Quality Assurance Project Plan for
National Coastal Condition Assessment (NCCA)
2020 Great Lakes Human Health Fish Sample Preparation
July 15, 2020
Prepared for:
United States Environmental Protection Agency
Office of Water
Office of Science and Technology (OST)
Standards and Health Protection Division
Prepared with support from:
Tetra Tech, Inc.
under
OST Engineering and Analysis Division
Contract No. EP-C-17-024
EPA-822-B-24-003
-------
NCCA 2020 Great Lakes Human Health Fish Sample Preparation QAPP
Revision 0
Date: July 15, 2020
Page 1 of 40
Quality Assurance Project Plan for National Coastal Condition Assessment
(NCCA) 2020 Great Lakes Human Health Fish Sample Preparation
A. PROJECT MANAGEMENT
The U.S. Environmental Protection Agency's (EPA's) Office of Science and Technology (OST)
within the Office of Water (OW) prepared this Quality Assurance Project Plan (QAPP) with
support from Tetra Tech under EPA Contract No. EP-C-17-024. It presents objectives,
procedures, performance requirements, and acceptance criteria for the preparation of fish fillet
tissue samples from whole fish composite samples collected by field crews during the 2020
sampling season of the National Coastal Condition Assessment (NCCA). It does not address fish
sample collection because that information is included in separate documents (USEPA 2020a
and USEPA 2020b) prepared by the Office of Wetlands, Oceans, and Watersheds (OWOW).
This QAPP was prepared in accordance with the most recent version of EPA QA/R-5, EPA
Requirements for Quality Assurance Project Plans (USEPA 2001), that was reissued in 2006. It
is a dynamic document that is subject to change as project activities progress. Changes to
procedures in this QAPP must be reviewed by the OST Project Manager and the OST Standards
and Health Protection Division (SHPD) Quality Assurance Coordinator to determine whether the
changes will impact the technical and quality objectives of the project. If so, the QAPP is
revised accordingly, circulated for approval, and forwarded to all project participants listed in the
QAPP distribution list (Section A3). Key project personnel and their roles and responsibilities
are discussed in the QAPP section to follow (Section A4), and information on project
background and description is provided in Sections A5 and A6, respectively.
The following information will be added to this QAPP when it becomes available:
the laboratory designated for mercury analysis of rinsates and blanks (page 27, Section B4.3;
page 29, Section B5.2)
the laboratory designated for PCB analysis of rinsates and blanks (page 27, Section B4.4;
page 30, Section B5.3; page B-9, step 27)
the analytical method used to analyze rinsates and blanks for PCBs (page B-15, Section C)
the laboratory designated forPFAS analysis of rinsates and blanks (page 33, Section Cl.l;
page B-9, step 27)
the analytical method used for PFAS analysis of rinsates and blanks (page 28, Section B4.5)
the laboratory designated for triplicate and single lipid analysis (page 27, Section B4.2; page
28, Section B5.1; page B-10, step 28)
the analytical method used for lipid analysis (page 27, Section B4.2; page B-8, step 24)
the QC acceptance criteria for PCB analysis of aqueous QC samples (page 30, Table 4)
the QC acceptance criteria for PFAS analysis of aqueous QC samples (page 31, Section
B5.4)
the laboratories designated for analysis of fish tissue aliquots (page B-6, Table 1; page B-7,
step 20; page B-8, step 24)
-------
NCCA 2020 Great Lakes Human Health Fish Sample Preparation QAPP
Revision 0
Date: July 15, 2020
Page 2 of 40
Al. Approvals
07-15-2020
Leanne Stahl, OST Project Manager, EPA
Date
JOHN HEALEY
Digitally signed by JOHN HEALEY
Date: 2020.07.21 13:04:02 -04'00'
John Healey, OST Fish Sample Preparation Technical Leader, EPA Date
C IJ A RI D A D A CLI Digitally signed by SHARI BARASH
Dm Mix I DnnnJll Date: 2020.07.21 12:04:53 -04'00'
Shari Barash, Chief, National Branch, EPA
Date
II A DDV |/DAyCD Digitally signed by HARRY KRAMER
rlMnn Y rvnMIVIIirv Date:2020.07.16 14:01:30-04'00'
Bill Kramer, SHPD QA Coordinator, EPA
MARION KELLY
Date
Digitally signed by MARION KELLY
Date: 2020.07.16 16:04:12 -04'00'
Marion Kelly, OST QA Officer, EPA
Date
jLjuulM
7/21/2020
Brian Lenell, GLNPO Project Manager, EPA
Date
Louis Blume, GLNPO QA Manager, EPA
- .a
Blaine Snyder, Tetra Tech Project Leader
July 15, 2020
Date
07/17/2020
Date
Susan Lanberg, Tetra Tech QA Officer
Date
-------
NCCA 2020 Great Lakes Human Health Fish Sample Preparation QAPP
Revision 0
Date: July 15, 2020
Page 3 of 40
A2. Table of Contents
A. PROJECT MANAGEMENT 1
Al. Approvals 2
A2. Table of Contents 3
A3. Di stributi on Li st 6
A4. Project/Task Organization 7
A5. Problem Definition/Background 14
A6. Project/Task Description 14
A7. Quality Objectives and Criteria 16
A8. Special Training/Certification 20
A9. Documents and Records 20
B. DATA GENERATION AM) ACQUISITION 21
B1. Sampling Process Design (Experimental Design) 21
B2. Sampling Methods 23
B3. Sample Receipt and Inspection 24
B4. Fish Sample Preparation and Analytical Methods 25
B4.1 Fish Sample Preparation 26
B4.2 Lipid Analysis 27
B4.3 Mercury Rinsate Analysis 27
B4.4 PCB Rinsate Analysis 27
B4.5 PFAS Rinsate Analysis 28
B5. Fish Sample Preparation Quality Control Requirements 28
B5.1 Homogenized Fillet Samples 28
B5.2 Mercury Analysis of Rinsate Samples 29
B5.3 PCB Analysis of Rinsate Samples 30
B5.4 PFAS Analysis of Rinsate Samples 31
B6. Instrument/Equipment Testing, Inspection, and Maintenance 31
B7. Instrument/Equipment Calibration and Frequency 31
B8. Inspection/Acceptance of Supplies and Consumables 32
B9. Non-direct Measurements 32
B10. Data Management 32
C. ASSESSMENT AM) OVERSIGHT 33
CI. Assessments and Response Actions 33
CI. 1 Fish Sample Preparation 33
C1.2 Performance Audits 34
C1.3 System Audits 34
C2. Surveillance 34
C2.1 Whole Fish Sample Shipment 34
C2.2 Fish Sample Preparation 35
C3. Reports to Management 35
I). DATA VALIDATION AM) USABILITY 36
Dl. Data Review, Verification, and Validation 36
D1.1 Data Re vie w 36
D1.2 Data Verification 36
D1.3 Data V alidation 37
D2. Verification and Validation Methods 38
D3. Reconciliation with User Requirements 39
-------
NCCA 2020 Great Lakes Human Health Fish Sample Preparation QAPP
Revision 0
Date: July 15, 2020
Page 4 of 40
References 39
TABLES
Table 1. Types of Laboratory Data to Be Collected in Association with Great Lakes Human
Health Fish Fillet Sample Preparation for the 2020 NCCA 17
Table 2. Primary Target Fish Species and Secondary Alternative Fish Species for the 2020
GLHHFFTS and ORD-Duluth 2020 Great Lakes Special Study 23
Table 3. QC Samples and Acceptance Criteria for Mercury Analysis of Rinsates 29
Table 4. QC Samples and Acceptance Criteria for PCB Analysis of Rinsates 30
FIGURES
Figure 1. 2020 GLHHFFTS project team organization for fish fillet sample preparation 8
Figure 2. NCCA 2020 Great Lakes human health whole fish sampling locations (226 nearshore
sites are blue dots and 50 Lake Michigan enhancement sites are green triangles) 16
APPENDICES
Appendix A Target Lists of NCCA 2020 Great Lakes Human Health Whole Fish Sampling
Locations
Appendix B NCCA 2020 Great Lakes Human Health Fish Fillet Sample Preparation,
Homogenization, and Distribution Procedures
Appendix C 2020 Great Lakes Human Health Fish Fillet Tissue Study Fish Sample
Preparation Laboratory Bench Sheet
Appendix D ORD-Duluth 2020 Great Lakes Special Study Human Health Fish Sample
Preparation Laboratory Bench Sheet
-------
NCCA 2020 Great Lakes Human Health Fish Sample Preparation QAPP
Revision 0
Date: July 15, 2020
Page 5 of 40
LIST OF ACRONYMS AND ABBREVIATIONS
C Celsius
DI Deionized
DQO Data quality objectives
EPA Environmental Protection Agency
FDA Food and Drug Administration
g Gram
GLHHFFTS Great Lakes Human Health Fish Fillet Tissue Study
GLNPO Great Lakes National Program Office
HDPE High density polyethylene
ID Identification
IDL Instrument Detection Limit
IM Information Management
IR Infrared
MDL Method detection limit
mL Milliliter
NARS National Aquatic Resource Survey
NCCA National Coastal Condition Assessment
NRSA National Rivers and Streams Assessment
ORD Office of Research and Development
OST Office of Science and Technology
OW Offi ce of Water
OWOW Office of Wetlands, Oceans, and Watersheds
PCB Polychlorinated biphenyl
PDF Portable Document Format
PFAS Per- and polyfluoroalkyl substances
PTFE Polytetrafluoroethylene
QA Quality assurance
QAPP Quality Assurance Project Plan
QC Quality control
RSD Relative standard deviation
SD Standard deviation
SHPD Standards and Health Protection Division
SOP Standard operating procedure
TBD To be determined
-------
NCCA 2020 Great Lakes Human Health Fish Sample Preparation QAPP
Revision 0
Date: July 15, 2020
Page 6 of 40
A3. Distribution List
Shari Barash
USEPA/OW/OST (4305T)
1200 Pennsylvania Avenue, N.W.
Washington, DC 20460
202-566-0996
barash.shari@epa.gov
Bill Kramer
USEPA/OW/OST (4305T)
1200 Pennsylvania Avenue, N.W.
Washington, DC 20460
202-566-0385
kramer.bill@epa.gov
David Bolgrien
USEPA/ORD/Great Lakes Toxicology and
Ecology Division
6201 Congdon Boulevard
Duluth, MN 55804
218-529-5216
bolgrien.dave@epa.gov
Susan Lanberg
Tetra Tech, Inc.
10306 Eaton Place, Suite 340
Fairfax, VA 22030
703-385-6000
susan. 1 anb erg@tetratech. com
Louis Blume
USEPA Great Lakes National Program Office
77 West Jackson Boulevard (LAB-10C)
Chicago, IL 60604
312-353-2317
blume.louis@epa.gov
Sarah Lehmann
USEPA/OW/OWOW (4503T)
1200 Pennsylvania Avenue, N.W.
Washington, DC 20460
202-566-1379
lehmann.sarah@epa.gov
Marcus Bowersox
Tetra Tech, Inc.
10711 Red Run Blvd., Suite 105
Owings Mills, MD 21117
410-902-3142
marcus. b ower sox@tetratech. com
Brian Lenell
USEPA Great Lakes National Program Office
77 W. Jackson Blvd. (G-9J)
Chicago, IL 60604
312-353-4891
lenell.brian@epa.gov
Yildiz Chambers-Velarde
CSRA/GDIT
6361 Walker Lane, Suite 300
Alexandria, VA 22310
703-254-0061
yildiz.chambers@gdit.com
Harry McCarty
CSRA/GDIT
6361 Walker Lane, Suite 300
Alexandria, VA 22310
703-254-0093
harry. mccarty @gdit. com
John Healey
USEPA/OW/OST (4305T)
1200 Pennsylvania Avenue, N.W.
Washington, DC 20460
202-566-0176
healey.john@epa.gov
Mari Nord
USEPA Region 5 Office
77 W. Jackson Blvd. (WS-15J)
Chicago, IL 60604
312-886-3017
nord.mari@epa.gov
-------
NCCA 2020 Great Lakes Human Health Fish Sample Preparation QAPP
Revision 0
Date: July 15, 2020
Page 7 of 40
Elizabeth Hinchey
USEPA Great Lakes National Program Office
77 W. Jackson Blvd. (G-9J)
Chicago, IL 60604
312-886-3451
hinchey.elizabeth@epa.gov
Blaine Snyder
Tetra Tech, Inc.
10711 Red Run Blvd., Suite 105
Owings Mills, MD 21117
410-902-3158
blaine.snyder@tetratech.com
Marion Kelly
USEPA/OW/OST (4303T)
1200 Pennsylvania Avenue, N.W.
Washington, DC 20460
202-566-1045
kelly.marion@epa.gov
Leanne Stahl
USEPA/OW/OST (4305T)
1200 Pennsylvania Avenue, N.W.
Washington, DC 20460
202-566-0404
stahl.leanne@epa.gov
A4. Project/Task Organization
This current study of contaminants in Great Lakes fish is referred to as the 2020 Great Lakes
Human Health Fish Fillet Tissue Study (GLHHFFTS). The EPA project team for the 2020
GLHHFFTS consists of managers, scientists, and QA personnel in OST and the Great Lakes
National Program Office (GLNPO) and statisticians in the Pacific Ecological Systems Division
within the Center for Public Health and Environmental Assessment (Corvallis, Oregon) in the
Office of Research and Development (ORD). The EPA project team receives scientific,
technical, and logistical support from contractors at Tetra Tech and at CSRA General Dynamics
Information Technology (GDIT). Tetra Tech provides primarily fisheries support (e.g., fish
sampling and fish sample preparation) and CSRA/GDIT provides analytical support for the
proj ect team.
Members of the project team responsible for fish fillet sample preparation include the OST
Project Manager, the OST Fish Sample Preparation Technical Leader, the GLNPO Project
Manager, the OST QA Officer, the SHPD QA Coordinator, the GLNPO QA Manager, the Tetra
Tech Project Leader, the Tetra Tech QA Officer, and Tetra Tech staff providing scientific,
technical, and logistical support for this activity. The project team organization provides the
framework for conducting fish sample preparation to meet study objectives. The organizational
structure and function also facilitate project performance and adherence to quality control (QC)
procedures and quality assurance (QA) requirements. The project organizational chart is
presented in Figure 1. It identifies individuals serving in key roles and the relationships and lines
of communication among these project team members. Responsibilities for key members of the
project team are described below.
-------
NCCA 2020 Great Lakes Human Health Fish Sample Preparation QAPP
Revision 0
Date: July 15, 2020
Page 8 of 40
Figure 1. 2020 GLHHFFTS project team organization for fish fillet sample preparation
-------
NCCA 2020 Great Lakes Human Health Fish Sample Preparation QAPP
Revision 0
Date: July 15, 2020
Page 9 of 40
Leanne Stahl of OST is the OST Project Manager who is providing overall direction for
planning and implementation of the 2020 GLHHFFTS being conducted under the NCCA. This
role involves the following responsibilities related to the 2020 GLHHFFTS:
developing technical information for whole fish sample collection for fillet analysis that
includes preparation of the fish sampling protocols and coordination with the NCCA
Project Leader in OWOW to integrate field sampling technical information for the 2020
GLHHFFTS into NCCA documents and training materials (This technical information
also applies to the ORD-Duluth 2020 Great Lakes special study.)
providing technical support to conduct training on the 2020 GLHHFFTS field sampling
requirements in coordination with the NCCA Project Leader in OWOW (This training
also applies to the ORD-Duluth 2020 Great Lakes special study.)
developing the fish sample preparation procedures and requirements in coordination with
the 2020 GLHHFFTS Fish Sample Preparation Technical Leader (These fish sample
preparation procedures and requirements also apply to the ORD-Duluth 2020 Great
Lakes special study.)
managing analysis of fish fillet samples for target chemicals and subsequent activities,
including obtaining technical support for chemical analysis of fish fillet tissue samples,
directing development of the initial NCCA 2020 Great Lakes human health fish fillet
tissue sample analysis QAPP and subsequent QAPP revisions, providing for QA review
of the analytical results, developing the data files for statistical analysis of the data,
reviewing and approving the final analytical QA report, and providing oversight for
development of the database to store 2020 GLHHFFTS fish tissue results (This series of
fillet sample analysis and related activities (including fillet sample analysis QAPP
development, fillet sample chemical analysis, analytical data quality review, and data
analysis and management activities) also apply to the ORD-Duluth 2020 Great Lakes
special study.)
facilitating communication among 2020 GLHHFFTS project team members and
coordinating with all of these individuals to ensure technical quality and adherence to
QA/QC requirements (This responsibility for communicating and coordinating with
project team members also applies to the ORD-Duluth 2020 Great Lakes special study.)
developing and managing work assignments and task orders under OST or other EPA
contracts to provide technical support for the 2020 GLHHFFTS, providing oversight of
contractor activities, and reviewing and approving study deliverables for each work
assignment and task order (Contractor support for 2020 Great Lakes human health fish
sample collection and analysis activities and associated activities also applies to the
ORD-Duluth 2020 Great Lakes special study.)
scheduling and leading meetings and conference calls with 2020 GLHHFFTS project
team members for planning study activities, reporting progress on study tasks, and
discussing and resolving technical issues related to the study (This responsibility also
applies to the ORD-Duluth 2020 Great Lakes special study.)
-------
NCCA 2020 Great Lakes Human Health Fish Sample Preparation QAPP
Revision 0
Date: July 15, 2020
Page 10 of 40
working with QA staff to identify corrective actions necessary to ensure that study
quality objectives are met for both Great Lakes studies involving human health fish
sample collection and analysis
managing the development of and/or reviewing and approving all major work products
associated with the 2020 GLHHFFTS and various other fish tissue studies, including
products prepared by OWOW
leading the Fish Tissue Study Team for reporting the 2020 GLHHFFTS human health
fish fillet indicator results and various other fish tissue study results in technical journal
articles and federal technical reports (This responsibility includes collaborating with the
ORD-Duluth 2020 Great Lakes special study project team for reporting 2020 Great Lakes
human health fish fillet analysis results.)
coordinating with John Healey (TOCOR) to obtain Tetra Tech support through the task
order for preparing fish study briefings and presentations and for providing general
technical support; concurring on approval of task order deliverables
presenting 2020 GLHHFFTS and other fish tissue study briefings for EPA managers and
delivering fish tissue study presentations in various forums (e.g., scientific conferences,
government meetings, and webinars)
John Healey of OST is the OST Fish Sample Preparation Technical Leader who is providing
support for planning and implementation of the 2020 GLHHFFTS being conducted under the
NCCA. This role involves the following responsibilities related to the 2020 GLHHFFTS:
developing and managing a task order to provide technical support for preparation of
NCCA 2020 Great Lakes human health fish fillet tissue samples for chemical analysis,
which includes ensuring training for laboratory processing of NCCA fish samples,
providing technical direction for and oversight of fish sample preparation activities (e.g.,
providing oversight for analysis of fish sample preparation QC samples and single lipid
analysis of fillet tissue samples), and reviewing and approving task order deliverables
(e.g., fish sample preparation weekly progress reports and results for analysis of QC
samples) with OST Project Manager concurrence (This responsibility also applies to the
ORD-Duluth 2020 Great Lakes special study.)
participating in developing, reviewing, and approving the NCCA 2020 Great Lakes
Human Health Fish Sample Preparation QAPP (This responsibility also applies to the
ORD-Duluth 2020 Great Lakes special study.)
developing and managing a task order to provide Tetra Tech support for preparing 2020
GLHHFFTS and other fish tissue study presentations and briefings and reviewing and
approving task order deliverables with OST Project Manager concurrence and EPA
management approval
coordinating with OST QA staff and 2020 GLHHFFTS project team members to ensure
technical quality and adherence to QA/QC requirements for task order deliverables
obtaining training on the 2020 Great Lakes human health fish sampling requirements in
coordination with the OST Project Manager
-------
NCCA 2020 Great Lakes Human Health Fish Sample Preparation QAPP
Revision 0
Date: July 15, 2020
Page 11 of 40
participating in meetings and conference calls with Fish Tissue Study Team members for
planning 2020 GLHHFFTS and other fish tissue study activities, reporting progress on
various fish tissue study tasks, and discussing and resolving technical issues related to the
2020 GLHHFFTS and other fish tissue studies
attending OWOW weekly NARS meetings and reporting information presented in the
meeting (particularly information related to the NCCA and NRSA) to the OST Project
Manager and SHPD managers
managing the development of and/or reviewing and approving all major work products
associated with the 2020 GLHHFFTS and various other fish tissue studies, including
products prepared by OWOW
collaborating with Fish Tissue Study Team members for reporting 2020 GLHHFFTS
results and results for other fish tissue studies in technical journal articles and federal
technical reports
coordinating with the OST Project Manager to obtain CSRA/GDIT support for preparing
materials for fish tissue study briefings and presentations
participating in presenting 2020 GLHHFFTS and other fish tissue study briefings for
EPA managers and delivering fish tissue study presentations in various forums (e.g.,
scientific conferences, government meetings, and webinars)
Brian Lenell of GLNPO is the 2020 GLHHFFTS GLNPO Project Manager who is providing
support for planning and implementation of this regional Great Lakes study being conducted
under the NCCA. This role involves the following responsibilities related to the 2020
GLHHFFTS:
reviewing and concurring on technical information developed for 2020 GLHHFFTS fish
sample collection
participating in training on the 2020 Great Lakes human health fish sampling
requirements in coordination with OST
arranging additional support for 2020 GLHHFFTS fish sample collection through
GLNPO fisheries contacts
participating in the review of the fish sample preparation and sample analysis QAPPs for
the 2020 GLHHFFTS
managing analysis of fish tissue samples for the fatty acids, including identification of a
laboratory for analysis of fillet samples for fatty acids and oversight of fatty acid analysis
and reporting results
coordinating with 2020 GLHHFFTS project team members to ensure technical quality
and adherence to QA/QC requirements
participating in conference calls with project team members for planning study activities,
reporting progress on study tasks, and discussing and resolving technical issues related to
the study
-------
NCCA 2020 Great Lakes Human Health Fish Sample Preparation QAPP
Revision 0
Date: July 15, 2020
Page 12 of 40
reviewing and concurring on all major work products associated with the 2020
GLHHFFTS
collaborating with the 2020 GLHHFFTS project team for reporting the study results in
technical journal articles and federal technical reports
participating in preparing and/or reviewing fish tissue study presentations and presenting
them in various forums (e.g., scientific conferences, government meetings, and webinars)
Marion Kelly is the OST Quality Assurance Officer who is responsible for reviewing and
approving all QAPPs that involve scientific work being conducted by OST. Bill Kramer is the
Standards and Health Protection Division (SHPD) QA Coordinator who is responsible for
reviewing and recommending approval of all QAPPs that include scientific work being
conducted by SHPD within OST. The OST QA Officer and SHPD QA Coordinator are also
responsible for the following QA/QC activities:
reviewing and approving this QAPP
reviewing and evaluating the QA/QC requirements and data for all the 2020 GLHHFFTS
activities and procedures
conducting external performance and system audits of the procedures applied for all 2020
GLHHFFTS activities
participating in Agency QA reviews of the study
Louis Bloom is the GLNPO Quality Assurance Manager who is responsible for reviewing and
approving all QAPPs that involve scientific work being conducted by GLNPO. The GLNPO QA
Manager is also responsible for the following QA/QC activities:
reviewing and approving this QAPP
reviewing and evaluating the QA/QC requirements and data for all the 2020 GLHHFFTS
activities and procedures
participating in external performance and system audits of the procedures applied for all
2020 GLHHFFTS activities
participating in Agency QA reviews of the study
Blaine Snyder is the Tetra Tech Project Leader who is responsible for managing all aspects of
the technical and logistical support being provided by Tetra Tech staff for the 2020 GLHHFFTS.
