. EPA (U.S. Environmental Protection Agency). 1979b. Methods for the
Chemical Analysis of Water and Wastes. EPA-600/4-79-020. Environmental
Monitoring and Support Laboratory, Cincinnati, OH.
U.S. EPA (U.S. Environmental Protection Agency). 1980a. Ambient Water
Quality Criteria for Endrin. EPA-440/5-80-047. Office of Water Regulations
and Standards, Washington, DC.
U.S. EPA (U.S. Environmental Protection Agency). 1980b. Interim Guidelines
and Specifications for Preparing Quality Assurance Project Plans.
QAMS-005/80. Quality Assurance Management Staff, Washington, DC.
10-19
-------
10. LITERATURE CITED
U.S. EPA (U.S. Environmental Protection Agency). 1981 a. An Exposure and
Risk Assessment for Mercury, EPA-440/4-85-011. Office of Water
Regulations and Standards, Washington, DC.
U.S. EPA (U.S. Environmental Protection Agency). 1981b. Interim Methods for
the Sampling and Analysis of Priority Pollutants in Sediments and Fish
Tissue. EPA-600/4-81-055. Environmental Monitoring and Support
Laboratory, Cincinnati, OH.
U.S. EPA (U.S. Environmental Protection Agency). 1982a. Methods for the
Chemical Analysis of Municipal and Industrial Wastewater.
EPA-600/4-82-057. Environmental Monitoring and Support Laboratory,
Cincinnati, OH.
U.S. .EPA (U.S. Environmental Protection Agency). 1983a. Analyses of the
Risks and Benefits of Seven Chemicals Used for Subterranean Termite
Control. EPA-540/9-83-005. Office of Pesticide Programs, Washington, DC.
U.S. EPA (U.S. Environmental Protection Agency). 1983b. Pesticide Fact
Sheet—Dicofol. Office of Pesticide Programs, Washington, DC.
U.S. EPA (U.S. Environmental Protection Agency). 1984a. Internal
memorandum from G. LaRocca to B. Burnam et al., August 16,1984. Office
of Pesticide Programs, Washington, DC.
U.S. EPA (U.S. Environmental Protection Agency). 1984b. Policy and Program
Requirements to Implement the Quality Assurance Program. EPA Order
5360.1. Quality Assurance Management Staff, Washington, DC.
U.S. EPA (U.S. Environmental Protection Agency). 1984c. Pesticide Fact
Sheet—Carbophenothion. Office of Pesticides and Toxic Substances, Office
of Pesticide Programs, Washington, DC.
U.S. EPA (U.S. Environmental Protection Agency). 1984d. Pesticide Fact
Sheet—Chlorpyrifos. Office of Pesticides and Toxic Substances, Office of
Pesticide Programs, Washington, DC.
U.S. EPA (U.S. Environmental Protection Agency). 1984e. Pesticide Fact
Sheet—Disulfoton. Office of Pesticides and Toxic Substances, Office of
Pesticide Programs, Washington, DC.
U.S. EPA (U.S. Environmental Protection Agency). 1985a. Bioaccumulation
Monitoring Guidance: 3. Recommended Analytical Detection Limits. EPA-
503/6-90-001. Office of Marine and Estuarine Protection, Washington, DC.
10-20
-------
10. LITERATURE CITED
U.S. EPA (U.S. Environmental Protection Agency). 1985b. Development of
Statistical Distribution for Ranges of Standard Factors Used in Exposure
Assessment. EPA-600/8-85-010. Office of Health and Environmental
Assessment, Washington, DC.
U.S. EPA (U.S. Environmental Protection Agency). 1985c. Guidance for the
Registration of Pesticide Products Containing Lindane as the Active
Ingredient. EPA-540/RS-86-121. Office of Pesticide Programs, Washington,
DC.
U.S. EPA (U.S. Environmental Protection Agency). 1985d. Pesticide Fact
Sheet—Terbufos. Office of Pesticides and Toxic Substances, Office of
Pesticide Programs, Washington, DC.
U.S. EPA (U.S. Environmental Protection Agency). 1986a. Ambient Water
Quality Criteria for Chlorpyrifos. EPA-440/5-86-005. Office of Water
Regulations and Standards, Washington, DC.
U.S. EPA (U.S. Environmental Protection Agency). 1986b. Bioaccumulation
Monitoring Guidance: 4. Analytical Methods for U.S. EPA Priority Pollutants
and 301(h) Pesticides in Tissues from Marine and Estuarine Organisms.
EPA-503/6-90-002. Office of Marine and Estuarine Protec.tion, Washington,
DC.
U.S. EPA (U.S. Environmental Protection Agency). 1986c. Health Assessment
Document for Polychlorinated Dibenzofurans. Draft. EPA 600/8-86-018A.
Environmental Criteria and Assessment Office, Cincinnati, OH.
U.S. EPA (U.S. Environmental Protection Agency). 1986d. Pesticide Fact
Sheet—Diazinon. Office of Pesticides and Toxic Substances, Office of
Pesticide Programs, Washington, DC.
U.S. EPA (U.S. Environmental Protection Agency). 1986e. Research and
Development Methodology for Evaluating Potential Carcinogenicity in
Support of Reportable Quality Adjustments to CERCLA Section 102. OHEA-
C-073 Draft. Carcinogen Assessment Group Office of Environmental
Assessment, Washington, DC.
U.S. EPA (U.S. Environmental Protection Agency). 1986f. Test Methods for the
Evaluation of Solid Waste, Physical/Chemical Methods. SW-846; 3rd Edition
(with 1990 updates). Office of Solid Waste and Emergency Response,
Washington, DC.
U.S. EPA (U.S. Environmental Protection Agency). 1987a. Bioaccumulation
Monitoring Guidance: 2. Selection of Target Species and Review of
Available Data. EPA-430/9-86-005. Office of Marine and Estuarine
Protection, Washington, DC.
10-21
-------
10. LITERATURE CITED
U.S. EPA (U.S. Environmental Protection Agency). 1987b. Bioaccumulation
Monitoring Guidance: 5. Strategies for Sample Replication and Composit-
ing. EPA-430/9-87-003. Office of Marine and Estuarine Protection,
Washington, DC.
U.S. EPA (U.S. Environmental Protection Agency). 1987c. Cadmium Health
Advisory Draft. Office of Drinking Water, Washington, DC.
U.S. EPA (U.S. Environmental Protection Agency). 1987d. National Dioxin
Study. Tiers 3, 5, 6, and 7. EPA-440/4-87-003. Office of Water
Regulations and Standards, Washington, DC.
U.S. EPA (U.S. Environmental Protection Agency). 1987e. Quality
Assurance/Quality Control (QA/QC) for 301 (h) Monitoring Programs:
Guidance on Field and Laboratory Methods. EPA-430/9-86-004. Office of
Marine and Estuarine Protection, Washington, DC.
U.S. EPA (U.S. Environmental Protection Agency). 1987f. The Risk
Assessment Guidelines of 1986. EPA/600/8-87/045. Office of Health and
Environmental Assessment, Washington, DC.
U.S. EPA (U.S. Environmental Protection Agency). 1988a. Drinking Water
Criteria Document for Polychlorinated Biphenyls (PCBs). ECAO-CIN-414.
Prepared by Environmental Criteria and Assessment Office for Office of
Drinking Water, Washington, DC.
U.S. EPA (U.S. Environmental Protection Agency). 1988b. Public Health Risk
Evaluation Database. Office of Emergency and Remedial Response,
Washington, DC.
U.S. EPA (U.S. Environmental Protection Agency). 1989a. Analytical
Procedures and Quality Assurance Plan for the Determination of Mercury in
Fish. Draft. Environmental Research Laboratory, Duluth MN.
U.S. EPA (U.S. Environmental Protection Agency). 1989b. Analytical
Procedures and Quality Assurance Plan for the Determination of
PCDD/PCDF in Fish. EPA-600/3-90-022. Environmental Research
Laboratory, Duluth, MN.
U.S. EPA (U.S. Environmental Protection Agency). 1989c. Analytical
Procedures and Quality Assurance Plan for the Determination ofXenobiotic
Chemical Contaminants in Fish. EPA-600/3-90-023. Environmental
Research Laboratory, Duluth, NM.
U.S. EPA (U.S. Environmental Protection Agency). 1989d. Assessing Human
Health Risks from Chemically Contaminated Fish and Shellfish: A Guidance
Manual. EPA-503/8-89-002. Office of Water Regulations and Standards,
Office of Marine and Estuarine Protection, Washington, DC.
10_22
-------
10. LITERATURE CITED
U.S. EPA (U.S. Environmental Protection Agency). 1989e. Pesticide Fact
Sheet—Ethion. Office of Pesticides and Toxic Substances, Office of
Pesticide Programs, Washington, DC.
U.S. EPA (U.S. Environmental Protection Agency). 1990a. Exposure Factors
Handbook. EPA-600/8-89/043. Office of Health and Environmental
Assessment, Washington, DC.
U.S. EPA (U.S. Environmental Protection Agency). 1990b. Test Methods for
Evaluating Solid Waste, Physical/Chemical Methods. SW-846, 3rd edition,
proposed Update II. Office of Solid Waste and Emergency Response,
Washington, DC.
U.S. EPA (U.S. Environmental Protection Agency). 1990c. Tetrachlorodibenzo-
p-Dioxins and-Dibenzofurans in Edible Fish Tissue at Selected Sites in
Arkansas and Texas. Water Quality Management Branch and Surveillance
Branch, Region 6, Dallas, TX.
U.S. EPA (U.S. Environmental Protection Agency). 1990d. Work Plan for FY
91 Regional Ambient Fish Tissue Monitoring Program Activity No. ELR 80.
Environmental Monitoring and Compliance Branch, Region 7, Kansas City,
KS.
U.S. EPA (U.S. Environmental Protection Agency). 1991 a. Assessment and
Control of Bioconcentratable Contaminants in Surface Waters. Draft. Office
of Research and Development, Washington, DC.
U.S. EPA (U.S. Environmental Protection Agency). 1991b. Contract Laboratory
Program Statement of Work for Inorganic Analysis, Multi-Media,
Multi-Concentration. SOW 788, July. Washington, DC.
U.S. EPA (U.S. Environmental Protection Agency). 1991c. Contract Laboratory
Program Statement of Work for Organic Analysis. Washington, DC.
U.S. EPA (U.S. Environmental Protection Agency). 1991d. Environmental
Monitoring and Assessment Program (EMAP) Near Coastal Program
Laboratory Methods for Filleting and Compositing Fish for Organic and
Inorganic Contaminant Analyses. Draft. U.S. EPA Office of Research and
Development, Environmental Research Laboratory Narragansett, Rl.
U.S. EPA (U.S. Environmental Protection Agency). 1991e. Environmental
Monitoring and Assessment Program (EMAP) Near Coastal Virginian
Province Quality Assurance Project Plan. Draft. Office of Research and
Development, Environmental Research Laboratory, Narragansett, Rl.
U.S. EPA (U.S. Environmental Protection Agency). 1991f. Environmental
Monitoring Methods Index, Version 1.0 Software, User's Manual, EMMIUser
Support. Office of Water, Sample Control Center, Alexandria, VA.
10-23
-------
10. LITERATURE CITED
U.S. EPA (U.S. Environmental Protection Agency). 1991g. Methods for the
Determination of Metals in Environmental Samples. EPA-600/4-91/010.
Environmental Monitoring Systems Laboratory, Office of Research and
Development, Cincinnati, OH.
U.S. EPA (U.S. Environmental Protection Agency). 1991 h. National
Bioaccumulation Study. Draft. Office of Water Regulations and Standards,
Washington, DC.
U.S. EPA (U.S. Environmental Protection Agency). 1991L Proposed Water
Quality Guidance for the Great Lakes System. Office of Science and
Technology, Washington, DC.
U.S. EPA (U.S. Environmental Protection Agency). 1991J. Workshop Report on
Toxicity Equivalency Factors for Polychlorinated Biphenyl Congeners.
EPA/625/3-91/020. Risk Assessment Forum, Washington, DC.
U.S. EPA (U.S. Environmental Protection Agency). 1992a. Classification List
of Chemicals Evaluated for Carcinogenicity Potential. Office of Pesticide
Programs, Washington, DC.
U.S. EPA (U.S. Environmental Protection Agency). 1992b. Consumption
Surveys for Fish and Shellfish: A Review and Analysis of Survey Methods.
EPA-822/R-92-001. Office of Water, Washington, DC.
U.S. EPA (U.S. Environmental Protection Agency). 1992c. National Study of
Chemical Residues in Fish. Volume I. EPA-823/R-92-008a. Office of
Science and Technology, Washington, DC.
U.S. EPA (U.S. Environmental Protection Agency). 1992d. National Study of
Chemical Residues in Fish. Volume II. EPA-823/R-92-008b. Office of
Science and Technology, Washington, DC.
U.S. EPA (U.S. Environmental Protection Agency). 1992e. 304(a) Criteria and
Related Information for Toxic Pollutants. Spreadsheet. Water Quality
Standards Unit, Water Management Division, Region 4, Atlanta, GA.
U.S. EPA (U.S. Environmental Protection Agency). 1993a. Fate One Liner
Database. Office of Pesticide Programs, Washington, DC.
U.S. EPA (U.S. Environmental Protection Agency). 1993b. Reference Dose
List. Office of Pesticide Programs, Health Effects Division, Washington, DC.
U.S. EPA (U.S. Environmental Protection Agency). 1993c. Workshop Report
on Developmental Neurotoxic Effects Associated with Exposure to PCBs.
September 14-15, 1992, Research Triangle Park, NC. Risk Assessment
Forum, Washington, DC.
10-24
-------
10. LITERATURE CITED
U.S. EPA (U.S. Environmental Protection Agency). In preparation. Proceedings
from National Workshop on PCBs in Fish Tissue. May 11-12, 1993,
Washington, DC. Office of Water, Washington, DC.
U.S. FDA (U.S. Food and Drug Administration). 1990. Pesticide Analytical
Manual, Volumes I and II. Report No. FDA/OMO-90/15A. U.S. Department
of Health and Human Services, Washington, DC.
USGS (U.S. Geological Survey). 1987. Methods for the determination of
organic substances in water and fluvial sediments. Chapter A3. In:
Techniques of Water-Resources Investigations of the United States
Geological Survey. R.L. Wershaw, M.J. Fishman, R.R. Grable, and L.E.
Lowe (eds). Books and Open-File Reports Section, Denver, CO.
Varanasi, U., W.L Reichert, J.E. Stein, et al. 1985. Bioavailability and
biotransformation of aromatic hydrocarbons in benthic organisms exposed
to sediment from an urban estuary. Environ. Sci. Technol. 19:836-841.
Versar, Inc. 1982. Sampling Protocols for Collecting Surface Water, Bed
Sediment, Bivalves and Fish for Priority Pollutant Analysis-Final Draft
Report. EPA Contract 68-01-6195. Prepared for Office of Water
Regulations and Standards, U.S. Environmental Protection Agency.
Springfield, VA.
Versar, Inc. 1984. Sampling Guidance Manual for the National Dioxin
Study-Final Draft Report. EPA Contract 68-01 -6160. Prepared for Office of
Water Regulations and Standards, U.S. Environmental Protection Agency.
Springfield, VA.
Voiland, M.P., K.L. Gall, D.J. Lisk, and D.B. MacNeill. 1991. Effectiveness of
Recommended Fat-Trimming Procedures on the Reduction of PCB and
Mirex Levels in Brown trout (Salmo trutta) from Lake Ontario. J. Great
Lakes Res. 17(4):454-460.
Wanderstock, J., W. Iskat, W. Gutenmann, and D. Lisk. 1971. Effect of several
cooking methods on concentration of DDT residues in lake trout and coho
salmon. New York Fish and Game J18:70-72.
Ware, G.W. 1978. The Pesticide Book. W.H. Freeman and Company, San
Francisco, CA.
WDNR (Wisconsin Department of Natural Resources). 1988. Fish Contaminant
Monitoring Program-Field and Laboratory Guidelines. Report No. 1005.1.
Madison, Wl.
