Quality Assurance Project Plan for Analysis of the National Coastal Condition Assessment 2020 Great Lakes Fish Fillet Samples for Mercury, Per- and Polyfluoroalkyl Substances (PFAS), Polychlorinated Biphenyl (PCB) Congeners, Aroclors, and Fatty Acids Revision 4 November 19, 2021 Prepared by: United States Environmental Protection Agency Office of Water Office of Science and Technology Standards and Health Protection Division Prepared with support from: CSRA, LLC, a General Dynamics Information Technology Company under: Office of Water Engineering and Analysis Division Contract No. EP-C-17-024 EPA-822-B-24-004 ------- NCCA 2020 Great Lakes Fish Fillet Sample Analysis QAPP Revision 4 Date: November 19, 2021 Page 2 of 45 Revision History November 19, 2021 Revision 4 This revision includes edits to describe the procedures for omega-3 and omega-6 fatty acid analyses of National Coastal Condition Assessment 2020 Great Lakes fillet samples and add them to the mercury, PFAS, PCB congener, and Aroclor analyses that are currently underway. A more detailed list of edits follows below: • The revision history was updated. • Section A was updated. • The fatty acid analysis laboratory was added to Section A3 and to Figure 1. • Section A7 was updated to refer to the fatty acid QC criteria. • The details of the analytical procedure for fatty acids and the QC criteria were added in Sections B4.5 and B5.5. • Section B7 was updated to include the calibration information for fatty acid analysis. • Sections CI, Cl.l, CI.3, and CI.4 were updated to include the information for fatty acids. • Sections D1.2, D1.3, and D3 were updated to include the fatty acid results in the list of analytical data that will be verified and validated, respectively. • The Reference section was updated with the citation for the QA manual from the fatty acid analysis laboratory. • Appendix B was updated to include the fatty acid MDLs and MLs from the laboratory selected for fatty acid analysis. April 26, 2021 Revision 3 This revision includes edits to describe the procedures for Aroclor analyses of National Coastal Condition Assessment 2020 Great Lakes fillet samples, as planned for the future, and add them to the mercury, PFAS, and PCB congener analyses that are currently underway. A more detailed list of edits follows below: • The revision history was updated. • Section A was updated. • The Aroclor analysis laboratory was added to Section A3 and to Figure 1. • Section A6 was updated to include shipping fillet tissue samples from the 2020 GLHHFFTS for Aroclor analyses. • Section A7 was updated to refer to the Aroclor QC criteria. • The placeholder text for Aroclors in Sections B4.4 and B5.4 was replaced with the actual details. • Section B7 was updated to include the calibration information for Aroclor analysis. • Sections CI, Cl.l, CI.3, and CI.4 were updated to include the information for Aroclors. • Section D1.2 and D1.3 were updated to include the Aroclor results in the list of analytical data that will be verified and validated, respectively. • The Reference section was updated with the citation for the QA manual from the Aroclor analysis laboratory and the citations for the Aroclor methods. • Appendix B was updated to include the Aroclor MDLs and MLs from the laboratory selected for Aroclor analysis. ------- NCCA 2020 Great Lakes Fish Fillet Sample Analysis QAPP Revision 4 Date: November 19, 2021 Page 3 of 45 March 12, 2021 Revision 2 This revision includes edits to describe the procedures for PCB congener analyses of National Coastal Condition Assessment 2020 Great Lakes fillet samples, as planned for the future, and add them to the mercury and PFAS analyses that are currently underway. A more detailed list of edits follows below: • The revision history was updated. • Section A was updated. • The PCB congener analysis laboratory was added to Section A3 and to Figure 1. • Section A7 was updated to refer to the PCB congener QC criteria. • The placeholder text for PCB congeners in Sections B4.3 and B5.3 was replaced with the actual details. • Section B7 was updated to include the calibration information for PCB congener analysis. • Sections CI, CI. 1, CI.3, and CI.4 were updated to include the information for PCB congeners. • The Reference section was updated with the citation for the QA manual from the PCB congener analysis laboratory and the citations for the PCB congener method. • Appendix B was updated to include the PCB congener MDLs and MLs from the laboratory selected for PCB congener analysis. • Appendix D, with the PCB congener QC acceptance criteria, was added. February 26, 2021 Revision 1 This revision includes edits to describe the procedures for PFAS analyses of National Coastal Condition Assessment 2020 Great Lakes fillet samples and aqueous QC samples, as planned for the future, and add them to the mercury analyses that are currently underway. A more detailed list of edits follows below: • The revision history was added. • Section A was updated. • The PFAS laboratory was added to Section A3 and to Figure 1. • Section A7 was updated to refer to the PFAS QC criteria. • The placeholder text for PFAS in Sections B4.2 and B5.2 was replaced with the actual details. • Section B7 was updated to include the calibration information for PFAS. • Sections CI, CI.1, CI.3, CI.4, and C2 were updated to include the information for PFAS. • The Reference section was updated with the citation for the QA manual from the PFAS laboratory and the citations for the PFAS methods. • Appendix B was updated to include the PFAS MDLs and MLs from the laboratory selected for these analyses. • Appendix C was added to include the QC acceptance criteria for the PFAS analyses. December 16, 2020 - Original QAPP (Revision 0) signed ------- NCCA 2020 Great Lakes Fish Fillet Sample Analysis QAPP Revision 4 Date: November 19, 2021 Page 4 of 45 Quality Assurance Project Plan for Analysis of the National Coastal Condition Assessment 2020 Great Lakes Fish Fillet Samples for Mercury, Per- and Polyfluoroalkyl Substances (PFAS), Polychlorinated Biphenyl (PCB) Congeners, Aroclors, and Fatty Acids 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 CSRA under EPA Contract No. EP-C-17-024. It presents objectives, performance requirements, and acceptance criteria for the analyses of National Coastal Condition Assessment (NCCA) 2020 Great Lakes Human Health Fish Fillet Tissue Study (GLHHFFTS) and Office of Research and Development (ORD)-Duluth 2020 Great Lakes special study fillet samples for mercury (Revision 0), fillet samples and aqueous QC samples for per- and polyfluoroalkyl substances (PFAS) (Revision 1), fillet samples for the full complement of 209 polychlorinated biphenyl (PCB) congeners (Revision 2), fillet samples for PCBs as Aroclors (Revision 3), and fillet samples for omega-3 and omega-6 fatty acids (Revision 4). This QAPP does not address fish sample preparation because OST developed a separate QAPP in July 2020 that presents objectives, procedures, performance requirements, and acceptance criteria for the preparation of fillet tissue samples from whole fish composite samples collected from designated Great Lakes nearshore sites for the 2020 GLHHFFTS and from Lake Michigan enhancement sites for the ORD-Duluth 2020 Great Lakes special study by field crews during the NCCA 2020 field sampling season (USEPA 2020a). (Note: Some field crews will need to continue the NCCA 2020 Great Lakes field sampling season in 2021 due to travel restrictions related to the coronavirus pandemic in 2020.) This QAPP also does not address fish sample collection because that information is included in separate documents (USEPA 2020a and USEPA 2020b) prepared by EPA's OW/Office of Wetlands, Oceans, and Watersheds (OWOW) with support from OST. This QAPP was prepared in accordance with the most recent version of EPA QA/R-5, EPA Requirements for Quality Assurance Project Plans (USEPA 2001a), which was reissued in 2006. In accordance with EPA QA/R-5, this QAPP 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 for the 2020 NCCA and by the EPA 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 will be 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. ------- NCCA 2020 Great Lakes Fish Fillet Sample Analysis QAPP Revision 4 Date: November 19, 2021 Page 5 of 45 Al. Approvals J-JUL+i+UL November 19, 2021 Leanne Stall 1. OST Project Manager, EPA Date rij a pi n a n A C L-J Digitally signed by SHARI BARASH 3 n A X\ 1 DMnMj n Date: 2021.12.02 08:23:29 -05'00' Digitally signed by Harry McCarty Date: 2021.12.02 10:03:22 -05'00' Digitally signed by Marguerite E. Jones Date: 2021.12.02 09:41:44 -05'00' Shari Barash. Chief. National Branch. EPA Date Digitally signed by HARRY HARRY KRAMER KRAMER Date: 2021.12.02 07:31:43 -05'00' Bill Kramer. SHPD QA Coordinator. EPA Date Digitally signed by JOSEPH JOSEPH BEAMAN BEAMAN Date: 2021.12.02 08:30:03 -05'00' Joe Beam an. OST QA Officer. EPA Date Harry McCarty Harry McCarty, CSRA Project Leader Date YilHi7 Thamhprq Vplardp Di9ital|ysi9ned by YildizChambers.velarde i iiuiz v_ridmueib_veidiue Date:2021 .i2.0209:47:42-os'oo1 Yildiz Chambers-Velarde, CSRA Work Assignment Manager Date Marguerite EJones Marguerite Jones. CSRA QA Manager Date ------- NCCA 2020 Great Lakes Fish Fillet Sample Analysis QAPP Revision 4 Date: November 19, 2021 Page 6 of 45 A2. Table of Contents A. PROJECT MANAGEMENT 4 Al. Approvals 5 A2. Table of Contents 6 A3. Di stributi on Li st 9 A4. Project/Task Organization 10 A5. Problem Definition/Background 17 A6. Project/Task Description 17 A7. Quality Objectives and Criteria 19 A8. Special Training/Certification 20 A9. Documents and Records 21 B. DATA GENERATION AM) ACQUISITION 21 B1. Sampling Process Design (Experimental Design) 21 B2. Fish Sampling and Fillet Sample Preparation Methods 23 B2.1 Fish Sampling Methods 23 B2.2 Fillet Sample Preparation Methods 24 B3. Sample Receipt and Inspection 25 B4. Analytical Methods 26 B4.1 Mercury Analysis of Fillet Tissue 26 B4.2 PFAS Analysis of Fillet Tissue and Rinsate Samples 26 B4.3 PCB Congener Analysis of Fillet Tissue 27 B4.4 PCBs as Aroclors Analysis of Fillet Tissue 28 B4.5 Fatty Acid Analysis of Fillet Tissue 28 B5. Analytical Quality Control 29 B5.1 Mercury Analysis QC Criteria 29 B5.2 PFAS Analysis QC Criteria 30 B5.3 PCB Congener Analysis QC Criteria 31 B5.4 PCBs as Aroclors Analysis QC Criteria 33 B5.5 Fatty Acid Analysis QC Criteria 35 B6. Instrument/Equipment Testing, Inspection, and Maintenance 35 B7. Instrument/Equipment Calibration and Frequency 35 B8. Inspection/Acceptance of Supplies and Consumables 36 B9. Non-direct Measurements 36 B10. Data Management 36 C. ASSESSMENT AM) OVERSIGHT 37 CI. Assessments and Response Actions 37 Cl.l Surveillance 37 CI.2 Product Review 38 CI.3 Quality Systems Audit 39 CI.4 Readiness Review 39 C1.5 Techni cal Sy stem s Audit 39 C1.6 Data Quality Assessment 40 C2. Reports to Management 40 ------- NCCA 2020 Great Lakes Fish Fillet Sample Analysis QAPP Revision 4 Date: November 19, 2021 Page 7 of 45 D. DATA VALIDATION AND USABILITY 40 Dl. Data Review, Verification, and Validation 41 Dl.l Data Review 41 D1.2 Data Verification 41 D1.3 Data Validation 41 D2. Verification and Validation Methods 41 D2.1 Verification Methods 41 D2.2 Validation Methods 42 D3. Reconciliation with User Requirements 43 REFERENCES 44 TABLES Table 1. Primary Target Fish Species and Secondary Alternative Fish Species for the 2020 GLHHFFTS and ORD-Duluth 2020 Great Lakes Special Study 23 Table 2. QC Samples and Acceptance Criteria for Mercury Analysis of Fish Tissue 30 Table 3. QC Samples and Acceptance Criteria for PFAS Analysis of Tissue and Rinsates .... 