His specific responsibilities include the following:
providing direct technical and logistical support for the following 2020 GLHHFFTS
activities or providing leadership and oversight for Tetra Tech staff supporting these
activities:
developing procedures for fish sampling and fish sample preparation
preparing 2020 GLHHFFTS training materials and project information to incorporate
into NCCA documents developed by OWOW
-------
NCCA 2020 Great Lakes Human Health Fish Sample Preparation QAPP
Revision 0
Date: July 15, 2020
Page 13 of 40
preparing 2020 GLHHFFTS documents specific to the Great Lakes human health fish
fillet indicator (including this QAPP)
providing fish sampling and fish sample preparation training
planning and implementing 2020 GLHHFFTS logistics
collecting fish samples at whole fish sampling Great Lakes nearshore sites designated
by the OST Project Manager
obtaining and performing QC reviews of Great Lakes human health field sampling
data related to the human health fish fillet indicator
assigning batches for fish sample preparation and preparing fish sample preparation
instructions for whole fish samples collected from designated NCCA 2020
GLHHFFTS nearshore sites and ORD-Duluth 2020 Great Lakes special study
enhancement sites
managing implementation of the fish sample preparation procedures, including
obtaining laboratory services for analysis of QC samples generated during fish
sample preparation and lipid samples for all Great Lakes human health fillet samples
preparing weekly fish sample processing reports and evaluating the reports for
adherence to the technical and quality requirements in the fish sample preparation
procedures
packing and shipping fish fillet tissue and any related samples (e.g., rinsate samples)
to analytical laboratories designated for mercury, PCB, PFAS, lipid, and fatty acid
analyses
preparing project information and graphics for development of project fact sheets,
briefings, presentations, and other EPA meeting and outreach materials
providing technical support for planning and reviewing statistical analysis of 2020
GLHHFFTS and ORD-Duluth 2020 special study fillet tissue data and reporting the
final results
monitoring the performance of Tetra Tech staff participating in this study to ensure that
they are following all QA procedures described in this QAPP that are related to Tetra
Tech tasks being performed to support this study
ensuring completion of high-quality deliverables within established budgets and time
schedules
participating in meetings and conference calls with project team members for planning
study activities, reporting progress on study tasks, and discussing and resolving technical
issues related to the study
Susan Lanberg is the Tetra Tech QA Officer whose primary responsibilities include the
following:
assisting Tetra Tech's Project Leader with the review of this QAPP
approving this QAPP
-------
NCCA 2020 Great Lakes Human Health Fish Sample Preparation QAPP
Revision 0
Date: July 15, 2020
Page 14 of 40
providing oversight for the implementation of QA procedures related to Tetra Tech tasks
that are described in this QAPP
reporting deviations from this QAPP to the Tetra Tech Project Leader and assisting in
implementing corrective actions to resolve these deviations
A5. Problem Definition/Background
Obtaining statistically representative occurrence data on multiple contaminants in fish tissue is a
priority area of interest for EPA. Since 2010, OST has collaborated with the Great Lakes
National Program Office (GLNPO), Office of Wetlands, Oceans, and Watersheds (OWOW)
within the Office of Water (OW), and with the Office of Research and Development (ORD) to
conduct a series of regional-scale assessments of chemical contaminants in Great Lakes fish
from nearshore areas as part of EPA's National Coastal Condition Assessment (NCCA). This
current study of contaminants in Great Lakes fish is referred to as the 2020 Great Lakes Human
Health Fish Fillet Tissue Study (GLHHFFTS). It is the third study of Great Lakes fish
contamination conducted by OST and GLNPO under the NCCA. The two previous OST/
GLNPO studies of contaminants in Great Lakes fish were the 2010 Great Lakes Human Health
Fish Tissue Study and the 2015 Great Lakes Human Health Fish Fillet Tissue Study.
Overall, the 2020 NCCA is a probability-based survey designed to assess the condition of coastal
waters of the United States, which includes coastal waters of the Great Lakes. Building on
EPA's experience from the 2010 NCCA and the 2015 NCCA, it includes collection and analysis
of physical, chemical, and biological indicator data that will allow a statistically valid
characterization of the condition of the Nation's coastal waters. EPA used an unequal
probability design to select 725 estuarine sites along the coasts of the contiguous United States,
226 freshwater sites from nearshore areas throughout the Great Lakes, and 50 enhancement sites
in Lake Michigan. OWOW within OW is responsible for managing the planning and
implementation of the NCCA.
A6. Project/Task Description
OST and GLNPO began planning and mobilizing for the 2020 GLHHFFTS in 2019. An
important new decision during the planning phase for the 2020 GLHHFFTS was to expand
human health fish sample collection to the full set of 226 Great Lakes nearshore sites (45 sites
per Great Lake except 46 sites in Lake Superior) randomly selected by ORD (Figure 2 and
Appendix A). During the previous two Great Lakes human health fish tissue studies in 2010 and
2015, fish sample collection was limited to approximately 150 nearshore sites (about 30 sites per
Great Lake). Mobilizing activities for the 2020 GLHHFFTS included updating fish sampling
and handling protocols for the NCCA 2020 Field Operation Manual (USEPA 2020b) and Field
Sampling QAPP (USEPA 2020a), along with assembling and shipping human health fish
sampling kits to the NCCA central supply distribution center in Traverse City, Michigan. During
the mobilization phase, OWOW had to develop and implement a significantly different approach
for NCCA 2020 field sampling training due to the coronavirus pandemic. Rather than
conducting a series of up to 14 onsite training workshops across the U.S., OWOW provided
NCCA 2020 field sampling training through a series of virtual training workshops that began in
late March and continued until late May 2020.
-------
NCCA 2020 Great Lakes Human Health Fish Sample Preparation QAPP
Revision 0
Date: July 15, 2020
Page 15 of 40
OST and GLNPO also coordinated with EPA scientists at the ORD facility in Duluth, Minnesota
(abbreviated as ORD-Duluth) to add collection and analysis of human health fish samples from
50 enhancement sites in Lake Michigan as part of the ORD-Duluth 2020 Great Lakes special
study. These enhancements sites include 38 island nearshore sites in northern Lake Michigan
and 12 National Park nearshore sites in southern Lake Michigan (Figure 2 and Appendix A).
Collection and preparation of human health whole fish samples from the Lake Michigan
enhancement sites will involve procedures that are identical to the fish sample collection and
preparation procedures for the 2020 GLHHFFTS.
NCCA 2020 Great Lakes human health whole fish sample collection and preparation for both the
2020 GLHHFFTS and ORD-Duluth 2020 Great Lakes special study involve the following key
components:
Collecting human health whole fish samples at 226 randomly selected Great Lakes
nearshore sites and 50 Lake Michigan enhancement sites in the nearshore regions
(Appendix A) during 2020 and possibly into 2021 if field crews are subject to travel
restrictions in 2020 due to the coronavirus pandemic. Both types of sites have depths up
to 30 meters or distances up to 5 kilometers from the shore.
Obtaining one fish composite sample from each Great Lakes nearshore site and Lake
Michigan enhancement site designated for human health fish sampling, which ideally
consists of five similarly sized adult fish of the same species that are commonly
consumed by humans.
Shipping Great Lakes human health whole fish samples to freezers at Microbac
Laboratories in Baltimore, MD for interim storage.
Transferring the whole fish samples to the Tetra Tech facility in Owings Mills, MD for
fish sample preparation.
Preparing fillet tissue samples for chemical analysis by scaling and filleting each fish in
the composite sample, homogenizing the fillets from all the fish in the sample, and
dividing the fillet tissue into aliquots for various chemical analyses and for long-term
storage of archived samples in a freezer.
Shipping fillet tissue samples to laboratories contracted to analyze these samples for
mercury, PFAS, PCB congeners, PCBs as Aroclors, lipids, and fatty acids.
This QAPP focuses on fish sample preparation activities for the NCCA 2020 Great Lakes human
health whole fish samples, which include the last three study components listed above. Specific
fish sample preparation procedures and requirements are described in Appendix B.
-------
NCCA 2020 Great Lakes Human Health Fish Sample Preparation QAPP
Revision 0
Date: July 15, 2020
Page 16 of 40
Figure 2. NCCA 2020 Great Lakes human health whole fish sampling locations (226 nearshore
sites are blue dots and 50 Lake Michigan enhancement sites are green triangles)
A7. Quality Objectives and Criteria
Data of known and documented quality are essential to the success of any sampling program.
Data quality objectives (DQOs) are qualitative and quantitative statements that clarify the
intended use of the data, define the type of data needed to support the decision, identify the
conditions under which the data should be collected, and specify tolerable limits on the
probability of making a decision error due to uncertainty in the data. DQOs are developed by
data users to specify the data quality needed to support specific decisions. Sources of error or
uncertainty include the following:
Sampling error: The difference between sample values and in situ true values from
unknown biases due to collection methods and sampling design.
Measurement error: The difference between sample values and in situ true values
associated with the measurement process.
-------
NCCA 2020 Great Lakes Human Health Fish Sample Preparation QAPP
Revision 0
Date: July 15, 2020
Page 17 of 40
Natural variation: Natural spatial heterogeneity and temporal variability in population
abundance and distribution.
Error sources or biases associated with compositing, sample handling, storage, and
preservation.
This QAPP addresses activities associated with Great Lakes human health fish sample
preparation, so the relevant quality objectives are related to issues involving fillet tissue sample
preparation and handling in the laboratory. Table 1 lists the types of fillet tissue sample
preparation data needed for the 2020 GLHHFFTS and ORD-Duluth 2020 Great Lakes special
study. Methods and procedures described in this document are intended to reduce the magnitude
of the sources of uncertainty and their frequency of occurrence by applying the following
approaches:
Use of standardized fish sample preparation procedures (Appendix B)
Use of trained scientists to perform the fish sample preparation activities
Table 1. Types of Laboratory Data to Be Collected in Association with Great Lakes Human Health
Fish Fillet Sample Preparation for the 2020 NCCA
Data Type
Measurement Endpoint(s) or Units
Fish weight
Grams (g)
Unhomogenized fillet weight
Grams (g)
Homogenized fillet weight
Grams (g)
Tissue homogenate recovery
Percent (%)
Tissue aliquot weight
Grams (g)
Tissue archive weight
Grams (g)
Measurement performance criteria are quantitative statistics that are used to interpret the degree
of acceptability or utility of the data to the user. These criteria, also known as data quality
indicators, include the following:
Precision
Accuracy
Representativeness
Completeness
Comparability
Precision
Precision is a measure of internal method consistency. It is demonstrated by the degree of
agreement between individual measurements (or values) of the same property of a sample
measured under similar conditions. The only analytical testing that is within the scope of this
QAPP is the analysis of fish preparation rinsate and solvent blank samples for mercury and PCB
-------
NCCA 2020 Great Lakes Human Health Fish Sample Preparation QAPP
Revision 0
Date: July 15, 2020
Page 18 of 40
congeners (to ensure that the preparation laboratory environment and equipment are not an
extraneous source of mercury or PCBs) and lipid analysis to test the homogeneity of the prepared
fish fillet tissue samples and to provide lipid results for the full complement of fish composite
samples in each fish sample preparation batch (usually 20 fish samples per batch).
The sample preparation laboratory will prepare two sets of rinsate samples (each consisting of
one deionized [DI] water equipment rinsate sample and one DI water blank sample) for mercury
and PFAS analysis, one set of rinsate samples (consisting of one hexane equipment rinsate
sample and one hexane blank sample) for PCB analysis, and one homogenized fillet sample for
triplicate lipid determinations per fish sample preparation batch, as described in Steps 25 through
29 of Appendix B. The batch-specific homogeneity and paired rinsate and solvent blank results
are reviewed by EPA against the QC specifications detailed in Section B5.1 (for homogenates),
Section B5.2 (for mercury rinsates), and Section B5.3 (for PCB rinsates). The QC requirements
for PFAS analysis of rinsate and solvent blank samples will be specified in the NCCA 2020
Great Lakes human health fish fillet sample analysis QAPP, which will be developed at a later
date.
Accuracy
Accuracy is defined as the degree of agreement between an observed value and an accepted
reference or true value. Accuracy is a combination of random error (precision) and systematic
error (bias) introduced during sampling and analytical operations. Bias is the systematic
distortion of a measurement process that causes errors in one direction, so that the expected
sample measurement is always greater or lesser to the same degree than the sample's true value.
Proper sample handling procedures will be followed to minimize sample contamination during
fish sample preparation (Section B4.1) and QC analyses (Sections B4.2 through B4.5).
Representativeness
Representativeness expresses the degree to which data represent a characteristic of a population,
a parameter, a process condition, an environmental condition, or variations at a sampling point.
The representativeness goal for the 2020 GLHHFFTS and ORD-Duluth 2020 Great Lakes
special study was addressed during the study design phase and will be satisfied by using
experienced field biologists to ensure that the samples collected are actually of the type (species)
specified for this study and are from the randomly selected sites specifically identified in the
statistical (i.e., probabilistic) study design. Representativeness in the fish sample preparation
phase centers on the fish fillet composite samples or fillet tissue aliquots and how accurately they
represent the fillet mass for analysis of mercury, PCBs, PFAS, or fatty acids in the fish. Batch-
specific rinsate and homogeneity sample analyses (Sections B5.1 through B5.4) are included as
QC steps in the fish preparation process to ensure that the fish fillet homogenates are free from
any laboratory sources of contamination and are thoroughly mixed, so they are therefore a
representative sample of the fillet tissue.
Completeness
Completeness is defined as the percentage of measurements made that are judged to be valid
according to specific criteria and entered into the data management system. To optimize
-------
NCCA 2020 Great Lakes Human Health Fish Sample Preparation QAPP
Revision 0
Date: July 15, 2020
Page 19 of 40
completeness, every effort is made to avoid sample and/or data loss. Accidents during sample
transport or lab activities that cause the loss of the original samples will result in irreparable loss
of data, which will reduce the ability to perform analyses, integrate results, and prepare reports.
Whole fish samples are packed in unbreakable (plastic) shipping containers (i.e., insulated ice
chests) to avoid damage during shipment of the samples to the sample repository (i.e. Microbac
Laboratories in Baltimore, MD) or during transport of the samples to the fish sample preparation
laboratory in Owings Mills, MD. Fillet homogenates are also packed in unbreakable shipping
containers for shipment to each analytical laboratory.
Percent completeness (%C) for measurement parameters can be defined as follows:
%C = X 100
T
Where v = the number of measurements judged valid and
T= the total number of measurements.
Completeness for the NCCA 2020 Great Lakes human health fish fillet indicator sample
preparation effort is the number of samples processed relative to the number of samples that are
collected and identified as valid samples for analysis. The completeness goal for the sample
preparation phase of this study is 100% because processing all valid samples is critical for
maintaining the integrity of the statistical design for the study. In some cases, whole fish
samples may contain small fish and/or less than five individual specimens, and therefore may not
provide sufficient tissue for analysis of the full list of target chemicals. In those cases, the OST
Project Manager and the OST Fish Sample Preparation Technical Leader must be notified before
sample preparation activities begin, and EPA's priority order for preparing fillet aliquots for
analysis must be followed (i.e., highest to lowest priority order is mercury, PFAS, PCB
congeners, single lipids, PCBs as Aroclors, and fatty acids). The completeness goals for
individual fillet aliquots listed in the bullets below reflect these priorities for analyzing the target
chemicals. It should be noted that the total number of sampling locations may change over the
course of each Great Lakes study based on location conditions (e.g., accessibility of target
locations) and the availability of target species (e.g., natural biological abundance or
distribution). Any changes must be approved by the OST Project Manager, and approved
changes must be considered when assessing completeness. The completeness goal is achieved
when the following requirements are met:
Fillet samples are collected from each fish identified by the OST Project Manager as a
valid specimen for inclusion in the composite sample, and those fillets are homogenized
to prepare fish tissue aliquots for mercury, PFAS, PCB congener, PCBs as Aroclors, lipid
and fatty acid analyses (completeness goal is 100% for mercury, 100% for PFAS, 90%
for PCB congeners, 80% for PCBs as Aroclors and single lipids, and 75% for fatty acids).
All homogenized fillet aliquots are shipped with no errors in documentation or sample
handling procedures, which facilitates timely delivery of every shipment of samples and
arrival of the samples at the analytical laboratory in good condition.
-------
NCCA 2020 Great Lakes Human Health Fish Sample Preparation QAPP
Revision 0
Date: July 15, 2020
Page 20 of 40
Comparability
Comparability is an expression of the confidence with which one data set can be compared with
another. Comparability is dependent on the proper design of the sampling program and on
adherence to accepted sampling techniques, procedures, and quality assurance guidelines. For
fish sample preparation, comparability of data is accomplished by standardizing the sample
preparation methods and the laboratory training as follows:
All homogenized fillet samples are prepared by sample preparation laboratory personnel
according to the procedures contained in this QAPP (Appendix B).
All laboratory personnel involved with fish sample preparation will have adequate
training and appropriate fillet tissue sample preparation experience (Section A8).
A8. Special Training/Certification
All laboratory staff involved in the preparation of fish fillet tissue samples must be proficient in
the associated tasks, as required by the NCCA 2020 Great Lakes Human Health Fish Fillet
Sample Preparation, Homogenization, and Distribution Procedures (Appendix B). Specialized
training is being provided for laboratory technicians who will be preparing homogenized fillet
tissue samples from the whole fish samples collected for the 2020 GLHHFFTS and the ORD-
Duluth 2020 Great Lakes special study. This training will be conducted at the Tetra Tech
Biological Research Facility in Owings Mills, MD for all laboratory staff involved with fillet
tissue sample preparation to accomplish the following objectives:
Present homogenized fillet tissue sample preparation and distribution procedures as
described in Appendix B,
Demonstrate filleting and homogenizing techniques with practice fish provided by the
Tetra Tech laboratory, and
Provide hands-on opportunities for Tetra Tech laboratory staff to develop proficiency
with filleting and homogenizing fish fillet samples, including equipment cleaning
procedures and collection of equipment rinsate and solvent blank samples.