10-25
-------
10. LITERATURE CITED
Weber, C.I. (ed.) 1973. Biological Field and Laboratory Methods for Measuring
the Quality of Surface Waters and Effluents. EPA/670/4-73-001. Office of
Research and Development, U.S. Environmental Protection Agency,
Cincinnati, OH.
WHO (World Health Organization). 1976. Environmental Health Criteria. 1.
Mercury. Geneva, Switzerland.
WHO (World Health Organization). 1990. Environmental Health Criteria 101:
Methylmercury. Geneva, Switzerland.
Wiener, J.G. 1987. Metal contamination of fish in low-pH lakes and potential
implications for piscivorous wildlife. Trans N. Am. Wildl. Nat. Res. Conf.
52:654-657.
Williams, S. (ed.). 1984. Official Methods of Analysis of the Association of
Official Analytical Chemists. Fourteenth edition. The Association of Official
Analytical Chemists, Inc., Arlington, VA.
Williams, C.D., D.M. Nelson, M.E. Monaco, S.L. Stone, C. lancu, L. Coston-
Clements, LR. Settle, and E.A. Irlandi. 1990. Distribution and Abundance
of Fishes and Invertebrates in Eastern Gulf of Mexico Estuaries. ELMR
Report No. 6. Strategic Assessment Branch, National Oceanic and
Atmospheric Administration, U.S. Department of Commerce, Rockville, MD.
Wolf, R.J., and R.J. Walker. 1987. Economies in Alaska: Productivity,
geography, and development impacts. Arctic Anthropology 24:56-81.
Wood, J.M. 1974. Biological cycles for toxic elements in the environment.
Science 183:1049-1052.
Worthing, C.R. 1991. The Pesticide Manual: A World Compendium. 9th
edition. British Crop Protection Council, Croydon, England.
Zabik, M., C. Hoojjat, and D. Weaver. 1979. Polychlorinated biphenyls, dieldrin,
and DDT in lake trout cooked by broiling, roasting, or microwave. Bull.
Environ. Contamin. Toxicol. 21:136-143.
Zweig, G., and J. Sherma (eds.). 1980. Updated General Techniques and
Additional Pesticides. Volume 11. In: Analytical Methods for Pesticides and
Plant-Growth Regulators. Academic Press, New York.
10-26
-------
APPENDIX A
FISH AND SHELLFISH SPECIES FOR WHICH STATE
CONSUMPTION ADVISORIES HAVE BEEN ISSUED
-------
APPENDIX A
APPENDIX A
FISH AND SHELLFISH SPECIES FOR WHICH STATE CONSUMPTION
ADVISORIES HAVE BEEN ISSUED
FRESHWATER FINFISH SPECIES FOR WHICH STATE
CONSUMPTION ADVISORIES HAVE BEEN ISSUED
AL catfish (unspecified), fish species (unspecified), bigmouth buffalo, brown
bullhead, channel catfish, white bass
AK no consumption advisories
AS no consumption advisories
AZ fish species (unspecified)
AR fish species (unspecified)
CA goldfish, Sacramento blackfish, brown bullhead, crappie (unspecified),
hitch, common carp, largemouth bass, smallmouth bass, channel catfish,
white catfish, rainbow trout, croaker (unspecified), orangemouth corvina,
sargo, tilapia (unspecified), squawfish, sucker (unspecified), trout (unspeci-
fied), fish species (unspecified)
CO rainbow trout, yellow perch, northern pike, walleye, smallmouth bass,
largemouth bass, black crappie, kokanee salmon, channel catfish, trout
(unspecified), fish species (unspecified)
CT common carp and fish species (unspecified)
DE white catfish, channel catfish, fish species (unspecified)
DC channel catfish, common carp, American eel
PL largemouth bass, gar, bowfin, warmouth, yellow bullhead, Mayan cichlid,
oscar, spotted sunfish
GA common carp, largemouth bass, catfish (unspecified), fish species
(unspecified)
GU no consumption advisories
_
-------
APPENDIX A
HI no consumption advisories
ID no consumption advisories
IL lake trout, coho salmon, Chinook salmon, brown trout, common carp,
catfish (unspecified), bigmouth buffalo, channel catfish, flathead catfish,
smallmouth buffalo, shovelnose sturgeon, bluegill, crappie (unspecified),
freshwater drum, largemouth bass, spotted bass, alewife
IN common carp, catfish (unspecified), coho salmon, brown trout, lake trout,
Chinook salmon, channel catfish, fish species (unspecified)
IA channel catfish, common carp, carpsucker (unspecified), fish species
(unspecified)
KS buffalo (unspecified), catfish (unspecified), common carp, freshwater drum,
sturgeon (unspecified), carpsucker (unspecified)
KY channel catfish, paddlefish, white bass, common carp, fish species
(unspecified)
LA bass (unspecified), fish species (unspecified)
ME fish species (unspecified)
MD channel catfish, American eel, black crappie, common carp, bullhead
(unspecified), sunfish (unspecified)
MA brown trout, yellow perch, white sucker, American eel, smallmouth bass,
largemouth bass, lake trout, channel catfish, brown bullhead, common
carp, white catfish, fish species (unspecified)
Ml common carp, rock bass, crappie (unspecified), yellow perch, largemouth
bass, smallmouth bass, walleye, northern pike, muskellunge, sauger, white
bass, longnose sucker, white perch, carpsucker (unspecified), brown
bullhead, bullhead (unspecified), bluegill, freshwater drum, sturgeon
(unspecified), brown trout, ciscowet, lake trout, coho salmon, Chinook
salmon, splake, catfish (unspecified), rainbow trout, brook trout, sucker
(unspecified), gizzard shad, freshwater drum, white sucker, lake whitefish
MN yellow perch, brown bullhead, black bullhead, yellow bullhead, quillback
carpsucker, brown trout, brook trout, lake trout, Chinook salmon, ciscowet,
walleye, northern pike, brook trout, muskellunge, splake, smallmouth bass,
largemouth bass, rock bass, white bass, rainbow trout, white sucker, tulli-
bee, bluegill, black crappie, white crappie, shorthead redhorse, silver
redhorse, common carp, smallmouth buffalo, redhorse sucker, sauger,
bigmouth buffalo, channel catfish, lake whitefish, freshwater drum,
pumpkinseed, chub bloater, lake herring, flathead catfish, bowfin
-------
APPENDIX A
MS fish species (unspecified), catfish (unspecified), buffalo (unspecified)
MO sturgeon (unspecified), common carp, channel catfish, buffalo
(unspecified), flathead catfish, sucker (unspecified), paddlefish, catfish
(unspecified), redhorse, freshwater drum
MT fish species (unspecified)
NE common carp, channel catfish
NV fish species (unspecified)
NH fish species (unspecified)
NJ striped bass, American eel, white perch, white catfish, fish species
(unspecified)
NM white crappie, channel catfish, common carp, brown trout, river
carpsucker, kokanee salmon, largemouth bass, bluegill, white bass,
walleye, white sucker, yellow perch, black bullhead, black crappie, bass
(unspecified), crappie (unspecified), rainbow trout, longnose dace, walleye,
northern pike, trout (unspecified), carpsucker (unspecified), bullhead
(unspecified), black bass
NY common carp, lake trout, brown trout, yellow perch, smallmouth bass,
splake, American eel, goldfish, striped bass, white perch, bluefish,
largemouth bass, brown bullhead, white catfish, walleye, rainbow smelt,
tiger muskellunge, white sucker, northern pike, Chinook salmon, coho
salmon, rainbow trout
NC largemouth bass, fish species (unspecified)
ND walleye, white bass, yellow perch, northern pike, bigmouth buffalo,
common carp, crappie (unspecified), bullhead (unspecified), white sucker,
channel catfish, goldeye, chinook salmon, sauger, carpsucker
(unspecified), sunfish (unspecified), smallmouth bass
OH common carp, catfish (unspecified), white bass, sucker (unspecified), fish
species (unspecified)
OK channel catfish, largemouth bass, fish species (unspecified)
OR fish species (unspecified), crayfish
PA white sucker, white perch, common carp, American eel, channel catfish,
goldfish, largemouth bass, green sunfish, quillback carpsucker, white bass,
lake trout, walleye, smallmouth bass, shorthead redhorse, sucker
(unspecified), fish species (unspecified)
A-5
-------
APPENDIX A
PR no fish consumption advisories
Rl striped bass
SC fish and shellfish species (unspecified)
SD no fish consumption advisories
IN catfish (unspecified), largemouth bass, crappie (unspecified), common
carp, rainbow trout, striped bass, sauger, white bass, smallmouth buffalo,
fish species (unspecified)
IX catfish (unspecified), fish species (unspecified)
UT fish species (unspecified)
VT brown trout, lake trout, walleye
VA fish species (unspecified)
VI no fish consumption advisories
WA no fish consumption advisories
WV channel catfish, brown bullhead, common carp, sucker (unspecified), fish
species (unspecified)
Wl lake trout, coho salmon, Chinook salmon, brown trout, common carp,
catfish (unspecified), splake, rainbow trout, brook trout, lake trout,
ciscowet, northern pike, white bass, white sucker, walleye, yellow perch,
muskellunge, flathead catfish, freshwater drum, channel catfish, bullhead
(unspecified), bluegill, black crappie, crappie (unspecified), rock bass,
smallmouth bass, redhorse (unspecified), largemouth bass, lake sturgeon,
buffalo (unspecified), fish species (unspecified)
WY no fish consumption advisories
Source: RTI, 1993. National Listing of State Fish and Shellfish Consumption
Advisories and Bans. (Current as of July 22,1993.) Research Triangle
Institute, Research Triangle Park, NC.
A-6
-------
APPENDIX A
ESTUARINE/MARINE FISH AND SHELLFISH SPECIES FOR WHICH STATE
CONSUMPTION ADVISORIES HAVE BEEN ISSUED
AL no consumption advisories
AK no consumption advisories
AS fish and shellfish species (unspecified)
CA white croaker, black croaker, corbina, surf perch, queenfish, sculpin,
rockfish, kelp bass, striped bass, fish and shellfish species (unspecified)
CT striped bass, bluefish
DE no consumption advisories
DC channel catfish, American eel
FL shark (unspecified)
GA no consumption advisories
GU no consumption advisories
HI no consumption advisories
LA fish and shellfish species (unspecified)
ME no consumption advisories
MD channel catfish, American eel
MA American eel, flounder, American lobster, bivalves (unspecified), fish
species (unspecified)
MS no consumption advisories
NH no consumption advisories
NJ striped bass, bluefish, American eel, white perch, white catfish, blue crab,
fish and shellfish species (unspecified)
NY American eel, striped bass, bluefish, white perch, white catfish, rainbow
smelt, Atlantic needlefish, blue crab
NC fish species except herring, shad, striped bass, and shellfish species
(unspecified)
_
-------
APPENDIX A
OR no consumption advisories
PA white perch, channel catfish, American eel
PR no consumption advisories
Rl striped bass, bluefish
SC fish and shellfish species (unspecified)
TX blue crab, catfish (unspecified), fish species (unspecified)
VA fish species (unspecified)
VI no consumption advisories
WA no consumption advisories
Source: RTI, 1993. National Listing of State Fish and Shellfish Consumption
Advisories and Bans. (Current as of July 22,1993.) Research Triangle
Institute, Research Triangle Park, NC.
A-8
-------
APPENDIX B
TARGET ANALYTES ANALYZED IN NATIONAL OR
REGIONAL MONITORING PROGRAMS
-------
APPENDIX B
Table B-1. Target Analytes Analyzed In National or
Regional Monitoring Programs
Analyte
Monitoring program
a b c d1 e f g h i
Metafs
Aluminum (Al) • •
Antimony (Sb) • • •
Arsenic (As) • • • • • •
Barium (Ba) •
Beryllium (Be) • •
Cadmium (Cd) O • • • • •
Chromium (Cr) • • • • •
"Copper (Cu) • • • V~~~ V
Cyanide •
Iron (Fe) . • •
Lead(Pb) O • •• •" •
Manganese (Mn) • • •
Mercury (Hg) • • • • • •
Methylmercury O •
Molybdenum •
"Nickel (NO V ~" • • V
Selenium (Se) • • • • •
Silicon (Si) •
Silver (Ag) O ~" """• ~V
"fhallium (fl) V •
fin (Sn) • ~ ~
fributyltin •
Vanadium •
Zinc (Zn) » V V V
Pesticides
Aldrin O • • • O
Butachlor •
Chlordane (cis & trans) O • • • •'* • O •
Chlorpyrifos • •
Danitof O
(continued)
B-3
-------
APPENDIX B
Table B-1 (continued)
Monitoring program
Analyte abode f g h i
DCPA (chlorthal) • •
'"6Df~(tota7f ................... • • •
2,4'-DbD~(2,4'-fb¥) ~~ ~" "• ¥ ~V •
4,4'-DDD (4,4'-TDE) • • • • • • •
2~4~DDE ....... • • • •
4,4'-DDE ••••• •••
Demeton •
Dicofol • • •
Dieldrin ••••• •••
Diphenyl disulfide •
Endosulfan
cx-Endosulfan (endosulfan I) • •
3-Endosulfan (endosulfan II) • •
Endosulfan sulfate • •
Endrin" " "" "• • V~V ~V V
Endrin aldehyde •
Ethyl-p-nitrophenylphenylphosphorothioate (EPN) •
Fonofos •
Guthion •
Heptachlor ...... " • —- — — ^— --^-~-^
Heptachlor epoxide • • ~ • • •••
Hexachlorocyclohexane (HCH) also known
as Benzene hexachloride (BHC)
a-Hexachlorocyclohexane • • • • • • •
G-Hexachlorocyclohexane • • • •
5-Hexachlorocyclohexane • • •
y-Hexachlorocyclohexane (lindane) ••••• •••
Technical-hexachlorocyclohexane •
Hexachlorophene •
Isopropalin • •
Kepone • • ,
Malathion •
(continued)
B-4
-------
APPENDIX B
Table B-1 (continued)
Monitoring program
Analyte
a b c d1 e f g h i
Methoxychlor G • • •
Mirex ••••• • « «
Nitrofen •
cis-Nonachlor • • • •
trans-Nonachlor • • • • •
Oxychlordane • • • •
Parathion O
Toxaphene (mixture) O • • • O
Triazine herbicides 93
Trichloronate •
frifiuralin • • V
Base/Neutral Organic Compounds
Acenaphthene O • •
Acenaphthylene O • •
Anthracene • • •
Benzidine •
Benzo(a)anthracene O • •
Benzo(a)pyrene O • •
Benzo(e)pyrene •
Benzo(b)fluoranthene 9 • •
Benzo(k)fluoranthene • • •
Benzo(g,h,i)perylene €> • •
Benzyl butyl phthalate 9
Biphenyl • •
4-Bromophenyl ether •
bis(2-Chloroethoxy)methane O
bis(2-Chloroethyl)ether «
bis(2-Chloroisopropyl)ether •
"Isisfi-'Ethylhexyl^V
Chlorinated benzenes •
2-Chloronaphthalene O
4-Chlorophenyl ether •>
Chrysene • • •
(continued)
B-5
-------
APPENDIX B
Table B-1 (continued)
Analyte
Monitoring program
c d1 e
Dibenzo(a,h)anlhracene • • •
Di-n-bulyl phthalate •
1,2-Dichlorobenzene •
1,3-Dichlorobenzene •
1,4-Dichlorobenzene •
3,3'-Dichlorobenzidine • •
Diethyl phthalate •
2,6-Dimethylnaphthalene • •
2,3,5-Trimethylnaphthalene •
Dimethyl phthalate •
2,4-Dinitrotoluene •
2,6-DinitrotoIuene •
Di-n-octyl phthalate •
1,2-Diphenylhydrazine •
bis(2-Ethyihexyl) phthaTate •
Fluoranthene • • •
Fluorene • • •
Heptachlorostyrene •
Hexachlorostyrene •
Hexachlorobenzene • • • • • • •
Hexachlorobutadiene • •
Hexachlorocyclopentadiene • •
Hexachloroethane •
lndeno(1,2,3-cd)pyrene • •
Isophorone •
4,4'-Methylene bisfN.N'-dimethylJaniline •
1 -Methylnaphthalene •
2-Methylnaphthalene •
1-Methylphenanthrene •
Naphthalene • • •
Nitrobenzene •
N-Nitroso-di-n-butylamine •
N-Nitrosodimethylamine •
(continued)
B-6
-------
APPENDIX B
Table B-1 (continued)
Monitoring program
Analyte
c d1 e f g
N-Nitrosodiphenylamine •
N-Nitrosodipropylamine •
Octachlorostyrene • • •
PAHs (polycyclic aromatic hydrocarbons) *3
PBBs (polybrominated biphenyls) •
PCBs (polychlorinated biphenyls) • • • • •
Aroclor 1016 (mixture) • •
Aroclor 1221 (mixture) • •
Aroclor 1232 (mixture) • •
Aroclor 1242 (mixture) • • •
Aroclor 1248 (mixture) • • •
Aroclor 1254 (mixture) • • •
Aroclor 1260 (mixture) • • •
Selected individual congeners •
Pentachloroanisole (PCA) • • •
Pentachlorobenzene • • •
Pentachloronttrobenzene (PCNB) • •
Pentachlorophenyl methyl ether •
Pentachlorophenyl methyl sulfide •
Pentachlorostyrene •
Perthane • •
Perylene •
Phenanthrene • • •
Pyrene • • •
Terphenyl •
1,2,3,4-Tetrachlorobenzene • •
1,2,3,5-Tetrachlorobenzene • •
1,2,4,5-Tetrachlorobenzene • • •
1,2,3-Trichlorobenzene •
1,2,4-Trichlorobenzene • • •
1,3,5-Trichlorobenzene • •
Triphenyl phosphate •
(continued)
B-7
-------
APPENDIX B
Table B-1 (continued)