31 Table 4. QC Samples and Acceptance Criteria for PCB Analysis of Fish Tissue 32 Table 5. QC Samples and Acceptance Criteria for Aroclor Analysis of Fish Tissue 33 Table 6. QC Samples and Acceptance Criteria for Fatty Acid Analysis of Fish Tissue 35 FIGURES Figure 1. 2020 GLHHFFTS project team organizations from fish fillet sample analyses 11 Figure 2. NCCA 2020 Great Lakes human health whole fish sampling locations 19 APPENDICES Appendix A Target List of NCCA 2020 Great Lakes Human Health Whole Fish Sampling Locations Appendix B NCCA 2020 Great Lakes Human Health Detection and Quantitation Limits for Tissue Analyses Appendix C 2020 NCCA Quality Control (QC) Acceptance Criteria for PFAS Analyses of Great Lakes Fish Fillet Tissue Samples and QC Rinsate Samples Appendix D 2020 NCCA Quality Control (QC) Acceptance Criteria for PCB Congener Analysis of Great Lakes Fish Fillet Tissue Samples ------- NCCA 2020 Great Lakes Fish Fillet Sample Analysis QAPP Revision 4 Date: November 19, 2021 Page 8 of 45 LIST OF ACRONYMS AND ABBREVIATIONS ccv Continuing calibration verification EPA Environmental Protection Agency GLHHFFTS Great Lakes Human Health Fish Fillet Tissue Study HRGC High resolution gas chromatography HRMS High resolution mass spectrometry ID Identification LCS Laboratory control sample (also known as an OPR) MDL Method detection limit ML Minimum level (also referred to as the quantitation limit) MS Matrix spike sample MSD Matrix spike duplicate sample NCCA National Coastal Condition Assessment OPR Ongoing precision and recovery sample OST Office of Science and Technology OW Office of Water OWOW Office of Wetlands, Oceans, and Watersheds PCB Polychlorinated biphenyl PFAS Per- and polyfluoroalkyl substances QA Quality assurance QAPP Quality Assurance Project Plan QC Quality control QCS Quality control sample QSA Quality system audit RPD Relative percent difference RSD Relative standard deviation SHPD Standards and Health Protection Division SOP Standard operating procedure SPE Solid-phase extraction VER Verification ------- NCCA 2020 Great Lakes Fish Fillet Sample Analysis QAPP Revision 4 Date: November 19, 2021 Page 9 of 45 A3. Distribution List Shari Barash Bill Kramer USEPA/OW/OST (4305T) USEPA/OW/OST (4305T) 1200 Pennsylvania Ave., N.W. Washington, DC 20460 202-566-0996 1200 Pennsylvania Avenue, N.W. Washington, DC 20460 202-566-0385 barash.shari@epa.gov kramer.bill@epa. gov Joe Beaman Tom Kincaid USEPA/OW/OST (4303T) U SEPA/ORD/CPHEA/PESD 1200 Pennsylvania Avenue, N.W. 200 S.W. 35th Street Washington, DC 20460 202-566-0420 Corvallis, OR 97333 541-754-4479 beaman.joe@epa.gov kincaid.tom@epa.gov Louis Blume Sarah Lehmann USEPA Great Lakes National Program Office USEPA/OW/OWOW (4503T) 77 West Jackson Boulevard (LAB-10C) Chicago, IL 60604 312-353-2317 1200 Pennsylvania Avenue, N.W. Washington, DC 20460 202-566-1379 blume.louis@epa.gov lehmann. sarah@epa. gov David Bolgrien USEPA/ORD/CCTE/WWRB (Alll) Brian Lenell USEPA Great Lakes National Program Office 6201 Congdon Blvd Duluth, MN 55804 218-529-5216 77 W. Jackson Blvd. (G-9J) Chicago, IL 60604 bolgrien.dave@epa.gov 312-353-4891 lenell.brian@epa.gov Yildiz Chambers-Velarde CSRA Harry McCarty CSRA 6361 Walker Lane, Suite 300 6361 Walker Lane, Suite 300 Alexandria, VA 22310 Alexandria, VA 22310 703-254-0061 703-254-0093 yildiz.chambers@gdit.com harry.mccarty@gdit.com Tara Cohen Mari Nord Tetra Tech, Inc. USEPA Region 5 Office 10711 Red Run Blvd., Suite 105 77 W. Jackson Blvd. (WS-15J) Owings Mills, MD 21117 410-902-3143 Chicago, IL 60604 312-886-3017 cohen.tara@tetratech.com nord.mari@epa.gov John Healey USEPA/OW/OST (4305T) Tony Olsen U SEPA/ORD/CPHEA/PESD 1200 Pennsylvania Ave., N.W. Washington, DC 20460 202-566-0176 200 S.W. 35th Street Corvallis, OR 97333 541-754-4790 healey.john@epa.gov olsen.tony@epa.gov Elizabeth Hinchey USEPA Great Lakes National Program Office Blaine Snyder Tetra Tech, Inc. 77 W. Jackson Blvd. (G-9J) 10711 Red Run Blvd., Suite 105 Chicago, IL 60604 312-886-3451 Owings Mills, MD 21117 410-902-3158 hinchey. elizabeth@epa.gov blaine. sny der@tetratech. com Marguerite Jones CSRA Leanne Stahl USEPA/OW/OST (4305T) 6361 Walker Lane, Suite 300 1200 Pennsylvania Ave., N.W. Alexandria, VA 22310 Washington, DC 20460 703-254-0081 202-566-0404 maggie .j ones@gdit. com stahl.leanne@epa.gov ------- NCCA 2020 Great Lakes Fish Fillet Sample Analysis QAPP Revision 4 Date: November 19, 2021 Page 10 of 45 Hugh Sullivan USEPA/OW/OWOW (4504T) 1200 Pennsylvania Avenue, N.W. Washington, DC 20460 202-566-1763 sullivan.hugh@epa.gov Vista Analytical Laboratory 1104 Windfield Way El Dorado Hills, CA 95762 (Contact Harry McCarty at CSRA) ALS Environmental 1317 South 13th Avenue Kelso, WA 98626 (Contact Harry McCarty at CSRA) Eurofins-TestAmerica 301 Alpha Drive Pittsburgh, PA 15238 (Contact Harry McCarty at CSRA) SGS-AXYS Analytical Services, Ltd. 2045 Mills Road Sidney, BC Canada V8L 5X2 (Contact Harry McCarty at CSRA) Clarkson University Dept. Civil and Environmental Engineering 8 Clarkson Avenue Potsdam, NY 13699 (Contact Harry McCarty at CSRA) 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), along with statisticians in the Pacific Ecological Systems Division within EPA's ORD Center for Public Health and Environmental Assessment (Corvallis, Oregon). The EPA project team receives scientific, technical, and logistical support from contractors at Tetra Tech and at CSRA, a General Dynamics Information Technology company. Tetra Tech provides primarily fisheries support (e.g., fish sampling and fish sample preparation) and CSRA provides analytical support for the project team. Members of the project team technically and/or financially responsible for fish fillet sample analysis include the OST Project Manager and Work Assignment Contracting Officer Representative (WACOR), the OST Alternate WACOR (Alt-WACOR), the OST Quality Assurance (QA) Officer, the SHPD QA Coordinator, the GLNPO Project Manager, the GLNPO QA Manager, the CSRA Work Assignment Manager, the CSRA Project Leader, and the CSRA QA Manager who collectively provide scientific, technical, logistical, and quality control (QC) support for the study. The project team organization provides the framework for conducting fish sample analysis to meet study objectives. The organization structure and function also facilitate project performance and adherence to QC procedures and 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 Fish Fillet Sample Analysis QAPP Revision 4 Date: November 19, 2021 Page 11 of 45 Figure 1. 2020 GLHHFFTS project team organizations from fish fillet sample analyses ------- NCCA 2020 Great Lakes Fish Fillet Sample Analysis QAPP Revision 4 Date: November 19, 2021 Page 12 of 45 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. She is also serving as the Fish Sample Analysis Technical Leader to provide technical and work assignment management support for 2020 Great Lakes fish fillet sample analysis and related analytical activities. Both roles involve the following 2020 GLHHFFTS responsibilities: • 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 John Healey who serves additionally as the 2020 GLHHFFTS Fish Sample Preparation Technical Leader as described in the Quality Assurance Project Plan for National Coastal Condition Assessment (NCCA) 2020 Great Lakes Human Health Fish Sample Preparation (USEPA 2020c). (These fish sample preparation procedures and requirements also apply to the ORD-Duluth 2020 Great Lakes special study.) • managing analysis of 2020 Great Lakes fish fillet samples for target chemicals and related analytical support activities, including developing and managing a work assignment to provide CSRA support for analyzing the 2020 Great Lakes fillet samples, directing development of the initial NCCA 2020 Great Lakes fillet 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 separate databases to store 2020 GLHHFFTS and ORD-Duluth 2020 Great Lakes special study fillet sample analysis results (this series of fillet sample analysis and related analytical activities also applies 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 other work assignments and/or 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 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 Fish Fillet Sample Analysis QAPP Revision 4 Date: November 19, 2021 Page 13 of 45 • 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 Great Lakes special study project team for reporting 2020 Great Lakes human health fish fillet analysis results) • coordinating with John Healey (Task Order Contracting Officer Representative or 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 serving as the Fish Sample Analysis Deputy Technical Leader to assist in providing technical and work assignment management support for 2020 Great Lakes fish fillet sample analysis and related analytical activities. He is also serving as the Fish Sample Preparation Technical Leader. Both roles involve the following 2020 GLHHFFTS responsibilities: • providing support for development of the initial QAPP for analysis of the NCCA 2020 Great Lakes fillet samples and preparation of the subsequent QAPP revisions (this responsibility also applies to the ORD-Duluth 2020 Great Lakes special study) • providing assistance in managing analysis of 2020 Great Lakes fish fillet samples for target chemicals and related analytical support activities, including assistance for developing and managing a work assignment to provide CSRA support for analyzing the 2020 Great Lakes fillet samples, participating in review of work assignment deliverables, and providing data management and analysis support (this series of fillet sample analysis and related analytical activities also applies to the ORD-Duluth 2020 Great Lakes special study) • 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 Great Lakes human health fish samples, providing technical direction for and oversight of fish sample preparation activities (e.g., providing oversight for fish sample processing and analysis of fish sample preparation QC samples and single lipid 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) ------- NCCA 2020 Great Lakes Fish Fillet Sample Analysis QAPP Revision 4 Date: November 19, 2021 Page 14 of 45 • 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 • 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 • providing support for 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 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 QAPP for the 2020 GLHHFFTS ------- NCCA 2020 Great Lakes Fish Fillet Sample Analysis QAPP Revision 4 Date: November 19, 2021 Page 15 of 45 • managing analysis of fish tissue samples for the fatty acids • 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 • 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) Joe Beaman 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 Yildiz Chambers-Velarde is the CSRA Work Assignment Manager who is responsible for managing all aspects of the technical support being provided by CSRA staff for the 2020 GLHHFFTS fish fillet indicator. Her specific responsibilities include the following: • monitoring the performance of CSRA staff participating in this study to ensure