A9. Documents and Records
Thorough documentation of all NCCA 2020 Great Lakes human health fish sample preparation
activities is necessary for proper sample processing in the laboratory and, ultimately, for the
interpretation of study results. The Tetra Tech Biological Research Facility in Owings Mills,
MD is serving as the fish sample preparation laboratory, and Tetra Tech is responsible for
producing and maintaining the following documents and records:
The Tetra Tech laboratory must prepare and submit a weekly progress report to the OST
Fish Sample Preparation Technical Leader and the OST Project Manager (based on fish
processing information recorded on each 2020 GLHHFFTS Fish Sample Preparation
-------
NCCA 2020 Great Lakes Human Health Fish Sample Preparation QAPP
Revision 0
Date: July 15, 2020
Page 21 of 40
Laboratory Bench Sheet in Appendix C and ORD-Duluth 2020 Great Lakes Special
Study Human Health Fish Sample Preparation Laboratory Bench Sheet in Appendix D)
to document the status of fish sample preparation activities and provide information
specified in the procedures described in Appendix B.
The Tetra Tech laboratory must report the results for the paired rinsate and solvent blank
sample analyses for mercury and PCBs (PFAS rinsates and solvent blanks will be
analyzed by the laboratory selected for analysis of NCCA 2020 Great Lake human health
fish fillet samples for PFAS) and for the triplicate lipid results associated with each fish
sample preparation batch (generally 20 fish samples per batch) to the OST Fish Sample
Preparation Technical Leader and the OST Project Manager. The Tetra Tech laboratory
is also responsible for reporting the full set of lipid analysis results to the OST Fish
Sample Preparation Technical Leader and the OST Project Manager. This includes the
single lipid analysis results for 19 of the 20 fish samples in a typical batch and the
average triplicate lipid results for one fish sample in the batch.
The Tetra Tech laboratory must provide shipping information (e.g., tracking number,
airbills, and shipping forms) to the OST Fish Sample Preparation Technical Leader and
the OST Project Manager for aqueous QC samples and fillet tissue samples sent to
designated analytical laboratories.
All documents and records prepared for the NCCA 2020 Great Lakes human health fish fillet
sample preparation will be maintained by Tetra Tech for the duration of the study and retained
for a period of five years following completion of the study (unless otherwise directed by EPA).
B. DATA GENERATION AND ACQUISITION
Bl. Sampling Process Design (Experimental Design)
The target population for the 2020 GLHHFFTS consists of all 226 of the Great Lakes nearshore
sites randomly selected for 2020 NCCA sampling. Additionally, there are 50 ORD-Duluth 2020
Great Lakes special study enhancement sites in Lake Michigan identified for human health fish
collection. Together these 226 nearshore sites and 50 enhancement sites are designated as the
NCCA 2020 Great Lakes human health whole fish sampling sites. The design for selecting the
human health whole fish sampling sites incorporated the following objectives:
Statistically representative data on the concentrations of mercury, PCBs, and PFAS in
Great Lakes fish commonly consumed by humans
Information on the potential for PFAS to bioaccumulate in fish fillet tissue.
Data to answer questions concerning the occurrence of PFAS in the fillets of fish and the
potential for human exposure through fish consumption.
-------
NCCA 2020 Great Lakes Human Health Fish Sample Preparation QAPP
Revision 0
Date: July 15, 2020
Page 22 of 40
Species-specific information on fatty acid content of Great Lakes fish that are commonly
targeted by fishermen and consumed by humans (analysis of fatty acids will be limited to
fillet samples from the 2020 GLHHFFTS).
Fish fillet tissue data from the 2020 GLHHFFTS will also provide EPA with the opportunity to
evaluate changes in the levels of Great Lakes fish contamination over time by comparing 2020
GLHHFFTS fillet tissue results to the fillet tissue data generated during the 2015 GLHHFFTS
and the 2010 GLHHFTS.
The details of the sampling process design, sampling methods, and sample handling and custody
procedures are described in EPA's National Coastal Condition Assessment 2020 Field
Operations Manual prepared by OWOW (USEPA 2020b).
Sampling at the 2020 GLHHFFTS nearshore sites and ORD-Duluth 2020 Great Lakes special
study enhancement sites in Lake Michigan involves collection of whole fish samples for analysis
of fillet tissue samples for mercury, PFAS, PCBs, lipids, and fatty acids (note that analysis of
fillet samples from Lake Michigan enhancement sites will not include fatty acids). To meet the
study objectives, one fish sample is collected from each nearshore and enhancement site.
Ideally, each fish sample is a routine fish composite sample that consists of five fish of adequate
size to provide a minimum of 75 grams of edible tissue for analysis. Fish are selected for each
composite sample by applying the following criteria:
All are of the same species.
All satisfy legal requirements of harvestable size (or weight) for the sampled site, or at
least be of consumable size if no legal harvest requirements are in effect.
All are of similar size, so that the smallest fish specimen in a composite sample is no less
than 75% of the total length of the largest specimen.
All are collected at the same time, i.e., collected as close to the same time as possible, but
no more than one week apart. (Note: Individual fish may have to be frozen until all fish
to be included in the composite sample are available for delivery to the designated
laboratory.)
Accurate taxonomic identification is essential for preventing the mixing of closely related target
species. Under no circumstances are specimens from different species used in a human health
fish composite sample.
The sample collection goal at each NCCA 2020 Great Lakes site designated for whole fish
sample collection is to obtain a composite sample of fish that are adequate in size to provide a
minimum of 75 grams of fillet tissue for chemical analysis. Field crews are collecting human
health fish samples between June and September during the 2020 field season unless additional
time is required to complete fish sampling due to constraints imposed on field crews because of
the coronavirus pandemic.
-------
NCCA 2020 Great Lakes Human Health Fish Sample Preparation QAPP
Revision 0
Date: July 15, 2020
Page 23 of 40
B2. Sampling Methods
Sampling method procedures and requirements for collection of human health fish samples are
detailed in EPA's National Coastal Condition Assessment 2020 Quality Assurance Project Plan
(USEPA 2020a) and National Coastal Condition Assessment 2020 Field Operations Manual
(USEPA 2020b). These sampling procedures and requirements, which apply to human health
whole fish sample collection at both the 2020 GLHHFFTS nearshore sites and the ORD-Duluth
2020 Great Lakes special study Lake Michigan enhancement sites, are summarized below.
The sampling objective is for field crews to obtain one representative human health whole fish
composite sample from each nearshore and enhancement site. Collecting fish composite samples
is a cost-effective means of estimating average chemical concentrations in the tissue of target
species, and compositing fish ensures adequate sample mass for analysis of multiple chemicals.
The sampling procedures specify that each human health fish composite sample should consist of
five similarly sized adult fish of the same species. OST developed a recommended fish species
list with GLNPO concurrence that contains 25 priority target fish species and 18 alternative fish
species. Field teams use this list as the basis for selecting appropriate fish species for the NCCA
2020 Great Lakes human health fish samples. The method applied for fish collection is left to
the discretion of the field team, but it typically involves angling or gillnetting and occasionally
trawling.
Table 2. Primary Target Fish Species and Secondary Alternative Fish Species for the 2020
GLHHFFTS and ORD-Duluth 2020 Great Lakes Special Study
Primary Fish Species
Scientific Name*
Primary Fish
Species Common
Name
Ambloplites rupestris
Rock bass
Micropterus dolomieu
Smallmouth bass
Micropterus salmoides
Largemouth bass
Pomoxis annularis
White crappie
Pomoxis nigromaeulatus
Black crappie
Cyprinus carpio
Common carp
Esox lucius
Northern pike
Esox masquinongy
Muskellunge
Esox niger
Chain pickerel
Ictalurus punctatus
Channel catfish
Lota lota
Burbot
Morone americana
White perch
Morone chrysops
White bass
Perca flavescens
Yellow perch
Sander canadensis
Sauger
Sander vitreus
Walleye
Coregonus clupeaformis
Lake whitefish
Oncorhynchus gorbuscha
Pink salmon
Secondary Fish Species
Scientific Name*3
Secondary Fish
Species Common
Name
Carpiodes cyprinus
Quillback
Catostomus catostomus
Longnose sucker
Catostomus commersonii
White sucker
Hypentelium nigracans
Northern hogsucker
Ictiobus cyprinellus
Bigmouth buffalo
Ictiobus niger
Black buffalo
Lepomis cyanellus
Green Sunfish
Lepomis gibbosus
Pumpkinseed
Lepomis gulosus
Warmouth
Lepomis macrochirus
Bluegill
Lepomis megalotis
Longear Sunfish
Ameiurus melas
Black bullhead
Ameiurus natalis
Yellow bullhead
Ameiurus nebulosus
Brown bullhead
Coregonus artedi
Cisco/ lake herring
Coregonus hoyi
Bloater
Prosopium cylindraceum
Round whitefish
Salvelinus fontinalis
Brook trout
-------
NCCA 2020 Great Lakes Human Health Fish Sample Preparation QAPP
Revision 0
Date: July 15, 2020
Page 24 of 40
Primary Fish Species
Scientific Name*
Primary Fish
Species Common
Name
Oncorhynchus kisutch
Coho salmon
Oncorhynchus tshcrwytscha
Chinook salmon
Oncorhynchus mykiss
Rainbow trout
Salmo salar
Atlantic salmon
Salmo trutta
Brown trout
Salvelinus namaycush
Lake trout
Aplodinotus grunniens
Freshwater drum
Secondary Fish Species
Scientific Name*3
Secondary Fish
Species Common
Name
* Minimum acceptable length is 190 mm, TL
a Only send if preferred species are not available
* Minimum acceptable length is 190 mm, TL
In preparing Great Lakes human health fish samples for shipping, field teams record sample
number, species name, specimen length, sampling location, and sampling data and time on an
electronic Human Health Fish Collection Form in the NCCA 2020 app. Each fish is wrapped in
solvent-rinsed, oven-baked aluminum foil, with the dull side in using foil sheets provided by
EPA. Individual foil-wrapped specimens are placed into a length of food-grade polyethylene
tubing, each end of the tubing is sealed with a plastic cable tie, and a fish specimen label is
affixed to the outside of the food-grade tubing with clear tape. All of the wrapped fish in the
sample from each site are placed in a large plastic bag and sealed with another cable tie, then
placed immediately on dry ice for shipment to Microbac Laboratories in Baltimore, Maryland.
Field crews are directed to pack fish samples on dry ice in sufficient quantities to keep samples
frozen for up to 48 hours (i.e., 50 pounds), and to ship them via priority overnight delivery
service (i.e., FedEx), so that they arrive at Microbac Laboratories in less than 24 hours from the
time of sample collection. Alternatively, field crews may transport Great Lakes human health
whole fish samples on wet or dry ice (depending on the distance) to an interim facility where the
fish samples are frozen and stored for up to two weeks before overnight shipping to Microbac
Laboratories on dry ice as described above.
B3. Sample Receipt and Inspection
This section describes the procedures that apply once the Great Lakes human health whole fish
(HTIS) samples are shipped from the field to Microbac Laboratories. The Great Lakes human
health fish samples are being collected by various organizations participating with EPA in this
study, including state and tribal agencies and contractors. Although samples are shipped frozen
on dry ice, they must be inspected promptly on receipt. As samples are received, a Microbac
Laboratories representative will:
Check that each cooler has arrived undamaged and verify that samples are still frozen and
in good condition.
Check the temperature of one of the samples in the cooler using a thermometer that reads
to at least -20 degrees Celsius (°C), or an infra-red (IR) temperature "gun" and report the
reading to CSRA/GDIT via email.
Notify CSRA/GDIT via email about receipt of samples on the day of delivery and report
if each sample arrives frozen with dry ice remaining in the cooler.
-------
NCCA 2020 Great Lakes Human Health Fish Sample Preparation QAPP
Revision 0
Date: July 15, 2020
Page 25 of 40
Store the coolers in the onsite freezer.
Microbac Laboratories will notify CSRA/GDIT on the day of delivery about any problems
encountered upon receipt of samples (e.g., no dry ice left in cooler, fish partly or completely
thawed, etc.). CSRA/GDIT subsequently reports sample receipt and inspection issues to the
OST Project Manager and OST Fish Sample Preparation Technical Leader, then coordinates
with EPA to resolve the issues.
Generally within two weeks of Great Lakes human health whole fish sample deliveries, a
CSRA/GDIT staff member acting as a sample custodian will:
Retrieve coolers containing NCCA 2020 Great Lakes human health whole fish samples
from the freezer at Microbac Laboratories.
Remove whole fish samples from each cooler and record the Site ID and EPA Sample
Number.
Transfer the whole fish samples to trays in the freezer for interim storage and leave
empty coolers outside the freezer for Tetra Tech staff to retrieve.
CSRA/GDIT reports any discrepancies in individual fish label information and receipt of
fish samples from sampling sites to the OST Project Manager for resolution.
After completing whole fish sample check-in, CSRA/GDIT stores the whole fish samples in the
freezer at Microbac Laboratories where they are kept at temperatures less than or equal to -20°C
until they are ready to transfer to the Tetra Tech laboratory in Owings Mills, MD for fish sample
preparation. (The freezers are maintained by Microbac Laboratories under a separate agreement
with CSRA/GDIT and are continuously monitored by an automated temperature monitoring
system.)
B4. Fish Sample Preparation and Analytical Methods
The laboratory at Tetra Tech's Biological Research Facility in Owings Mills, MD is the fish
sample preparation laboratory (prep lab) for the NCCA 2020 Great Lakes human health fish
samples. In this role, Tetra Tech is responsible for removing scales from each valid fish in the
whole fish sample, filleting valid fish in the whole fish samples, homogenizing the fillet tissue,
preparing the required number of fish fillet tissue aliquots for analysis and archive, shipping the
fillet tissue aliquots for each type of analysis to the designated analytical laboratories, and
providing short-term storage for archived fillet tissue samples in a freezer at their facility. The
specific procedures for NCCA 2020 Great Lakes human health fish sample preparation activities
are described in Appendix B.
This section describes NCCA 2020 Great Lakes human health fish sample preparation methods
(i.e., methods for preparing homogenized fillet tissue samples). It also describes methods for
analysis of lipids in ground fillet tissue samples for homogeneity testing and lipid content and for
analysis of mercury, PCBs, and PFAS in equipment rinsate and solvent blank samples to test the
adequacy of equipment cleaning. Lipid analysis of ground fillet tissue samples for homogeneity
testing and mercury, PCB, and PFAS analysis of equipment rinsate and solvent blank samples
for testing sufficient equipment cleaning are conducted as part of the QC procedures for fish
sample preparation. Analytical method requirements for analysis of homogenized fillet samples
-------
NCCA 2020 Great Lakes Human Health Fish Sample Preparation QAPP
Revision 0
Date: July 15, 2020
Page 26 of 40
for target chemicals (e.g., mercury, PFAS, PCB congeners, PCBs as Aroclors, and fatty acids)
are described in the NCCA 2020 Great Lakes human health fish fillet tissue sample analysis
QAPP, which will be developed at a later date.
B4.1 Fish Sample Preparation
Trained laboratory staff at Tetra Tech's Biological Research Facility in Owings Mills, MD are
responsible for preparation of homogenized fillet tissue samples from NCCA 2020 Great Lakes
human health whole fish samples. Preparing the homogenized fillet tissue samples involves
removing scales from each valid fish specimen in a sample, filleting the individual fish to be
included in the tissue composite sample, homogenizing the fillet tissue from each whole fish
sample, preparing the required number of fish fillet tissue aliquots for analysis and archive,
shipping the fillet tissue aliquots for each type of analysis to the designated analytical
laboratories, and storing archived fillet tissue samples on an interim basis in a freezer at their
facility. The specific procedures for fillet tissue sample preparation activities are summarized
below and fully described in Appendix B.
Homogenized Fillet Sample Preparation
The filleting process involves removing the fillet (with skin on and "belly flap" or ventral muscle
attached) from both sides of each valid fish in the whole fish sample. The combined fillets from
all valid fish in the whole fish sample are weighed to the nearest gram (wet weight) before they
are homogenized together. An electric meat grinder is used to prepare each homogenized fillet
sample. The entire set of fillets (with skin and belly flap) from both sides of every valid fish in
the whole fish sample (i.e., ideally 5 fish per sample) are homogenized, and the entire
homogenized volume is used to prepare the fillet tissue sample aliquots. Grinding of the fillet
tissue is repeated until the tissue consists of a uniform color and finely ground texture.
Homogeneity is confirmed by conducting triplicate analyses of the lipid content in one fish
sample from each fish sample preparation batch (generally one in 20 fish samples). The
collective weight of the homogenized fillet tissue from the whole fish sample is recorded to the
nearest gram (wet weight) after processing. Tetra Tech lab technicians prepare fillet tissue
sample aliquots for chemical analysis and archive according to specifications in Table 1 of
Appendix B.
Fish Sample Preparation Batches
Each NCCA 2020 Great Lakes human health fish sample preparation batch generally consists of
20 whole fish samples. The number of whole fish samples in the final fish sample preparation
batch (or two) may be adjusted to include a few more than 20 or fewer than 20, depending on
what fraction of 20 whole fish samples are left near the end of fish processing for assignment to a
batch. Tetra Tech staff will develop fish sample preparation instructions that include all valid
fish samples available for processing. Processing may not begin until the OST Fish Sample
Preparation Technical Leader and the OST Project Manager review the draft instructions and the
OST Fish Sample Preparation Technical Leader approves the final instructions and batch
assignments with OST Project Manager concurrence.
-------
NCCA 2020 Great Lakes Human Health Fish Sample Preparation QAPP
Revision 0
Date: July 15, 2020
Page 27 of 40
B4.2 Lipid Analysis
Tetra Tech will procure the services of an analytical laboratory (to be determined or TBD) to
conduct one set of triplicate lipid analyses per fish sample preparation batch (see definition in
Section B4.1) as described in Steps 28 and 29 of Appendix B. A single homogenized fillet tissue
aliquot is analyzed for lipid content for each of the remaining samples in the batch (usually 19).
Lipids are extracted from each fillet tissue sample using a method proposed by the TBD
analytical laboratory. (This QAPP will be amended to include a description of this method after
the analytical laboratory is selected and their proposed method for lipid analysis is approved.)
Percent lipid content is calculated by dividing the lipid weight by the initial fillet tissue aliquot
weight and multiplying that result by 100. The batch-specific lipid results for homogeneity
evaluations are reviewed initially by Tetra Tech and independently by CSRA/GDIT against the
QC specifications detailed in Section B5.1.
B4.3 Mercury Rinsate Analysis
Tetra Tech laboratory technicians prepare one set of paired rinsate and solvent blank samples per
fish sample preparation batch (see definition in Section B4.1) for mercury analysis as described
in Steps 25 through 27 of Appendix B. This set of paired QC samples consists of one deionized
(DI) water equipment rinsate sample and one DI water blank sample. Tetra Tech will procure
the services of an analytical laboratory (TBD) to conduct mercury analysis of these QC samples
using EPA Method 245.1 (USEPA 1994). This method was developed to measure total mercury
(organic and inorganic) in aqueous samples. The flameless atomic absorption procedure is a
physical method based on the absorption of radiation at 253.7 nanometers by mercury vapor.
Mercury is first reduced to its elemental state using a potassium permanganate digestion
procedure. The samples/standards and a stannous chloride reagent are then pumped into the
mercury analyzer and mixed. Argon gas is introduced into the solution stream. Absorbance
(peak height) is measured as a function of mercury concentration and recorded as ppb of
mercury. Results of the batch-specific mercury rinsate and solvent blank analyses are reviewed
initially by Tetra Tech and independently by CSRA/GDIT against the QC specifications detailed
in Section B5.2.
B4.4 PCB Rinsate Analysis
Tetra Tech will procure the services of an analytical laboratory (TBD) to conduct analyses of
paired rinsate and solvent blank samples for a subset of PCB congeners that contain at least PCB
congeners 52, 66, 105, 118, 141, 146, 170, 174, 177, and 187. These congeners represent those
that EPA has found frequently and at relatively high concentrations in other fish tissue studies.
Laboratories responding to Tetra Tech's solicitation will propose a PCB method that can meet
the analytical specifications in the solicitation. This QAPP will be amended to include a
description of the method after the analytical laboratory is selected and their proposed method
for PCB analysis is approved.
-------
NCCA 2020 Great Lakes Human Health Fish Sample Preparation QAPP
Revision 0
Date: July 15, 2020
Page 28 of 40
B4.5 PFAS Rinsate Analysis
The PFAS paired rinsate and solvent blank samples will not be analyzed during the fish sample
preparation process. Instead, they will be analyzed by the laboratory selected for PFAS analysis
of the 2020 GLHHFFTS and ORD-Duluth 2020 Great Lakes special study fish fillet tissue
samples. A description of the method used for analysis of the aqueous PFAS rinsate and solvent
blank samples will be provided in the NCCA 2020 Great Lakes human health fish fillet tissue
sample analysis QAPP, which will be developed at a later date.