Monitoring program
Analyte
a b c d1 e fgh
Dtoxfru*
1,2,3,7,8-Pentachlorodibenzodioxin (PeCDD) •
2,3,7,8-Tetrachlorodibenzodioxin fTCDD) • • • •
1,2,3,4,6,7,8-Heptachlorodibenzodioxin (HpCDD) •
1,2,3,4,7,8-Hexachlorodibenzodioxin (HxCDD) •
1,2,3,6,7,8-Hexachlorodibenzodioxin (HxCDD) •
1,2,3.7,8,9-Hexachlorodibenzodioxin (HxCDD) •
| Dlbertzoturans
1,2,3,4,6,7,8-Heptachlorodibenzofuran (HpCDF) •
1,2,3,4,7,8,9-Heptachlorodibenzofuran (HpCDF) •
1,2,3,4,7,8-Hexachlorodibenzofuran (HxCDF) •
1,2,3,6,7,8-Hexachlorodibenzofuran (HxCDF) •
1,2,3,7,8,9-Hexachlorodibenzofuran (HxCDF) •
2,3,4,6,7,8-Hexachlorodibenzofuran (HxCDF) •
1,2,3,7,8-Pentachlorodibenzofuran (PeCDF) •
2,3,4,7,8-Pentachlorodibenzofuran (PeCDF) •
2,3,7,8-Tetrachlorodibenzofuran (TCDF) •
[Acidic Organic Compounds
Chlorinated phenols *3
4-Chloro-3-cresol •
2-Chlorophenol •
2,4-Dichlorophenol •
2,4-Dimethylphenol •
4,6-Dinitro-2-cresol •
2-4-Dinitrophenol •
___•*.•— — — «»«. ___________ «.._______ ^.._^^^_^__ _________ «_ _ _ ^^_____ _ _ _^ _ _ .
2-Nitrophenol •
4-Nitrophenol •
Pentachlorophenol (PCP) •
Phenol •
2,4,6-Trichlorophenol •
| VolatHa Organic Compounds
Acrolein •
Acrylonitrile •
•
v
_.
V
(continued)
B-8
-------
APPENDIX B
Table B-1 (continued)
Analyte
Monitoring program
a b c d1 e
g h i
Benzene •
Bromodichloromethane •
Bromoform •
Bromomethane •
Carbon tetrachloride •
Chlorobenzene •
Chloroethane •
2-Chloroethylvinyl ether •
Chloroform •
Chloromethane •
Dibromochloromethane •
1,1 -Dichloroethane •
1,2-Dichloroethane •
1,1-Dichloroethene •
trans-1,2-Dichloroethene •
1,2-Dichloropropane •
cis-1,3-Dichloropropene •
trans-1,3-Dichloropropene •
Ethylbenzene •
Methylene chloride •
1,1,2,2-Tetrachloroethane •
Tetrachloroethene •
Toluene •
1,1,1-Trichloroethane •
1,1,2-Trichloroethane •
Trichloroethene •
Vinyl chloride •
1 Contaminants listed were monitored by at least one Great Lakes State. NOTE: Contaminants monitored
exclusively by the Canadian Province of Ontario were not included.
2 Only the cis-isomer is monitored.
3 FDA recommends method development/improvement for this analysis.
a 301 (h) Monitoring Program. Source: U.S. EPA. 1985. Bioaccumulation Monitoring Guidance: 1. Estimating
the Potential for Bioaccumulation of Priority Pollutants and 301 (h) Pesticides Discharged into Marine and
Estuarine Waters. EPA 503/3-90-001. Office of Marine and Estuarine Protection, Washington, DC.
B-9
-------
APPENDIX B
Table B-1 (continued)
b Food and Drug Administration recommendations. Source: Michael Bolger, FDA, personal communication, 1990.
c National Study of Chemical Residues in Fish. Source: U.S. EPA. 1992. National Study of Chemical Residues
in Fish. Volumes I and II. EPA 823/R-92-008a and 008b. Office of Science and Technology, Washington, DC.
d Great Lakes Sport Fish Contaminant Advisory Program. Source: Hesse, J. L 1990. Summary and Analyses
of Existing Sportfish Consumption Advisory Programs in the Great Lakes Basin—the Great Lakes. Fish
Consumption Advisory Task Force, Michigan Department of Hearth, Lansing, Ml.
8 NOAA Status and Trends Program. Source: NOAA. 1989. National Status and Trends Program for Marine
Environmental Quality-Progress Report: A Summary of Selected Data on Tissue Contamination from the First
Three Years (1986-1988) of the Mussel Watch Project. NOAA Technical Memorandum NOS OMA 49. U.S.
Department of Commerce, Rockville, MD.
f EPA National Dioxin Study. Source: U.S. EPA. 1987. National Dioxin Study. Tiers 3, 5, 6 and 7. EPA
440/4-87-003. Office of Water Regulations and Standards, Washington, DC.
9 U.S. Fish and Wildlife Service National Contaminant Biomonitoring Program. Sources: C. J. Schmitt, J. L.
Zajicek, and P. H. Peterman, 1990, National Contaminant Biomonitoring Program: Residues of organochlorine
chemicals in U.S. freshwater fish, 1976-1984, Arch. Environ. Contam. Toxicol. 19:748-781; and T. P. Lowe, T.
W. May, W. G. Brumbaugh, and D. A. Kane, 1985, National Contaminant Biomonitoring Program: Concentrations
of seven elements in freshwater fish, 1978-1981, Arch. Environ. Contam. Toxicol. 14:363-388.
h U.S. EPA. 1991. Assessment and Control of Bioconcentratable Contaminants in Surface Waters. Draft. Office
of Water, Office of Research and Development, Washington, DC.
1 U.S. Geological Survey National Water-Quality Assessment Program. Source: J.K. Crawford and S.N. Luoma.
1993. Guidelines for Studies of Contaminants in Biological tissues for the National Water-Quality Assessment
Program. USGS Open-File Report 92-494. U.S. Geological Survey, Lemoyne, PA.
B-10
-------
APPENDIX C
PESTICIDES AND HERBICIDES RECOMMENDED
AS TARGET ANALYTES
-------
Table C-1. Pesticides and Herbicides Recommended as Target Analytes
Pesticide
Family
Use
Registration
EPA
Toxictty carcinogenicity
class" classification6
Orgartochlorines
Chlordane
Chlorinated
cyclodiene
Termite control. Historically used
for control of fire ants, cutworms,
grasshoppers, and on other
insects on corn, grapes, straw-
berries, and other crops and as a
dip for nonfood roots and tips of
plants (Hartley and Kidd, 1987).
In March 1978, the EPA issued a
cancellation proceeding on chlor-
dane, allowing only limited use on
certain crops and pests until July
1983, but no use thereafter
except for underground termite
control (43 FR 12372). All uses
except subsurface ground inser-
tion for termite control were
canceled November 30, 1987. All
chlordane/heptachlor products
were voluntarily canceled by the
registrant, Velsicol. All other
chlordane/heptachlor products are
either voluntarily canceled or
suspended for failure to meet
EPA data requirements. The only
commercial use of chlordane/
heptachlor products still permitted
is for fire ant control in power
transformers (U.S. EPA, 1990).
The sale, distribution, and ship-
ment of existing stocks of all
canceled chlordane/heptachlor
products is prohibited in the U.S.
as of April 15, 1988. The use of
existing stocks of termiticide
products in the possession of
homeowners is also permitted (53
FR 11798; 54 FR 20194).
B2
TJ
m
g
x
o
o
CO
See notes and references at end of table.
(continued)
-------
Table C-1 (continued)
Pesticide
Family
Use
Registration
EPA
Toxictty carcinogenicity
class* classification1*
DDT
Chlorinated
hydrocarbon
Dicofol
Chlorinated
hydrocarbon
Insecticide
Acaricide on many fruit,
vegetable, ornamental and fried
crops. Historically used to control
mites on cotton and citrus (60%).
Other major uses included control
of mites on apples (10%),
ornamental plants and turf (10%)
and 20% on a variety of other
agricultural products (pears,
apricots and cherries), seed crop
soil treatment, vegetables, (e.g.,
beans and corn) and shade trees
(51 FR 19515) (U.S. EPA, 1986b).
All uses in U.S. were canceled as
of January 1, 1973, except for
emergency public health uses.
Effective December 31, 1988, all
uses were canceled unless
registered formulas contain less
than 0.1% DDT (51 FR 19508).
Active since 1957; however all
uses are to be canceled after
January 1989 unless registered
formulas contain less than 0.1%
DDT and related contaminants
(51 FR 19508).c
B2
rn
g
x
o
o
See notes and references at end of table.
(continued)
-------
Table C-1 (continued)
Pesticide
Family
Use
Registration
EPA
Toxicrty carcinogen icity
class" classification11
Dieldrin
Chlorinated
cyclodiene
Endosulfan (I and II)
Chlorinated
bicyclid sulfite
Control of locusts, tropical disease
carriers (e.g., mosquitoes), and
termites, use as wood
preservative, and moth proofing
for woolen clothes and carpets
(Worthing, 1991).
Insecticide and acaricide on citrus,
deciduous, small fruits, coffee,
tea, fiber crops, forage crops,
forest, grains, nuts, oil crops,
tobacco, ornamentals, and
vegetables.
All uses on food products were
suspended in 1974 (ATSDR,
1987a). All uses in the U.S. were
banned in 1985 except for
subsurface termite control,
dipping of nonfood roots and
tops, and moth proofing in a
closed system (U.S. EPA, 1985b).
These uses have been voluntarily
canceled by industry (ATSDR,
1987a).
Active since 1954. Used for
control of aphids, thrips, beetles,
foliar feeding larvae, mites,
borers, cutworms, bollworms,
bugs, whiteflies, leafhoppers and
slugs on citrus, deciduous, small
fruits, coffee, tea, fiber crops,
forage crops, forest, grains, nuts,
oil crops, ornamentals, tobacco,
and vegetables.0
B2
•o
"0
m
z
a
x
o
o
en
See notes and references at end of table.
(continued)
-------
Table C-1 (continued)
Pesticide
Family
Use
Registration
EPA
Toxicfty carcinogenicity
class" classification11
Endrin
Chlorinated
cyclodiene
Heptachlor epoxide
Chlorinated
cyclodiene
Historically used to control cotton
bollworms, as a foliar treatment
for citrus, potatoes, small grains,
apple orchards, sugarcane, and
as flower and bark treatment on
trees. Endrin has also been used
to control populations of birds and
rodents (U.S. EPA, 1980).
Heptachlor epoxide is an oxidation
product of heptachlor. It is a
contaminant of both heptachlor
and chlordane. Heptachlor was
widely used as a termiticide and
insecticide, primarily for ant
control (Hodges, 1977).
In 1964, endrin persistence in
soils led to cancellation of its use
on tobacco (U.S. EPA, 1980). By
1979, specified uses on cotton,
small grains, apple orchards,
sugarcane and ornamentals were
also restricted (44 PR 43632). In
1984, the sole producer of endrin
voluntarily requested cancellation
of all endrin products (U.S. EPA,
1984a).
Termide (chlordane) sales halted
per Velsicol and EPA agreement
pending results of specific
application tests. Restrictions on
heptachlor were first instituted in
1978 and heptachlor can no
longer be sold in the U.S. as of
August 1987 but remaining stocks
can be used in some States by
commercial exterminators for
termite control. All uses have
been banned in Minnesota,
Massachusetts, and New York
(ATSDR, 1987b).
NA
B2
•o
TJ
m
g
x
o
o
I
o>
See notes and references at end of table.
(continued)
-------
Table C-1 (continued)
Pesticide
Family
Use
Registration
EPA
Toxicfty carcinogen icity
class' classification1*
Hexachlorobenzene
Chlorinated
benzene
Primary use prior to 1985 was as
a fungicide seed protectant in
small grain crops, particularly
wheat.
Registration of
hexachlorobenzene as a pesticide
was voluntarily canceled in 1984
(Morris and Cabral, 1986).
IV
B2
Lindane Chlorinated
(y-hexachlorocyclohexane) hydrocarbon
Mirex
Chlorinated
cyclodiene
Toxaphene
Chlorinated
camphene
Seed treatments, soil treatments
for tobacco transplants, foliage
applications on fruit and nut trees,
vegetables, and wood and timber
protection.
Historically used primarily in fire
ant control in southeastern States
(Kutz et al., 1985) and was used
industrially as a fire retardant and
polymerizing agent in plastics
under the name dechlorane.
Historically used extensively
on cotton (Farm Chemicals
Handbook, 1989). Note: A
toxaphene-like compound can be
a byproduct of the paper industry
and has been identified in the
Great Lakes region (J. Hesse,
Michigan Department of Public
Health, personal communication,
1992).
Use of lindane in smoke
fumigation devices for indoor
domestic purposes was banned in
1985 (48 FR 48512, 50 FR 5424).
Use in dog dips permitted only for
veterinary use (U.S. EPA, 1985a).
Application permitted only under
supervision of certified applicator
(U.S. EPA, 1985a).
All registered uses of mirex were
canceled.in 1977 (41 FR 56703).
All existing stocks were not to be
sold, distributed, or used after
June 30, 1978 (NAS, 1978).
Registration for all uses was
canceled in the U.S. in November
1982 (47 FR 53784; U.S. EPA,
1990).
B2/C
d.e
B2
o
See notes and references at end of table.
(continued)
o
X
o
-------
Table C-1 (continued)
Pesticide
Family
Use
Registration
EPA
Toxicrty carcinogeniclty
class* classification*
Carbophenothion
Chlorpyrifos
Organophosphate
Heterocyclic
organophosphate
Insecticide and acaricide on wide
variety of vegetable, fruit, nut,
forage, ornamental and forestry
sites. The majority of pesticide
use is on citrus.6
Insecticide primarily used to
control soil and foliar insect pests
on cotton, peanuts, and sorghum
(Worthing, 1983; U.S. EPA,
1986a). In addition, it is used to
control root-infesting and boring
insects on a variety of fruits (e.g.,
citrus crops, apples, bananas,
peaches, grapes, nectarines), nuts
(e.g., almonds, walnuts), vege-
tables (e.g., beans, broccoli,
brussel sprouts, cauliflower,
soybeans, cabbage, peas) and
field crops (e.g., alfalfa and corn)
(U.S. EPA, 1984d) and to control
ticks on cattle and sheep
(Thomson, 1985). As a house-
hold insecticide it has been used
to control ants, cockroaches,
fleas, and mosquitoes (Worthing,
1983) and is registered for use in
controlling subsurface termites in
California (U.S. EPA, 1983).
Compound discontinued by
Stauffer Chemical Company in
1987 and reregistration is not
being supported by registrant.6-'
Active since 1965 (U.S. EPA,
1984b).c
Dd
Dd
•o
"0
m
g
x
o
o
CO
See notes and references at end of table.