that they are following all the technical and QA procedures described in this QAPP that are related to CSRA tasks being performed to support this study • ensuring completion of high-quality deliverables within established budgets and time schedules • developing monthly progress and financial reports for support provided by CSRA • 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 ------- NCCA 2020 Great Lakes Fish Fillet Sample Analysis QAPP Revision 4 Date: November 19, 2021 Page 16 of 45 Harry McCarty is the CSRA Project Leader who is primarily providing technical support for the 2020 GLHHFFTS fish fillet tissue indicator. His specific responsibilities include the following: • providing direct technical support for the following 2020 GLHHFFTS fish fillet tissue indicator activities: preparing information related to technical and quality assurance requirements for chemical analysis of homogenized fish fillet tissue samples for target analytes (e.g., mercury, PFAS, and PCB congeners, Aroclors, and fatty acids), verification and validation of analytical data (data quality review), and development of 2020 GLHHFFTS fish fillet indicator documents (including this QAPP) or characterization of this indicator in other 2020 GLHHFFTS documents obtaining laboratory services to analyze 2020 Great Lake human health fish fillet tissue samples for target analytes (e.g., mercury, PFAS, and PCB congeners, Aroclors, and fatty acids), and providing technical and QA oversight of laboratory operations completing review of the fillet tissue analytical data and developing the analytical data QA report compiling fish fillet tissue analytical data files for statistical analysis and for public release developing and maintaining separate project-specific databases for storing 2020 GLHHFFTS human health fish sample collection information from Great Lakes nearshore sites and fillet sample analysis data and for storing ORD-Duluth 2020 Great Lakes special study human health fish sample collection information from Lake Michigan enhancement sites and fillet sample analysis data, and initiating queries of these databases to respond to data requests from Agency and external users preparing summary project information and graphics for development of project fact sheets, presentations, and other EPA meeting and outreach materials supporting development of text and graphics for technical journal articles and final project reports for reporting 2020 GLHHFFTS data and data from other EPA fish tissue studies obtaining freezer space that meets the requirements for long-term storage of archived fish tissue samples, organizing the archived fish tissue samples by project to facilitate retrieval of the samples, and developing and maintaining an inventory of the archived samples, as required • 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 • serving as the project team member providing technical expertise on any issues related to analytical chemistry and analytical methods for the 2020 GLHHFFTS and other EPA fish tissue studies ------- NCCA 2020 Great Lakes Fish Fillet Sample Analysis QAPP Revision 4 Date: November 19, 2021 Page 17 of 45 Marguerite Jones is the CSRA QA Manager, whose primary responsibilities include the following: • approving this QAPP • providing oversight for the implementation of QA procedures related to CSRA tasks that are described in this QAPP • reporting deviations from this QAPP to the CSRA Project Leader and recommending 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 U.S. 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 Sampling QAPP (USEPA 2020a) and the Field Operations Manual (USEPA 2020b), 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 ------- NCCA 2020 Great Lakes Fish Fillet Sample Analysis QAPP Revision 4 Date: November 19, 2021 Page 18 of 45 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. 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. 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 at 50 Lake Michigan enhancement sites (Appendix A) during 2020 and into 2021 because field crews were 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 composite 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 from both studies to laboratories contracted to analyze these samples for mercury, PFAS, PCB congeners, and lipids. (The fish sample preparation laboratory at the Tetra Tech facility in Owings Mills, Maryland is responsible for this activity in coordination with CSRA to conform to contract analytical laboratory fillet sample analysis schedules.) • Shipping fillet tissue samples from the 2020 GLHHFFTS to laboratories contracted to analyze those samples for Aroclors and fatty acids, which are not part of the ORD-Duluth 2020 Great Lakes special study. • Obtaining laboratory services to analyze 2020 Great Lakes fillet samples for target chemicals and monitoring analytical laboratory performance. ------- NCCA 2020 Great Lakes Fish Fillet Sample Analysis QAPP Revision 4 Date: November 19. 2021 Page 19 of 45 • Conducting data quality reviews for fish fillet tissue analytical and QC data and assigning data qualifiers when applicable. • Developing project-specific databases (i.e., separate databases) for storage and retrieval of biological and analytical data generated during the 2020 GLHHFFTS and the ORD- Duluth 2020 Great Lakes special study. • Compiling data files for each target chemical or group of related target chemicals for statistical analysis and for public release. • Preparing summary project information and graphics for meeting materials, public outreach materials, and interim and final data reporting. This QAPP focuses on fish fillet sample analyses activities for the 2020 GLHHFFTS and ORD- Duluth 2020 Great Lakes special study, which involve the last 5 study components listed above. A7. Quality Objectives and Criteria The overall quality objective for the analysis of the 2020 GLHHFFTS and ORD-Duluth 2020 Great Lakes special study fish fillet tissue samples for mercury, PFAS, PCB congeners, PCBs as Aroclors, and fatty acids (2020 GLHHFFTS fillet samples only) is to obtain a complete set of data for each chemical or chemical group and to produce data of known and documented quality. Analytical completeness is defined as the percentage of valid samples collected in the study for which usable analytical results are produced. The goal for analytical completeness is 95% and it 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) ------- NCCA 2020 Great Lakes Fish Fillet Sample Analysis QAPP Revision 4 Date: November 19, 2021 Page 20 of 45 is calculated at the sample-analyte level, such that an issue with the quality of one analyte out of many does not invalidate the entire sample. OST specified the use of Method 163 IE (USEPA 2002) and its quality control acceptance criteria for analyses of 2020 GLHHFFTS and ORD-Duluth 2020 Great Lakes special study fish fillet tissue samples for mercury. The information describing the analytical method is provided in Section B4 of this QAPP. Data usability for each analysis will be assessed using QC criteria summarized in Section B5. Because EPA has not formally validated methods for PFAS analyses of fish tissue samples, the laboratory selected for this work proposed the analytical procedures and quality control acceptance criteria that they would use for these analyses. The information describing the fish tissue and rinsate analytical methods is provided in Section B4 of this QAPP. Data usability for each analysis will be assessed using QC criteria summarized in Section B5. OST specified the use of Method 1668C (USEPA 2010) and its quality control acceptance criteria for analyses of 2020 GLHHFFTS and ORD-Duluth 2020 Great Lakes special study fish fillet tissue samples for PCB congeners. The information describing the analytical method is provided in Section B4 of this QAPP. Data usability for each analysis will be assessed using QC criteria summarized in Section B5. There are few EPA methods for the analysis of Aroclors in fish tissue. OST had employed EPA Method 1656A in the National Lake Fish Tissue Study because that method was available through an existing laboratory contracting program in the Engineering and Analysis Division (EAD). By the time of the current study, no commercial laboratories were offering that method for analysis of fish tissue samples, so OST reviewed and accepted a series of sample preparation, extraction, cleanup, and determinative procedures from the SW-846 methods manual proposed by the laboratory that will be performing those analyses. The information describing these methods is provided in Section B4 of this QAPP. Data usability for each analysis will be assessed using QC criteria summarized in Section B5. Because they are not environmental contaminants, EPA has not developed methods for fatty acid analyses of fish tissue samples. Therefore, the laboratory selected for this work proposed the analytical procedures and quality control acceptance criteria that they would use for these analyses. The information describing their fish tissue analytical method is provided in Section B4 of this QAPP. Data usability for each analysis will be assessed using QC criteria summarized in Section B5. A8. Special Training/Certification All laboratory staff involved in the analyses of fish tissue samples (and of rinsate samples, which applies to PFAS analyses only) must be proficient in the associated tasks, as required by each analytical laboratory's existing quality system. All contractor staff involved in analytical data review and assessment will be proficient in data review, and no specialized training is required for data reviewers for this project. ------- NCCA 2020 Great Lakes Fish Fillet Sample Analysis QAPP Revision 4 Date: November 19, 2021 Page 21 of 45 A9. Documents and Records The Statements of Work (SOWs) for the analytical subcontracts provide the specific requirements for laboratory deliverables. The major points are summarized below: • The laboratory must provide reports of all results required from analyses of environmental and QC samples. • Summary level data must be submitted in electronic format and must include the following information: EPA sample number, analyte name and CAS number, laboratory sample ID, measured amount, reporting units, sample preparation date, and analytical batch ID (if applicable). • The laboratory shall provide raw data in the form of direct instrument readouts with each data package. Raw data include: Copy of traffic report, chain-of-custody records, or other shipping information Instrument readouts and quantitation reports for analysis of each sample, blank, standard and QC sample, and all manual worksheets pertaining to sample or QC data or the calculations thereof Copies of bench notes, including preparation of standards and instrumental analyses The laboratories will maintain records and documentation associated with these analyses for a minimum of three years after completion of the study. Additional copies will be maintained by CSRA for at least five years after completion of the study, and they will be transferred to EPA on request. 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 sample collection goal is to collect enough specimens to provide a composite sample consisting of a minimum of 75 grams of fillet tissue from each NCCA 2020 Great Lakes human health whole fish sampling site. 