B5. Fish Sample Preparation Quality Control Requirements
The procedures associated with the NCCA 2020 Great Lakes human health fish sample
preparation process include the following: preparation of homogenized fillet tissue samples,
which includes triplicate lipid analyses of homogenized fillet tissue samples to test for
homogeneity (Section B5.1) and analytical testing for mercury, PCBs, and PFAS in equipment
rinsate and solvent blank samples (Sections B5.2, B5.3, and B5.4, respectively) for QC purposes.
The QC procedures are performed for one whole fish sample in each fish sample preparation
batch (usually one in a batch of 20 fish samples).
B5.1 Homogenized Fillet Samples
Lipid content analysis is used as a surrogate to confirm homogeneity of the homogenized fish
fillet samples that are prepared in the Tetra Tech Biological Research Facility laboratory. A
laboratory (TBD) under contract to Tetra Tech will conduct triplicate lipid analyses of ground
fillet tissue aliquots from one whole fish sample in each fish sample preparation batch and use
the lipid content of those 3 fillet tissue aliquots to confirm that the ground fillet tissue is
homogeneous. Laboratory technicians prepare this triplicate lipid aliquot (30 to 35 g) of
homogenized fillet tissue mass for lipid analysis following the specific procedures described in
Appendix B for placing the aliquot in a container, labeling it, and storing it in a freezer (refer to
Step 19 in Appendix B). All homogenized fillet tissue aliquots for lipid analysis are shipped on
dry ice and under chain of custody to the designated analytical laboratory. The results of the
homogeneity testing are delivered to Tetra Tech for their initial review and forwarded to the OST
Fish Sample Preparation Technical Leader and OST Project Manager for independent review by
CSRA/GDIT and EPA.
From the triplicate lipid results, CSRA/GDIT calculates the mean lipid content (in percent), the
standard deviation (SD), and the relative standard deviation (RSD) using the formulae below, or
the corresponding functions in Excel, and reports the results to the OST Fish Sample Preparation
Technical Leader and the OST Project Manager.
3
2 (%lipids)-
mean % lipids =
-------
NCCA 2020 Great Lakes Human Health Fish Sample Preparation QAPP
Revision 0
Date: July 15, 2020
Page 29 of 40
I Z(%lipidSi meanlipids)2
SD = .
mean
If the RSD of the triplicate lipid results is less than or equal to 15% for mean % lipid
measurements that are greater than or equal to 2.5% or if the RSD of the triplicate lipid results is
less than or equal to 20% for mean % lipid measurements less than 2.5%, then EPA will notify
Tetra Tech that the homogenization effort is sufficient for all of the homogenized fillet tissue
samples (usually 20) in each analysis batch (refer to Step 29 in Appendix B).
Tetra Tech laboratory staff may continue to process up to two additional fish sample preparation
batches. However, the laboratory may not continue to process batches beyond that third fish
sample preparation batch until receiving notification from the OST Fish Sample Preparation
Technical Leader (after OST Project Manager concurrence) that review of homogeneity test
results from the initial batch is complete and the results are deemed satisfactory.
B5.2 Mercury Analysis of Rinsate Samples
The Tetra Tech laboratory prepares one set of DI water equipment rinsate and DI water blank
samples during processing of each fish sample preparation batch and a subcontracted laboratory
(TBD) analyzes each set of rinsate and blank samples for total mercury using EPA Method
245.1, which is a cold-vapor atomic absorption procedure applicable to water samples (Section
B4.3). The pair of blank and rinsate samples are analyzed individually, not in batches of up to
20, in order to provide timely feedback of the cleanliness of the homogenization equipment.
EPA Method 245.1 requires daily instrument calibration and analysis of two quality control
samples, an instrument blank and a laboratory control sample. The rinsates are prepared in
reagent water, so there is little chance of a "matrix effect." Each laboratory control sample,
which is also prepared in reagent water, provides sufficient information on the performance of
the method and the laboratory. The QC sample requirements, including the acceptance criteria
and corrective actions, are summarized in Table 3 below.
Table 3. QC Samples and Acceptance Criteria for Mercury Analysis of Rinsates
Quality Control Sample
Frequency
Acceptance Criteria
Instrument blank
With each rinsate
sample
Result must be less than the MDL. Otherwise, redigest
and reanalyze the rinsate sample.
Laboratory control
sample
With each rinsate
sample
80 - 120% recovery of mercury. Otherwise, correct
instrumental problems, and redigest and reanalyze the
rinsate sample.
The batch-specific rinsate results are reviewed initially by Tetra Tech and forwarded to the OST
Fish Sample Preparation Technical Leader and the OST Project Manager for independent review
by CSRA/GDIT. The rinsate results are evaluated based on the mass of mercury detected and
the assumption that all of the apparent contamination could be transferred to a nominal 465-g
-------
NCCA 2020 Great Lakes Human Health Fish Sample Preparation QAPP
Revision 0
Date: July 15, 2020
Page 30 of 40
mass of homogenized tissue. If review of the results shows that the rinsate samples are below
the acceptance limit for mercury, i.e., 0.2 |ig/L for total mercury based on the method detection
limit for an aqueous sample, then the equipment cleaning effort is sufficient for all samples in
that fish sample preparation batch.
Rinsate results for mercury above the reporting limit mentioned above may cause a need for
corrective actions by the Tetra Tech laboratory. These corrective actions may include revisions
to the laboratory's equipment cleaning procedures, followed by a successful demonstration of the
revised cleaning procedures through preparation and analysis of additional rinsate samples.
Tetra Tech laboratory staff may continue to process up to two additional fish sample preparation
batches. However, laboratory staff may not continue to process batches beyond that third fish
sample preparation batch until receiving notification from the OST Fish Sample Preparation
Technical Leader (with OST Project Manager concurrence) that review of rinsate test results
from the initial fish sample preparation batch is complete and the results are deemed satisfactory.
B5.3 PCB Analysis of Rinsate Samples
The Tetra Tech laboratory prepares one set of hexane equipment rinsate and solvent blank
samples during processing of each fish sample preparation batch. A subcontracted laboratory
(TBD) analyzes each set of rinsate and blank samples for a selected subset of PCB congeners
using a PCB method proposed and approved during the laboratory solicitation process.
For the NCCA 2020 Great Lakes human health fish preparation process, PCB rinsate analyses
will focus on at least PCB congeners 52, 66, 105, 118, 141, 146, 170, 174, 177, and 187. These
congeners represent those that EPA has found frequently and at relatively high concentrations in
other fish tissue studies. As with the mercury rinsate analysis, the PCB rinsate samples are
prepared and analyzed individually, not in batches of up to 20, in order to provide timely
feedback of the cleanliness of the homogenization equipment. Therefore, the quality control
samples associated with the rinsate samples analyzed for PCBs are usually analyzed with each
rinsate sample. When PCB method information becomes available, this QAPP will be amended
to identify QC samples and acceptance criteria in Table 4 below.
Table 4. QC Samples and Acceptance Criteria for PCB Analysis of Rinsates
QC Sample
Frequency
Acceptance Criteria
The batch-specific rinsate results are reviewed initially by Tetra Tech and forwarded to the OST
Fish Sample Preparation Technical Leader and the OST Project Manager for independent review
by CSRA/GDIT. If review of the results shows that rinsate samples are below the acceptance
limit for PCBs, i.e., 0.5 ng/mL per congener (based on the instrument detection limit for a 1.0-
mL final volume of solvent concentrated from the original 100-mL rinsate sample), then the
equipment cleaning effort is sufficient for all samples in that fish sample preparation batch.
Rinsate results for PCBs above the reporting limit mentioned above may cause a need for
corrective actions by the Tetra Tech laboratory. These corrective actions may include revisions
-------
NCCA 2020 Great Lakes Human Health Fish Sample Preparation QAPP
Revision 0
Date: July 15, 2020
Page 31 of 40
to the laboratory's equipment cleaning procedures, followed by a successful demonstration of the
revised cleaning procedures through preparation and analysis of additional rinsate samples.
The Tetra Tech laboratory may continue to process up to two additional fish sample preparation
batches. However, the laboratory may not continue to process batches beyond that third fish
sample preparation batch until receiving notification from the OST Fish Sample Preparation
Technical Leader that review of PCB rinsate results from the initial fish sample preparation batch
is complete and the results are deemed satisfactory.
B5.4 PFAS Analysis of Rinsate Samples
The QC requirements for PFAS analysis of rinsate samples will be specified in the NCCA 2020
Great Lakes human health fish fillet tissue sample analysis QAPP, which will be developed at a
later date.
B6. Instrument/Equipment Testing, Inspection, and Maintenance
There are no analytical instruments used in the preparation of the fillet tissue samples. However,
the balances used to weigh the whole fish and the fillet tissue sample aliquots are inspected daily
and the homogenization equipment (meat grinder) is inspected when it is reassembled after
cleaning between samples.
All analytical instruments associated with fish sample preparation operations are inspected and
maintained as described in the respective analytical methods and laboratory Standard Operating
Procedures (SOPs). This includes the instruments involved with the fish fillet homogeneity
(lipid) testing and with analyses of aqueous rinsate and solvent blank samples for mercury,
PCBs, and PFAS and fillet tissue samples for percent lipids.
B7. Instrument/Equipment Calibration and Frequency
The balances used to weigh the whole fish and the fillet tissue during the various stages of
homogenization and aliquot preparation are calibrated on a regular schedule and calibrations are
verified at the beginning of each day on which the balances are used.
All analytical instrumentation associated with the rinsate analyses and with fillet homogeneity
(lipid) testing and percent lipid analyses will be calibrated as described in the respective
analytical methods. The methods cited in Sections B4 and B5 for the rinsate analyses require
multi-point initial calibrations and periodic calibration verifications, and all the methods contain
QC acceptance criteria for calibration. The mercury analysis method for the rinsate samples,
Method 245.1, specifies calibration with five calibration standards. The PCB analysis method
for the rinsate samples specifies calibration with three to five calibration standards depending on
the proposed method.
-------
NCCA 2020 Great Lakes Human Health Fish Sample Preparation QAPP
Revision 0
Date: July 15, 2020
Page 32 of 40
B8. Inspection/Acceptance of Supplies and Consumables
Careful and thorough planning is necessary to ensure the efficient and effective completion of
the fillet tissue sample preparation tasks. Fish preparation laboratory equipment and supplies are
described in Appendix B. All fish sample packaging and shipping supplies are provided by Tetra
Tech. It is the responsibility of the Tetra Tech laboratory technicians to procure, compile, and
inspect the necessary fillet sample preparation equipment and supplies prior to commencement
of fillet tissue sample preparation activities, and to inspect packaging and shipping supplies
before fillet tissue samples are shipped to the respective analytical laboratories for analysis.
B9. Non-direct Measurements
Non-direct measurements are not required for this project. The analytical results from the
2010 GLHHFTS or the 2015 GLHHFFTS to which any new data are to be compared are primary
data that EPA generated under an approved QAPP for each study.
B10. Data Management
Data management practices employed in this study are based on standard data management
practices used for EPA's National Lake Fish Tissue Study and other OST fish contamination
studies (e.g., the 2008-09 NRSA, 2010 Great Lakes Human Health Fish Tissue Study,
2013-14 NRSA, 2015 Great Lakes Human Health Fish Fillet Tissue Study, and 2018-19 NRSA).
The data management (i.e., sample tracking, data tracking, data inspection, data quality
assessment, and database development) procedures have been regularly applied to other technical
studies by Tetra Tech and CSRA/GDIT.
Fish Sample Collection Data
Samples are documented and tracked through the use of standardized human health fish tissue
(HTIS) fields in the 2020 NCCA app, Whole Fish Sample Identification Labels, and 2020 NCCA
HTIS tracking spreadsheets. Specific fish sample collection data requirements are detailed in the
Field Operations Manual (USEPA 2020b) prepared by OWOW. Whole fish samples are shipped
to Microbac Laboratories (Baltimore, MD) by an overnight air delivery service that provides
constant tracking of shipments (i.e., FedEx).
The Tetra Tech laboratory retains a copy of the 2020 NCCA HTIS tracking spreadsheet sample
information that is received electronically from the NARS IM group upon shipment of each 2020
NCCA whole fish sample. Tetra Tech staff perform a data QC check on each sample tracking
spreadsheet, compile each individual sample tracking spreadsheet into a combined current master
sample tracking spreadsheet, and forward it to the OST Project Manager and the OST Fish
Sample Preparation Technical Leader, along with documentation reporting the field data QC
review (consistent with field data QC documentation provided for previous EPA fish tissue
studies). All electronic files related to fish sample collection that are produced and retained by
Tetra Tech are maintained in a project file during the active phase of this project, and for a
period of 5 years following completion of the project (unless otherwise directed by EPA).
-------
NCCA 2020 Great Lakes Human Health Fish Sample Preparation QAPP
Revision 0
Date: July 15, 2020
Page 33 of 40
Upon completion of sampling activities, Tetra Tech develops a Fish Sample Master Spreadsheet
based on information recorded by all field sampling teams in the 2020 NCCA app and provided
by the NARS IM group in sample tracking spreadsheets. This field data is entered into Excel
spreadsheets to create separate master spreadsheets for the NCCA 2020 GLHHFFTS and ORD-
Duluth 2020 Great Lakes special study. All data entries are checked for errors in transcription
and computer input by qualified persons (minimum of two) who did not originally enter the data.
If there is any indication that requirements for sample integrity or data quality have not been met,
the Tetra Tech QA Officer is notified immediately (with an accompanying explanation of the
problems encountered) for discussion and resolution of quality issues before delivery of the Fish
Sample Master Spreadsheet to the OST Project Manager and the OST Fish Sample Preparation
Technical Leader.
Fish Sample Preparation Data
The Tetra Tech laboratory is required to maintain all records and documentation associated with
the preparation of 2020 GLHHFFTS and ORD-Duluth 2020 Great Lakes special study human
health fish samples (e.g., weekly reports containing fillet tissue sample preparation data and fillet
tissue aliquot documentation), the analyses of fillet tissue samples for lipids for homogeneity
testing and lipid content, and the analyses of rinsate samples for mercury and PCBs. All required
analytical laboratory reports and documentation, including raw data, must be sequentially
paginated and clearly labeled with the laboratory name, and associated sample numbers. Any
electronic media submitted must be similarly labeled. The sample preparation laboratory and
analytical laboratories contracted for homogeneity testing and rinsate analyses will adhere to a
comprehensive data management plan that is consistent with the principles set forth in Good
Automated Laboratory Practices, EPA Office of Administration and Resources Management
(USEPA 1995) or with commonly employed data management procedures approved by the
National Environmental Laboratory Accreditation Conference (NELAC).
Data Retention
All computer files associated with the NCCA 2020 GLHHFFTS and ORD-Duluth 2020 Great
Lakes special study are stored in a project subdirectory by Tetra Tech and are copied to network
storage for archive for the 5 years subsequent to project completion (unless otherwise directed by
the OST Project Manager).
C. ASSESSMENT AND OVERSIGHT
CI. Assessments and Response Actions
Cl.l Fish Sample Preparation
The Tetra Tech laboratory supporting fish sample preparation for this study and the analytical
laboratories responsible for lipid testing and rinsate analyses each have a comprehensive QA
program in place and operating at all times. In performing fish sample preparation and QC and
lipid sample analyses for this study, each laboratory will adhere to the requirements of those
respective QA programs. Copies of those plans are maintained on file at Tetra Tech.
-------
NCCA 2020 Great Lakes Human Health Fish Sample Preparation QAPP
Revision 0
Date: July 15, 2020
Page 34 of 40
If any technical problems are encountered during operations at the Tetra Tech laboratory, the
Tetra Tech Project Leader will consult with the Tetra Tech Laboratory Manager, the OST Fish
Sample Preparation Technical Leader, and the OST Project Manager to identify corrective
actions. The Tetra Tech Project Leader is responsible for ensuring that the corrective actions are
successfully implemented. Section B5 of this QAPP identifies corrective actions for any lipid,
mercury, or PCB analysis results generated by the analytical laboratory (or laboratories) that do
not meet the QC acceptance criteria. The Tetra Tech Project Leader is responsible for ensuring
that each analytical laboratory implements the required corrective actions.
Analysis of the QC rinsate samples for PFAS will be conducted by a TBD laboratory selected for
PFAS analysis of the 2020 GLHHFFTS and ORD-Duluth 2020 Great Lakes special study
homogenized fillet tissue samples. PFAS analyses of both the QC rinsate samples (from the fish
sample preparation process) and the fillet tissue samples are included under a separate 2020
Great Lakes human health fish fillet tissue sample analysis QAPP that will be developed at a
later date.
C1.2 Performance Audits
Performance audits are qualitative checks on different segments of project activities. For the
2020 GLHHFFTS, performance audit techniques include checks on post-collection review of
field measurements and the use of triplicate lipid analyses of one homogenized sample in every
fish sample preparation batch as a check on homogeneity. The Tetra Tech Project Leader is
responsible for overseeing work as it is performed and for periodically conducting QC checks
during fillet sample preparation for this project. Results of these checks are reported to the Tetra
Tech Quality Assurance Officer, the OST Fish Sample Preparation Technical Leader, and the
OST Project Manager.
CI.3 System Audits
System audits are qualitative reviews of project activities to check that the overall quality
program is functioning and that the appropriate QC measures identified in the QAPP are being
implemented. If the results of the performance audits described in Section CI.2 indicate
problems, the Tetra Tech QA Officer will conduct an internal system audit during the project and
report the results to the OST Fish Sample Preparation Technical Leader and the OST Project
Manager. If QA/QC deficiencies are discovered, additional internal system audits are conducted
until the Tetra Tech QA Officer, the OST Fish Sample Preparation Technical Leader, and the
OST Project Manager conclude that overall project quality requirements are being met.
C2. Surveillance
C2.1 Whole Fish Sample Shipment
When NCCA 2020 Great Lakes human health whole fish samples are shipped to Microbac
Laboratories (Baltimore, MD), the NARS Information Management (IM) Group contacts the
OST Project Manager, OST Fish Sample Preparation Technical Leader, Tetra Tech Project
Leader, and CSRA/GDIT via email distribution of the sample tracking spreadsheet to notify
them of the upcoming fish sample deliveries, and CSRA/GDIT contacts the OST Project
-------
NCCA 2020 Great Lakes Human Health Fish Sample Preparation QAPP
Revision 0
Date: July 15, 2020
Page 35 of 40
Manager and the OST Fish Sample Preparation Technical Leader when the samples arrive.
Within 24 hours of sample receipt, CSRA/GDIT notifies the OST Project Manager and the OST
Fish Sample Preparation Technical Leader of sample condition. If problems with the shipment
are noted, Tetra Tech and CSRA/GDIT will work with the OST Project Manager to resolve any
problems as quickly as possible to minimize data integrity problems.
C2.2 Fish Sample Preparation
The content of fish sample preparation batches and the process for forming the batches are
described in Section B4.1. The Tetra Tech laboratory may not begin processing any
2020 GLHHFFTS and ORD-Duluth 2020 Great Lakes special study whole fish samples until this
QAPP is approved and the laboratory personnel responsible for fish sample preparation have
been trained on the fish sample preparation procedures and requirements described in this QAPP.
The Tetra Tech Project Leader coordinates with the OST Fish Sample Preparation Technical
Leader and the OST Project Manager regarding fish tissue sample shipments to other
laboratories (i.e., the analytical laboratories responsible for mercury, PCB, and PFAS analysis of
fish sample preparation equipment rinsate and solvent blank QC samples and for lipid analysis of
homogenized fillet tissue samples, including triplicate lipid analysis of one fish sample per fish
sample preparation batch for homogeneity testing) once analysis contracts are in place. Tetra
Tech communicates periodically with laboratory staff by telephone or email to monitor the
progress of lipid, mercury, and PCB rinsate analyses. If technical problems are encountered
during fish sample preparation or during lipid, mercury, or PCB analyses, the Tetra Tech Project
Leader will identify a technical expert within Tetra Tech to assist in resolving the problem, and
work with the OST Fish Sample Preparation Technical Leader and the OST Project Manager to
identify and implement a solution to the problem. The Tetra Tech laboratory is permitted to
work two batches ahead of the delivery and review of batch-specific QC results that indicate if
the homogenization and equipment cleaning procedures for each fish sample preparation batch
are adequate.
If the fish sample preparation (Tetra Tech) or analytical (TBD) laboratories fail to deliver QC
data on time, or if an analytical laboratory notifies Tetra Tech of anticipated reporting or sample
processing delays, Tetra Tech notifies the OST Fish Sample Preparation Technical Leader and
the OST Project Manager of the situation. To the extent possible, Tetra Tech will adjust
schedules and shift resources as necessary to minimize the impact of laboratory delays on EPA
schedules. Tetra Tech will immediately notify the OST Fish Sample Preparation Technical
Leader and the OST Project Manager of any laboratory delays that are anticipated to impact EPA
schedules.