(continued)
-------
Table C-1 (continued)
Pesticide
Family
Use
Registration
EPA
ToxicRy carcinogenicity
class" classification6
Diazinon
Heterocyclic
organophosphate
DisuKoton
Aliphatic
organophosphate
Insecticide and nematicide for
control of soil insects and pests of
fruits, vegetables, tobacco, forage,
field crops, range, pasture,
grasslands and ornamentals.
Used to control cockroaches and
other household insects; grubs
and nematodes in turf; as a seed
treatment and for fly control (Farm
Chemicals Handbook, 1989).
Systemic insecticide and a
caricide on grain, nut, cole and
root crops; pome, strawberry and
pineapple fruits; forage, field and
vegetable crops, sugarcane, seed
crops, forest plantings,
ornamentals and potted plants
(houseplants) (U.S. EPA, 1984c).
Active since 1952 (U.S. EPA,
1986c).c
Dd
Active since 1958 (U.S. EPA,
1984c).c
Ethion
Organothiophosphate
Insecticide (nonsystemic) for
control of leaf-feeding insects,
mites, and scale insects. Citrus
accounts for 86%-89% of total
pounds of ethion used in the U.S.
with the remaining 11%-14%
applied to cotton, a variety of fruit
trees, nut trees, and vegetables
(U.S. EPA, 1989).
Active since 1965 (U.S. EPA,
1989).c
TJ
•o
m
•z.
g
x
o
9
to
See notes and references at end of table.
(continued)
-------
Table C-1 (continued)
Pesticide
Family
Use
Registration
EPA
Toxicrty carcinogenicity
class8 classification1*
Terfoufos
Organophosphate
Systemic insecticide and
nematicide on corn, sugar beets,
and grain sorghum (U.S. EPA,
1985c).
Active since 1974; however,
granular end-use products
containing 15% or more terbufos
were classified as "Restricted
Use" after September 1985 (U.S.
EPA, 1985c).c
Dd
Chlorophenoxy Herbicides
Oxyfluorfen
Diphenyl ether Pre- and postemergence herbicide
for a wide spectrum of annual
broadleaf weeds and grasses in
apples, artichokes, corn, cotton,
tree fruit, grapes, nuts, spearmint,
peppermint, certain topical
plantation, and ornamental crops
(Farm Chemicals Handbook,
1989)
Active since 1979.°
a Designations are from Farm Chemicals Handbook (1 989):
I = Oral LDgQ up to and including 50 mg/kg in laboratory animals.
IV
NA
Oral LDgQ from 50 through 500 mg/kg in laboratory animals.
Oral LDgQ from 500 through 5,000 mg/kg in laboratory animals.
Oral LDgQ greater than 5,000 mg/kg in laboratory animals.
No value available.
Designations are from IRIS (1992) unless otherwise noted: NA = not evaluated; A = human carcinogen; B1, B2 = probable human carcinogen; C = possible human
carcinogen; D = inadequate evidence of animal carcinogenicity; E = no evidence of carcinogenicity for humans; R = under review by EPA.
O
o
(continued)
TJ
TJ
m
g
x
o
-------
Table C-1 (continued)
c This pesticide has an active registration for agricultural use. The EPA Office of Pesticide Programs is responsible for registration and reregistration of pesticides.
The 1988 Amendment of FIFRA requires EPA to reregister each "registered pesticide containing any active ingredient contained in any pesticide first registered
before November 1,1984, except for any pesticide as to which the Administration has determined, after November 1,1984 that—(1) there are no outstanding
data requirements; and (2) the requirements of section 3(c)(5) have been satisfied" (U.S. EPA, 1988). The Agency will review all relevant data submitted by the
registrant for each pesticide reregistration and will use the data to conduct a risk assessment. Any subsequent regulatory action will be based on the results of
the risk assessment. If the data submitted are incomplete at the predetermined review time, the pesticide may be suspended.
d EPA carcinogenicily classification based on Classification List of Chemicals Evaluated for Carcinogenicrty Potential (U.S. EPA, 1992).
6 Previously classified by EPA as B2 (IRIS, 1989).
References:
ATSDR (Agency for Toxic Substances and Disease Registry). 1987a. Draft Toxicological Profile for Aldrin/Dieldrin. U.S. Public Health Service, Washington, DC.
ATSDR (Agency for Toxic Substances and Disease Registry). 1987b. Draft Toxicological Profile for Heptachlor. U.S. Public Health Service, Washington, DC.
Farm Chemicals Handbook. 1989. Meister Publishing Company, Willoughby, OH.
Hartley, D., and H. Kidd (eds.). 1987. Agrochemicals Handbook. Royal Society of Chemistry, Nottingham, England.
Hodges, L. 1977. Environmental Pollution. Holt, Rinehart and Winston, New York, NY.
IRIS (Integrated Risk Information System). 1989. U.S. Environmental Protection Agency, Duluth, MN.
IRIS (Integrated Risk Information System). 1992. U.S. Environmental Protection Agency, Duluth, MN.
Kutz F.W., S.C. Strassman, C.R. Stroup, J.C. Carra, C.C. Leininger, D.L. Watts, and C.M. Sparacino. 1985. The human body burden of mirex in the southeastern
United States. J. Toxicol. and Environ. Health 15:385-394.
Morris, C.R., and J.R.P. Cabral (eds.). 1986. Hexachlorobenzene: Proceedings of an International Symposium. IARC Scientific Publication No. 77. World Health
Organication, Lyon, France.
NAS (National Academy of Sciences). 1978. Kepone/Mirex/Hexachlorocyclopentadiene: An Environmental Assessment. National Academy of Sciences, National
Research Council, Washington, DC.
z
(continued)
O
-------
Table C-1 (continued)
Thomson, W.T. 1985. Agricultural Chemicals Book I - Insecticide, 1985 revision. Thomas Publication, Davis, CA.
U.S. EPA (U.S. Environmental Protection Agency). 1980. Ambient Water Quality Criteria for Endrin. EPA-440/5-80-047. Office of Water Regulations and Standards,
Washington, DC.
U.S. EPA (U.S. Environmental Protection Agency). 1983. Analyses of the Risks and Benefits of Seven Chemicals Used for Subterranean Termite Control. EPA-
540/9-83-005. Office of Pesticide Programs, Washington, DC.
U.S. EPA (U.S. Environmental Protection Agency). 1984a. Internal memorandum from G. LaRocca to B. Burnam et al., August 16, 1984. Office of Pesticide
Programs, Washington, DC.
U.S. EPA (U.S. Environmental Protection Agency). 1984b. Pesticide Fact Sheet—Chlorpyrifos. Office of Pesticides and Toxic Substances, Office of Pesticide
Programs, Washington, DC.
U.S. EPA (U.S. Environmental Protection Agency). 1984c. Pesticide Fact Sheet—Disulfoton. Office of Pesticides and Toxic Substances, Office of Pesticide
Programs, Washington, DC.
U.S. EPA (U.S. Environmental Protection Agency). 1984d. Registration Standard for Chlorpyrifos. Office of Pesticides and Toxic Substances, Office of Pesticide
Programs, Washington, DC.
U.S. EPA (U.S. Environmental Protection Agency). 1985a. Guidance for the Registration of Pesticide Products Containing Lindane as the Active Ingredient. EPA-
540/RS-86-121. U.S. EPA Office of Pesticide Programs. Washington, DC.
U.S. EPA (U.S. Environmental Protection Agency). 1985b. Suspended, Cancelled, and Restricted Pesticides. U.S. EPA Office of Pesticides and Toxic Substances,
Washington, DC.
U.S. EPA (U.S. Environmental Protection Agency). 1985c. Pesticide Fact Sheet—Terbufos. Office of Pesticides and Toxic Substances, Office of Pesticide
Programs, Washington, DC.
U.S. EPA (U.S. Environmental Protection Agency). 1986a. Ambient Water Quality Criteria for Chlorpyrifos. EPA-440/5-86-005. Office of Water Regulations and
Standards, Washington, DC.
U.S. EPA (U.S. Environmental Protection Agency). 1986b. Computer Retrieval from Office of Pesticides for Registered Sites and Number of Products Registered.
Office of Pesticide Programs, Washington, DC. ^
TJ
m
z
(continued) 2
O
-------
Table C-1 (continued)
o
CO
U.S. EPA (U.S. Environmental Protection Agency). 1986c. Pesticide Fact Sheet—Diazinon. Office of Pesticides and Toxic Substances, Office of Pesticide Programs,
Washington, DC.
U.S. EPA (U.S. Environmental Protection Agency). 1988. The Federal Insecticide, Fungicide, and Rodenticide Act as Amended. EPA-540/09-89-012. Office of
Pesticide Programs, Washington, DC.
U.S. EPA (U.S. Environmental Protection Agency). 1989. Pesticide Fact Sheet—Ethion. Office of Pesticides and Toxic Substances, Office of Pesticide Programs,
Washington, DC.
U.S. EPA (U.S. Environmental Protection Agency). 1990. Suspended, Cancelled, and Restricted Pesticides. Document 20T-1002, Office of Pesticides and Toxic
Substances, Washington, DC.
U.S. EPA (U.S. Environmental Protection Agency). 1992. Classification List of Chemicals Evaluated for Carcinogenicity Potential. Office of Pesticide Programs,
Washington, DC.
Worthing, C.R. 1991. The Pesticide Manual: A World Compendium. 9th edition. British Crop Protection Council, Croydon, England.
TJ
m
z
g
x
-------
APPENDIX D
TARGET ANALYTE DOSE-RESPONSE
VARIABLES AND ASSOCIATED INFORMATION
-------
Table D-1. Target Analyte Dose-Response Variables and Associated Information
Noncarclnogens
Target analyte
Metals
Cadmium
Mercury (as methyl
mercury)
Selenium9
Orqanochlortne
Pesticides
Chlordane (sum of cis-
and trans-chtordane. ds-
RfO"
(degree of
confidence;
uncertainty factor)
IxKT3
(high; 10)
6x10-*'
(medium; 10)
SxlO-3
(high; 3)
6x10-*
(tow; 1000)
Critical toxic effect
Significant proteinurea in
humans
Central nervous system
effects (e.g., ataxia,
parathesia) in humans
Selenosis in humans
Regional fiver lesions
(hypertrophy) in one
SF"
(discussion of
confidence)
NA
NA
NA
1.3
(Adequate number of
Carcinogens
EPA
Critical carcinogenic cardnogenlclty
effect0 classification*
- B1e
— NA
— D
Hepatocellular carcinomas 62
in two strains of mice (male
and trans-nonachlor, and
oxychtordane)
strain of female rats
DDT (sum of 4.4'- and
2,4'- isomers of DDT.
DDE. and DDD)
Liver lesions in rats
(medium; 100)
animals observed. SF
is the geometric mean
of SFs for four data
sets from two studies.
This SF is consistent
with SF= 1.1 derived
from less sensitive rat
species)
3.4 xlO"1
(SF is geometric mean
of SFs from 10 data
sets. SF from mouse
data only «* 4.8 x 10"1;
SF from rat data only
«=1.5x 10'1)
and female)
DDT: Uver tumors in six
studies in two
mouse strains and
two studies in two
rat strains
B2
O
OJ
See notes and references at end of table.
(continued)
•o
m
o
x
o
-------
Table D-1 (continued)
Target analyte
Noncarclnogens
RID"
(degree of
confidence;
uncertainty factor) Critical toxic effect
SFb
(discussion of
confidence)
Carcinogens
Critical carcinogenic
effect6
EPA
carclnogenlctty
classification*
3.4 x 10'1
(Adequate number of
animals observed. SF
for mouse studies
alone is within a factor
of 2 for mouse and
hamster data
combined)
2.4 x10'1
(Adequate number of
animals observed. SF
calculated using tumor
incidence data from
only one dose)
DDE: Liver tumors B2
(including
carcinomas) in two
strains of mice and
in hamsters
DDD: Liver tumors in one 62
strain of mice (males
only)
Dicofol
Dieldrin
Endosulfan (sum of
endosulfan I and II)
Endrin
1x10-3h
(NA, 1000)
5 x 10'5
(medium; 100)
1.5x103"
(medium; 100)
3 x 10'4
(medium; 100)
Increase liver to body
weight ratios observed in
2-yr rat feeding study.11
Liver lesions (focal
proliferation and focal
hyperplasia) in one strain
of female rats
Kidney toxicity in one
strain of male ratsh
Mild histological lesions
in livers in dogs (both
sexes)
NA —
16 Liver carcinomas in five
(SF is the geometric strains of mice (male and
mean of SFs from 13 female)
data sets. Individual
SFs ranged within a
factor of 8)
NA —
NA —
C1
B2
E1
D
See notes and references at end of table.
(continued)
•D
•o
m
g
x
o
-------
Table D-1 (continued)
Noncarclnogens
Target analyte
Heptachlor epoxide
RID"
(degree of
confidence;
uncertainty factor)
1.3x 10'5
(low; 1000)
Critical toxic effect
Increased liver-to-body
weight ratios in male and
SFb
(discussion of
confidence)
9.1
(Adequate number of
Carcinogens
Critical carcinogenic
effect0
Hepatocellular carcinomas
in two strains of mice (male
EPA
carclnogenlclty
classification"
B2
Hexachlorobenzene
8 x 10'"
(medium; 100)
Lindane (y-BHC)
3 x 10'4
(medium; 1000)
Mirex
2 x 104
(high; 300)
female dogs
Liver effects (hepatic
centrilobular basophilic
chromogenesis) in one
strain of rats (both sexes)
Liver and kidney toxicity
(liver hypertrophy, kidney
tubular degeneration,
hyaline droplets, tubular
distension, interstitial
nephritis, and basiophilic
tubules) in both sexes of
one strain of rats
Liver cytomegaly, fatty
metamorphosis,
angiectasis and thyroid
cystic follicles in one
strain of rats.
animals observed in
both studies, but
survival in one study
was low. This SF is
consistent with SF =
5.8 for one strain of
seven rats.)
1.6
(Significant increases
in malignant tumors
observed among an
adequate number of
animals observed for
their lifetime)
1.3'
NA1
and female)
Hepatocellular carcinomas
in one strain of rats
(females only)
B2
B2/Cilk
O
cn
See notes and references at end of table.
(continued)
rn
g
x
o
-------
Table D-1 (continued)
Target analyte
Toxaphene
Noncarclnogens
RID"
(degree of
confidence;
uncertainty factor) Critical toxic effect
2.5 xKT411'"1 Slight fiver
SF"
(discussion of
confidence)
1.1
Carcinogens
Critical carcinogenic
effect0
Hepatocellular carcinomas
EPA
carclnogenlctty
classification"
B2
(NA, 1000)
Organophosphate
Pesticides
Carbophenothion
Chlorpyrifos
Diazinon
Disulfoton
1.3 xlO"4
(NA, 1000)
3 x 10 3
(medium, 10)
9X10-5"
(NA, 100)
4 x 10'5
(medium, 1000)
degeneration—granularity
and vacuolization of
hepatocytes.h
Decreased plasma and
brain chofinesterase
(ChE) observed in 2-yr
chronic dog feeding
study."
Decreased plasma ChE
activity observed in 20-
day human feeding
study.11
Inhibition of plasma ChE
observed in 90-day rat
feeding study.h
ChE inhibition and
degeneration of the optic
nerve observed in 2-yr
dog feeding study.*1
(Adequate number of
animals observed. A
dose-response effect
was seen in a study
with three non-zero
dose levels)
NA
and neoplastic nodules in
one strain of mice (males
only)
NA
NA
NA
D1
D1
D'
D1
O
o>
See notes and references at end of table.
(continued)
TJ
m
o
X
o
-------
Table D-1 (continued)
Target analyte
Ethion
Terbulos
Chlorophenoxy
Herbicides
Oxyfluorten
PCBs
Total PCBs (sum of
Aroclors)
Dloxlns/dlbenzofurans
See notes and references
Noncarclnogens
RfD8
(degree of
confidence;
uncertainty factor) Critical toxic effect
5 x 10~* Plasma ChE inhibition
(medium, 100) and inhibition of brain
ChE observed in 21 -day
human feeding study.*1
1 .3 x 1 0"4 h Inhibition of plasma ChE
(NA, 10) observed in 28-day dog
feeding study .h
3 x 10*3 Increased absolute fiver
(high, 100) weight and nonneoplastic
lesions were observed in
20-month mouse feeding
study."