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 Fish Fillet Sample Analysis QAPP Revision 4 Date: November 19, 2021 Page 22 of 45 • Species-specific information on fatty acid content of Great Lakes fish that are commonly targeted by fishermen and consumed by humans. Fatty acid analyses are being limited to the 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. 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, PCB congeners, PCBs as Aroclors, lipids, and fatty acids. (Analyses of fillet samples from Lake Michigan enhancement sites will not include PCBs as Aroclors or fatty acids. The lipid analyses of all samples are being conducted as part of the fillet sample preparation under USEPA 2020c.) 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 the required amount of fillet tissue for analysis (USEPA 2020c). 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 in 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 provides a minimum of 75 grams of fillet tissue for chemical analysis. Field crews collected the majority of human health fish samples between June and September during the 2020 field season, but they require additional time in 2021 to complete fish sampling due to logistical constraints imposed on field crews in 2020 because of the coronavirus pandemic. ------- NCCA 2020 Great Lakes Fish Fillet Sample Analysis QAPP Revision 4 Date: November 19, 2021 Page 23 of 45 B2. Fish Sampling and Fillet Sample Preparation Methods B2.1 Fish 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 primary (priority) target fish species and 18 secondary alternative fish species (see Table 1). 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. In preparing Great Lakes human health whole 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. Table 1. 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 Secondary Fish Species Scientific Name*3 Secondary Fish Species Common Name Ambloplites rupestris Rock bass Carpiodes cyprinus Quillback Micropterus dolomieu Smallmouth bass Catostomus catostomus Longnose sucker Micropterus salmoides Largemouth bass Catostomus commersonii White sucker Pomoxis annularis White crappie Hypentelium nigracans Northern hogsucker Pomoxis nigromaculatus Black crappie Ictiobus cyprinellus Bigmouth buffalo Cyprinus carpio Common carp Ictiobus niger Black buffalo Esox lucius Northern pike Lepomis cyanellus Green Sunfish Esox masquinongy Muskellunge Lepomis gibbosus Pumpkinseed Esox niger Chain pickerel Lepomis gulosus Warmouth Ictalurus punctatus Channel catfish Lepomis macrochirus Bluegill Lota lota Burbot Lepomis megalotis Longear Sunfish Morone americana White perch Ameiurus me las Black bullhead ------- NCCA 2020 Great Lakes Fish Fillet Sample Analysis QAPP Revision 4 Date: November 19, 2021 Page 24 of 45 Table 1. 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 Secondary Fish Species Scientific Name*3 Secondary Fish Species Common Name Morone chrysops White bass Ameiurus natalis Yellow bullhead Perca flavescens Yellow perch Ameiurus nebulosus Brown bullhead Sander canadensis Sauger Coregonus artedi Cisco/ lake herring Sander vitreus Walleye Coregonus hoyi Bloater Coregonus clupeaformis Lake whitefish Prosopium cylindraceum Round whitefish Oncorhynchus gorbuscha Pink salmon Salvelinus fontinalis Brook trout Oncorhynchus kisutch Coho salmon * Minimum acceptable length is 190 mm, TL Oncorhynchus tshawytscha Chinook salmon a Only send if preferred species are not available Oncorhynchus mykiss Rainbow trout a Only send if preferred species are not available Salmo salar Atlantic salmon Salmo trutta Brown trout Salvelinus namaycush Lake trout Aplodinotus grunniens Freshwater drum * Minimum acceptable length is 190 mm, TL 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. B2.2 Fillet Sample Preparation 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 and all of the sample preparation methods described here are governed by a separate QAPP (USEPA 2020c). Prior to initiating fish sample preparation, Tetra Tech coordinates with CSRA for transfer of NCCA 2020 GLHHFFTS and ORD-Duluth 2020 whole fish samples from Microbac Labs (Baltimore, Maryland) to the Tetra Tech lab, where a sample custodian checks in the whole fish samples before storing them in a freezer at a temperature of < -20° Celsius (C). Whole fish sample check-in procedures involve (1) verifying that all associated paperwork stored with the samples is complete, legible, and accurate, (2) comparing the information on the label for each fish specimen to the fish sample preparation batch spreadsheet containing fish sample processing instructions, (3) reporting problems involving sample paperwork, sample integrity, or fish sample label and processing instruction information inconsistencies to the Fish Sample Preparation Technical Leader and the OST Project Manager via email, and (4) coordinating with the Fish Sample Preparation Technical Leader and OST Project Manager to resolve any discrepancies before beginning fish sample processing. Fish Sample Preparation Batches Each NCCA 2020 Great Lakes human health fish sample preparation batch generally consists of 20 whole fish samples. Whole fish samples from the 2020 GLHHFFTS and the ORD-Duluth ------- NCCA 2020 Great Lakes Fish Fillet Sample Analysis QAPP Revision 4 Date: November 19, 2021 Page 25 of 45 2020 Great Lakes special study are assigned to separate batches (labeled Batch 1, Batch 2, etc. for the 2020 GLHHFFTS and Batch ORD1, Batch ORD2, etc., for the ORD-Duluth 2020 Great Lakes special study). The number of whole fish samples in the final fish sample preparation batch (or two) for each of these series 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 for assignment to a batch. Tetra Tech staff develop fish sample preparation instructions that include all valid fish samples available for processing. Processing may not begin until the Fish Sample Preparation Technical Leader and the OST Project Manager review the draft instructions and the Fish Sample Preparation Technical Leader approves the final instructions and batch assignments and the OST Project Manager concurs with the approvals. Homogenized Fillet Sample Preparation The homogenized fillet sample preparation process begins with 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 separately 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 of the NCCA 2020 Great Lakes Human Health Fish Sample Preparation QAPP (USEPA 2020c). B3. Sample Receipt and Inspection This section describes the sample receipt and inspection procedures that apply to the shipment of 2020 NCCA Great Lakes human health homogenized fillet tissue samples to the analytical laboratories selected for analysis of these samples. In coordination with CSRA, Tetra Tech staff initiate packing and shipping the 2020 NCCA Great Lakes human health homogenized fillet tissue samples from their fish sample preparation laboratory in Owings Mills, Maryland, to the analytical laboratories designated for analysis of these fillet samples, following procedures described in Appendix B of the NCCA 2020 Great Lakes Human Health Fish Sample Preparation QAPP (USEPA 2020c). CSRA staff prepare sample tracking paperwork that is included in each shipment, notify the laboratories in advance of each shipment, track the progress of each shipment, and identify and resolve any delays that arise during shipment of the fillet samples. When coolers are received at each analytical laboratory, the fillet tissue samples are inspected for damage, logged into the laboratory, and placed into freezers immediately after the laboratory measures and records the temperature of each cooler. Homogenized fillet tissue samples are ------- NCCA 2020 Great Lakes Fish Fillet Sample Analysis QAPP Revision 4 Date: November 19, 2021 Page 26 of 45 stored frozen at < -20° C until analyzed. Because the samples are shipped frozen, typical temperature blanks consisting of a bottle of water are not practical (they may break due to expansion), so they are not required. The laboratory measures and records the temperature of the coolers containing fillet samples on receipt using an infrared temperature sensor or other suitable device. Each laboratory notifies the CSRA Project Leader about the receipt of the fillet tissue samples by email, and the CSRA Project Leader advises the OST Project Manager of fillet sample receipt on the day of delivery. Any questions from the analytical laboratory regarding sample paperwork or sample condition are sent to CSRA and routed to OST or Tetra Tech, as appropriate, before CSRA sends the answers back to the laboratory. B4. Analytical Methods B4.1 Mercury Analysis of Fillet Tissue ALS Environmental Lab prepares (a process involving tissue digestion and oxidation prior to tissue analysis) and analyzes fillet tissue samples using Procedure I from "Appendix to Method 1631, Total Mercury in Tissue, Sludge, Sediment, and Soil by Acid Digestion and BrCl Oxidation" from Revision B of Method 1631 (163 IB) for sample preparation (USEPA 2001b), and Revision E of Method 1631 (163 IE) for the analysis of mercury in fish tissue samples (USEPA 2002). This method requires approximately 1 g of tissue for the analysis. The sample is digested with a combination of nitric and sulfuric acids. The mercury in the sample is oxidized with bromine monochloride (BrCl) and analyzed by cold-vapor atomic fluorescence spectrometry. Tissue sample results are reported based on the wet weight of the tissue sample, in nanograms per gram (ng/g). The mercury method detection limit (MDL) and Minimum Level (quantitation limit) are listed in Appendix B. B4.2 PFAS Analysis of Fillet Tissue and Rinsate Samples There are no formal analytical methods from EPA or any voluntary consensus standards bodies for the PFAS analyses of tissue samples. Therefore, fish tissue samples will be analyzed by SGS-AXYS Analytical Services, Ltd. (Sidney, BC, Canada), using procedures developed, tested, and documented in that laboratory. CSRA reviewed the SOP during the solicitation process and will maintain a copy of the SGS-AXYS SOP on file that will be made available to EPA for review on request. The analytical procedures are briefly described below, based on information in the SOP. The 40 target PFAS analytes are shown in Appendix B. The concentration of each PFAS analyte is determined using the responses from one of the 13C- or Deuterium-labeled standards added prior to sample extraction, applying the technique known as isotope dilution. As a result, all of the target analyte concentrations are corrected for the recovery of the labeled standards, thus accounting for extraction efficiencies and losses during cleanup. ------- NCCA 2020 Great Lakes Fish Fillet Sample Analysis QAPP Revision 4 Date: November 19, 2021 Page 27 of 45 Approximately 2 g of fish tissue are required for analysis. (If matrix-related analytical problems are identified during the analysis of a given fish tissue sample, a sample aliquot of 1 g may be used to minimize those problems.) The sample is spiked with 24 isotopically labeled standards and extracted by shaking the tissue in a caustic solution of methanol, water, potassium hydroxide, and acetonitrile. The hydroxide solution breaks down the tissue and allows the PFAS analytes to be extracted into the solution. After extraction, the solution is centrifuged to remove the solids, and the supernatant liquid is diluted with reagent water and processed