C3. Reports to Management
Upon completion of weekly fish sampling and sample preparation activities, the Tetra Tech
Project Leader provides the OST Project Manager and OST Fish Sample Preparation Technical
Leader with reports of fish sampling team and Tetra Tech laboratory progress for the preceding
week when these activities are occurring. These weekly progress reports include specific details
about the fish sample collection and fillet sample preparation activities and note any concerns
about sample quality and resolution of those concerns. Following completion of fish sampling
-------
NCCA 2020 Great Lakes Human Health Fish Sample Preparation QAPP
Revision 0
Date: July 15, 2020
Page 36 of 40
and fillet sample preparation activities, Tetra Tech prepares a fish collection effort summary
(which details all sampling participants, sampling locations, and specimens collected) and a
sample preparation summary (which lists all samples processed and identifies all fillet tissue
aliquots prepared) for review by the OST Project Manager and the OST Fish Sample Preparation
Technical Leader.
D. DATA VALIDATION AND USABILITY
Dl. Data Review, Verification, and Validation
The processes for data review, verification, and validation provide an approach for standardized
data quality assessment. These processes are also important for determining the usability and
limitations of the whole fish sample collection and fillet tissue sample preparation data generated
for the NCCA 2020 GLHHFFTS and the ORD-Duluth 2020 Great Lakes special study.
Processes for each step in the data quality assessment are described below.
Dl.l Data Review
Fish Sample Collection
Tetra Tech reviews data entries in the 2020 GLHHFFTS whole fish and ORD-Duluth 2020 Great
Lakes special study whole fish sample tracking spreadsheets and individual fish sample labels
for completeness, correctness, and consistency among the fish sampling records. Any errors or
omissions identified during this review are reported to the OST Project Manager and corrected
by contacting the 2020 NCCA Field Logistics Coordinator (Chris Turner of GLEC) or the field
crew leader who initially made the data entries, if necessary. The Tetra Tech Project Leader is
responsible for ensuring that all errors or omissions are addressed in the fish sampling records
before the fish samples are transferred from Microbac Laboratories to the Tetra Tech Laboratory
for fish sample preparation.
Analysis of Lipid and Fish Sample Preparation QC Samples
The Laboratory Managers at each analytical laboratory designated for analysis of lipid and fish
sample preparation QC samples review all laboratory results and calculations prior to submission
of a data package. Any errors identified during this peer review are returned to the lab analyst
for correction. Following correction of the errors, each Laboratory Manager verifies that the
final data package is complete and compliant with the contract, signs each data submission to
certify that the package was reviewed and determined to be in compliance with the terms and
conditions of the contract, and submits the data package to the Tetra Tech Project Leader.
D1.2 Data Verification
The basic goal of data verification is to ensure that project participants know what data were
produced, if these data are complete, if the data are contractually compliant, and if the data meet
the objectives of the study and the QA requirements described in this QAPP.
-------
NCCA 2020 Great Lakes Human Health Fish Sample Preparation QAPP
Revision 0
Date: July 15, 2020
Page 37 of 40
Fish Sample Collection
Tetra Tech staff independent of fish sampling teams verify fish sample collection data reviewed
and submitted by each field team leader. This involves verifying that all data entries in the
sample tracking spreadsheets for NCCA 2020 Great Lakes human health whole fish samples and
whole fish sample labels are complete, correct, and consistent among the fish sampling records.
The data verifier reports any discrepancies identified during this process to the Tetra Tech
Project Leader. The Tetra Tech Project Leader is responsible for reconciling any discrepancies
reported during data verification with the appropriate associated field personnel and for notifying
the data verifier about the resolution of these discrepancies. The data verifier is responsible for
documenting resolution of these data entry discrepancies.
Analysis of Lipid and Fish Sample Preparation QC Samples
The Tetra Tech Laboratory Manager conducts initial reviews of the fish sample preparation QC
sample analysis results for each Great Lakes human health fish sample preparation batch and for
the single lipid analysis results associated with each fish sample to verify the completeness and
accuracy of these data and their compliance with QC acceptance criteria in Section B5 of this
QAPP. Verification of these analytical results involves review of data for percent lipid
measurements (including the triplicate lipid analysis results for the homogeneity testing of one
fish sample per fish sample preparation batch, along with lipid analysis results for the remaining
fish samples) and review of sample processing equipment rinsate and corresponding solvent
blank QC sample data. The Tetra Tech Project Leader verifies the summary level results for
these QC samples and remaining lipid samples, determines if they meet the project objectives in
this QAPP, and reports the verification findings to the OST Fish Sample Preparation Technical
Leader and the OST Project Manager. The CSRA/GDIT analytical chemist supporting the 2020
GLHHFFTS conducts an independent review of analytical results for lipids and for fish sample
preparation QC samples and reports the verification findings to the OST Fish Sample Preparation
Technical Leader and the OST Project Manager. The OST Project Manager and OST Fish
Sample Preparation Technical Leader work with the Tetra Tech Project Leader and the
CSRA/GDIT analytical chemist to resolve any differences in their respective verification
findings.
D1.3 Data Validation
Data validation is the process of evaluating the quality of the results relative to their intended
use. This process is applied to fish sample collection and fish sample preparation as described
below.
Fish Sample Collection
Evaluating the quality of fish sample collection results involves comparing these results to the
fish sampling requirements described in the NCCA 2020 Field Operations Manual (USEPA
2020b) prepared by OWOW. These requirements include collecting fish samples from specific
waterbodies and obtaining particular fish species and numbers of fish to meet study objectives.
-------
NCCA 2020 Great Lakes Human Health Fish Sample Preparation QAPP
Revision 0
Date: July 15, 2020
Page 38 of 40
Fish Sample Preparation
The data validation process for fillet tissue sample preparation is more limited than the process
applied during validation of analytical data from analysis of fillet tissue samples for the study-
specific target chemicals. It focuses on evaluating the clarity and accuracy of required
information in the fish sample preparation weekly progress reports (e.g., percentage of total fillet
mass compared to the total body mass, tissue mass requirements for specified tissue aliquots,
etc.).
D2. Verification and Validation Methods
Fish Sample Collection Data
The initial step in the process for data verification involves Tetra Tech staff independent of fish
sampling operations conducting reviews of all data related to fish sample collection as a means
of identifying any discrepancies among sampling data and related information entered in the
sample tracking spreadsheets for 2020 Great Lakes human health whole fish samples and whole
fish sample labels. Results from each review are documented in a series of data verification
forms and compiled in a data review file that is submitted to the OST Project Manager after the
end of the final field sampling season. There will be separate fish sample collection data QC
files for the 2020 GLHHFFTS and the ORD-Duluth 2020 Great Lakes special study. Each fish
sample collection data QC file includes the study-specific collective Great Lakes human health
sample tracking spreadsheet for whole fish samples, as well as detailed results of the QC review
of the study-specific fish sampling data and sample description information that are recorded on
standard forms (e.g., the Data Review Form and the Sample Description Review Form). Any
discrepancies among the fish sampling records for each of the 2020 Great Lakes studies and
resolution of these discrepancies are reported to the OST Project Manager.
Lipid Data and Fish Sample Preparation QC Sample Data
The first stage in the data verification process involves the Tetra Tech Laboratory Manager
performing a completeness check in which all elements in each analytical laboratory submission
are evaluated to verify that results for all specified samples are provided, that data are reported in
the correct format, and that all relevant information, such as preparation and analysis logs, are
included in the data package. Corrective action procedures will be initiated if deficiencies are
noted. The Tetra Tech Lab Manager will transmit the analytical laboratory submission to the
OST Fish Sample Preparation Technical Leader, the OST Project Manager, and the CSRA/GDIT
data review chemist for independent review.
The second stage of the data verification process focuses on an instrument performance check in
which the CSRA/GDIT data review chemist verifies that calibrations, calibration verifications,
standards, and calibration blanks, as they apply to either lipid analysis or analysis of fish sample
preparation QC samples, were analyzed at the appropriate frequency and met method or study
performance specifications. If errors are noted at this stage, CSRA/GDIT will identify corrective
action procedures for Tetra Tech to initiate with the analytical laboratory immediately.
-------
NCCA 2020 Great Lakes Human Health Fish Sample Preparation QAPP
Revision 0
Date: July 15, 2020
Page 39 of 40
Stage three of the data verification process focuses on a laboratory performance check in which
the CSRA/GDIT data review chemist verifies that the laboratory correctly performed the
required analytical procedures and was able to demonstrate a high level of precision and
accuracy. This stage includes evaluation of QC elements, such as laboratory control samples and
blanks, as they apply to either analysis of lipid samples or fish sample preparation QC samples.
CSRA/GDIT will provide corrective action procedures for Tetra Tech to initiate with the
analytical laboratory to resolve any deficiencies identified.
D3. Reconciliation with User Requirements
Fish Sample Collection Data
As soon as possible following completion of fish sampling operations during the
2020 NCCA field season, the Tetra Tech Project Leader assesses fish sample collection data for
completeness, precision, and representativeness by comparing these data with the criteria
discussed for each of these measures in Section A7 of this QAPP. This represents the final
determination of whether the fish samples collected for the 2020 GLHHFFTS and the ORD-
Duluth 2020 Great Lakes special study are of the correct type, quantity, and quality to support
their intended use for this study. The Tetra Tech Project Leader will report any problems
encountered in meeting the performance criteria (or uncertainties and limitations in the use of the
data) to the OST Project Manager, and work with the OST Project Manager to reconcile the
problems, if possible.
Lipid Data and Fish Sample Preparation QC Sample Data
The QC results for lipids from the homogeneity testing and for the mercury and PCB rinsate
analyses from homogenization of fillet tissue samples for each fish sample preparation batch are
assessed by the CSRA/GDIT data review chemist against the QC acceptance criteria in Section
B5 of this QAPP. Although the Tetra Tech laboratory will be permitted to work two fish sample
preparation batches ahead of the delivery of the batch-specific QC results, the Tetra Tech Project
Leader will track laboratory performance, notify the OST Fish Sample Preparation Technical
Leader and the OST Project Manager of any issues, initiate corrective actions, and track progress
by the fish sample preparation laboratory.
References
USEPA. 1994. Method 245.1, Determination of Mercury in Water by Cold Vapor Atomic
Absorption Spectrometry, Revision 3.0. U.S Environmental Protection Agency, Office of
Research and Development, Environmental Monitoring Systems Laboratory, Cincinnati, OH.
USEPA. 1995. Good Automated Laboratory Practices. EPA Manual 2185. U.S. Environmental
Protection Agency, Office of Administration and Resources Management, Washington, DC,
August 1995.
-------
NCCA 2020 Great Lakes Human Health Fish Sample Preparation QAPP
Revision 0
Date: July 15, 2020
Page 40 of 40
USEPA. 2001. EPA Requirements for Quality Assurance Project Plans. EPA QA/R-5. U.S.
Environmental Protection Agency, Office of Environmental Information, Washington, DC.
EPA/240/B-01/003.
USEPA. 2020a. National Coastal Condition Assessment 2020 Quality Assurance Project Plan.
EPA-841-F-19-003. U.S. Environmental Protection Agency, Office of Water, Washington, DC.
USEPA. 2020b. National Coastal Condition Assessment 2020 Field Operations Manual. EPA-
841-F-19-005. U.S. Environmental Protection Agency, Office of Water, Washington, DC.
-------
Appendix A
Target Lists of NCCA 2020 Great Lakes Human
Health Whole Fish Sampling Locations
-------
2020 Great Lakes Human Health Fish Fillet Tissue Study
Nearshore Target Sampling Locations (226)1
LAKE
STATE
SITE ID
LATITUDE
LONGITUDE
Lake Erie
MI
NGL20 MI-10001
41.855488
-83.371811
Lake Erie
MI
NGL20 MI-10002
41.978389
-83.226068
Lake Erie
MI
NGL20 MI-10003
41.775408
-83.424598
Lake Erie
MI
NGL20 MI-10004
41.928531
-83.250994
Lake Erie
MI
NGL20 MI-10005
41.920233
-83.297047
Lake Erie
MI
NGL20 MI-10006
41.739698
-83.421714
Lake Erie
NY
NGL20 NY-10001
42.732124
-78.970972
Lake Erie
NY
NGL20 NY-10002
42.538288
-79.275335
Lake Erie
NY
NGL20 NY-10003
42.681456
-79.086131
Lake Erie
NY
NGL20 NY-10004
42.504435
-79.383478
Lake Erie
NY
NGL20 NY-10005
42.753035
-78.929682
Lake Erie
NY
NGL20 NY-10006
42.645265
-79.138888
Lake Erie
NY
NGL20 NY-10007
42.771058
-78.910310
Lake Erie
NY
NGL20 NY-10008
42.318041
-79.692515
Lake Erie
NY
NGL20 NY-10009
42.358800
-79.581425
Lake Erie
NY
NGL20 NY-10010
42.566027
-79.158291
Lake Erie
NY
NGL20 NY-10011
42.730634
-79.073758
Lake Erie
OH
NGL20 OH-10001
41.746250
-83.379173
Lake Erie
OH
NGL20 OH-10002
41.510475
-82.139115
Lake Erie
OH
NGL20 OH-10003
41.500632
-82.214543
Lake Erie
OH
NGL20 OH-10004
41.975683
-80.615216
Lake Erie
OH
NGL20 OH-10005
41.633941
-83.168247
Lake Erie
OH
NGL20 OH-10006
41.428897
-82.581778
Lake Erie
OH
NGL20 OH-10007
41.488651
-81.741749
Lake Erie
OH
NGL20 OH-10008
41.566691
-82.765201
Lake Erie
OH
NGL20 OH-10009
41.779146
-81.271315
Lake Erie
OH
NGL20 OH-10010
41.919821
-80.859975
Lake Erie
OH
NGL20 OH-10011
41.712038
-83.249070
Lake Erie
OH
NGL20 OH-10012
41.552818
-82.700842
Lake Erie
OH
NGL20 OH-10013
41.472286
-82.692721
Lake Erie
OH
NGL20 OH-10014
41.837711
-81.051557
Lake Erie
OH
NGL20 OH-10015
41.426038
-82.438730
Lake Erie
OH
NGL20 OH-10016
41.549394
-82.924082
Lake Erie
OH
NGL20 OH-10017
41.961265
-80.629612
Lake Erie
OH
NGL20 OH-10018
41.516712
-81.951277
Lake Erie
OH
NGL20 OH-10019
41.569624
-82.744472
Lake Erie
OH
NGL20 OH-10020
41.668449
-81.492888
Lake Erie
OH
NGL20 OH-10021
41.495360
-81.733241
Lake Erie
OH
NGL20 OH-10022
41.422845
-82.470374
Lake Erie
OH
NGL20 OH-10023
41.994433
-80.541792
Target Lists of NCCA 2020 Great Lakes Human Health Whole Fish Sampling Locations
A-l
-------
2020 Great Lakes Human Health Fish Fillet Tissue Study
Nearshore Target Sampling Locations (226)1
LAKE
STATE
SITE ID
LATITUDE
LONGITUDE
Lake Erie
OH
NGL20 OH-10024
41.693706
-83.281403
Lake Erie
OH
NGL20 OH-10025
41.773549
-81.262087
Lake Erie
OH
NGL20 OH-10026
41.629780
-81.589718
Lake Erie
PA
NGL20 PA-10001
42.216060
-79.908285
Lake Erie
PA
NGL20 PA-10002
42.248563
-79.833229
Lake Huron
MI
NGL20 MI-10020
43.661172
-83.813745
Lake Huron
MI
NGL20 MI-10021
43.246154
-82.464205
Lake Huron
MI
NGL20 MI-10022
45.750362
-84.563951
Lake Huron
MI
NGL20 MI-10023
44.839345
-83.242500
Lake Huron
MI
NGL20 MI-10024
43.691878
-83.607161
Lake Huron
MI
NGL20 MI-10025
45.963069
-84.714304
Lake Huron
MI
NGL20 MI-10026
45.378104
-83.647971
Lake Huron
MI
NGL20 MI-10027
44.005051
-83.227579
Lake Huron
MI
NGL20 MI-10028
45.939652
-84.669392
Lake Huron
MI
NGL20 MI-10029
45.006597
-83.359057
Lake Huron
MI
NGL20 MI-10030
43.879908
-83.436643
Lake Huron
MI
NGL20 MI-10031
44.013500
-82.767393
Lake Huron
MI
NGL20 MI-10032
45.960709
-84.419150
Lake Huron
MI
NGL20 MI-10033
45.186507
-83.333887
Lake Huron
MI
NGL20 MI-10034
44.262727
-83.475720
Lake Huron
MI
NGL20 MI-10035
45.365339
-83.558984
Lake Huron
MI
NGL20 MI-10036
43.933393
-83.375104
Lake Huron
MI
NGL20 MI-10037
44.058141
-82.890460
Lake Huron
MI
NGL20 MI-10038
44.975283
-83.442236
Lake Huron
MI
NGL20 MI-10039
43.987738
-83.643622
Lake Huron
MI
NGL20 MI-10040
43.762320
-82.615251
Lake Huron
MI
NGL20 MI-10041
45.700899
-84.357468
Lake Huron
MI
NGL20 MI-10042
44.380129
-83.305156
Lake Huron
MI
NGL20 MI-10043
43.998848
-82.682268
Lake Huron
MI
NGL20 MI-10044
45.637261
-84.224843
Lake Huron
MI
NGL20 MI-10045
44.220559
-83.436314
Lake Huron
MI
NGL20 MI-10046
44.560749
-83.269783
Lake Huron
MI
NGL20 MI-10047
43.325180
-82.522957
Lake Huron
MI
NGL20 MI-10048
45.931044
-84.370602
Lake Huron
MI
NGL20 MI-10049
43.747200
-83.482180
Lake Huron
MI
NGL20 MI-10050
43.069218
-82.418883
Lake Huron
MI
NGL20 MI-10051
45.510818
-84.053375
Lake Huron
MI
NGL20 MI-10052
44.995087
-83.402484
Lake Huron
MI
NGL20 MI-10053
43.763117
-83.450645
Lake Huron
MI
NGL20 MI-10054
43.866042
-83.384851
Target Lists of NCCA 2020 Great Lakes Human Health Whole Fish Sampling Locations
A-2
-------
2020 Great Lakes Human Health Fish Fillet Tissue Study
Nearshore Target Sampling Locations (226)1
LAKE
STATE
SITE ID
LATITUDE
LONGITUDE
Lake Huron
MI
NGL20 MI-10055
45.399225
-83.640746
Lake Huron
MI
NGL20 MI-10056
44.881911
-83.263937
Lake Huron
MI
NGL20 MI-10057
43.672811