NA —
NA —
on next page.
Carcinogens
SF* EPA
(discussion of Critical carcinogenic carclnogenlctty
confidence) effect0 classification*1
NA — D1
NA — D1
1 .3 x 10 ' ' Evidence of cartinogenitity C1
(fiver tumors) in mice.*1
7.7" Trabecular B2
(Adequate number of cartinomas/adenocartino-
animals observed for mas, neoplastic nodules in
their normal fifespan. one strain of rats (females
Only one non-zero only)
test dose used)
1.56x105° NA B2
(continued) >
TJ
TO
m
a
x
a
-------
Table D-1 (continued)
co
NA = Not available in IRIS (1992).
a RfD = Oral reference dose (mg/kg/day); from IRIS (1992) unless otherwise noted (see Section 5.1.1).
b SF = Oral slope factor (mg/kg/day)~1; from IRIS (1992) unless otherwise noted (see Section 5.1.2).
c The critical effect is the effect observed in oral dose response studies used to determine the SF.
d Except where noted, all EPA carcinogenicity classifications are taken from IRIS (1992):
A = Human carcinogen based on sufficient evidence from epidemiologic studies.
B1 = Probable human carcinogen based on at least limited evidence of carcinogenicity to humans.
B2 = Probable human carcinogen based on a combination of sufficient evidence in animals and inadequate data in humans.
C = Possible human carcinogen based on limited evidence of carcinogenicity in animals in the absence of human data.
D = Not classifiable based on lack of data or inadequate evidence of carcinogenicity from animal data.
E = No evidence of carcinogenicity for humans (no evidence of carcinogenicity in at least two adequate animal tests in different species or in
both epidemiologic and animal studies).
R = Currently under review by EPA.
6 Based on limited evidence from human occupational epidemiologic studies where the primary route of exposure was by inhalation, and on sufficient
evidence from studies in which rats and mice were exposed by inhalation and intramuscular and subcutaneous injection. However, data are inadequate
to conclude that cadmium is carcinogenic via ingestion. The EPA Office of Drinking Water classifies cadmium as a Group D carcinogen in the health
advisory for cadmium (U.S. EPA, 1987).
' For the purpose of calculating an SV, the RfD for methylmercury currently available in the EPA IRIS database (3 x 10~4 mg/kg/d) has been lowered by a
factor of 5 to a value of 6 x 10'5 mg/kg/d. The EPA is reevaluating the RfD for methylmercury and is especially concerned about evidence that the
fetus, and possibly pregnant women, are at increased risk of adverse neurological effects from exposure to methylmercury (WHO, 1976, 1990;
Piotrowski and Inskip, 1981; Marsh et al., 1987). In the general adult population, blood methylmercury concentrations of 200 ug/L (corresponding to
approximately 50 jig/g in hair) have been associated with a 5 percent risk of parasthesia; whereas for the fetus, a 5 percent risk of neurological and
developmental abnormalities is associated with peak mercury concentrations of 10-20 ng/g in the maternal hair (WHO, 1990). These findings suggest a
possible fivefold increase in fetal sensitivity to methylmercury exposure. Consequently, the EPA has chosen to apply an uncertainty factor of 5 to the
current IRIS RfD for methylmercury. This approach was deemed to be the most prudent as an interim measure until the current revaluation of the
methylmercury RfD is completed. The degree of confidence and uncertainty factor listed are for the current oral RfD for methylmercury (RfD=3x10~4)
reported in IRIS (1992).
(continued)
-------
Table D-1 (continued)
8 The oral RfD is for selenious acid (IRIS, 1992). The evidence of carcinogenicity for various selenium compounds in animals and mutagenicity studies is
conflicting and difficult to interpret. However, evidence for selenium sulfides is sufficient for a B2 classification (IRIS, 1992).
h Reference dose information is taken from the Reference Dose List (U.S. EPA, 1993).
1 EPA carcinogenicity classifications are taken from Classification List of Chemicals Evaluated for Carcinogenicity Potential (U.S. EPA, 1992a).
' IRIS (1992) has not provided an SF for lindane. The SF value listed for lindane was calculated from the water quality criteria (0.063 ng/L) (U.S. EPA,
1992d) and is comparable to the SF of 1.33 mg/kg/d"1 from the Public Health Risk Evaluation Database (U.S. EPA, 1988b).
k Previously classified by EPA as B2 (IRIS, 1989). Available data need to be reviewed further, but at a minimum lindane will be classified as a C
carcinogen (U.S. EPA, 1992a).
1 The National Study of Chemical Residues in Fish (U.S. EPA, 1992b, 1992c) used a value of SF = 1.8 for mirex from HEAST (1989).
m The RfD value is the Office of Pesticide Programs value; this value was never submitted for verification.
n The SF is based on a carcinogenicity assessment of Aroclor 1260. The SF of Aroclor 1260 is intended to represent the upper bound risk for all PCB
mixtures (IRIS, 1992).
0 The SF value listed is for 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)(U.S. EPA, 1986). The National Study of Chemical Residues in Fish used a value
of RfD = 1x10"9 for 2,3,7,8-TCDD from ATSDR (1987). It is recommended that, in both screening and intensive studies, the tetra- through octa-
chlorinated dibenzo-p-dioxins (PCDDs) and dibenzofurans (PCDFs) be determined and a toxicity-weighted total concentration be calculated for each
sample for comparison with the recommended SV, using the revised interim method for estimating Toxicity Equivalency Concentration (TECs) (Barnes
and Bellin, 1989; U.S. EPA, 1991). If resources are limited, the 2,3,7,8-TCDD and 2,3,7,8-TCDF congeners should be determined, at a minimum.
References:
ATSDR (Agency for Toxic Substances and Disease Registry). 1987. Toxicological Profile for 2,3,7,8-TCDD (Dioxin). Draft. U.S. Public Health Service in
collaboration with the U.S. Environmental Protection Agency, Washington, DC.
•o
m
o
(continued)
O
-------
Table D-1 (continued)
o
o
Barnes, D.G., and J.8. Bellin. 1989. Interim Procedures for Estimating Risks Associated with Exposures to Mixtures of Chlorinated Dibenzo-p-Dioxins
and -Dibenzofurans (CDDs and CDFs). Risk Assessment Forum, U.S. Environmental Protection Agency, Washington, DC.
HEAST. 1989. Health Effects Summary Tables. Office of Emergency and Remedial Response, U.S. Environmental Protection Agency, Washington, DC.
IRIS (Integrated Risk Information System). 1989. U.S. Environmental Protection Agency, Duluth, MN.
IRIS (Integrated Risk Information System). 1992. U.S. Environmental Protection Agency, Duluth, MN.
Marsh, D.O., T.W. Clarkson, C. Cox, G.J. Meyers, L Amin-Zaki and S. AI-Tikriti. 1987. Fetal methylmercury poisoning: relationship between
concentration in single strands of maternal hair and child effects. Archives of Neurology 44:1017-1022.
Piotrowski, J.K., and M.J. Inskip. 1981. Health Effects of Mercury; A Technical Report (1981). MARC Report Number 24. Chelsea College, University of
London. 82 pp.
U.S. EPA (U.S. Environmental Protection Agency). 1986. Health Assessment Document for Polychlorinated Dibenzofurans. Draft. EPA 600/8-86-018A.
Environmental Criteria and Assessment Office, Cincinnati, OH.
U.S. EPA (U.S. Environmental Protection Agency). 1987. Quality Assurance/Quality Control (QA/QC) for 301(h) Monitoring Programs: Guidance on Field
and Laboratory Methods. EPA-430/9-86-004. Office of Marine and Estuarine Protection, Washington, DC.
U.S. EPA (U.S. Environmental Protection Agency). 1988. Public Health Risk Evaluation Database. Office of Emergency and Remedial Response,
Washington, DC.
U.S. EPA (U.S. Environmental Protection Agency). 1991. National Bioaccumulation Study. Draft. Office of Water Regulations and Standards,
Washington, DC.
U.S. EPA (U.S. Environmental Protection Agency). 1992a. Classification List of Chemicals Evaluated for Carcinogenicity Potential. Office of Pesticide
Programs, Washington, DC.
U.S. EPA (U.S. Environmental Protection Agency). 1992b. National Study of Chemical Residues in Fish. Volume I. EPA-823/R-92-008a. Office of
Science and Technology, Washington, DC.
(continued)
•u
•a
m
o
x
o
-------
Table 0-1 (continued)
U.S. EPA (U.S. Environmental Protection Agency). 1992c. National Study of Chemical Residues in Fish. Volume II. EPA-823/R-92-008b. Office of
Science and Technology, Washington, DC.
U.S. EPA (U.S. Environmental Protection Agency). 1992d. 304(a) Criteria and Related Information for Toxic Pollutants. Spreadsheet. Water Quality
Standards Unit, Water Management Division, Region 4, Atlanta, GA.
U.S. EPA (U.S. Environmental Protection Agency). 1993. Reference Dose List. Office of Pesticide Programs, Health Effects Division, Washington, DC.
WHO (World Health Organization). 1976. Environmental Health Criteria. 1. Mercury. Geneva, Switzerland.
WHO (World Health Organization). 1990. Environmental Health Criteria 101: Methylmercury. Geneva, Switzerland.
•o
fn
O
X
o
-------
APPENDIX E
QUALITY ASSURANCE AND
QUALITY CONTROL GUIDANCE
-------
APPENDIX E
APPENDIX E
QUALITY ASSURANCE(QA) AND
QUALITY CONTROL (QC) GUIDANCE
E.1 GENERAL QA AND QC CONSIDERATIONS
The primary objective of the specific QA and QC guidance provided in this
document is to ensure that
Appropriate data quality objectives or requirements are established prior
to sample collection and analysis
Samples are collected, processed, and analyzed according to scientifically
valid, cost-effective, standardized procedures
The integrity and security of samples and data are maintained at all times
Recordkeeping and documentation procedures are adequate to ensure the
traceability of all samples and data from initial sample collection through
final reporting and archiving, and to ensure the verifiability and defensibility
of reported results
Data quality is assessed, documented, and reported properly
Reported results are complete, accurate, and comparable with those from
other similar monitoring programs.
E.2 QA PLAN REQUIREMENTS
To ensure the quality, defensibility, and comparability of the data used to
determine exposure assessments and fish consumption advisories, it is essential
that an effective QA program be developed as part of the overall design for each
monitoring program. The QA program should be documented in a written QA
plan or in a combined Work/QA Project Plan and should be implemented strictly
throughout all phases of the monitoring program. The QA plan should include
the following information either in full or by reference to appropriate standard
operating procedures (SOPs):
1. A clear statement of program objectives
E-3
-------
APPENDIX E
2. A description of the program organization and personnel roles and
responsibilities, including responsibility for ensuring adherence to the QA
plan
3. Specification of data quality objectives in terms of accuracy, precision,
representativeness, and completeness, for data generated from each type
of measurement system
4. Detailed descriptions of field sample collection and handling procedures,
including documentation of
• Target species and size (age) class
• Sampling site locations
• Target contaminants
• Sampling times/schedules
• Numbers of samples and sample replication strategy
• Sample collection procedures
• Sample processing procedures, including sample identification, labeling,
preservation, and storage conditions
• Sample shipping procedures
5. A detailed description of chain-of-custody procedures, including specifi-
cation of standard chain-of-custody forms and clear assignment of field and
laboratory personnel responsibilities for sample custody
6. Detailed descriptions of laboratory procedures for sample receipt, storage,
and preparation, including specification of the kinds of samples to be
prepared for analyses (e.g., composite vs. individual, whole body vs. fillet,
replicates)
7. Detailed descriptions of the analytical methods used for quantitation of
target contaminants, and percent lipid determination including
• Specification and definition of method detection limits
• Method validation procedures for verification of specifications for
method accuracy, precision, and detection limits prior to analysis of field
samples
8. Detailed descriptions of methods routinely used to assess data accuracy,
precision, and completeness, including
__
-------
APPENDIX E
Internal QC checks using field, reagent, or method blanks; spiked
samples; split samples; QC samples prepared from standard reference
materials; and replicate analyses
• Calibration checks
• Data quality assessments
9. Detailed descriptions of calibration procedures for all measurement
instruments, including specification of reference materials used for
calibration standards and calibration schedules
10. Detailed descriptions of preventive maintenance procedures for sampling
and analysis equipment
11. Detailed description of health and safety procedures
12. Detailed descriptions of recordkeeping and documentation procedures,
including requirements for
Maintaining field and laboratory logs and notebooks
• Use of standard data collection and reporting forms
• Making changes to original records
• Number of significant figures to be recorded for each type of data
• Units of reporting
• Routine procedures to assess the accuracy and completeness of
records
13. Detailed descriptions of data analysis procedures, including
• Statistical treatment of data
Data summary formats (e.g., plots, tables)
14. Detailed descriptions of data management and reporting procedures,
including requirements for
• Technical reports
• QA and QC reports
Data coding procedures
• Database specifications
• QA review of reported data
• Data storage and archiving procedures
E-5
-------
APPENDIX E
15. Detailed descriptions of procedures for internal QC performance and/or
systems audits for sampling and analysis programs.
16. Detailed descriptions of procedures for external QA performance and/or
systems audits for sampling and analysis programs, including participation
in certified QA proficiency testing or interlaboratory comparison programs.
17. Detailed descriptions of corrective action procedures in both sampling and
analysis programs, including
• Criteria and responsibility for determining the need for corrective action
• Procedures for ensuring that effective corrective action has been taken
• Procedures for documenting and reporting corrective actions
18. A description of procedures for documenting deviations from standard
procedures, including deviations from QA or QC requirements
19. A description of the procedure for obtaining approval for substantive
changes in the monitoring program.
Guidance for addressing each of the QA or QC elements outlined above,
including a list of recommended standard reference materials and external QA
or interlaboratory comparison programs for the analyses of target analytes, is
incorporated in the appropriate sections of this guidance document.
E-6
-------
APPENDIX F
RECOMMENDED PROCEDURES FOR PREPARING
WHOLE FISH COMPOSITE HOMOGENATE SAMPLES
-------
APPENDIX F
APPENDIX F
RECOMMENDED PROCEDURES FOR PREPARING WHOLE
FISH COMPOSITE HOMOGENATE SAMPLES
F.1 GENERAL GUIDELINES
Laboratory processing to prepare whole fish composite samples (diagrammed
in Figure F-1) involves
Inspecting individual fish for foreign material on the surface and rinsing if
necessary
Weighing individual fish
Examining each fish for morphological abnormalities (optional)
Removing scales or otoliths for age determination (optional)
Determining the sex of each fish (optional)
Preparing individual whole fish homogenates
Preparing a composite whole fish homogenate.
Whole fish should be shipped on wet or blue ice from the field to the sample
processing laboratory if next-day delivery is assured. Fish samples arriving in
this manner (chilled but not frozen) should be weighed, scales and/or otoliths
removed, and the sex of each fish determined within 48 hours of sample
collection. The grinding/homogenization procedure may be carried out more
easily and efficiently if the sample has been frozen previously (Stober, 1991).
Therefore, the samples should then be frozen (<20 °C) in the laboratory prior to
being homogenized.
If the fish samples arrive frozen (i.e., on dry ice) at the sample processing
laboratory, precautions should be taken during weighing, removal of scales
and/or otoliths, and sex determination to ensure that any liquid formed in thawing
remains with the sample. Note: The liquid will contain target analyte
contaminants and lipid material that should be included in the sample for
analysis.