by solid-phase extraction (SPE) on a weak anion exchange sorbent. The PFAS analytes are eluted from the SPE cartridge, and the eluant is spiked with additional labeled recovery standards and analyzed by high performance liquid chromatography with tandem mass spectrometry. The aqueous rinsate samples will be analyzed for the 40 PFAS analytes using the same isotope dilution procedure as used for the fish tissue samples, but with an extraction step based on EPA Method 537 from the Office of Ground Water and Drinking Water (USEPA 2009). The 250-mL aqueous rinsate sample is spiked with the labeled standards and processed by SPE, in a similar manner as is used for the tissue samples. The PFAS analytes are eluted from the SPE cartridge and the eluant is spiked with additional labeled recovery standards and analyzed by high performance liquid chromatography with tandem mass spectrometry. Tissue sample results are reported based on the wet weight of the tissue sample, in nanograms per gram (ng/g). Method detection limits and Minimum Levels (quantitation limits) for PFAS analytes are listed in Appendix B. Aqueous rinsate results are reported based on the volume of the rinsate sample, in nanograms per liter (ng/L). B4.3 PCB Congener Analysis of Fillet Tissue Fish tissue samples are being prepared and analyzed by Vista Analytical Laboratory (El Dorado Hills, California), in general accordance with Revision C of EPA Method 1668, Chlorinated Biphenyl Congeners in Water, Soil, Sediment, Biosolids, and Tissue by HRGC/HRMS (USEPA 2010). The samples are being analyzed for all 209 PCB congeners and reported as either individual congeners or coeluting groups of congeners. The following method modifications have been reviewed, found to be within the allowance for flexibility in Section 9.1.2 of Method 1668C, supported by performance data that are maintained on file at the laboratory, and have been approved for use in this study: • Section 7.6.4: Vista uses sodium sulfate as the reference matrix for QC samples associated with tissue analyses rather than vegetable oil because they have not found a source of vegetable oil that did not have traces of PCBs in it. • Sections 7.10.1 and 15.4.2.1: Vista uses a CS-3 (mid-level calibration) standard that contains all 209 of the PCB congeners, rather than the subset of congeners listed in the method. Therefore, they do not run a separate standard containing all 209 congeners during the calibration verification process in Section 15.4.2.1. • Section 12.5: Vista uses sodium hydroxide to adjust the pH of the solution in the back- extraction procedure, rather than potassium hydroxide. ------- NCCA 2020 Great Lakes Fish Fillet Sample Analysis QAPP Revision 4 Date: November 19, 2021 Page 28 of 45 • Table 3: Vista adds 44 13C-labeled compounds to each sample, 17 more than the 27 labeled compounds specified in the method, and monitors the recoveries of all of these standards in each sample. Note: Given the large number of target analytes involved, the final list of PCB congeners and coelutions is provided in Appendix B of this QAPP, along with their MDLs and MLs. Tissue sample results are reported based on the wet weight of the tissue sample in units of picograms per gram (pg/g). B4.4 PCBs as Aroclors Analysis of Fillet Tissue Fish fillet tissue samples are being prepared and analyzed by Eurofins-TestAmerica (Pittsburgh, PA), using sample extraction, cleanup and determinative procedures from the SW-846 methods manual. The laboratory will extract approximately 10 g of tissue for the analysis. The samples are being analyzed for seven common Aroclor mixtures (i.e., 1016 to 1260), plus Aroclor 1268. The laboratory proposed and EPA accepted the following analytical scheme: • Extraction by Method 3541 (automated Soxhlet) • Cleanups by Methods 3660B (sulfur), 3665A (sulfuric acid-permanganate), and 3640A (gel-permeation chromatography) • Analysis by Method 8082A (gas chromatography with electron capture detection) Tissue sample results are reported based on the wet weight of the tissue sample in units of nanograms per gram (ng/g). Method detection limits and Minimum levels (quantitation limits) for each of the Aroclors are listed in Appendix B. B4.5 Fatty Acid Analysis of Fillet Tissue Because they are not environmental contaminants, there are no formal EPA methods for the analysis of fatty acids in any matrix. Therefore, fish tissue samples will be analyzed by Clarkson University (Potsdam, NY), using procedures developed, tested, and documented in that laboratory and currently employed under GLNPO Grant No. GL 00E02957. The laboratory's SOP was reviewed by CSRA during the solicitation process, along with the supporting materials. A copy of the Clarkson SOP will be maintained on file at CSRA and will be made available to EPA for review on request. The analytical procedures are briefly described below. The 38 target fatty acid analytes are shown in Appendix C. Approximately 2 g of homogenized fish tissue is spiked with a surrogate solution (nonadecacanoic acid, CI9:0) and extracted with 2:1 mixture of chloroform and methanol using ultrasonic extraction. The extract is centrifuged to separate the water from the chloroform, and concentrated to approximately 20 mL. A 10-|iL aliquot of the extract is transferred to a clean autosampler vial, purged for 30 seconds with nitrogen, capped, and then placed on the instrument for derivatization and injection. ------- NCCA 2020 Great Lakes Fish Fillet Sample Analysis QAPP Revision 4 Date: November 19, 2021 Page 29 of 45 The automated instrument adds 100 |iL of deuterated C18:0 (as an internal standard) and 250 |iL of 12% boron trifluoride (BF3) in methanol. The solution is mixed and heated to 70 °C for 50 minutes. After heating, 25 |iL of water is added to quench the derivatization reaction and the derivatized extract is mixed, followed by the addition of 0.65 mL of hexane and further mixing to separate the fatty acid methyl esters from the aqueous solution. An aliquot of the hexane extract is analyzed by gas chromatography, with flame ionization detection (GC/FID), using a 100 m x 250 [j,m x 0.2 [j,m HP-88 column. The concentration of each fatty acid is calculated based on a multi-point calibration curve and reported based on the wet weight of the tissue sample, in micrograms per gram (jug/g). Method detection limits and quantitation limits for the fatty acids are listed in Appendix B. B5. Analytical Quality Control The analytical procedures being applied by the laboratories designated for analysis of 2020 NCCA Great Lakes human health homogenized fillet tissue samples include many of the traditional EPA analytical quality control (QC) activities. For example, all samples are analyzed in batches and each batch includes: • up to 20 field samples and the associated QC samples • blanks - at least 5% of the samples within a batch are method blanks (with higher percentages specified in some analytical methods) Other common quality control activities vary by the analysis type. The QC activities associated with the chemical analyses of fillet samples for target chemicals are described in Subsection B5.1 for mercury, Subsection B5.2 for PFAS, Subsection B5.3 forPCB congeners, and Subsection B5.4 for PCBs as Aroclors. B5.1 Mercury Analysis QC Criteria Quality control samples associated with each batch of fillet tissue samples analyzed for mercury are summarized in Table 2 below. The cold-vapor atomic fluorescence instrument is calibrated daily, as described in Method 163 IE and the laboratory's SOP. At least five calibration standards and a blank are used for calibration, and the variability in the calibration factors for the five standards must have a relative standard deviation (RSD) less than or equal to 15%. The calibration is verified after every 20 samples by the analysis of the ongoing precision and recovery (OPR) standard, or the laboratory control sample (LCS). The results for the OPR/LCS standard must fall within the limits in Table 2. ------- NCCA 2020 Great Lakes Fish Fillet Sample Analysis QAPP Revision 4 Date: November 19, 2021 Page 30 of 45 Table 2. QC Samples and Acceptance Criteria for Mercury Analysis of Fish Tissue QC Operation Frequency* Acceptance Limit Corrective Action Bubbler blank or System blank (depending on instrument configuration) 3 blanks run during calibration and with each analytical batch of up to 20 field samples 50 picograms (pg) of mercury If the bubbler or system blank is above 50 pg, take corrective action to reduce the blank level to below 50 pg, and reanalyze any samples in the affected batch. OPR/LCS Prepared once per batch of up to 20 field samples, analyzed once prior to the analysis of any field samples, and again at the end of each analytical batch, spiked at 4.0 ng 70 - 130% recovery (5.6-10.4 ng/g) If the OPR recovery is not within the QC acceptance limits, • take corrective action and repeat the OPR analysis, beginning with a fresh aliquot, reanalyze all samples in the affected analytical batch. Method blank 3 method blanks per batch of up to 20 field samples, with analyses interspersed among the samples in the analysis batch 0.4 nanograms (ng) (400 pg) of mercury, or Less than one tenth the concentration of an associated sample If any of the three method blank results is above 0.4 nanograms, • take corrective action to reduce the blank level to below 0.4 ng, • reanalyze any samples in the affected batch with results less than 10 times the observed results for any of the three blanks, and • flag sample results greater than 10 times the observed blank level to advise the data user of the potential contamination. QC sample Once per batch of up to 20 field samples Per the provider of the QCS or 75 - 125% recovery if no criteria provided by the supplier If the QCS results are not within the provider's acceptance limits, • take corrective action and repeat the QCS analysis, beginning with a fresh aliquot, • reanalyze all samples in the affected analytical batch. MS/MSD Once per every 10 field samples (e.g., twice per 20 samples in a preparation batch) See note below this table regarding spiking levels and the use of a sample from a previous analysis batch for preparation of the MS/MSD aliquots. 