-83.905286
Lake Huron
MI
NGL20 MI-10058
44.082561
-82.990724
Lake Huron
MI
NGL20 MI-10059
44.563966
-83.305080
Lake Huron
MI
NGL20 MI-10060
43.915173
-83.840608
Lake Huron
MI
NGL20 MI-10061
43.857134
-82.597289
Lake Huron
MI
NGL20 MI-10062
44.062607
-83.575569
Lake Huron
MI
NGL20 MI-10063
45.079141
-83.300411
Lake Huron
MI
NGL20 MI-10064
45.961854
-84.254088
Lake Michigan
IL
NGL20 IL-10001
42.141568
-87.745414
Lake Michigan
IN
NGL20 IN-10001
41.663612
-87.266719
Lake Michigan
IN
NGL20 IN-10002
41.626684
-87.268386
Lake Michigan
MI
NGL20 MI-10088
45.937953
-84.994048
Lake Michigan
MI
NGL20 MI-10089
45.769089
-86.742179
Lake Michigan
MI
NGL20 MI-10090
43.375394
-86.462772
Lake Michigan
MI
NGL20 MI-10091
45.000710
-85.477045
Lake Michigan
MI
NGL20 MI-10092
44.944033
-85.840715
Lake Michigan
MI
NGL20 MI-10093
44.396029
-86.308820
Lake Michigan
MI
NGL20 MI-10094
45.888090
-86.257438
Lake Michigan
MI
NGL20 MI-10095
43.918899
-86.457437
Lake Michigan
MI
NGL20 MI-10096
42.944195
-86.246774
Lake Michigan
MI
NGL20 MI-10097
45.797503
-84.792273
Lake Michigan
MI
NGL20 MI-10098
43.102397
-86.271767
Lake Michigan
MI
NGL20 MI-10099
45.934508
-85.719973
Lake Michigan
MI
NGL20 MI-10100
45.097762
-85.699194
Lake Michigan
MI
NGL20 MI-10101
44.312549
-86.299545
Lake Michigan
MI
NGL20 MI-10102
46.051344
-85.241341
Lake Michigan
MI
NGL20 MI-10103
44.678209
-86.261122
Lake Michigan
MI
NGL20 MI-10104
45.128687
-87.553117
Lake Michigan
MI
NGL20 MI-10105
45.772779
-86.809326
Lake Michigan
MI
NGL20 MI-10106
42.327680
-86.334775
Lake Michigan
MI
NGL20 MI-10107
46.023363
-85.195092
Lake Michigan
MI
NGL20 MI-10108
45.708931
-87.015603
Lake Michigan
MI
NGL20 MI-10109
45.107563
-85.596550
Lake Michigan
MI
NGL20 MI-10110
42.629003
-86.255722
Lake Michigan
MI
NGL20 MI-10111
45.667295
-86.518665
Lake Michigan
MI
NGL20 MI-10112
46.068962
-85.381571
Lake Michigan
MI
NGL20 MI-10113
45.590173
-87.221718
Lake Michigan
MI
NGL20 MI-10114
43.778261
-86.456345
Target Lists of NCCA 2020 Great Lakes Human Health Whole Fish Sampling Locations
A-3
-------
2020 Great Lakes Human Health Fish Fillet Tissue Study
Nearshore Target Sampling Locations (226)1
LAKE
STATE
SITE ID
LATITUDE
LONGITUDE
Lake Michigan
MI
NGL20 MI-10115
41.884623
-86.630807
Lake Michigan
WI
NGL20 WI-10001
45.029147
-87.093383
Lake Michigan
WI
NGL20 WI-10002
42.614701
-87.810577
Lake Michigan
WI
NGL20 WI-10003
43.328918
-87.864071
Lake Michigan
WI
NGL20 WI-10004
44.948027
-87.698607
Lake Michigan
WI
NGL20 WI-10005
45.336088
-86.956190
Lake Michigan
WI
NGL20 WI-10006
43.719069
-87.657049
Lake Michigan
WI
NGL20 WI-10007
43.331739
-87.865814
Lake Michigan
WI
NGL20 WI-10008
44.513063
-87.479199
Lake Michigan
WI
NGL20 WI-10009
44.863781
-87.481783
Lake Michigan
WI
NGL20 WI-10010
44.132949
-87.500974
Lake Michigan
WI
NGL20 WI-10011
45.188723
-86.979138
Lake Michigan
WI
NGL20 WI-10012
43.653379
-87.685584
Lake Michigan
WI
NGL20 WI-10013
42.798074
-87.717127
Lake Michigan
WI
NGL20 WI-10014
44.621561
-87.961059
Lake Ontario
NY
NGL20 NY-10032
43.968268
-76.115404
Lake Ontario
NY
NGL20 NY-10033
43.913595
-76.183412
Lake Ontario
NY
NGL20 NY-10034
43.358199
-78.702733
Lake Ontario
NY
NGL20 NY-1003 5
43.337974
-77.673215
Lake Ontario
NY
NGL20 NY-10036
43.506222
-76.487717
Lake Ontario
NY
NGL20 NY-10037
43.912188
-76.284124
Lake Ontario
NY
NGL20 NY-1003 8
43.311974
-78.888999
Lake Ontario
NY
NGL20 NY-10039
43.254800
-77.488727
Lake Ontario
NY
NGL20 NY-10040
43.587591
-76.250649
Lake Ontario
NY
NGL20 NY-10041
44.075882
-76.376997
Lake Ontario
NY
NGL20 NY-10042
43.381275
-78.085324
Lake Ontario
NY
NGL20 NY-10043
43.431453
-76.627178
Lake Ontario
NY
NGL20 NY-10044
43.803382
-76.251820
Lake Ontario
NY
NGL20 NY-10045
44.005321
-76.185961
Lake Ontario
NY
NGL20 NY-10046
43.361377
-77.930974
Lake Ontario
NY
NGL20 NY-10047
43.319131
-76.879006
Lake Ontario
NY
NGL20 NY-10048
43.334502
-78.808970
Lake Ontario
NY
NGL20 NY-10049
43.256623
-77.569462
Lake Ontario
NY
NGL20 NY-10050
43.470100
-76.579153
Lake Ontario
NY
NGL20 NY-10051
43.380115
-78.595466
Lake Ontario
NY
NGL20 NY-10052
43.294845
-77.351292
Lake Ontario
NY
NGL20 NY-10053
43.684236
-76.239633
Lake Ontario
NY
NGL20 NY-10054
43.313321
-78.918995
Lake Ontario
NY
NGL20 NY-1005 5
43.985167
-76.060517
Lake Ontario
NY
NGL20 NY-10056
43.822188
-76.317569
Target Lists of NCCA 2020 Great Lakes Human Health Whole Fish Sampling Locations
A-4
-------
2020 Great Lakes Human Health Fish Fillet Tissue Study
Nearshore Target Sampling Locations (226)1
LAKE
STATE
SITE ID
LATITUDE
LONGITUDE
Lake Ontario
NY
NGL20 NY-10057
43.340606
-77.703826
Lake Ontario
NY
NGL20 NY-1005 8
43.331946
-78.852610
Lake Ontario
NY
NGL20 NY-10059
43.684749
-76.220903
Lake Ontario
NY
NGL20 NY-10060
43.972557
-76.335932
Lake Ontario
NY
NGL20 NY-10061
43.288035
-77.553622
Lake Ontario
NY
NGL20 NY-10062
43.373254
-78.324593
Lake Ontario
NY
NGL20 NY-10063
43.546523
-76.314592
Lake Ontario
NY
NGL20 NY-10064
44.144023
-76.327305
Lake Ontario
NY
NGL20 NY-10065
43.516782
-76.433882
Lake Ontario
NY
NGL20 NY-10066
43.330161
-78.766097
Lake Ontario
NY
NGL20 NY-10067
43.913117
-76.242845
Lake Ontario
NY
NGL20 NY-10068
43.278913
-77.545576
Lake Ontario
NY
NGL20 NY-10069
43.332665
-76.863334
Lake Ontario
NY
NGL20 NY-10070
43.794633
-76.246004
Lake Ontario
NY
NGL20 NY-10071
43.986909
-76.206235
Lake Ontario
NY
NGL20 NY-10072
43.436884
-76.619956
Lake Ontario
NY
NGL20 NY-10073
43.387118
-77.986445
Lake Ontario
NY
NGL20 NY-10074
43.347117
-77.755232
Lake Ontario
NY
NGL20 NY-10075
44.059480
-76.369895
Lake Ontario
NY
NGL20 NY-10076
43.298707
-77.100180
Lake Superior
MI
NGL20 MI-10130
47.388639
-87.924763
Lake Superior
MI
NGL20 MI-10131
46.532907
-87.389569
Lake Superior
MI
NGL20 MI-10132
46.887188
-88.324722
Lake Superior
MI
NGL20 MI-10133
47.283798
-88.517410
Lake Superior
MI
NGL20 MI-10134
46.685295
-86.169696
Lake Superior
MI
NGL20 MI-10135
46.924509
-87.843784
Lake Superior
MI
NGL20 MI-10136
46.793415
-85.233590
Lake Superior
MI
NGL20 MI-10137
47.042887
-88.981274
Lake Superior
MI
NGL20 MI-10138
46.512006
-87.148603
Lake Superior
MI
NGL20 MI-10139
46.589141
-85.020580
Lake Superior
MI
NGL20 MI-10140
46.487506
-86.740911
Lake Superior
MI
NGL20 MI-10141
46.720289
-85.762836
Lake Superior
MI
NGL20 MI-10142
46.730769
-89.968199
Lake Superior
MI
NGL20 MI-10143
46.846226
-89.573089
Lake Superior
MI
NGL20 MI-10144
46.686942
-85.506656
Lake Superior
MI
NGL20 MI-10145
46.582070
-90.406321
Lake Superior
MI
NGL20 MI-10146
46.898957
-89.388481
Lake Superior
MI
NGL20 MI-10147
46.708512
-85.708076
Lake Superior
MI
NGL20 MI-10148
46.918146
-89.272481
Lake Superior
MI
NGL20 MI-10149
47.393787
-87.701592
Target Lists of NCCA 2020 Great Lakes Human Health Whole Fish Sampling Locations
A-5
-------
2020 Great Lakes Human Health Fish Fillet Tissue Study
Nearshore Target Sampling Locations (226)1
LAKE
STATE
SITE ID
LATITUDE
LONGITUDE
Lake Superior
MI
NGL20 MI-10150
47.296755
-88.548888
Lake Superior
MI
NGL20 MI-10151
46.508743
-87.146502
Lake Superior
MI
NGL20 MI-10152
46.604595
-90.378582
Lake Superior
MI
NGL20 MI-10153
46.839705
-88.262902
Lake Superior
MI
NGL20 MI-10154
46.485322
-84.958745
Lake Superior
MI
NGL20 MI-10155
46.689560
-86.114568
Lake Superior
MI
NGL20 MI-10156
46.967084
-89.234385
Lake Superior
MI
NGL20 MI-10157
46.876538
-87.730033
Lake Superior
MI
NGL20 MI-10158
46.790886
-85.026494
Lake Superior
MI
NGL20 MI-10159
47.259431
-88.643379
Lake Superior
MI
NGL20 MI-10160
46.494254
-86.608618
Lake Superior
MN
NGL20 MN-10001
47.141140
-91.450357
Lake Superior
MN
NGL20 MN-10002
47.556276
-90.867735
Lake Superior
MN
NGL20 MN-10003
46.790489
-92.044779
Lake Superior
MN
NGL20 MN-10004
47.771646
-90.180874
Lake Superior
MN
NGL20 MN-10005
47.480891
-90.983105
Lake Superior
MN
NGL20 MN-10006
47.973099
-89.635062
Lake Superior
MN
NGL20 MN-10007
46.793050
-91.995742
Lake Superior
MN
NGL20 MN-10008
47.062637
-91.592607
Lake Superior
MN
NGL20 MN-10009
47.727800
-90.426800
Lake Superior
WI
NGL20 WI-10022
46.770510
-91.622243
Lake Superior
WI
NGL20 WI-10023
46.672803
-90.816963
Lake Superior
WI
NGL20 WI-10024
46.729253
-91.787980
Lake Superior
WI
NGL20 WI-10025
46.680720
-90.632506
Lake Superior
WI
NGL20 WI-10026
46.754410
-91.731229
Lake Superior
WI
NGL20 WI-10027
46.666176
-90.692698
1 This list of sites is subject to change as the project proceeds. For example, access to some sites may not be
granted by property owners. Other sites may not yield fish of suitable size or species. OST maintains the list of
valid sites, and this QAPP will not be revised just to address changes in the list of sites.
Target Lists of NCCA 2020 Great Lakes Human Health Whole Fish Sampling Locations
A-6
-------
ORD-Duluth 2020 Great Lakes Special Study
Lake Michigan Enhancement Target Sampling Locations (50)1
LAKE
STATE
SITE ID
LATITUDE
LONGITUDE
Lake Michigan
MI
ISA20-01
45.022576
-85.955361
Lake Michigan
MI
ISA20-02
44.992944
-86.143602
Lake Michigan
MI
ISA20-03
45.810411
-85.536973
Lake Michigan
MI
ISA20-04
45.732114
-85.586710
Lake Michigan
MI
ISA20-05
45.100510
-86.075701
Lake Michigan
MI
ISA20-06
45.754047
-85.396307
Lake Michigan
MI
ISA20-07
45.744411
-85.518414
Lake Michigan
MI
ISA20-08
45.406036
-85.865609
Lake Michigan
MI
ISA20-09
45.373191
-86.950673
Lake Michigan
MI
ISA20-10
45.806034
-85.336755
Lake Michigan
MI
ISA20-11
45.690690
-85.460965
Lake Michigan
MI
ISA20-12
45.457843
-85.912753
Lake Michigan
MI
ISA20-13
45.386882
-86.829426
Lake Michigan
MI
ISA20-14
45.821972
-85.334852
Lake Michigan
MI
ISA20-15
45.627184
-85.633541
Lake Michigan
MI
ISA20-16
45.161108
-87.409663
Lake Michigan
MI
ISA20-17
44.994629
-86.163159
Lake Michigan
MI
ISA20-18
45.740626
-85.583030
Lake Michigan
MI
ISA20-19
45.071318
-86.101963
Lake Michigan
MI
ISA20-20
45.733368
-85.438622
Lake Michigan
MI
ISA20-21
45.780002
-85.549848
Lake Michigan
MI
ISA20-22
45.367833
-85.835910
Lake Michigan
MI
ISA20-23
45.418739
-86.835769
Lake Michigan
MI
ISA20-24
45.803657
-85.395611
Lake Michigan
MI
ISA20-25
45.600707
-85.478461
Lake Michigan
MI
ISA20-26
45.243421
-87.297889
Lake Michigan
MI
ISA20-27
45.333378
-86.809511
Lake Michigan
MI
ISA20-28
45.798403
-85.577589
Lake Michigan
MI
ISA20-29
45.748655
-85.684087
Lake Michigan
MI
ISA20-30
45.164952
-87.285285
Lake Michigan
MI
ISA20-31
45.032575
-86.005719
Lake Michigan
MI
ISA20-32
45.758626
-85.313298
Lake Michigan
MI
ISA20-33
45.791583
-85.527481
Lake Michigan
MI
ISA20-34
45.577667
-85.641267
Lake Michigan
MI
ISA20-35
45.037425
-85.997924
Lake Michigan
MI
ISA20-36
45.742621
-85.459929
Lake Michigan
MI
ISA20-37
45.321337
-86.892611
Lake Michigan
MI
ISA20-38
45.829346
-85.393017
Target Lists of NCCA 2020 Great Lakes Human Health Whole Fish Sampling Locations
A-7
-------
ORD-Duluth 2020 Great Lakes Special Study
Lake Michigan Enhancement Target Sampling Locations (50)1
LAKE
STATE
SITE ID
LATITUDE
LONGITUDE
Lake Michigan
MI
NPA20-01
44.914807
-86.099659
Lake Michigan
MI
NPA20-02
44.799368
-86.090591
Lake Michigan
IN
NPA20-03
41.646178
-87.217218
Lake Michigan
IN
NPA20-04
41.699221
-87.039672
Lake Michigan
MI
NPA20-05
44.859089
-86.088796
Lake Michigan
MI
NPA20-06
44.749306
-86.096894
Lake Michigan
IN
NPA20-07
41.632908
-87.216226
Lake Michigan
IN
NPA20-08
41.699639
-87.078886
Lake Michigan
MI
NPA20-09
44.772349
-86.171933
Lake Michigan
MI
NPA20-10
44.971318
-85.896002
Lake Michigan
IN
NPA20-11
41.635451
-87.266269
Lake Michigan
IN
NPA20-12
41.699258
-87.087526
1 This list of sites is subject to change as the project proceeds. For example, access to some sites may not be
granted by property owners. Other sites may not yield fish of suitable size or species. OST maintains the list of
valid sites, and this QAPP will not be revised just to address changes in the list of sites.
Target Lists of NCCA 2020 Great Lakes Human Health Whole Fish Sampling Locations
A-8
-------
Appendix B
NCCA 2020 Great Lakes Human Health
Fish Fillet Sample Preparation, Homogenization, and
Distribution Procedures
-------
Appendix B
NCCA 2020 Great Lakes Human Health Fish Fillet Sample Preparation,
Homogenization, and Distribution Procedures
I. PURPOSE
This document describes the procedures that the fish sample preparation laboratory (Tetra Tech
laboratory at Owings Mills, MD) follows when preparing fish fillet tissue samples for EPA's National
Coastal Condition Assessment (NCCA) 2020 Great Lakes Human Health Fish Fillet Tissue Study
(GLHHFFTS) and the ORD-Duluth 2020 Great Lakes special study. Adherence to these procedures
ensures that fillet tissue sample preparation activities at the Tetra Tech laboratory are performed
consistently across all study samples and in a manner consistent with previous EPA fish tissue studies.
The effort is divided into two primary components:
Fish fillet tissue sample preparation and distribution procedures, including quality control steps (e.g.,
triplicate lipid analysis of homogenized fillet tissue aliquots), for all fish sample preparation batches.
Preparation and analyses of rinsate and solvent blank samples for mercury, PCBs, and PFAS for each
fish sample preparation batch.
Each of these components is described in detail below.
II. FISH FILLET TISSUE PROCESSING AND DISTRIBUTION PROCEDURES
The procedures for processing 2020 GLHHFFTS whole fish samples and distributing fillet tissue samples
for mercury, PFAS, PCB, lipid, and fatty acid analyses are described below. This process description is
organized into the following components, including the quality control (QC) procedures:
A. Sample Receipt and Storage
B. Sample Handling
C. Filleting and Homogenization Procedures
D. Aliquoting and Distribution Procedures
E. Equipment Cleaning between Fish Samples
F. Lipid Determination for Every Homogenized Fillet Sample
G. Quality Control (QC) Procedures
H. Reporting Requirements
I. Sample Shipping Procedures
The individual tasks in the overall process are presented as a series of numbered steps across the nine
components listed above.
Fillet Tissue Processing Definitions
Whole Fish Composite Sample: A whole fish composite sample for the 2020 GLHHFFTS and
the ORD-Duluth 2020 Great Lakes special study consists of 5 fish (ideally) of the same species
that are similar in size and typically consumed by humans (See Section B1). One human health
whole fish composite sample is collected from each viable Great Lakes nearshore site and Lake
Michigan enhancement site whole fish sampling location. There are 226 Great Lakes nearshore
sites that are designated as whole fish sampling locations for the 2020 GLHHFFTS, as well as 50
enhancement sites in Lake Michigan that are designated as whole fish sampling locations for the
ORD-Duluth 2020 Great Lakes special study.
NCCA 2020 Great Lakes Human Health Fish Fillet Tissue Sample Preparation Procedures
B-l
-------
Fish sample preparation batch: Each fish sample preparation batch consists of 20 whole fish
composite samples. The number of whole fish composite samples in the final fish sample
preparation batch (or two) may be adjusted to include a few more than 20 or fewer than 20,
depending on what fraction of 20 whole fish composite samples remain for assignment to a batch.
Note that Tetra Tech will assign separate fish sample preparation batches for the 2020
GLHHFFTS and the ORD-Duluth 2020 Great Lakes special study.
Analytical batch: An analytical batch consists of the 20 fillet tissue aliquots generated for each
target chemical (i.e., mercury, PFAS, PCB congeners, PCBs as Aroclors, lipids and fatty acids)
during processing of a fish sample preparation batch. The number of fillet tissue samples in the
final analytical batch (or two) may be adjusted to include a few more than 20 or fewer than 20,
depending on what fraction of 20 fillet sample aliquots remain for assignment to a batch. Note
that analytical batches correspond to fish sample preparation batches, so there will be separate
analytical batches for the 2020 GLHHFFTS and the ORD-Duluth 2020 Great Lakes special study.
II.A. Sample Receipt and Storage
Field crews are collecting NCCA 2020 Great Lakes human health fish samples from June through
September (or possibly through October or early November) during 2020. A total of 226 Great Lakes
nearshore sites and 50 enhancement sites in Lake Michigan are designated as human health whole fish
sampling locations (Appendix A).
Whole fish samples are shipped by priority overnight delivery service from field locations in the Great
Lakes to the sample repository at Microbac Laboratories in Baltimore, MD, where they are held in
freezers for interim storage at a temperature of less than or equal to -20 °C. All samples are subsequently
hand-delivered to the Tetra Tech laboratory in Owings Mills, MD for preparation of fillet tissue samples
and interim storage of fillet samples until sample shipment to designated analytical laboratories. The
Tetra Tech laboratory must have sufficient freezer space to store at least 3 batches of unprocessed fish
samples at a temperature of less than or equal to -20 °C from the time of receipt until completion of
sample processing and sufficient freezer space to store homogenized fillet tissue aliquots from up to 60
processed samples (e.g., up to 600 homogenized tissue jars) prior to distribution.
1. Although whole fish samples are delivered frozen, on dry ice, they must be inspected promptly on
receipt. As samples are received at Microbac Laboratories, a laboratory representative must:
Check that each shipping container has arrived undamaged and verify that samples are still frozen
and in good condition.
Check the temperature of one of the samples in the cooler using a thermometer that reads to at
least -20 °C, or an infra-red (IR) temperature "gun," and record the reading.
Transfer the samples to the freezer for long-term storage.
2. Notify CSRA/GDIT immediately about any problems encountered upon receipt of samples.
CSRA/GDIT will communicate these problems to the OST Project Manager for resolution.
Section B3 of the QAPP contains details about sample inspection by the CSRA/GDIT sample
custodian before whole fish samples are transferred to the Tetra Tech laboratory for fillet tissue
sample preparation. Following fillet sample processing, the Tetra Tech laboratory must store
homogenized fillet tissue sample aliquots frozen to less than or equal to -20 °C until they are
distributed to the laboratories designated for fillet tissue analysis.