F-3
-------
APPENDIX F
Log in fish samples using COC procedures
Unwrap individual fish, weigh, and record weight (g)
Examine fish for morphological abnormalities (optional)
Remove scales and/orotoliths forage determination (optional)
Determine sex (optional)
Fish <1,000 g
Fish 21,000 g
Partially thaw
Grind whole fish in a hand crank
meat grinder (<300 g) or a food
processor (300-1000 g)
Partially thaw
Chop sample into -2.5-cm
cubes
Pass entire chopped sample
through a meat grinder
Divide ground sample into
quarters, mix opposite quarters
and then mix halves
Repeat from * two more times
Composite equal weights (g) of
homogenized tissues from the
selected number of fish (200-g)
Seal and label (200-g)
homogenate in appropriate
containers) and store at £20 °C
until analysis (see Table F-1 for
recommended container materials
and holding times)
Optional
Save remainder of
homogenate from each
individual fish; seal and label
in appropriate container and
archive at £20 °C until
analysis (see Table F-1 for
recommended container
materials and holding times)
COC = Chain of Custody
Figure F-1. Laboratory sample preparation and handling for
whole fish composite homogenate samples.
F-4
-------
APPENDIX F
Table F-1. Recommendations for Container Materials,
Preservation Temperature, and Holding Times for Fish Tissues
from Receipt at Central Processing Laboratory to Analysis
Analyte
Mercury
Other metals
Organics
Metals and
organics
Matrix
Tissue (whole
specimens,
homogenates)
Tissue (whole
specimens,
homogenates)
Tissue (whole
specimens,
homogenates)
Tissue (whole
specimens,
homogenates)
Sample
container
Plastic,
borosilicate
glass, quartz,
and PTFE
Plastic,
borosilicate
glass, quartz,
and PTFE
Borosilicate
glass, quartz,
PTFE, and
aluminum foil
Borosilicate
glass, quartz,
and PTFE
Storage
Preservation Holding
time
Freeze at <-20 °C 28 days
Freeze at £-20 °C 1 year
Freeze at <-2Q °C 1 year
Freeze at <-2Q °C 28 days
(mercury
analysis) and
1 year (other
metals and
organics)
PTFE = polytetrafluoroethylene; Teflon.
The thawed or partially thawed whole fish should then be homogenized
individually, and equal weights of each homogenate should be combined to form
the composite sample. Individual homogenates and/or composite homogenates
may be frozen; however, frozen individual homogenates must be rehomogenized
before compositing, and frozen composite homogenates must be rehomogenized
before aliquotting for analysis. The maximum holding time from sample
collection to analysis for mercury is 28 days at <-20 °C; for all other analytes, the
holding time is 1 year at <-20 °C (Stober, 1991). Recommended container
materials, preservation temperatures, and holding times are given in Table F-1.
F.2 SAMPLE PROCESSING PROCEDURES
Fish sample processing procedures are discussed in more detail in the sections
below. Each time custody of a sample or set of samples is transferred from one
person to another during processing, the Personal Custody Record of the chain-
of-custody (COC) form that originated in the field (Figure 6-8) must be completed
and signed by both parties so that possession and location of the samples can
F-5
-------
APPENDIX F
be traced at all times (see Section 7.1). As each sample processing procedure
is performed, it should be documented directly in a bound laboratory notebook
or on standard forms that can be taped or pasted into the notebook. The use
of a standard form is recommended to ensure consistency and completeness of
the record. Several existing programs have developed forms similar to the
sample processing record for whole fish composite samples shown in Figure F-2.
F.2.1 Sample Inspection
Individual fish received for filleting should be unwrapped and inspected carefully
to ensure that they have not been compromised in any way (i.e., not properly
preserved during shipment). Any specimen deemed unsuitable for further
processing and analysis should be discarded and identified on the sample
processing record.
F.2.2 Sample Weighing
A wet weight should be determined for each fish. All samples should be
weighed on balances that are properly calibrated and of adequate accuracy and
precision to meet program data quality objectives. Balance calibration should be
checked at the beginning and end of each weighing session and after every 20
weighings in a weighing session.
Fish shipped on wet or blue ice should be weighed directly on a foil-lined
balance tray. To prevent cross contamination between individual fish, the foil
lining should be replaced after each weighing. Frozen fish (i.e., those shipped
on dry ice) should be weighed in clean, tared, noncontaminating containers if
they will thaw before the weighing can be completed. Liquid from the thawed
sample must be kept in the container as part of the sample because it will
contain lipid material that has separated from the tissue (Stober, 1991).
All weights should be recorded to the nearest gram on the sample processing
record and/or in the laboratory notebook.
F.2.3 Age Determination
Age provides a good indication of the duration of exposure to pollutants (Versar,
1982). A few scales or otoliths (Jearld, 1983) should be removed from each fish
and delivered to a fisheries biologist for age determination. For most warm
water inland gamefish, 5 to 10 scales should be removed from below the lateral
line and behind the pectoral fin. On softrayed fish such as trout and salmon, the
scales should be taken just above the lateral line (WDNR, 1988). For catfish
and other scaleless fish, the pectoral fin spines should be clipped and saved
(Versar, 1982). The scales, spines, or otoliths may be stored by sealing them
in small envelopes (such as coin envelopes) or plastic bags labeled with, and
cross-referenced by, the identification number assigned to the tissue specimen
F-6
-------
APPENDIX F
Sample Processing Record for Fish Contaminant Monitoring Program—Whole Fish Composites
Project No..
Sampling Date and Time:.
STUDY PHASE: Screening [_];
SITE LOCATION
Site Name/Number
County/Parish:
Intensive: Phase 11 I Phase IILJ
State Waterbody Segment Number.
LatAong.:
Waterbody Type:.
Bottom Feeder - Species Name:.
Composite Sample #:
Number of Individuals:
Fish*
001
002
003
004
005
006
007
008
009
010
Analyst
Initials/Data.
Weight (g)
Scales/Otoliths
Removed (/)
Sex
(M,F)
Homogenate
Prepared (/)
Weight of homogenato
taken for composite (g)
Total Composite Homogenate Weight
Predator - Species Name:
Composite Sample #:
Fish*
001
002
003
004
005
006
007
008
009
010
Analyst
Initials/Data.
Weight (g)
Scales/Otoliths
Removed (/)
Number of Individuals:
Sex Homogenate
(M,F) Prepared (/)
Weight of hpmogenoto
taken for composite (g)
Total Composite Homogenate Weight
Notes:
Figure F-2. Example of a sample processing record for fish contaminant monitoring
program—whole fish composites.
F-7
-------
APPENDIX F
(Versar, 1982). Removal of scales, spines, or otoliths from each fish should be
noted (by a check mark) on the sample processing record.
F.2.4 Sex Determination (Optional)
To determine the sex of a fish, an incision should be made on the ventral
surface of the body from a point immediately anterior to the anus toward the
head to a point immediately posterior to the pelvic fins. If necessary, a second
incision should be made on the left side of the fish from the initial point of the
first incision toward the dorsal fin. The resulting flap should be folded back to
observe the gonads. Ovaries appear whitish to greenish to golden brown and
have a granular texture. Testes appear creamy white and have a smooth texture
(Texas Water Commission, 1990). The sex of each fish should be recorded on
the sample processing record.
F.2.5 Assessment of Morphological Abnormalities (Optional)
Assessment of gross morphological abnormalities in finfish is optional. This
assessment may be conducted in the field (see Section 6.3.1.5) or during initial
inspection at the central processing laboratory prior to filleting. States interested
in documenting morphological abnormalities should consult Sinderman (1983)
and review recommended protocols for fish pathology studies used in the Puget
Sound Estuary Program (1990).
F.2.6 Preparation of Individual Homogenates
To ensure even distribution of contaminants throughout tissue samples, whole
fish must be ground and homogenized prior to analyses.
Smaller whole fish may be ground in a hand crank meat grinder (fish < 300 g)
or a food processor (fish 300-1,000 g). Larger (>1,000 g) fish may be cut into
2.5-cm cubes with a food service band saw (e.g., Hobart Model 5212) and then
ground in either a small (e.g., Hobart, 1/4 hp, Model 4616) or large (e.g., Hobart,
1 hp, Model 4822) homogenizer. To avoid contamination by metals, grinders,
and homogenizers used to grind and blend tissue should have tantalum or
titanium blades and/or probes. Stainless steel blades and probes have been
found to be a potential source of nickel and chromium contamination (due to
abrasion at high speeds) and should be avoided.
Grinding and homogenization of biological tissue, especially skin from whole fish
samples, is easier when the tissue is partially frozen (Stober, 1991). Chilling the
grinder/homogenizer briefly with a few chips of dry ice will reduce the tendency
of the tissue to stick to the grinder.
The ground sample should be divided into quarters, opposite quarters mixed
together by hand, and the two halves mixed back together. The grinding,
quartering, and hand mixing should be repeated two more times. If chunks of
tissue are present at this point, the grinding/homogenizing should be repeated.
_
-------
APPENDIX F
No chunks of tissue should remain because these may not be extracted or
digested efficiently. If the sample is to be analyzed for metals only, the ground
tissue may be mixed by hand in a polyethylene bag (Stober, 1991).
Homogenization of each individual fish should be noted on the sample
processing record. At this time, individual whole fish homogenates may be either
composited or frozen and stored at <-20 °C in cleaned containers that are
noncontaminating for the analyses to be performed (see Table F-1).
F.2.7 Preparation of Composite Homogenates
Composite homogenates should be prepared from equal weights of individual
homogenates. If individual whole fish homogenates have been frozen, they
should be thawed partially and rehomogenized prior to compositing. Any
associated liquid should be maintained as a part of the sample. The weight of
each individual homogenate that is used in the composite homogenate should
be recorded, to the nearest gram, on the sample processing record.
Each composite homogenate should be blended by dividing it into quarters,
mixing opposite quarters together by hand, and mixing the two halves together.
The quartering and mixing should be repeated at least two more times. If the
sample is to be analyzed only for metals, the composite homogenate may be
mixed by hand in a polyethylene bag (Stober, 1991). At this time, the composite
homogenate may be processed for analysis or frozen and stored at <-20 °C (see
Table F-1).
The remainder of each individual homogenate should be archived at <-20 °C with
the designation "Archive" and the expiration date recorded on the sample label.
The location of the archived samples should be indicated on the sample
processing record under "Notes."
It is essential that the weights of individual homogenates yield a composite
homogenate of adequate size to perform all necessary analyses. Weights of
individual homogenates required for a composite homogenate, based on the
number of fish per composite and the weight of composite homogenate
recommended for analyses of all screening study target analytes (see Table 4-1)
are given in Table F-2. The total composite weight required for intensive studies
may be less than in screening studies if the number of target analytes is reduced
significantly.
The recommended sample size of 200 g for screening studies is intended to
provide sufficient sample material to (1) analyze for all recommended target
analytes (see Table 4-1) at appropriate detection limits, (2) meet minimum QA
and QC requirements for the analyses of replicate, matrix spike, and duplicate
matrix spike samples (see Section 8.3.3.4), and (3) allow for reanalysis if the QA
and QC control limits are not met or if the sample is lost. However, sample size
requirements may vary among laboratories and the analytical methods used.
Therefore, it is the responsibility of each program manager to consult with the
analytical laboratory supervisor to determine the actual weights of composite
_
-------
APPENDIX F
Table F-2. Weights (g) of Individual Homogenates
Required for Screening Study Composite Homogenate Sample8
Number of
fish per
sample
3
4
5
6
7
8
9
10
Total composite weight
100 g
(minimum)
33
25
20
17
14
13
11
10
200 g
(recommended)
67
50
40
33
29
25
22
20
500 g
(maximum)
167
125
100
84
72
63
56
50
a Based on total number of fish per composite and the total composite weight required for
analysis in screening studies. The total composite weight required in intensive studies
may be less if the number of target analytes is reduced significantly.
homogenates required to analyze for all selected target analytes at appropriate
detection limits.
F.3 REFERENCES
Puget Sound Estuary Program. 1990 (revised). Recommended protocols for
fish pathology studies in Puget Sound. Prepared by PTI Environmental
Services, Bellevue, WA. In: Recommended Protocols and Guidelines for
Measuring Selected Environmental Variables in Puget Sound. Region 10,
U.S. Environmental Protection Agency, Seattle, WA. (Looseleaf)
Sinderman, C. J. 1983. An examination of some relationships between pollution
and disease. Rapp. P. V. Reun. Cons. Int. Explor. Mer. 182:37-43.
Stober, Q. J. 1991. Guidelines for Fish Sampling and Tissue Preparation for
Bioaccumulative Contaminants. Environmental Services Division, Region 4,
U.S. Environmental Protection Agency, Athens, GA.
Texas Water Commission. 1990. Texas Tissue Sampling Guidelines. Texas
Water Commission, Austin, TX.
F-10
-------
APPENDIX F
Versar, Inc. 1982. Sampling Protocols for Collecting Surface Water, Bed
Sediment, Bivalves and Fish for Priority Pollutant Analysis-Final Draft
Report. EPA Contract 68-01-6195. Prepared for U.S. EPA Office of Water
Regulations and Standards. Versar, Inc., Springfield, VA.
WDNR (Wisconsin Department of Natural Resources). 1988. Fish Contaminant
Monitoring Program-Field and Laboratory Guidelines (1005.1). Madison,
Wl.
F-11
-------
APPENDIX G
GENERAL PROCEDURES FOR REMOVING
EDIBLE TISSUES FROM SHELLFISH
-------
Heading, peeling and deveining shrimp
To head a shrimp, hold it in
one hand. With your thumb
behind shrimp head, push head
off. Be sure to push just the
head off so that you do not lose
any meat.
If using a deveiner, insert it
at head end, just above the
vein.
Push through shrimp to the tail
and split and remove shell.
This removes vein at the same
time.
If you prefer to use a paring
knife, shell shrimp with your
fingers or knife. Then use
knife to gently remove vein.
Source: UNC Sea Grant Publication UNC-SG-88-02
G-3
-------
Cleaning soft-shell crabs
Hold crab in one hand and cut
across body just behind eyes to
remove eyes and mouth.
Turn crab on its back. Lift
and remove apron and vein
attached to it.
Turn crab over and lift one
side of top shell.
With a small knife, scrape
off grayish-feathery gills.
Repeat procedure on other
side.
Source: UNC Sea Grant Publication UNC-SG-88-02
G-4
-------
Cleaning hard-shell crabs
Hold crab in one hand. Turn
crab over and stab straight
down at point of apron with a
knife.
Make two cuts from this
point to form a V-pattern
that will remove mouth.
Do not remove knife after
making second cut. Firmly
press crab shell to cutting
surface without breaking back
shell. With other hand, grasp
crab by legs and claws on the
side where you are holding
knife, and pull up. This should
pull crab body free from back
shell.
G-5
-------
Remove gray, feathery gills,
which are attached just above
legs. Cut and scrape upward to
remove gills.
Remove all loose
material—viscera and
eggs—from body cavity.
If apron did not come loose
with shell, remove it.
Source: UNC Sea Grant Publication UNC-SG-88-02
G-6
-------
Shucking oysters
Oyster shells are especially
sharp; be sure to wear gloves
to protect your hands. Chip off
a small piece of shell from the
thin lip of the oyster until
there is a small opening.
Insert knife blade into the
opening and cut muscle free
from top and bottom shells.
Remove oyster meat from the
shell.
Source: UNC Sea Grant Publication UNC-SG-88-02
G-7
-------
Shucking clams
In the back of clam near the
hinge is a black ligament.
Toward the front where
ligament ends is a weak spot.
Insert your knife at this spot.
Inside are two muscles.
Run the knife around the
shell to sever both
muscles.
Now insert the knife blade
into the front of the shell
and separate the two
shells.
Scrape the meat free
from the top and bottom
shell.