70 - 130%) recovery and RPD < 30% If either the MS or MSD recovery is not within the QC acceptance limits, • take corrective action and repeat the MS/MSD analysis, beginning with fresh aliquots, • reanalyze all samples in the affected analytical batch. If the RPD exceeds the acceptance limit, the laboratory will reanalyze the MS/MSD samples: • If the reanalysis results meet the RPD limit, then the laboratory will reanalyze all of the associated field and QC samples. * The term "field sample" refers to homogenized fillet tissue samples provided to the analytical laboratory for mercury analysis. Note: Provision of useful MS/MSD data is highly dependent on selection of an appropriate spiking level relative to the background concentration of mercury in the unspiked sample. After the first batch of samples, the MS/MSD sample may be prepared from excess volume of tissue from a sample in the previous batch, such that the background level is known. Spiking should be performed at approximately 3 to 5 times the background concentration. B5.2 PFAS Analysis QC Criteria The high performance liquid chromatograph/tandem mass spectrometer is calibrated as described in the laboratory's SOP. Seven calibration standards are used for calibration, using a weighted linear regression. The correlative coefficient for the regression must be > 0.95. The calibration is verified every 12 hours through the analysis of the calibration verification standard. The results for the calibration verification must meet the requirements in Appendix C of this QAPP. ------- NCCA 2020 Great Lakes Fish Fillet Sample Analysis QAPP Revision 4 Date: November 19, 2021 Page 31 of 45 Quality control samples associated with each batch of tissue samples or rinsate samples analyzed for PFAS are summarized in Table 3 below. Table 3. QC Samples and Acceptance Criteria for PFAS Analysis of Tissue and Rinsates QC Operation Frequency* Acceptance Limit Corrective Action Labeled compounds Spiked into every sample before extraction Per Appendix C of this QAPP Evaluate failure and impact on samples. If sample results are non-detects for analytes which have a high labeled compound recovery, report non-detect results with case narrative comment. For detected analytes with low labeled compound recovery, extract and analyze a smaller sample aliquot. Calibration Verification Every 12 hours, before sample analysis. Per Appendix C of this QAPP • Evaluate failure and impact on samples. If sample results are non-detects for analytes which have a high bias, report non- detect results with case narrative comment. or Immediately analyze two additional consecutive verification standards. If both pass, samples may be reported without reanalysis. If either fails, take corrective action(s) and re- calibrate; then reanalyze all affected samples since the last acceptable verification standard. Lab Control Sample (LCS) Once per batch of up to 20 field samples Per Appendix C of this QAPP • Reanalyze LCS once. If acceptable, report. Evaluate samples for detections, and LCS for high bias. If LCS has high bias, and sample results are non-detects, report with case narrative comment. If LCS has low bias, or if there are detected analytes with failures, evaluate and reprepare and reanalyze the LCS and all samples in the associated prep batch for failed analytes. Method blank Once per batch of up to 20 field samples Less than or equal to the MDLs in Appendix B of this QAPP All results, including blanks, are reported down to the method detection limit (MDL). • If the method blank result for any PFAS is above the MDL, but below the laboratory's nominal quantitation limit, the laboratory will flag all associated tissue sample and rinsate results as having a detectable method blank for that analyte. (Subsequent validation of the results by EPA or its contractors will evaluate the potential contribution of the blank to such sample results.) • If the method blank result is above the quantitation limit, the laboratory will reanalyze the method blank. - If the method blank reanalysis result is below the quantitation limit, then the laboratory will reanalyze all of the associated tissue or rinsate samples and QC samples. - If the method blank reanalysis result is still above the quantitation limit, then the laboratory will re-extract and reanalyze all tissue or rinsate samples with original results above the MDL. Laboratory duplicate Once per batch of up to 20 field samples The relative percent difference (RPD) of the duplicate measurements must be < 40% Evaluate the data, and re-extract and reanalyze the original sample and duplicate: • If the reanalysis results meet the RPD limit, then the laboratory will reanalyze all of the associated field and QC samples. • If the reanalysis result still does not meet the RPD limit, then the laboratory will re-extract and reanalyze all field samples with original results above the MDL. * The term "field sample" refers to homogenized fillet tissue samples provided to the analytical laboratory for PFAS analysis. B5.3 PCB Congener Analysis QC Criteria The high-resolution gas chromatograph/high-resolution mass spectrometer (HRGC/HRMS) is calibrated periodically as described in Method 1668C and the laboratory's SOP. At least five calibration standards are used for calibration, and the variability in the response factors for the five standards must have a relative standard deviation (RSD) less than or equal to 20%. The ------- NCCA 2020 Great Lakes Fish Fillet Sample Analysis QAPP Revision 4 Date: November 19, 2021 Page 32 of 45 calibration is verified every 12 hours by the analysis of the calibration verification (VER) standard. The results for the VER must meet the requirements in Appendix D. Quality control samples associated with each batch of tissue samples analyzed for PCBs are summarized in Table 4, below, and are based on the QC requirements of Method 1668C, with the project-specific addition of one laboratory duplicate sample per batch. Table 4. QC Samples and Acceptance Criteria for PCB Analysis of Fish Tissue QC Sample Frequency* Acceptance Limit Corrective Action Laboratory control sample One per sample batch Per Appendix D Per Method 1668C Calibration verification (VER) At the beginning of every 12-h analytical shift Per Appendix D Per Method 1668C Method blank Once per batch of up to 20 field samples 5x MDL for each congener (As noted elsewhere, all results, including blanks, are reported down to the MDL.) If the method blank result is above 5x MDL, the laboratory will reanalyze the method blank extract to confirm the presence of the blank contaminants. If the reanalysis result is still above 5x MDL, then the laboratory will compare the results in the method blank to the results in all of the associated field samples in the batch and take corrective action as follows: • If the result for a congener (or group of coeluting congeners) that is present in the method blank at 5x MDL or higher is not present in the field sample, then the result for that field sample may be reported without corrective actions. The result must be flagged with a "B" flag that indicates the presence of the analyte in the associated blank and the data package narrative must discuss the comparison of the blank and sample results for that sample. • If the result for the congener in the field sample is more than 10 times the level found in the method blank, then the result for that field sample also may be reported without corrective actions. The result must be flagged with a "B" flag that indicates the presence of the analyte in the associated blank and the data package narrative must discuss the comparison of the blank and sample results for that sample. • If the result for the congener in the field sample is less than or equal to 10 times the level found in the method blank, then re-extraction and reanalysis of the affected sample is required (but not samples that meet the conditions in #1 and #2 above) in conjunction with a new method blank and all other method-specified QC samples. CSRA will work with the laboratory to schedule any required reanalyses in a manner that does not delay analyses of subsequent batches of field samples. • If the results of the re-extraction and reanalysis of the field sample do not resolve the problem, i.e., the background levels in the method blank are still a concern, CSRA will require that the laboratory provide information on historical levels of blank contaminants for similar matrices. CSRA and EPA will evaluate those historical results and the reanalysis results on a case-by-case basis to determine if there is a pattern of blank contamination that is indicative of a broader problem and if any further corrective actions are required by the laboratory. ------- NCCA 2020 Great Lakes Fish Fillet Sample Analysis QAPP Revision 4 Date: November 19, 2021 Page 33 of 45 Table 4. QC Samples and Acceptance Criteria for PCB Analysis of Fish Tissue QC Sample Frequency* Acceptance Limit Corrective Action Laboratory Once per TheRPDofthe If the RPD exceeds the acceptance limit, the laboratory will duplicate batch of up to duplicate reanalyze the laboratory duplicate extract: 20 field measurements must samples be: • If the reanalysis result meets the RPD limit, then the laboratory will • < 50% for sample reanalyze all of the associated field and QC samples. concentrations greater than or • If the reanalysis result still does not meet the RPD limit, then the equal to 5 times the laboratory will re-extract and reanalyze all field samples with MDL, and original results above the MDL. • <100% for sample concentrations less than 5 times the MDL. (When comparing the sample concentration to the MDL, use the lower of the two concentrations in the paired samples.) * The term "field sample" refers to homogenized fillet tissue samples provided to the analytical laboratory for PCB congener analysis. B5.4 PCBs as Aroclors Analysis QC Criteria The gas chromatograph is calibrated periodically as described in Method 8082A and the laboratory's SOP. At least five calibration standards are used for calibration, and the variability in the response factors for the five standards must have a relative standard deviation (RSD) less than or equal to 20%. The calibration is verified at least once every 12-hour shift during which analyses are performed by the analysis of the calibration verification standard. The results for the calibration verification must meet the requirements in Table 5. Quality control samples associated with each batch of fillet tissue samples analyzed for Aroclors are summarized in Table 5 below. Table 5. QC Samples and Acceptance Criteria for Aroclor Analysis of Fish Tissue QC Operation Frequency* Acceptance Limit Corrective Action Lab Control Sample - Using Aroclor 1016 and 1260 Once per batch of up to 20 field samples 34-118% Reanalyze LCS once. If acceptable, report. Evaluate samples for detections, and LCS for high bias. If LCS has high bias, and samples non-detect, report with case narrative comment. If LCS has low bias, or if there are detections, evaluate and re-prepare and reanalyze the LCS and all samples in the associated prep batch for failed analytes Calibration Verification (CV) - Using Aroclor 1016 and 1260 Every 12 hours, before sample analysis. 