NCCA 2020 Great Lakes Human Health Fish Fillet Tissue Sample Preparation Procedures
B-2
-------
II.B. Sample Handling
The whole fish samples collected for the 2020 GLHHFFTS and the ORD-Duluth 2020 Great Lakes
special study must remain frozen at less than or equal to -20 °C until the Tetra Tech laboratory receives
direction from the OST Fish Sample Preparation Technical Leader to begin fish sample preparation. Fish
samples must be retrieved from the freezer, with their associated paperwork, and allowed to partially thaw
before they can be processed to prepare fillet tissue samples.
3. Prior to beginning fish sample processing, Tetra Tech prepares an Excel spreadsheet with draft fish
sample processing instructions and preparation batch assignments for each of the 2020 Great Lakes
studies and submits these spreadsheets to the OST Fish Sample Preparation Technical Leader and
OST Project Manager for review. The OST Fish Sample Preparation Technical Leader will approve
the fish sample preparation batch assignments and fish sample processing instructions with OST
Project Manager concurrence.
Processing for a fish sample preparation batch involves the following for each fish sample in the batch:
Preparation of one homogenized fillet tissue sample (consisting of fillets from both sides of each
fish in the sample) according to the sample processing instructions approved by the OST Fish
Sample Preparation Technical Leader with OST Project Manager concurrence.
Note: Processing a fish sample preparation batch produces a total of 20 homogenized fillet tissue
samples that are subdivided into aliquots for target chemical analyses and for long-term
storage of archived tissue (Section II.D). Each set of 20 target chemical aliquots (mercury,
PFAS, PCB congeners, PCBs as Aroclors, lipids, and fatty acids) constitutes an analysis batch
for the fish fillet indicator.
4. When retrieving samples from the freezer, the Tetra Tech sample custodian must:
Verify that all associated paperwork stored with the samples is complete, legible, and accurate.
Compare the information on the label on each fish specimen to the fish sample preparation batch
spreadsheet and notify the OST Fish Sample Preparation Technical Leader and the OST Project
Manager of any discrepancies between the sample labels and the Excel file of sample processing
instructions. Problems involving sample paperwork, sample integrity, or custody information
inconsistencies for all fish samples should be reported to the OST Fish Sample Preparation
Technical Leader and the OST Project Manager in writing (e.g., by email) within one business
day following sample retrieval and inspection. Do not proceed with sample processing until
discrepancies are resolved.
II.C. Filleting and Homogenization Procedures
The target chemical analyses for mercury, PFAS, PCB congeners, PCBs as Aroclors, lipids and fatty
acids are performed on aliquots of homogenized fillet tissue samples prepared from the NCCA 2020
human health whole fish samples. Steps 5-9 below must be completed before beginning processing and
preparing any fillet samples in the laboratory.
5. Prior to preparing any fillet samples, thoroughly clean utensils and cutting boards using the following
series of procedures:
Wash with a detergent solution (phosphate- and scent-free) and warm tap water
Rinse three times with warm tap water
Rinse three times with deionized (DI) water
Rinse with acetone
NCCA 2020 Great Lakes Human Health Fish Fillet Tissue Sample Preparation Procedures
B-3
-------
Rinse three times with DI water
Rinse with (not soak in) 5% nitric acid
Rinse three times with DI water
To control contamination, separate sets of utensils and cutting boards must be used for scaling
fish and for filleting fish.
6. Put on powder-free nitrile gloves before unpacking a whole fish sample for fillet removal and tissue
homogenization. After unwrapping, inspect each fish specimen in the sample carefully to verify that
it has not been damaged during collection or shipment. If damage (e.g., tearing the skin or puncturing
the gut) is observed, document it in the applicable Fish Sample Preparation Laboratory Bench Sheet
(Appendix C for 2020 GLHHFFTS nearshore sites and Appendix D for ORD-Duluth 2020 Great
Lakes special study Lake Michigan enhancement sites) and notify the OST Fish Sample Preparation
Technical Leader and the OST Project Manager before proceeding further.
7. Weigh each fish to the nearest gram (wet weight) prior to any sample processing. Enter weight
information for each specimen into the applicable Fish Sample Preparation Laboratory Bench Sheet
(Appendix C for nearshore sites and Appendix D for enhancement sites). Individual specimen
weights will be transferred to spreadsheets for submission to the OST Fish Sample Preparation
Technical Leader and the OST Project Manager.
8. Rinse each fish in the sample with DI water as a precautionary measure to treat for possible
contamination from sample handling in the field. Use HDPE wash bottles (not PTFE) for rinsing fish
and for cleaning homogenization equipment and utensils.
9. Before beginning the scaling process for each fish in the composite sample, put on new powder-free
nitrile gloves. (Gloves must be changed between whole fish composite samples.) Fish with scales
must be scaled (and any adhering slime should be removed) prior to filleting. Scale the first
designated fish by laying it flat on a clean glass cutting board and scraping from the tail to the head
using a stainless steel scaler or the blade-edge of a clean stainless steel knife.
10. Continue scaling all of the other fish in the whole fish sample as described in Step 9 above. Filleting
of the fish in the sample can proceed after all scales have been removed from the skin and a separate
clean cutting board and fillet knife are prepared or available.
11. Put on new powder-free nitrile gloves. Place each fish on a clean glass cutting board in preparation
for the filleting process. Note that filleting should be conducted under the supervision of an
experienced fisheries biologist. Ideally, fish should be filleted while ice crystals are still present in
the muscle tissue. Fish should be thawed only to the point where it becomes possible to make an
incision into the flesh. Remove both fillets (lateral muscle tissue with skin attached) from the fish
specimen using clean, high-quality stainless steel knives. Include the belly flap (ventral muscle and
skin) with each fillet. Care must be taken to avoid contaminating fillet tissues with material released
from inadvertent puncture of internal organs. In the event that an internal organ is punctured, rinse
the fillet with deionized water immediately after filleting and make a note on the laboratory bench
sheet that a puncture has occurred. Bones still present in the tissue after filleting should be carefully
removed using the tip of the fillet knife or a clean pair of forceps.
12. Whole fillet samples (consisting of the entire right and left fillets) are weighed to the nearest gram
(wet weight) and the weight is recorded on the bench sheet prior to homogenization. These samples
should be homogenized partially frozen for ease of grinding.
13. Process each whole fillet sample using a size-appropriate homogenization apparatus (e.g., automatic
grinder or high-speed blender). Entire fillets (with skin and belly flap) from both sides of the fish
must be homogenized. Mix the tissues thoroughly until they are completely homogenized as
NCCA 2020 Great Lakes Human Health Fish Fillet Tissue Sample Preparation Procedures
B-4
-------
evidenced by fillet tissue that consists of a uniform color and finely ground texture. Chunks of skin
or tissue will hinder extraction and digestion and, therefore, are NOT acceptable. Grinding of tissue
may be easier when tissues are partially frozen. Chilling the grinder briefly with a few small pieces
or pellets of dry ice may also keep the tissue from sticking to the equipment. Pellets of dry ice also
may be added to the tissue as it enters the grinder.
Note: The dry ice pellets used for homogenizing the fillet tissue are classified as food grade and meet
the specifications for substances Generally Regarded As Safe as a direct food ingredient in the
Food and Drug Administration regulation FDA 184.1240 (21 CFR 184.1240).
14. Grind the entire fillet sample a second time, using the same grinding equipment. This second
grinding should proceed more quickly. The grinding equipment does not need to be cleaned between
the first and second grinding of the sample. The final homogenized fillet sample must consist of
finely ground tissue of uniform color and texture. If there are obvious differences in color or texture,
grind the entire sample a third time or more to ensure uniform homogenization.
15. Measure the collective weight of the homogenized fillet tissue from each fish sample to the nearest
gram (wet weight) after processing and record the total homogenate weight on the laboratory bench
sheet. The total weights of the fillets and weights of the homogenized fillet tissue from each fish
sample are transferred to spreadsheets for submission to the OST Fish Sample Preparation Technical
Leader and the OST Project Manager. At least 465 g of homogenized tissue will be needed to fill all
of the containers in Table 1 below with their minimum acceptable masses. If a sample does not
yield at least 465 g of homogenized tissue, contact the OST Project Manager via email
immediately and await instructions. As appropriate, place any remaining homogenized fillet tissue
in the freezer while waiting for instructions, which are likely to involve preparing fewer archive
aliquots.
16. After the final (second, third, or higher number) grinding, clean the grinding equipment and all
other sample preparation equipment using the procedures described in Step 22.
17. Once in every fish sample preparation batch (generally containing 20 whole fish samples), verify the
continued absence of equipment contamination and uniformity of homogenization using the
procedures described in Steps 25 to 29.
II.D. Aliquoting and Distribution Procedures
18. The sample preparation laboratory prepares the bulk homogenate tissue from one whole fish sample
and uses it to fill the pre-cleaned sample containers specified for each type of aliquot listed in Table 1,
following the procedures described in Step 19. Except as noted in Table 1, all containers are
provided by the fish sample preparation laboratory. Documentation of their cleanliness provided
by the vendor (i.e., certificates of analysis) must be retained by the fish sample preparation laboratory
and provided to EPA on request. The target masses listed in Table 1 are designed to provide enough
tissue for multiple analyses of each sample, including tissue for QC purposes, as needed. The fish
sample preparation laboratory should not exceed those aliquot target masses when filling the
containers. The order of the containers and target masses in Table 1 are important (i.e., they indicate
priority order for aliquots) and are designed to ensure that adequate tissue is available for all analyses,
as well as for archiving.
NCCA 2020 Great Lakes Human Health Fish Fillet Tissue Sample Preparation Procedures
B-5
-------
Table 1. NCCA 2020 Great Lakes Human Health Fish Fillet Tissue Sample Aliquot Requirements
Analysis
Target Mass
Container Type
Destination
Mercury
5-10g
50-mL HDPE straight-sided jar, or conical HDPE tube
with snap top
TBD
PFAS
10 - 15 g
100-mL HDPE straight-sided jar with foil-lined lid, or
conical HDPE tube with snap top. PTFE lid liners not
allowed.
TBD
PCB congeners
20 - 25 g
125-mL straight-sided amber or clear glass jar with
PTFE-lined lid
TBD
Lipids, Fish 1
30-35 g (prepare 3
aliquots
containing at least
10 g each)
TBD laboratory's choice
TBD
Lipids, Fish 2-20
10 - 15 g
TBD laboratory's choice
TBD
PCBs as Aroclors
20 - 25 g
125-mL straight-sided amber or clear glass jar with
PTFE-lined lid
TBD
Fatty acids
10 - 15 g
125-mL straight-sided amber or clear glass jar with
PTFE-lined lid
TBD
Small Archive 1
50 - 60 g
125-mL straight-sided amber or clear glass jar with foil-
lined lid
CSRA/GDIT
Sample
Repository
Small Archive 2
50 - 60 g
125-mL straight-sided amber or clear glass jar with foil-
lined lid
CSRA/GDIT
Sample
Repository
Bulk Archive 1
240 - 250 g
500-mL straight-sided amber or clear glass jar with foil-
lined lid
CSRA/GDIT
Sample
Repository
Bulk Archive 2
All remaining
mass up to 250 g
500-mL straight-sided amber or clear glass jar with foil-
lined lid
CSRA/GDIT
Sample
Repository
Total (to the
nearest gram)3
465 - 740 g
Assumes at least 50 g of tissue is available for Bulk
Archive 2
a In the event that insufficient fish tissue mass exists to prepare the required number of aliquots, contact the OST
Project Manager for instructions as per Step 15.
19. Prepare the homogenized sample aliquots for mercury, PFAS, PCB congeners, lipids, PCBs as
Aroclors, and fatty acids (see Step 21 for lipid aliquot preparation). Weigh an appropriate clean
sample container (Table 1) to the nearest 0.5 g and record the weight. Transfer sufficient
homogenized fillet tissue to the container to achieve the target mass for that container in Table 1,
weigh the container again, record the weight, and determine the weight of the aliquot to the nearest
0.5 g by difference. The fish sample preparation laboratory must use foil-lined lids for jars
containing the fillet tissue aliquots for PFAS analysis and the archived fillet tissue samples, as
specified in Table 1.
Note: The archive sample jars are not filled until after sufficient volume for lipids determination,
PCBs as Aroclors, and fatty acids have been collected, as described in Step 21. The archive
jars are not filled until the triplicate lipid aliquot (30-35 g), the PCBs as Aroclors aliquot (20-
25 g), and the fatty acids aliquots (10-15 g) are collected (see Step 28 for triplicate lipid
aliquot preparation, which is used for homogeneity testing).
When filling jars, leave sufficient space at the top of each jar before sealing with the designated lid to
allow for expansion of the tissue as it freezes. In no case should jars be filled beyond 80% capacity,
NCCA 2020 Great Lakes Human Health Fish Fillet Tissue Sample Preparation Procedures
B-6
-------
as this may result in breakage on freezing. Wipe off the outside of the jars to remove any tissue
residue or moisture. Fill out a label for each container using a waterproof marker. Include the
following information (at a minimum) on each label:
sample identification number,
tissue sample type (i.e., homogenized fillet),
analysis type (e.g., mercury, PFAS, PCB congeners, lipids, PCBs as Aroclors, or fatty acids),
aliquot weight (to the nearest 0.5 gram),
preparation batch ID, and
preparation date (e.g., mm/dd/yyyy).
Affix the label to the container with clear wide tape. Place each container inside one heavy-weight
food-grade self-sealing plastic freezer bag to avoid sample loss due to breakage. Freeze the tissue
samples at -20 °C and maintain samples in the freezer until directed by the OST Fish Sample
Preparation Technical Leader with OST Project Manager concurrence to ship them to the analytical
laboratories. (The OST Fish Sample Preparation Technical Leader will not issue these instructions
until equipment rinsate and homogeneity tests described in Steps 24 to 29 have been completed,
reported, evaluated, and determined to be acceptable.)
20. After filling the containers with the tissue aliquots for mercury, PFAS, and PCB congeners, remove
30 to 35 g of homogenized fillet tissue from one sample in the batch (for triplicate lipid analysis) and
10 to 15 g of homogenized fillet tissue from all other samples in the batch (for single lipid analysis) to
be used to determine the lipid content of each fillet composite sample. Place these aliquots in clean
glass or plastic containers of suitable size (provided by the TBD analytical laboratory) and label each
of them with the sample ID number. Store the lipid aliquots in the freezer at -20 °C until they are
ready to be shipped to the designated analytical laboratory to perform the lipid determinations in
Steps 24, 28, and 29. After preparing the lipid aliquots, follow the procedures in Step 19 to prepare
the PCBs as Aroclors aliquot and the fatty acids aliquot.
21. The archive sample jars are not filled until after sufficient volume for determining lipids, PCBs as
Aroclors, and fatty acids have been collected. Once the aliquots for mercury, PFAS, PCB congeners,
lipids, PCBs as Aroclors, and fatty acids have been collected, the remaining tissue mass is used to
create the four archive samples. Begin by transferring 50 - 60 g of tissue to the first small archive
sample container. Continue by transferring a 50 - 60 g aliquot to the remaining small archive
container. Ideally, sufficient homogenized fillet tissue mass will remain to produce two bulk archive
containers. Therefore, transfer 240 - 250 g of tissue to the first bulk archive sample container.
Continue by transferring up to 250 g of tissue to the second bulk archive container. However, if less
than 250 g of tissue is available, transfer all of the remaining homogenized tissue to the bulk archive
container and weigh it to determine the tissue mass in the last archive container. Seal and label the
containers as described in Step 19 for the other aliquots.
Note: Step 15 requires that the laboratory contact the OST Project Manager whenever a
homogenized fillet sample does not yield at least 465 g of tissue. The OST Fish Sample
Preparation Technical Leader will provide direction to the laboratory with OST Project
Manager concurrence regarding samples yielding less than 465 g of tissue that must be
followed at this point in the procedure.
Any fillet tissue that remains after filling the second bulk archive jar may be discarded.
II.E. Equipment Cleaning between Composite Samples
22. All of the homogenization equipment must be thoroughly cleaned between each individual fish
sample. Once both of the fillets from the individual sample have been homogenized, disassemble the
NCCA 2020 Great Lakes Human Health Fish Fillet Tissue Sample Preparation Procedures
B-7
-------
homogenization equipment (i.e., blender, grinder, or other device) and thoroughly clean all surfaces
and parts that contact the sample. Similarly, clean all knives, cutting boards, and other utensils
used. At a minimum:
Wash with a detergent solution (phosphate- and scent-free) and warm tap water
Rinse three times with warm tap water
Rinse three times with deionized (DI) water
Rinse with acetone
Rinse three times with DI water
Rinse with (not soak in) 5% nitric acid
Rinse three times with DI water
Allow the components to air dry
23. Reassemble the homogenization equipment and proceed with homogenization of the next fish sample
in the batch (e.g., begin with Step 6 above).
II.F. Lipid Determination for Every Homogenized Fillet Sample
The first fish sample in each preparation batch is designated for triplicate lipid analysis for the
homogeneity testing process described in Steps 28 and 29. The procedures for determining the lipid
content of homogenized fillet tissue from all other fish samples in a fish sample preparation batch are
described in Step 24 below.
24. For samples 2 through 20 in each fish sample preparation batch, use the 10 to 15 g aliquot of
homogenized tissue collected in Step 20 to determine the lipid content of the sample. The analytical
laboratory (TBD) will extract the aliquot using an appropriate method (TBD) approved by EPA to
determine the lipid content of that aliquot, which is recorded in units of percent (i.e., grams of lipid
per gram of tissue x 100). This QAPP will be amended to include a description of this method after
the analytical laboratory is selected and their proposed method for lipid analysis is approved.
II.G. Quality Control (QC) Procedures
The QC procedures for fish sample preparation include preparation and testing of equipment rinsate
samples and homogeneity testing, using lipids as a surrogate.
During the fish sample preparation process, the Tetra Tech laboratory prepares three sets of aqueous
rinsate and solvent blank samples for mercury, PCB congener, and PFAS analyses, respectively,
(Attachment 1) and one homogenized fillet tissue aliquot for triplicate lipid determinations from each fish
sample preparation batch, as described in Steps 25 to 28 below. The batch-specific rinsate and
homogeneity results are reviewed by Tetra Tech and CSRA/GDIT. The Tetra Tech laboratory doing fish
sample preparation may continue to process up to 2 additional batches during the QC sample analysis and
review process. However, the Tetra Tech laboratory may not continue beyond the third batch of fish
samples until receiving notification from the OST Fish Sample Preparation Leader (with OST Project
Manager concurrence) that the review of initial batch rinsate and homogeneity test results is complete,
and the results were deemed satisfactory.
Continued sample processing is dependent on both the quality of the Tetra Tech laboratory's efforts and
on the timeliness of their delivery of QC results.
NCCA 2020 Great Lakes Human Health Fish Fillet Tissue Sample Preparation Procedures
B-8
-------
Rinsate and Blank Sample Production
25. Once per batch (of usually 20 fish samples) during the fish sample preparation operations, prepare
three sets of rinsate and blank solvent samples (see Attachment 1) prior to reassembling the
homogenization equipment (Step 23), as follows:
PCB rinsate and blank samples:
Prepare a hexane rinsate sample by pouring a 100-mL portion of pesticide-grade hexane over all
parts of homogenization equipment, including the cutting boards and knives, and collect it in a
clean glass container. Place an additional 100-mL aliquot of clean hexane in a similar glass
container for use as a solvent blank. Allow the solvent to evaporate from the equipment. This set
of rinsate and solvent blank samples will be analyzed for selected PCB congeners (see
Attachment 1). Label and store the PCB rinsate and blank samples as described Step 26.
Mercury rinsate and blank samples:
Once the hexane has evaporated from the equipment, prepare the first DI water rinsate using
250 mL of DI water. Collect the DI water rinsate in a clean glass or HDPE container. Place a
second aliquot of DI water in a separate similar clean container for use as a blank. Acidify these
two samples to pH < 2 with nitric acid. Label and store each sample as described in Step 26.
These rinsate and blank samples will be analyzed for mercury (see Attachment 1).
PFAS rinsate and blank samples:
Prepare the second DI water rinsate using an additional 250 mL of DI water. Collect this rinsate
in a clean glass container with a non-PTFE lid liner. Place a second aliquot of DI water in a
separate similar clean glass container for use as a blank. This set of rinsate and blank samples
will be analyzed by the laboratory selected at a later date to analyze fillet samples for PFAS, thus
the non-PTFE lid liners are essential. The OST Project Manager will provide the Tetra Tech
laboratory with the PFAS laboratory name and shipping information as soon as it is available.
Label and store these PFAS rinsate and blank samples as described in Step 26.
Note: In order to minimize the number of project samples that might be affected by cross contamination,
collect the rinsate and blank samples on the first day that fish samples in a sample preparation
batch of 20 are processed. Ideally, the laboratory will vary the point at which the rinsates are
collected on that first day over the course of the project (e.g., between the 1st and 2nd samples for
one batch, the 2nd and 3rd samples for another batch, etc.).