Source: UNC Sea Grant Publication UNC-SG-88-02
G-8
-------
APPENDIX H
COMPARISON OF TARGET ANALYTE SCREENING
VALUES (SVs) WITH DETECTION AND QUANTITATION LIMITS
OF CURRENT ANALYTICAL METHODS
-------
Table H-1. Comparison of Target Analyte Screening Values (SVs) with Detection and Quantltatlon Limits of
Current Analytical Methods8
Target Anelyte
Metals
Cadmium
Mercury
Selenium
Organochlorlne Pesticides
Chlordane (total)
cis-Chlordane
trans-Chlordane
cIs-Nonachlor
trans-Nonachlor
Oxychlordane
DDT (total)
4,4'-DDT
2,4 '-DDT
4,4'-DDD
2,4'-DDD
4,4'-DDE
2,4'-DDE
Dicolol
Dieldrin
Endosulfan (total)
Endosullan I
Endosullan II
Endrin
Heptachlor epoxlde
Hexachlorobenzene
Lindane
Mir ex
Toxaphene
SV"
10ppm
0.6 ppm
50 ppm
80 ppb
300 ppb
10,000 ppb
7ppb
20,000 ppb
3,000 ppb
10 ppb
70 ppb
80 ppb
2,000 ppb
100 ppb
Methods
Puget Sound
Protocols'
LOD1
0.01 ppm
0.01 ppm
N/l
1-5 ppb
N/l
N/l
N/l
N/l
0.1 -2 ppb
0.1 -2 ppb
0.1-2 ppb
0.1 -2 ppb
0.1 -2 ppb
0.1 -2 ppb
N/l
0.1 -2 ppb
N/l
N/l
N/l
N/l
0.1-2 ppb
0.1-2 ppb
N/l
3-15 ppb
POL"
N/R
N/R
N/l
20 ppb
N/l
N/l
N/l
N/l
4 ppb
4ppb
4pob
4ppb
4ppb
4ppb
N/l
4ppb
N/l
N/l
N/l
N/l
4ppb
4ppb
N/l
60 ppb
National Study of Chemical
Residues In Fish"
MLD1
N/l
1.3 ppb (LOD)1
N/l
N/R
N/R
N/R
N/R
N/R
N/l
N/l
N/l
N/l
N/R
N/l
N/R
N/R
N/l
N/l
N/R
N/R
N/R
N/R
N/R
N/l
TQLm
N/l
N/R
N/l
2.5 ppb
2.5 ppb
2.5 ppb
2.5 ppb
2.5 ppb
N/l
N/l
N/l
N/l
2.5 ppb
N/l
2.5 ppb
2.5 ppb
N/l
N/l
2.5 ppb
2.5 ppb
2.5 ppb
2.5 ppb
2 5 ppb
N/l
EMSL*
MDL"
0.02 ppm
0.1 ppm
0.6 ppm
N/l
National Contaminant
Blomonltorlng Program1
LOD°
0.005-0 046 ppm
0.01-0.05 ppm
0.01 7-0.1 5 ppm
<1.5ppb
<1 .5 ppb
<1 .5 ppb
<1.5ppb
<1.5ppb
<1 .5 ppb
<1.5ppb
<1 5ppb
<1.5ppb
<1.5ppb
<1.5ppb
N/l
<1.5ppb
N/l
N/l
<1-5ppb
< 1-5 ppb
<1-5ppb
<1-5ppb
<1-5ppb
60 ppb
LOO*
N/R
N/R
N/R
2-15 ppb
2-15 ppb
2-15 ppb
2-15 ppb
2-15 ppb
2-15 ppb
2-1 5 ppb
2-15 ppb
2-15 ppb
2-15 ppb
2-15 ppb
N/l
2-15 ppb
N/l
N/l
2-15 ppb
2-15 ppb
2-1 5 ppb
2-15 ppb
2-1 5 ppb
153 ppb
California
OEHHA9
MDL«
N/l
0.050 ppm
N/l
3-5 ppb
2-5 ppb
N/l
4-7 ppb
N/l
38 ppb
7-13 ppb
5-6 ppb
5-6 ppb
3-5 ppb
15-38 ppb
6-10 ppb
N/l
N/l
N/l
N/l
N/l
N/l
N/l
N/l
N/l
N/l
State of
California.
Dept. of Fish
and Game
Environmental
Services
Division"
LOO'
0.01-0.1 ppm
0.02 ppm
0.05 ppm
5ppb
5 ppb
Sppb
5ppb
Sppb
10 ppb
10 ppb
10 ppb
10 ppb
Sppb
10 ppb
100 ppb
Sppb
Sppb
70 ppb
15 ppb
N/l
N/R
2ppb
N/l
100 ppb
EPA
301(h)
Monitoring
Program
Detection Limits*
0.01 ppm (GFAA);
0.4 ppm (ICP)
0.01 ppm (CVAA)
0.02 ppm (GFAA)
0.1-5 ppbu
TJ
•a
m
z
a
x
X
See notes and references at end ol table.
(continued)
-------
Table H-1 (continued)
Target Anatyte
Organophoshate Pesticides
Carbophenothlon
Chlorpyrilos
Diazinon
Dlsultoton
Ethlon
Terbulos
Chlorophenoxy Herbicides
Oxyfluorfen
PCBs (total Aroclors)
Aroclor 1242
Aroclor 1248
Aroclor 1254
Aroclor 1260
Dloxlns/dlbenzo-
furans (total)"
TCDD/TCDF
PeCDD/PeCDF
HxCDD/HxCDF
HpCDD/HpCDF
OCDD/OCDF
sv"
1,000ppb
30.000 ppb
900 ppb
500 ppb
5,000 ppb
10,000 ppb
800 ppb
10 ppb
0.7 ppt
Methods
Puget Sound
Protocols"
LOO1
N/l
N/l
N/l
N/l
N/l
N/l
N/l
(1-5 ppb)'
N/l
N/l
N/l
N/l
HA
POL*
N/l
N/l
N/l
N/l
N/l
N/l
N/l
(20 ppb)'
N/l
N/l
N/1
N/l
N/l
National Study of Chemical
Residues In Fish"
MLD1
N/l
N/R
N/l
N/l
N/l
N/l
N/l
N/R
N/l
N/l
Ml
N/1
N/1
1ppt
2 ppt
4 ppt
10 ppt
N/l
TQLm
N/l
2.5 ppb
N/l
N/l
N/l
N/l
N/l
(1. 25-6.25 ppb)w
N/l
N/1
N/1
Ml
N/R
N/R
N/R
N/R
N/l
EMSL*
MDL"
N/l
N/l
N/l
National Contaminant
Blomonltorlng Program'
LOO"
N/l
N/I
N/1
N/1
N/l
N/l
N/l
N/R
62 ppb
41 ppb
61 ppb
N/l
LOO"
N/l
N/l
Ml
Ml
Ml
N/l
N/l
N/R
167 ppb
111 ppb
155 ppb
N/1
California
OEHHA9
MDLq
N/l
N/l
N/l
Ml
Ml
Ml
N/l
50 ppb
N/l
Ml
50 ppb
50 ppb
N/1
State of
California,
Dept. of Fish
and Game
Environmental
Services
Division1'
LODr
Ml
10 ppb
50 ppb
N/l
20 ppb
N/l
N/1
N/l
50 ppb
50 ppb
50 ppb
N/l
EPA
301(h)
Monitoring
Program
Detection Limits*
1-15 ppb
Ml
20 ppb
.f
N/l
CVAA = Cold vapor atomic absorption spectrophotometry.
GFAA = Graphite furnace atomic absorption spectrophotometry.
ICP = Inductively coupled plasma atomic emission spectrometry.
N/l = Target analyte not Included In monitoring program or recommended methods.
N/R - Not reported.
PCBs - Polychlorinated blphenyte.
(continued)
•o
•D
m
o
X
-------
Table H-1 (continued)
a All values for SVs, detection Emits, and quantitation limits are given in units of weight of analyte per wet weight of edtole fish/shellfish tissue.
b From Table 5-2. Except for mercury, SVs are for general adult population using oral RfOs or SFs available in the EPA IRIS database and assuming a consumption rate (CR) - 6.5 g/d,
average body weight (BW) = 70 kg, lifetime (70-yr) exposure, and lor carcinogens a risk level (RL) - 10'5. The IRIS RfD for methylmercury was lowered by a factor of 5 to calculate (he
recommended SV = 0.6 ppm In order to account for a possible fivefold increase in fetal sensitivity to methylmercury exposure (WHO, 1990). This approach is deemed to be most prudent as
an interim measure until the current reevaluation of the methylmercury RfO is completed (IRIS, 1993). Note: Increasing CR, decreasing BW and/or using an RL <10~5 will decrease the SV.
Program managers must ensure that detection and quantitation limits of analytical methods are sufficient to allow reliable quantitation of target analytes at or below selected SVs. If analytical
methodology is not sensitive enough to reliably quantitate target analytes at or below selected SVs (e.g., PCBs, dioxins/dibenzofurans), the program managers must determine appropriate fish
consumption guidance based on lowest detectable concentrations, or provide justification for adjusting SVs to values at or above achievable method detection or quantitation limits.
c Puget Sound Estuary Program (1990a,b). Analysis of cadmium and lead by GFAA. Analysis of mercury by CVAA. Analysis of organochlorine pesticides and PCBs by gas
chromatography/electron capture detection (GC/ECO). Analysis of PAHs by gas chromatography/mass spectrometry (GC/MS). Inorganic protocols based on U.S. EPA SW-846 methods (U.S.
EPA, 1986b) and U.S. EPA Contract Laboratory Methods (U.S. EPA, 1987a). Organic protocols based on Krahn et al. (1988), U.S. EPA (1984,1986b, 1988,1989d), Horwitz et al. (1980).
NUS (1985), MacLeod et al. (1985), and Brown et al. (1985), on a series of Puget Sound Estuary Program Workshops, and on a national QA Workshop sponsored by the National Oceanic .
and Atmospheric Administration (NOAA) and National Institute of Standards and Technology (NIST).
d National Study of Chemical Residues in Fish (U.S. EPA, 1992a, 1992b). Analysis of mercury by CVAA (U.S. EPA, 1989a). Analysis of organochlorine pesticides and PCBs by GC/MS (U.S.
EPA, 1989c). Analysis of dioxins/dibenzofurans by high-resolution gas chromatography/high-resolution mass spectrometry (HRGC/HRMS) (U.S. EPA, 19896).
• U.S. EPA (1991). Analysis of cadmium, lead, and selenium by ICP. Analysis of mercury by CVAA.
' U.S. Fish and Wildlife Service National Contaminant Biomonitoring Program (Schrritt and Brumbaugh, 1990; Schmitt et al., 1990). Analysis of cadmium and lead by GFAA. Analysis of
mercury by CVAA. Analysis of selenium by hydride generation atomic absorption (HAA). Analysis of organochlorine pesticides and PCBs by GC/ECD.
8 Pollock etal. (1991). Composited fish samples extracted and analyzed for organics by GC/ECD using FDA Method PAM 211.1 in the Pesticide Analytical Mawa/-Vol. I (U.S. FOA, 1978).
This method has been validated in interlaboratory studies and is an official method of the Association of Official Analytical Chemists (AOAC) for DDT, chlordane, and PCBs in fish. Mercury
was determined using the AOAC flametess atomic absorption method (Williams, 1984).
h California Department of Fish and Game. (1990). Metals methods based in part on EPA SW-846 methods (U.S. EPA, 1986b). Analysis of cadmium and lead by flame AA and GFAA.
Analysis of mercury by CVAA. Analysis of selenium by hydride generation AA. Organics methods based on FDA methods (U.S. FDA, 1975) and EPA 301 (h) methods (U.S. EPA, 1986a).
Analysis of organochlorine and organophosphate pesticides and PCBs by GC/ECD. Analysis of PAHs by gas chromatography/flame ionization detection (GC/FID).
1 U.S. EPA (1985,1986, 1987b). Analysis of cadmium and lead by GFAA or ICP. Analysis of selenium by GFAA. Analysis of mercury by CVAA. Analysis of organochlorine pesticides and
PCBs by GC/ECD. Analysis of organophosphate pesticides by GC/phosphorus specific flame photometric or alkali flame ionization detection. Analysis of PAHs by GC/MS. Extract cleanup
(e.g., removal of polar interferences by alumina column chromatography) assumed.
' LOD - Limit of detection. Method detection limit as defined in 40 CFR 136 using a minimum of three replicates.
TO
TJ
m
g
(continued) X
Oi
-------
Table H-1 (continued)
k PQL - Practical quantitation limit. Defined in the Puget Sound Estuary Program as the minimum concentration of an analyte required to be measured and allowed to be reported without
qualification as an estimated quantity for samples without substantial interferences. Based on the lowest concentration of the initial calibration curve (C, in ng/mL), the amount of sample
typically analyzed (W, in g), and the final extract volume (V, in ml):
' MLD - Minimum level of detection. Concentration predicted from ratio of baseline noise area to labeled internal standard plus three times the standard error of the estimate from the weighted
initial calibration curve.
m TQL - Target quantitation limit. Specific detection limits were not determined for individual samples, so were operationally set at zero.
" MDL - Method detection limit. Minimum concentration of an analyte that can be identified, measured, and reported with 99 percent confidence that the analyte concentration is greater than
zero. Determined according to the procedure in 40 CFR 136 using seven replicates.
0 LOD (for metals) - 3{Sb2 - Ss2), where Sb2 and S,z are variances of concentrations measured for procedural blanks and a low-level sample, respectively. LOD (for pesticides) = Mean method
blank plus three times the standard deviation. Determined according to Keith et al. (1983).
p LOQ - Limit of quantitation. Mean method blank plus 10 times the standard deviation. Determined according to Keith et al. (1983).
q MDL - Method detection limit. Determined according to procedure in 49 CFR 209.
' LOD - Limit of detection. The lowest concentration that is statistically different from a blank. Determined according to the IUPAC method in Long and Winefordner (1983).
s From U.S. EPA (1985). Based on detection levels normally achieved in methods commonly used for tissue analyses in environmental laboratories. These detection limits are generally
between the instrument detection limit (IDL) and method detection limit (MDL) (see Section 8.3.3.3) and are based on the expertise and best professional judgment of experienced analysts.
Detection limits for metals based on 5 g (wet weight) of muscle tissue digested and dDuted to 50 mL. Detection limits for organics based on 25 g (wet weight) of muscle tissue extracted,
concentrated to 0.5 mL after gel permeation chromatography cleanup, and 1 uL injected. Bonded, fused silica capillary GC columns, which provide better resolution than packed columns, are
assumed for analysis of semivolatile compounds.
' LOD •= Limit of detection. No procedure given for determining the LOD.
u The higher detection limits are appropriate for pesticides such as mirex, the DDTs, and endosulfans. Compounds such as lindane and hexachlorobenzene can be detected at the lower limits.
Toxaphene (a mixture) may require a higher detection limit than the other organochlorine pesticides.
v Aroclors not determined. Values given are for individual mono- through decachlorobtphenyls.
TJ
Aroclors not determined. PCBs reported by total congener at the following levels of chlorination (TQLS in parentheses): 1-3 (1.25 ppb); 4-6 (2.5 ppb); 7-8 (3.75 ppb); 9-10 (6.25 ppb). "0
m
z
g
(continued) X
-------
Table H-1 (continued)
* Detection and quantitation limits obtained from a survey of 10 laboratories with expertise in dioxin/dibenzofuran analyses by HRGC/HRMS ranged from 0.04-10 ppt and 0.2-100 ppt,
respectively.
References:
Brown, D.W., A.J. Friedman, and W.D. MacLeod, Jr. 1985. Quality Assurance Guidelines for Chemical Analysis of Aquatic Environmental Samples. Prepared for Seattle District, U.S. Army Corps
of Engineers, Seattle, Washington. National Analytical Facility, National Oceanographic and Atmospheric Administration, Seattle, WA.
California Department of Fish and Game. 1990. Laboratory Quality Assurance Program Plan. Environmental Services Division, Sacramento, CA.
Horwitz, W., L. Kamps, and K. Boyer. 1980. Quality assurance in the analysis of foods for trace costituents. Anal. Chem. 63:1344-1354.
IRIS (Integrated Risk Information System). 1993. U.S. Environmental Protection Agency, Duluth, MN.
Keith, L.H., W. Crommett. J. Deegan, Jr., R.A. Libby, J.K. Taylor, and G. Wentler. 1983. Principles of environmental analysis. Analyt. Chem. 55:1426-1435.
Krahn, M.M., C.A. Wigren, R.W. Pearce, L.K. Moore, R.G. Bogar, W.D. MacLeod, Jr., S.L. Chan, and D.W. Brown. 1988. Standard Analytical Procedures for the NOAA National Analytical
Facility. New HPLC Cleanup and Revised Extraction Procedures for Organic Contaminants. NOAA Tech. Memo NMFS F/NWC-153. National Oceanic and Atmospheric Administration, U.S.
Department of Commerce, Northwest and Alaska Fisheries Center, Seattle. WA. 52 pp.