80 - 120% Evaluate failure and impact on samples. If samples non-detect for analytes which have a high bias, report non-detect results with narrative comment. For analytes with low bias, or samples with detected analytes, use the approach below. Immediately analyze two additional consecutive CVs. If both pass, samples may be reported without reanalysis. If either fails, take corrective action(s) and re-calibrate; then reanalyze all affected samples since the last acceptable CV. ------- NCCA 2020 Great Lakes Fish Fillet Sample Analysis QAPP Revision 4 Date: November 19, 2021 Page 34 of 45 Table 5. QC Samples and Acceptance Criteria for Aroclor Analysis of Fish Tissue QC Operation Frequency* Acceptance Limit Corrective Action Method blank Once per batch of up to 20 field samples 3x MDL for each Aroclor (As noted elsewhere, all results, including blanks, are reported down to the MDL.) If the method blank result is above 3x MDL, the laboratory will reanalyze the method blank extract to confirm the presence of the blank contaminants. If the reanalysis result is still above 3x MDL, then the laboratory will compare the results in the method blank to the results in all of the associated field samples in the batch and take corrective action as follows: • If the result for an Aroclor that is present in the method blank at 3x MDL or higher is not present in the field sample, then the result for that field sample may be reported without corrective actions. The result must be flagged with a "B" flag that indicates the presence of the analyte in the associated blank and the data package narrative must discuss the comparison of the blank and sample results for that sample. • If the result for an Aroclor in the field sample is more than 10 times the level found in the method blank, then the result for that field sample also may be reported without corrective actions. The result must be flagged with a "B" flag that indicates the presence of the analyte in the associated blank and the data package narrative must discuss the comparison of the blank and sample results for that sample. • If the result for an Aroclor in the field sample is less than or equal to 10 times the level found in the method blank, then re-extraction and reanalysis of the affected sample is required (but not samples that meet the conditions in # 1 and #2 above) in conjunction with a new method blank and all other method-specified QC samples. CSRA will work with the laboratory to schedule any required reanalyses in a manner that does not delay analyses of subsequent batches of field samples. • If the results of the re-extraction and reanalysis of the field sample do not resolve the problem, i.e., the background levels in the method blank are still a concern, CSRA will require that the laboratory provide information on historical levels of blank contaminants for similar matrices. CSRA and EPA will evaluate those historical results and the reanalysis results on a case-by-case basis to determine if there is a pattern of blank contamination that is indicative of a broader problem and if any further corrective actions are required by the laboratory. MS/MSD - Using Aroclor 1016 and 1260 Once per batch of up to 20 field samples 34-118% recovery and RPD < 30% If either the MS or MSD recovery is not within the QC acceptance limits, • Take corrective action and repeat the MS/MSD analysis, beginning with fresh aliquots, • Reanalyze all samples in the affected analytical batch. If the RPD exceeds the acceptance limit, the laboratory will reanalyze the MS/MSD samples: • If the reanalysis results meet the RPD limit, then the laboratory will reanalyze all of the associated field and QC samples Surrogates Two surrogates added to each field and QC sample 20 - 125% Extract and analyze a smaller aliquot of tissue sample. • If the results of the re-extraction and reanalysis of the field sample resolve the problem, then report only the analytical results from the analysis with acceptable surrogate recovery. • Otherwise, report both sets of results and discuss in the narrative. * The term "field sample" refers to homogenized fillet tissue samples provided to the analytical laboratory for Aroclor analysis. ------- NCCA 2020 Great Lakes Fish Fillet Sample Analysis QAPP Revision 4 Date: November 19, 2021 Page 35 of 45 B5.5 Fatty Acid Analysis QC Criteria The gas chromatograph is calibrated periodically, as described in the laboratory's SOP. At least five calibration standards are used for calibration, and the variability in the response factors for the five standards must have a relative standard deviation (RSD) less than or equal to 20%. The calibration is verified at least once every 12-hour shift during which analyses are performed by the analysis of the calibration verification standard. The results for the calibration verification must meet the requirements in Table 6. Quality control samples associated with each batch of fillet tissue samples analyzed for fatty acids are summarized in Table 6 below. Table 6. QC Samples and Acceptance Criteria for Fatty Acid Analysis of Fish Tissue QC Operation Frequency* Acceptance Limit Corrective Action Method blank 1 per analysis batch (10 field samples, plus QC samples) Method detection limit (MDL) If any of the analytes are present in the method blank above the MDL, • take corrective action to reduce the blank level to below the MDL, • reanalyze any samples in the affected batch with results less than 10 times the observed results for the blank, and • flag sample results greater than 10 times the observed blank level to advise the data user of the potential contamination. Calibration linearity (5 points) Twice a year r2 > 0.95 Do not analyze samples until linear calibration is achieved Calibration verification Every 10 samples 70 to 130% recovery If the recovery of any analyte is outside the acceptance limits, recalibrate the instrument and reanalyze samples in the affected batch. Reference sample Analyze 1 aliquot of the Lake Superior reference tissue sample per analysis batch 50 to 150% of the certified value If the results are not within the acceptance limits, • take corrective action and repeat the reference sample analysis, beginning with a fresh aliquot, • reanalyze all samples in the affected analytical batch. Laboratory duplicate sample 1 per every two analysis batches (10 field samples, plus QC samples) RPD < 50% If the RPD exceeds the acceptance limit, the laboratory will reanalyze the duplicate sample: • If the reanalysis results meet the RPD limit, then the laboratory will reanalyze all of the associated field and QC samples. Surrogate Added to every field and QC sample 50 to 150%) recovery If the recovery of the surrogate is not within the acceptance criteria, reanalyze the affected samples. Internal Standard Added to every field and QC sample Not applicable Not applicable * The term "field sample" refers to homogenized fillet tissue samples provided to the analytical laboratory for fatty acid analysis. B6. Instrument/Equipment Testing, Inspection, and Maintenance All analytical instrumentation associated with the fillet tissue sample analyses will be inspected and maintained as described in the respective analysis methods and laboratory SOPs. B7. Instrument/Equipment Calibration and Frequency All analytical instrumentation associated with the fillet tissue sample analyses will be calibrated as described in the respective analysis methods. The mercury analysis method for tissue ------- NCCA 2020 Great Lakes Fish Fillet Sample Analysis QAPP Revision 4 Date: November 19, 2021 Page 36 of 45 samples, Method 163 IE, specifies calibration with at least five calibration standards and multiple blanks, as described in Section B5.1 above. The PFAS analysis procedures from SGS-AXYS for tissue and rinsate analyses specifies calibration with at least seven calibration standards, as described in Section B5.2 above. The PCB congener analysis method for tissue samples, Method 1668C, specifies calibration with at least five calibration standards (refer to Section B5.3 above). The Aroclor analysis method for tissue samples, Method 8082A, specifies calibration with at least five calibration standards (refer to Section B5.4 above). The fatty acid laboratory's SOP specifies calibration with at least five calibration standards (refer to Section B5.5 above). B8. Inspection/Acceptance of Supplies and Consumables The inspection and acceptance of any laboratory supplies and consumables associated with the fillet tissue sample analyses are addressed in the individual laboratory operating procedures to be used, and/or in the laboratory's existing overall quality system documentation. There are no additional requirements specific to this project, and therefore, none are described here. B9. Non-direct Measurements Non-direct measurements are not required for this project. (The analytical results from the previous fish tissue studies conducted under the NCCA (e.g., the 2010 NCCA and 2015 NCCA) to which any new data are to be compared are primary data that EPA generated under an approved QAPP for that study.) B10. Data Management Data management practices employed in this study will be based on standard data management practices used for EPA's National Lake Fish Tissue Study and other EPA fish contamination studies (e.g., NCCA 2015 GLHHFFTS). The data management (i.e., sample tracking, data tracking, data inspection, data quality assessment, database development) procedures have been regularly applied to other technical studies by CSRA. These procedures are being employed because they are effective, efficient, and have successfully withstood repeated internal and external audits, including internal review by EPA Quality Staff, public review and comment, judicial challenge, and an audit by the Government Accountability Office. These procedures, as implemented for the 2020 NCCA Great Lakes human health fish fillet indicator, are summarized below. • All laboratories performing analyses for this project are required to maintain all records and documentation associated with the analyses of the fish tissue samples for a minimum period of three years after completion of the study. • All required 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. • Each laboratory 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 ------- NCCA 2020 Great Lakes Fish Fillet Sample Analysis QAPP Revision 4 Date: November 19, 2021 Page 37 of 45 Accreditation Conference (NELAC). Each laboratory's data management plan is incorporated in its overall quality system documentation, e.g., its quality management plans, copies of which will be maintained on file at CSRA. C. ASSESSMENT AND OVERSIGHT CI. Assessments and Response Actions The laboratory contracts prepared to support analysis of Great Lakes human health homogenized fillet tissue samples for the 2020 GLHHFFTS and ORD-Duluth 2020 Great Lakes special study will stipulate that each laboratory has a comprehensive QA program in place and operating at all times during the performance of their contract, and that in performing laboratory work for this study, the laboratory shall adhere to the requirements of that QA program. These materials (ALS, 2020; SGS-AXYS, 2020; Vista, 2020; Eurofins-TestAmerica, 2020, and Clarkson, 2021) were reviewed by CSRA during the laboratory solicitations, as part of an assessment of laboratory capabilities. A copy of each QA plan will be maintained on file at CSRA and will be made available to EPA for review on request. Sections Cl.l through CI.6 describe other types of assessment activities and corresponding response actions identified to ensure that data gathering activities in the 2020 GLHHFFTS and ORD-Duluth 2020 Great Lakes special study are conducted as prescribed and that the performance criteria defined for this study are met. Cl.l Surveillance The CSRA Project Leader will schedule and track all analytical work performed by the laboratories designated for mercury, PFAS, PCB congener, Aroclor, and fatty acid analyses. The Project Leader will coordinate with Tetra Tech staff at the fish sample preparation laboratory regarding fillet tissue sample shipments to the analytical laboratory. When samples are shipped to the analytical laboratories for mercury, PFAS, PCB congener, Aroclor, or fatty acid analysis, the CSRA Project Leader will contact designated laboratory staff by email to notify them of the forthcoming shipment(s) and request that they contact CSRA on the scheduled day of delivery if the shipments do not arrive intact. Within 24 hours of scheduled sample receipt, CSRA will contact the laboratory to verify that the samples arrived in good condition, and if problems are noted, will work with the laboratory and EPA to resolve the problems