26. Label each container as either "rinsate -[insert the name of the solvent, either hexane or DI water]" or
"blank -[insert the name of the solvent, either hexane or DI water]," and include the date it was
prepared (mm/dd/yyyy), the analysis type (Hg, PCBs, or PFAS), and the preparation batch identifier.
Store the rinsate and blank samples in a refrigerator at a temperature of <6 °C.
Rinsate and Blank Sample Analyses
27. During the fish sample preparation operations, laboratories under contract to Tetra Tech (to be
determined for mercury and PCBs) and CSRA/GDIT (PFAS) will analyze one set of rinsate and blank
samples per batch for:
mercury using EPA Method 245.1, a cold-vapor atomic absorption procedure (Details for this
method are described in Attachment 1),
PCBs using an appropriate method proposed by the TBD laboratory and approved by EPA (Note
that this QAPP will be amended to include a description of this method after the analytical
laboratory is selected and their proposed method is approved), and
NCCA 2020 Great Lakes Human Health Fish Fillet Tissue Sample Preparation Procedures
B-9
-------
PFAS using an appropriate method proposed by the TBD laboratory and approved by EPA (Note
that the PFAS rinsate samples will be analyzed by the laboratory selected for PFAS analysis of
the 2020 GLHHFFTS fish fillet tissue samples).
Corrective Actions for Rinsates
The rinsate results will be evaluated based on the mass of each analyte detected, and assuming that all
of the apparent contamination could be transferred to a nominal 465-g mass of homogenized tissue.
Results for mercury and PCBs above the anticipated reporting limits for these analytes in
homogenized fillet tissue samples may be cause for corrective actions by the fish sample preparation
(Tetra Tech) laboratory. These corrective actions may include revisions to the laboratory's
equipment cleaning procedures, followed by a successful demonstration of the revised cleaning
procedures through preparation and analysis of additional rinsate samples.
Lipid Determination to Confirm Homogeneity
28. For one sample in each fish sample preparation batch of generally 20 samples, a laboratory under
contract to Tetra Tech will use the 30 - 35 g aliquot of homogenized fillet tissue to conduct triplicate
analyses of the lipid content of homogenized fillet tissue samples to confirm that they are
homogeneous. As with the collection of rinsate samples, the Tetra Tech laboratory should identify
and process the fish sample for homogeneity testing during the first day of fish sample preparation
operations for each batch.
Remove 30 to 35 g of fillet homogenate from the fish sample designated for homogeneity testing
before filling the archive sample containers. Place this aliquot in a glass or plastic container of
suitable size and label it with the sample ID number. Transfer the lipid aliquot to the TBD analytical
laboratory for triplicate lipid determination. This laboratory will use 5 to 10 g aliquots of fillet tissue
for each of the 3 lipid analyses.
29. From the lipid results, calculate the mean lipid content (in percent), the standard deviation (SD), and
the relative standard deviation (RSD) using the formulae below, or the corresponding functions in
Excel.
^](%lipids)1
mean % lipids =
2(%lipidsi mean lipids)2
SD
mean
If the RSD of the triplicate results is less than or equal to 15% for triplicate lipid samples with mean
lipid values at or above 2.5%, or if the RSD of the triplicate results is less than or equal to 20% for
triplicate lipid samples with mean lipid values below 2.5%, then the homogenization effort is judged
to be sufficient for all samples in that preparation batch. For this sample analyzed in triplicate, the
mean lipid content will be the lipid value reported for that sample, following the requirements
described in Step 24.
NCCA 2020 Great Lakes Human Health Fish Fillet Tissue Sample Preparation Procedures
B-10
-------
Corrective Actions for Homogeneity
If the RSD is greater than 15% for triplicate lipid samples with mean lipid values at or above 2.5%, or
if the RSD is greater than 20% for triplicate lipid samples with mean lipid values below 2.5%, then
corrective action is required for all samples in that preparation batch. Corrective actions will be
determined by EPA in direct consultation with the laboratory and Tetra Tech, but the default
corrective action consists of regrinding all of the aliquots from each whole fish sample in the affected
batch until the RSD criterion is met.
This may entail retrieving all sample aliquots (see Table 1) from the freezer, allowing them to
partially thaw, and homogenizing them again, beginning at Step 13. In these instances, all of the
equipment cleaning procedures will be repeated between each whole fish sample, new lipid results
will be determined for each fish sample, and a new homogenization QC determination (triplicate
lipids for one fish sample per batch) will be performed. New sample containers are required for any
rehomogenized samples.
II.H. Reporting Requirements
30. The fish sample preparation laboratory prepares a separate weekly progress report to document the
status of fish preparation activities for the 2020 GLHHFFTS and the ORD-Duluth 2020 Great Lakes
special study and forwards each report electronically to the OST Fish Sample Preparation Technical
Leader and the OST Project Manager. The format of each weekly progress report will be an Excel
spreadsheet using the 2015 GLHHFFTS fish sample preparation reports as a guide for organization of
each spreadsheet. For each homogenized sample processed during that period, include at least the
following information in the report:
site identification number,
sample identification number,
specimen numbers of the fish homogenized for the fillet composite sample,
common name for the fish species (provided to the laboratory in the processing instructions from
EPA),
field-determined length and lab-determined weight of each specimen in a whole fish sample,
total whole fillet (unhomogenized) weight (to the nearest gram),
total homogenized fillet composite sample (i.e., homogenate) weight (to the nearest gram),
analysis type (e.g., mercury, PFAS, PCB congeners, lipids, PCBs as Aroclors, fatty acids, and
archive samples),
fillet tissue aliquot weight (to the nearest 0.5 gram),
fish sample preparation batch ID,
preparation date (e.g., mm/dd/yyyy),
QC sample identifiers associated with the batch of homogenized fillet samples, and
lipid results for each fish sample.
Weekly progress reports will be due by COB Monday (or one day later in the case of holidays), and
each report will document fish sample preparation progress for the previous week.
In addition, the laboratory must report the results of the rinsate analyses for mercury, PCB congeners,
and the triplicate lipid results associated with the sample batch. Those results must be reported to the
OST Fish Sample Preparation Technical Leader and the OST Project Manager as soon after the
analyses as practical to facilitate timely review of the data from the QC samples and to minimize
delays in receiving approval from the OST Fish Sample Preparation Technical Leader (with OST
Project Manager concurrence) to process future batches.
NCCA 2020 Great Lakes Human Health Fish Fillet Tissue Sample Preparation Procedures
B-ll
-------
Note: As specified in the QC section of this QAPP (Section B5.1), the fish sample preparation
laboratory may not continue beyond the series of 3 fish sample preparation batches until
receiving notification from the OST Fish Sample Preparation Technical Leader (with OST
Project Manager concurrence) that the review of initial batch (in the series of 3 batches)
rinsate and homogeneity test results is complete, and the results were deemed satisfactory.
II.I. Shipping Samples
31. No samples (except fish sample preparation mercury, PCB, and triplicate lipid QC samples)
may be shipped until the OST Fish Sample Preparation Technical Leader and the OST Project
Manager have reviewed the fish sample preparation batch homogeneity testing and rinsate
results and authorized shipment of samples to designated analytical laboratories in writing.
The OST Project Manager will notify the Tetra Tech laboratory by email when specific batches of
samples may be shipped, and to whom.
Samples are shipped in batches (one batch per cooler) to each designated analytical laboratory. When
shipping batches of pre-frozen fillet tissue aliquots, keep the individual containers bagged in the food-
grade plastic freezer bags. Place these bags in a cooler with adequate space for the tissue containers,
packing materials, and dry ice blocks.
Secure each of the tissue containers with packing materials (e.g., bubble wrap or foam) before adding
the block dry ice. Place a layer of bubble wrap and a plastic cooler liner on top of the containers
before adding the dry ice, as this can prevent cracking the lids.
The amount of dry ice required for shipping will depend on the number of homogenized fillet tissue
samples in the cooler and the time of year. It should be an adequate supply to keep the tissue samples
frozen for 48 hours (i.e., a minimum of 30 pounds of dry ice per cooler for up to 10 pounds of fillet
tissue samples). Only blocks of dry ice are allowed for shipping fish tissue samples. Do not use dry
ice pellets for shipping fillet samples.
Record the samples contained in the cooler on a shipping form provided by CSRA/GDIT and place
the form in a plastic bag taped to the inside lid of the cooler. Secure the outside of the cooler with
sealing tape, address it to the sample recipient identified by the OST Project Manager, and attach a
dry ice (dangerous goods) label. Ship the cooler via an overnight express carrier on a date that will
allow delivery of the cooler to the analytical laboratory on a normal business day (e.g., no Saturday
deliveries and no deliveries on U.S. Federal holidays). Provide the air bill number for each
shipment to CSRA/GDIT, the OST Project Manager, and the OST Fish Sample Preparation Leader
via email on the day that the shipment occurs. CSRA/GDIT will provide the Tetra Tech
laboratory with a third-party FedEx account to which each shipment will be billed.
NCCA 2020 Great Lakes Human Health Fish Fillet Tissue Sample Preparation Procedures
B-12
-------
EPA and Tetra Tech Fish Sample Preparation Laboratory Contact Information
OST Project Manager
Leanne Stahl
OW/Office of Science & Technology
U.S. EPA, MC4305T
1200 Pennsylvania Avenue, NW
Washington, DC 20460
202-566-0404
stahl.leanne@epa.gov
Primary Tetra Tech Contact
Blaine Snyder
Tetra Tech Project Leader
Tetra Tech, Inc.
10711 Red Run Blvd., Suite 105
Owings Mills, MD 21117
410-902-3158
blaine. sny der@tetratech. com
OST Fish Sample Preparation Technical Leader
John Healey
OW/Office of Science and Technology
U.S. EPA, MC4305T
1200 Pennsylvania Avenue NW
Washington, DC 20460
202-566-0176
healey .j ohn@epa. gov
Alternative Tetra Tech Contact
Tara Cohen
Tetra Tech, Inc.
10711 Red Run Blvd., Suite 105
Owings Mills, MD 21117
410-902-3143
tara.cohen@tetratech.com
NCCA 2020 Great Lakes Human Health Fish Fillet Tissue Sample Preparation Procedures
B-13
-------
ATTACHMENT 1
ANALYSES OF RINSATES AND BLANKS FOR MERCURY AND PCBs
This attachment describes the analyses of rinsate samples and blank samples generated during the fish
sample preparation process. The results of those analyses are important in demonstrating that the Tetra
Tech laboratory's equipment cleaning procedures are effective at preventing cross-contamination between
fish tissue samples.
A. EQUIPMENT AND MATERIALS:
Mercury analyzer suitable for aqueous samples using cold-vapor atomic absorption (CVAA)
instruments compatible with EPA Method 245.1 (or other suitable analytical procedure and detector
system capable of achieving a method detection limit (MDL) of approximately 1 j^ig/L).
Gas chromatograph with an electron-capture detector (GC/ECD) and two dissimilar GC columns
suitable for analysis of PCB congeners, or other suitable analytical procedures and detector systems
(e.g., low-resolution or high-resolution GC/MS). The laboratory must be able to achieve an IDL for
each congener on the order of 0.5 ng/mL, for a 1-mL final volume.
Solvent concentration equipment suitable for reducing hexane rinsates to final volumes of
1.0 to 10 mL.
A PCB standard solution containing at least the following PCB congeners: 52, 66,105,118,141,146,
170,174,177, and 187, to be used to establish retention times and perform at least a 3-point
calibration of the GC/ECD, or to calibrate other detector systems that do not rely solely on retention
times for identification. (Additional congeners can be included by the laboratory. These congeners
represent those that EPA has found frequently, at relatively high concentrations, in other fish tissue
studies.)
Assorted glassware, syringes, etc.
B. RINSATE AND BLANK ANALYSES
The three sets of rinsate and blank samples include:
One deionized (DI) water rinsate sample and one DI water blank sample for mercury analysis.
One hexane rinsate sample and one hexane blank sample for analysis of PCB congeners.
One DI water rinsate sample and one DI water blank sample for PFAS analysis.
During fish sample preparation efforts, the Tetra Tech laboratory will prepare each set of rinsate and
blank samples at a frequency of one set for each batch of generally 20 fish samples prepared.
The analytical laboratory (to be determined) will digest and analyze the mercury rinsate and blank
samples by CVAA. For each analysis, the laboratory will determine the mass of mercury in the total
volume of each rinsate or blank sample, rather than the concentration of mercury. The analytical
laboratory will either perform a method detection limit (MDL) study for mercury in aqueous samples or
use existing aqueous MDL data for the CVAA instrument employed. The laboratory must be able to
achieve a MDL of approximately 1 (.ig/L. Mercury results will be reported down to the mass equivalent
to the mass at the MDL for aqueous samples.
For PCB rinsates and blanks, the analytical laboratory will concentrate the rinsates and blanks to a
suitable final volume for the analytical procedure and analyze the concentrated samples. Because the
PCB rinsates are not aqueous samples that are extracted, a traditional MDL study for aqueous samples
does not apply. Therefore, the laboratory must perform an instrument detection limit (IDL) study before
beginning any rinsate analyses. The IDL study will consist of analyzing 7 low-level standards containing
the PCBs listed above. The laboratory will determine the standard deviation of results for each PCB
across all 7 analyses, and multiply the standard deviation times 3.143, which is the Student's t-value for
NCCA 2020 Great Lakes Human Health Fish Fillet Tissue Sample Preparation Procedures
B-14
-------
7 replicates. The laboratory must achieve an IDL on the order of 0.5 ng/mL, for a 1-mL final volume, or
a total mass of 0.5 ng.
If using a GC/ECD procedure, PCB congeners will be identified based on retention time windows on both
GC columns (see EPA Methods 608 or 8000C for examples of procedures for determining retention time
windows). If using another proposed and approved procedure such as GC/MS, the congeners will be
identified based on the requirements in that procedure.
PCB results in the rinsates and blanks will be reported down to the mass equivalent of the IDL. If using a
GC/ECD procedure, any PCBs detected on one GC column must be confirmed by the analysis of the
sample on a second GC column with a different stationary phase. Alternatively, GC/ECD analyses may
be conducted on an instrument set up for simultaneous dual-column analyses. For each analysis, the
laboratory will determine the mass of each PCB congener in the total volume of each rinsate or blank
sample, rather than the concentration of each analyte.
The Tetra Tech laboratory will not be responsible for analysis of rinsate and blank samples for PFAS.
Tetra Tech will hold these samples in temporary storage until the OST Project Manager identifies the
laboratory selected to analyze 2020 GLHHFFTS fillet samples for PFAS, provides shipping information
for this laboratory, and notifies Tetra Tech when they can ship these samples.
C. QUALITY CONTROL
The quality control (QC) procedures required for the rinsate and blank analyses include:
MDL and IDL studies, as described above
Instrument calibration (see Method 245.1 and TBD PCB method for procedures and acceptance
criteria, or consult the specifications in any other procedures proposed)
Instrument blanks for mercury and PCB analyses
Calibration verification (once per analysis batch) for mercury and PCB analyses
Laboratory control sample (LCS) once per analysis batch (for mercury analysis only)
The MDL and IDL results are reviewed by CSRA/GDIT and EPA as soon as they become available for
each fish sample preparation batch, and the Tetra Tech laboratory will not be authorized to prepare fish
tissue samples beyond a series of 3 fish sample preparation batches until that review is complete and the
results are acceptable for the initial batch in each series of 3 fish sample preparation batches.
The matrix for the mercury rinsates is reagent (deionized) water, which should not adversely affect
method performance. Therefore, matrix spike samples are not required for mercury.
Because the PCB rinsates do not involve extraction of an environmental matrix, matrix spike samples are
not applicable. Likewise, laboratory control samples are not applicable to PCBs.
The instrument blanks for mercury and PCBs take the place of a traditional method blank that would be
extracted along with environmental samples.
D. DELIVERABLES
Summary data from the rinsate analyses are to be delivered to EPA in an Excel file. That file must
contain the following information, at a minimum:
Batch ID - assigned by EPA (numerical sequence beginning at 1)
Sample ID - as described in the instructions for preparing the rinsates (Step 26 in Appendix B)
Lab sample ID - unique internal identifier used by the laboratory, if any
NCCA 2020 Great Lakes Human Health Fish Fillet Tissue Sample Preparation Procedures
B-15
-------
Prep date - Date (MM/DD/YYYY) on which the rinsate and solvent blank samples were prepared
Analysis type - "Mercury" or "PCBs"
Analysis date - Date (MM/DD/YYYY) on which the rinsate and solvent blank samples were analyzed
Analyte name - Mercury (total) or PCBs
Mass of analyte found - in micrograms for mercury and nanograms for PCBs
Lab qualifiers - as needed to describe any analytical concerns. A complete list of the qualifiers and
their meanings must be included with each data submission (e.g., in a separate tab on the Excel file).
Reporting limit for mercury (i.e., the MDL for this study) and PCBs (i.e., the IDL for this study) - in
the same mass units used for the mercury and PCB results
Instrument calibration data - Submit as a separate tab in the Excel file. Must include results for the
initial calibrations for mercury and PCBs, as well as any relevant calibration verifications associated
with the analyses. Include calibration equations (e.g., regressions) and metrics (e.g., correlation
coefficient or calibration factor).
Provide Excel files for the mercury and PCB analysis results to the Tetra Tech Project Leader. The
laboratory may submit a separate Excel file for each type of analysis. Raw data supporting mercury and
PCB analyses (e.g., instrument printouts) must be retained by the laboratory and made available to EPA
when requested, at no additional cost. If requested, raw data may be submitted in hard copy, or as a PDF
file.
NCCA 2020 Great Lakes Human Health Fish Fillet Tissue Sample Preparation Procedures
B-16
-------
Appendix C
2020 Great Lakes Human Health
Fish Fillet Tissue Study
Fish Sample Preparation Laboratory Bench Sheet
-------
2020 GLHHFFTS Rsh Sample Preparation Laboratory Bench Sheet
Page of
Site ID:
Prep Date
HI1DC i ' s:
Sum of Fillet Mass fg):
Sample ID:
Filleter:
Homogenate ' issue Mass (g):
Ftah
EPA Batch ID
Fish Processer:
- e: & Her
:i=nate Mas
3 Difference (g):
Specimen
ID
Species
Fish
Length
(mm)
Fish
Mass (g)
Fillet
Mass (g)
Fillet
Tissue
Recovery
fxl
Notes
.01
.02
,03
.04
.05
.06
,£>?
JOB
M
.10
Sample
Jar
Hg
Mass
PFAS
PCB
Congeners
Lipids,
Fish 1
Lipids,
Fish
2-20
PCBs as
Aroclors
Fatty
Acids
Small
Archive
1
Small
Archive
2
Bulk
Archive 1
Bulk Archive
2
Target
Sample
Mass (g)
5-10g
10-15g
20 - 25 g
30 - 35 g
(prepare
mKKKk
aiiquots)
10 -15 g
20 - 25 g
10-15g
50 - 60 g
50 - 60 g
240-250 g
All remaining
mass up to 250 g
Sample
Mass (g)
1.
2.
3.
Tetra Tech, Inc.
Ecological Testing Facility Data Checked and Approved 2020
2020 GLHHFFTS Fish Sample Preparation Laboratory Bench Sheet
-------
Appendix D
ORD-Duluth 2020 Great Lakes Special Study
Human Health
Fish Sample Preparation Laboratory Bench Sheet
-------
2020 Great Lakes Special Study Hunan Health Fish Sanpie ^reparation Laboratory Bench Sheet Pa«eof
Site ID:
Prep Cate
il.lMCC" ' I
Sum of Fillet Mass (g):
Sample ID:
Fieter:
Homagenste Tissue Mass (g):
Fish
EPA Batch D
Fish Processer:
Filet & Homo-genate Mass Difference {§}:
Specimen
ID
Species
Fish
Length
(mm)
Fish
Mass (g)
Fillet
Mass (g)
Fillet
Tissue
Recovery
(X)
Notes
.01
,02
.05
,04
.05
.06
.0?
,00;
.09
.10
Sample
Jar
Hg
Mass
PFAS
PCB
Congeners
Lipids,
Fish 1
Lipids,
Fish
2-20
PCBs as
Aroclors
Fatty
Acids
Small
Archive
1
Small
Archive
2
Bulk
Archive 1
Bulk Archive
2
Target
Sample
Mass (g)
5 -10 g
10 -15 g
20-25g
30 - 35 g
(prepare
3
aliquots)
10 -15 g
20 - 25 g
10 -15 g
50 - 60 g
50 - 60 g
240- 250 g
All remaining i
mass up to 250 g i
Sample
Mass (g)
1.
2.
3.
Tetra lech, Inc.
Ecol og ica 11 esti ng F act I ity Data h eck ed and Approved 2020
ORD-Duluth 2020 Great Lakes Special Study Human Health Fish Sample Preparation Laboratory Bench Sheet
D-l
------- |