Long. G.L., and J.L. Winefordner. 1983. Limit of detection. A closer look at the IUPAC definition. Anal. Chem. 55(7):712A-724A.
MacLeod W., Jr., D. Brown, A. Friedman, O. Maynes, and R. Pierce. 1985. Standard Analytical Procedures of the NOAA National Analytical Facility. 1984-85. Extractable Toxic Organic
Compounds. NOAA Technical Memorandum NMFS F/NWC-64. Prepared for the National Status and Trends Program, National Oceanic and Atmospheric Administration, U.S. Department of
Commerce, Rockvfle, MD.
NUS. 1985. Laboratory data validation functional guidelines for evaluating organics analysis. Technical Directive Document No. HQ-8410-01. Prepared by the U.S. EPA Data Validation
Workgroup tor U.S. EPA Hazardous Site Control Division, Washington, DC.
Puget Sound Estuary Program. 1990a (revised). Recommended guidelines for measuring organic compounds in Puget Sound sediments and tissue samples. In: Recommended Protocols and
Guidelines tor Measuring Selected Environmental Variables in Puget Sound. Prepared by PTI Environmental Services, Bellevue, WA. Region 10, U.S. Environmental Protection Agency,
Seattle, WA. (Looseleaf)
Puget Sound Estuary Program. 1990b (revised). Recommended protocols for measuring metals in Puget Sound water, sediment, and tissue samples. In: Recommended Protocols and
Guidelines for Measuring Selected Environmental Variables in Puget Sound. Prepared by PTI Environmental Services, Bellevue, WA. Region 10, U.S. Environmental Protection Agency,
Seattle, WA. (Looseleaf)
Schmitt, C.J., and W.G. Brumbaugh. 1990. National Contaminant Biomonitoring Program: Concentrations of arsenic, cadmium, copper, lead, mercury, selenium, and zinc in U.S. freshwater fish,
1978-1984. Arch. Environ. Contam. Toxicol. 19:731-747.
m
o
(continued)
-------
Table H-1 (continued)
0>
Schmitt, C.J., J.L. Zajicek, and P.M. Peterman. 1990. National Contaminant Biomonitoring Program: Residues of organochlorine chemicals in U.S. freshwater fish, 1976-1984. Arch. Environ.
Contam. Toxicol. 19:748-781.
U.S. EPA (U.S. Environmental Protection Agency). 1984 (revised January 1985). Contract Laboratory Program Statement of Work for Organics Analysis, Multi-Media, Multi-Concentration. IFB
WA 85-T176, T177, T178. Washington, DC.
U.S. EPA (U.S. Environmental Protection Agency). 1985. Bioaccumulation Monitoring Guidance: 3. Recommended Analytical Detection Limits. EPA-503/6-90-001. Office of Marine and
Estuarine Protection, Washington, DC.
U.S. EPA (U.S. Environmental Protection Agency). 1986a. Bioaccumulation Monitoring Guidance: 4. Analytical Methods for U.S. EPA Priority Pollutants and 301 (h) Pesticides in Tissues from
Marine and Estuarine Organisms. EPA-503/6-90-002. Office of Marine and Estuarine Protection, Washington, DC.
U.S. EPA (U.S. Environmental Protection Agency). 19866. Test Methods for the Evaluation of Solid Waste, Physical/Chemical Methods. SW-846; 3rd Edition (with 1990 updates). Office of
Solid Waste and Emergency Response, Washington, DC.
U.S. EPA (U.S. Environmental Protection Agency). 1987a. Contract Laboratory Program Statement of Work, Inorganic Analysis, Multi-media, Multi-concentration. SOW No. 87 (Revised
December 1987). Washington, DC.
U.S. EPA (U.S. Environmental Protection Agency). 19876. Quality Assurance/Quality Control (QA/QC) for 301 (h) Monitoring Programs: Guidance on Field and Laboratory Methods. EPA-
430/9-86-004. Office of Marine and Estuarine Protection, Washington, DC.
U.S. EPA (U.S. Environmental Protection Agency). 1988. Laboratory Data Validation Functional Guidelines for Evaluating Organics Analysis. EPA R-582-5-5-01. U.S. EPA Sample Management
Office. Alexandria, VA.
U.S. EPA (U.S. Environmental Protection Agency). 1989a. Analytical Procedures and Quality Assurance Plan for the Determination of Mercury in Fish. Draft. Environmental Research
Laboratory. Duluth MN.
U.S. EPA (U.S. Environmental Protection Agency). 19896. Analytical Procedures and Quality Assurance Plan for the Determination of PCDD/PCDF in Fish. Draft. Environmental Research
Laboratory, Duluth MN.
U.S. EPA (U.S. Environmental Protection Agency). 1989c. Analytical Procedures and Quality Assurance Plan for the Determination of Xenobiotic Chemical Contaminants in Fish.
EPA-600/3-90-023. Environmental Research Laboratory. Duluth, NM.
U.S. EPA (U.S. Environmental Protection Agency). 1989d. Method 1624:' Volatile Organic Compounds by Isotope Dilution GC/MS. Method 1625: Semivolatile Organic Compounds by Isotope
Dilution GC/MS. Office of Water Regulations and Standards, Industrial Technology Division, Washington, DC. 75 pp.
U.S. EPA (U.S. Environmental Protection Agency). 1991. Methods for the Determination of Metals in Environmental Samples. EPA-600/4-91/D10. Environmental Monitoring Systems
Laboratory, Office of Research and Development, Cincinnati, OH. TJ
"TJ
U.S. EPA (U.S. Environmental Protection Agency). 1992a. National Study of Chemical Residues in Fish. Volume I. EPA-823/R-92-008a. Office of Science and Technology, Washington, DC. —
O
(continued)
I
-------
Table H-l (continued)
U.S. EPA (U.S. Environmental Protection Agency). 1992b. National Study of Chemical Residues in Fish. Volume II. EPA-823/R-92-008D. Office of Science and Technology, Washington, DC.
U.S. FDA (U.S. Food and Drug Administration). 1975. Pesticide Analytical Manual. Volume I, Methods Which Detect Multiple Residues. Section 2.32. Rockville, MD.
U.S. FDA (U.S. Food and Drug Administration). 1978. Pesticide Analytical Manual, Volumes I and II. Report No. FDA/ACA/79/76-3. U.S. Department of Health and Human Services,
Washington. DC.
WHO (World Hearth Organization). 1990. Environmental Health Criteria 101: Methyl mercury. World Health Organization, Geneva, Switzerland.
Williams, S. (ed.). 1984. Official Methods of Analysis of the Association of Official Analytical Chemists. Fourteenth edition. The Association of Official Analytical Chemists, Inc., Arlington, VA.
X
-------
APPENDIX I
SOURCES OF RECOMMENDED
REFERENCE MATERIALS AND STANDARDS
-------
APPENDIX I
APPENDIX I
SOURCES OF RECOMMENDED REFERENCE MATERIALS
AND STANDARDS
SOURCES OF ERA-CERTIFIED REFERENCE MATERIALS
EPA-certified analytical reference materials for priority pollutants and related
compounds are currently produced under five Cooperative Research and
Development Agreements (CRADAs) for: organic quality control samples;
organic solution standards; organic neat standards; inorganic quality control
standards; and solid matrix quality control standards. The CRADA cooperators
are listed below.
EPA-certified organic quality control samples, including standards for
pesticides in fish tissue, are produced by:
Supelco, Inc.
Supelco Park
Bellefonte, PA 16823-0048
TEL: 1 -800-247-6628 or 1 -814-359-3441
FAX: 1-814-359-3044
Contact: Linda Alexander
EPA-certified organic solution standards for toxic and hazardous materials
(formerly the EPA Toxic and Hazardous Materials Repository) are produced
by:
NSI Environmental Solutions, Inc.
P. 0. Box 12313
2 Triangle Drive
Research Triangle Park, NC 27709
TEL: 1 -800-234-7837 or 1 -919-549-8980
FAX: 1-919-544-0334
EPA-certified neat organic standards, including neat pesticide standards
(formerly the EPA Pesticide Repository), are produced by:
I-3
-------
APPENDIX
Ultra Scientific
250 Smith Street
North Kingston, Rl 02852
TEL: 1-401-294-9400
FAX: 1-401-295-2330
Contact: Dr. Bill Russo
EPA-certified inorganic quality control samples, including trace metals,
minerals, and nutrients, are produced by:
SPEX Industries, Inc.
3880 Park Avenue
Edison, NJ 08820
TEL: 1-201-549-7144 or 1-800-GET-SPEX
FAX: 1-201-549-5125
EPA-certified solid matrix quality control samples, including standards for
pesticides in fish tissue, are produced by:
Fisher Scientific
711 Forbes Avenue
Pittsburgh, PA 15219
The most recent information on EPA-certified materials is available on the EPA
Electronic Bulletin Board (Modum No. 513-569-7610). Names and addresses
of retailers of EPA-certified CRADA QA/QC samples or standards as of February
20, 1991, are given below. When ordering these materials, specify "EPA
Certified Materials."
I-4
-------
APPENDIX I
RETAILERS OF ERA-CERTIFIED ORGANIC QUALITY CONTROL SAMPLES
Accurate Chemical and Scientific
300 Shamee Drive
Westbury, NY 11590
TEL: 516-443-4900
FAX: 516-997-4938
Contact: Rudy Rosenberg
Accustandard
25 Science Park Road
New Haven, CT 06511
TEL: 203-786-5290
FAX: 203-786-5287
Contact: Mike Bolgar
Aldrich Chemical Company, Inc.
940 West Saint Paul Avenue
Milwaukee, Wl 53233
TEL: 414-273-3850
FAX: 800-962-9591
Contact: Roy Pickering
Alltech Associates/Applied
Science/Wescan Instruments
2051 Waukegan Road
Deerfield, IL 60015
TEL: 708-948-8600
FAX: 708-948-1078
Contact: Tom Rendl
Analytical Products Group
2730 Washington Boulevard
Belpre, OH 45714
TEL: 614-423-4200
FAX: 614-423-5588
Contact: Tom Coyner
Bodman Chemicals
P. 0. Box 2221
Aston, PA 19014
TEL: 215-459-5600
FAX: 215-459-8036
Contact: Kirk Lind
Chemical Research Supply
P. O. Box 888
Addison, IL 60101
TEL: 708-543-0290
FAX: 708-543-0294
Contact: Nelson Armstrong
Crescent Chemical Corporation
1324 Motor Parkway
Hauppauge, NY 11788
TEL: 516-348-0333
FAX: 516-348-0913
Contact: Eric Rudnick
Curtis Matheson Scientific
P. O. Box 1546
9999 Veterans Memorial Drive
Houston, TX 77251-1546
TEL: 713-820-9898
FAX: 713-878-2221
Contact: Mitchel Martin
Environmental Research Associates
5540 Marshall Street
Arvada, CO 80002
TEL: 303-431-8454
FAX: 303-421-0159
Contact: Mark Carter
Restek Corporation
110 Benner Circle
Bellefonte, PA 16823
TEL: 814-353-1300
FAX: 814-353-1309
Contact: Eric Steindle
Supelco
Supelco Park
Bellefonte, PA 16823-0048
TEL: 800-247-6628 or 814-359-3441
FAX: 814-359-3044
Contact: Linda Alexander
Ultra Scientific
250 Smith Street
North Kingston, Rl 02852
TEL: 401-294-9400
FAX: 401-295-2330
Contact: Dr. Bill Russo
I-5
-------
APPENDIX
RETAILERS OF EPA-CERTIFIED ORGANIC SOLUTION STANDARDS
(Formerly the EPA Toxic and Hazardous Materials Repository)
Absolute Standards
498 Russel Street
New Haven, CT 06513
TEL: 800-368-1131
FAX: 203-468-7407
Contact: JackCiscio
Accustandard
25 Science Park Road
New Haven, CT 06511
TEL: 203-786-5290
FAX: 203-786-5287
Contact: Mike Bolgar
Alltech Associates
2051 Waukegan Road
Deerfield, IL 60015
TEL: 708-948-8600
FAX: 708-948-1078
Contact: Tom Rendl
Alameda Chemical and Scientific
922 East Southern Pacific Drive
Phoenix, AZ 85034
TEL: 602-256-7044
FAX: 602-256-6566
Bodman Chemicals
P.O. Box 2221
Aston, PA 19014
TEL: 215-459-5600
FAX: 215-459-8036
Contact: Kirk LJnd
Cambridge Isotope Laboratories
20 Commerce Way
Woburn, MA 01801-9894
TEL: 800-322-1174 or 617-938-0067
FAX: 617-932-9721
NSI Environmental Solutions, Inc.
P.O. Box 12313
2 Triangle Drive
Research Triangle Park, NC 27709
TEL: 800-234-7837 or 919-549-8980
FAX: 919-544-0334
Contact: Zora Bunn
Promochem
Postfach 1246
D 4230 Wesel
West Germany
TEL: 0281/530081
FAX: 0281/89991-93
Ultra Scientific
250 Smith Street
North Kingston, Rl 02852
TEL: 401-294-9400
FAX: 401-295-2330
Contact: Dr. Bill Russo
I-6
-------
APPENDIX I
RETAILERS OF EPA-CERTIFIED NEAT ORGANIC STANDARDS
(Including the Former EPA Pesticide Repository Standards)
Absolute Standards Alltech Associates
498 Russel Street 2051 Waukegan Road
New Haven, CT 06513 Deerfield, IL 60015
TEL: 800-368-1131 TEL: 708-948-8600
FAX: 203-468-7407 FAX: 708-948-1078
Contact: Jack Ciscio Contact: Tom Rendl
Accustandard Ultra Scientific
25 Science Park Road 250 Smith Street
New Haven, CT 06511 North Kingston, Rl 02852
TEL: 203-786-5290 TEL: 401-294-9400
FAX: 203-786-5287 FAX: 401-295-2330
Contact: Mike Bolgar Contact: Dr. Bill Russo
RETAILERS OF EPA-CERTIFIED INORGANIC QUALITY CONTROL SAMPLES
SPEX Industries, Inc.
3880 Park Avenue
Edison, NJ 08820
TEL: 1-201-549-7144 or 1-800-GET-SPEX
FAX: 1-201-549-5125
RETAILERS OF EPA-CERTIFIED SOLID MATRIX QUALITY CONTROL SAMPLES
Fisher Scientific
711 Forbes Avenue
Pittsburgh, PA 15219
I-7
-------
APPENDIX I
RECOMMENDED PUBLICATIONS ON CERTIFIED STANDARDS
AND REFERENCE MATERIALS
Standard and Reference Materials for Marine Science (NOAA, 1992).
Available from
Dr. Adrianna Cantillo
National Ocean Service
National Oceanic and Atmospheric Administration
U.S. Department of Commerce
6001 Executive Blvd., Room 323
Rockville, MD 20852
This catalog lists approximately 2,000 reference materials from 16
producers and includes information on their use, sources, matrix type,
analyte concentrations, proper use, availability, and costs. Reference
materials are categorized as follows: ashes, gases, instrumental
performance, oils, physical properties, rocks, sediments, sludges, tissues,
and waters. This catalog has been published independently by both NOAA
and IOC/UNEP and is available in electronic form from the Office of Ocean
Resources, Conservation, and Assessment, NOAA/NOS.
• Biological and Environmental Reference Materials for Trace Elements,
Nuclides and Organic Microcontamlnants (Toro et al., 1990). Available
from
Dr. R.M. Parr
Section of Nutritional and Health-Related Environmental Studies
International Atomic Energy Agency
P.O. Box 100
A-1400 Vienna, Austria
This report contains approximately 2,700 analyte values for 117 analytes
in 116 biological and 77 nonbiological environmental reference materials
from more than 20 sources. Additional information on cost, sample size
available, and minimum amount of material recommended for analysis is
also provided.
REFERENCES
Toro, E. Cortes, R. M. Parr, and S. A. Clements. 1990. Biological and
Environmental Reference Materials for Trace Elements, Nuclides and
Organic Microcontaminants: A Survey. IAEA/RL/128(Rev. 1). International
Atomic Energy Agency, Vienna.
NOAA (National Oceanic and Atmospheric Administration). 1992. Standard and
Reference Materials for Marine Science. Third Edition. U.S. Department
of Commerce, Rockville, Maryland.
I-8
------- |