as quickly as possible to minimize data integrity problems. The laboratory designated for mercury analysis of 2020 GLHHFFTS and ORD-Duluth 2020 Great Lakes special study fillet tissue samples will be permitted to work one batch ahead of the CSRA-EPA review of the QC results associated with the fillet tissue sample analyses. CSRA will also immediately notify the OST Project Manager of any mercury laboratory delays that are anticipated to impact EPA schedules. The laboratory designated for PFAS analysis of 2020 GLHHFFTS and ORD-Duluth 2020 Great Lakes special study fillet tissue samples will be permitted to work two batches ahead of the CSRA/EPA review of the QC results associated with the fillet tissue sample analyses. CSRA ------- NCCA 2020 Great Lakes Fish Fillet Sample Analysis QAPP Revision 4 Date: November 19, 2021 Page 38 of 45 will also immediately notify the OST Project Manager of any PFAS laboratory delays that are anticipated to impact EPA schedules. The laboratory designated for PCB congener analysis of 2020 GLHHFFTS and ORD-Duluth 2020 Great Lakes special study fillet tissue samples will be permitted to work two batches ahead of the CSRA/EPA review of the QC results associated with the fillet tissue sample analyses. CSRA will also immediately notify the OST Project Manager of any PCB congener analysis laboratory delays that are anticipated to impact EPA schedules. The laboratory designated for Aroclor analysis of 2020 GLHHFFTS fillet tissue samples will be permitted to work two batches ahead of the CSRA/EPA review of the QC results associated with the fillet tissue sample analyses. CSRA will also immediately notify the OST Project Manager of any Aroclor analysis laboratory delays that are anticipated to impact EPA schedules. The laboratory designated for fatty acid analysis of 2020 GLHHFFTS fillet tissue samples will be permitted to work two batches ahead of the CSRA/EPA review of the QC results associated with the fillet tissue sample analyses. CSRA will also immediately notify the OST Project Manager of any fatty acid analysis laboratory delays that are anticipated to impact EPA schedules. Finally, the CSRA Project Leader will monitor the progress of the data quality audits (data reviews) and database development to ensure that the laboratory data submission is reviewed in a timely manner. In the event that dedicated staff are not able to meet EPA schedules, CSRA will identify additional staff who are qualified and capable of reviewing the data in a timely manner. If such resources cannot be identified, and if training new employees is not feasible, CSRA will meet with the OST Project Manager to discuss an appropriate solution. CI.2 Product Review Product reviews for validated analytical data packages will be performed within CSRA to verify that the CSRA data reviews are being performed consistently over time and across data reviewers, that the review findings are technically correct, and that the reviews are being performed in accordance with this QAPP. Product reviewers will be charged with evaluating the completeness of the original CSRA data review, the technical accuracy of the reviewer's findings, and the technical accuracy of the separate analytical databases developed to store results associated with the 2020 GLHHFFTS and ORD-Duluth 2020 Great Lakes special study data packages, respectively. Product reviews will be conducted on at least 10% of the data packages. Qualified product reviewers will include any staff members that have been trained in CSRA data review procedures, are experienced in reviewing data similar to those being reviewed and are familiar with the requirements of this QAPP. To ensure the findings of each data review are documented in a consistent and technically accurate manner, CSRA staff will review 100% of the data qualifier flags entered into each project database. The 2020 GLHHFFTS and ORD-Duluth 2020 Great Lakes special study data files prepared by CSRA for statistical analysis of the data will be reviewed internally by CSRA staff and independently by the OST Project Manager with support from Tetra Tech. ------- NCCA 2020 Great Lakes Fish Fillet Sample Analysis QAPP Revision 4 Date: November 19, 2021 Page 39 of 45 C1.3 Quality Systems Audit A quality system audit (QSA) is used to verify, by examination and evaluations of objective evidence, that applicable elements of the quality system are appropriate and have been developed, documented, and effectively implemented in accordance and in conjunction with specified requirements. The focus of these assessments is on the quality system processes - not on evaluating the quality of specific products or judging the quality of environmental data or the performance of personnel or programs. The SHPD QA Coordinator may perform a QSA of the 2020 GLHHFFTS Fish Tissue Study mercury, PFAS, PCB congener, Aroclor, or fatty acid analyses. CI.4 Readiness Review A readiness review of each analysis laboratory's capability to produce acceptable sample results begins with a review of materials submitted by the laboratory during the solicitation process and continues during a kick-off conference call with each laboratory (ALS Environmental for mercury, SGS-AXYS for PFAS, Vista Analytical Laboratory for PCB congeners, Eurofins- TestAmerica for Aroclors, and Clarkson University for fatty acids). The requested materials include information about the laboratory's capacity, past experience with tissue analyses, and accreditations or certifications for mercury, PFAS, PCB congener, Aroclor, or fatty acid analyses in tissue and other matrices. These materials are reviewed during the solicitation process to assess the laboratory's competency and will be kept on file by CSRA. Readiness reviews are performed by CSRA data reviewers. If problems are identified during these reviews, CSRA staff will work with the laboratory, to the extent possible, to resolve the problem prior to awarding an analysis contract. If the problem cannot be resolved within the time frame required by EPA, the CSRA Project Leader will notify the OST Project Manager immediately. Records of these reviews and any corrective actions are maintained by CSRA separate from the analytical results for the field samples. CSRA staff will document their findings and recommendations concerning the readiness review as part of a written analytical QA report to EPA. CI.5 Technical Systems Audit The laboratory contracts will require that the laboratory be prepared for and willing to undergo an on-site audit or technical systems audit of its facilities, equipment, staff, sample processing, tissue sample analysis, training, record keeping, data validation, data management, and data reporting procedures. An audit will be conducted only if the results of the readiness reviews, data quality audits, and surveillance suggest serious or chronic laboratory problems that warrant on-site examinations and discussion with laboratory personnel. If such an audit is determined to be necessary, a standardized audit checklist may be used to facilitate an audit walkthrough and document audit findings. Audit participants may include the OST Project Manager and/or the SHPD QA Coordinator (or a qualified EPA staff member designated by the OST QA Officer) and a CSRA staff member experienced in conducting laboratory audits. One audit team member will be responsible for leading the audit and conducting a post-audit debriefing to convey significant findings to laboratory staff at the ------- NCCA 2020 Great Lakes Fish Fillet Sample Analysis QAPP Revision 4 Date: November 19, 2021 Page 40 of 45 conclusion of the audit. Another audit team member will be responsible for gathering pre-audit documentation of problems that necessitated the audit, customizing the audit checklist as necessary to ensure that those problems are addressed during the audit, documenting audit findings on the audit checklist during the audit, and drafting a formal report of audit findings for review by EPA. CI.6 Data Quality Assessment Upon completion of data verification and validation procedures (see Section Dl), CSRA staff will create an analytical database that contains all fillet tissue and QC sample results from the 2020 GLHHFFTS and a separate analytical database that contains all fillet tissue and QC sample results from the ORD-Duluth 2020 Great Lakes special study. At selected intervals and upon completion of the study, the CSRA Project Leader will perform analyses to verify the accuracy of each database. The procedures will be directed at evaluating the overall quality of each database against data quality objectives established for the respective studies and in identifying trends in fillet tissue sample results derived from field samples and QC results obtained during each of the studies. CSRA staff will document their findings and recommendations concerning this data quality assessment and provide them to EPA. C2. Reports to Management CSRA will track the receipt of data submissions for the homogenized fish fillet tissue analyses (and aqueous sample analyses for PFAS only) and advise the OST Project Manager of progress on a monthly basis. Following data verification and validation of all project-specific analytical data, CSRA will apply data qualifier flags, where needed, to the fillet tissue results in each project database that describe data quality limitations and recommendations concerning data use. The data qualifier flags are based on those developed for the National Lake Fish Tissue Study and the complete list of qualifier flags used and their implications for data use will be summarized in a report to EPA at or near the end of the data assessment process. The CSRA Project Leader will provide a monthly report to the OST Project Manager that describes the status of all current analysis and data review activities, during each month in which analyses and data review are conducted. D. DATA VALIDATION AND USABILITY This QAPP addresses the generation of data from homogenized fish fillet tissue samples prepared from 2020 Great Lakes human health fish samples (and from aqueous QC samples for PFAS analyses only). Sections Dl, D2, and D3 of this QAPP apply to all of the analytical data generation for the 2020 GLHHFFTS and the ORD-Duluth 2020 Great Lakes special study. ------- NCCA 2020 Great Lakes Fish Fillet Sample Analysis QAPP Revision 4 Date: November 19, 2021 Page 41 of 45 Dl. Data Review, Verification, and Validation The data review, verification, and validation aspects of the homogenized fish fillet tissue analyses and aqueous QC sample analyses are described below for all of the analytical data generated for the 2020 GLHHFFTS and the ORD-Duluth 2020 Great Lakes special study. Dl.l Data Review All laboratory results and calculations will be reviewed by the Laboratory Manager prior to data submission. Any errors identified during this peer review will be returned to the analyst for correction prior to submission of the data package. Following correction of the errors, the Laboratory Manager will verify that the final package is complete and compliant with the contract, and will sign each data submission to certify that the package was reviewed and determined to be in compliance with the terms and conditions of the contract. D1.2 Data Verification The basic goal of data verification is to ensure that project participants know what data were produced, if they are complete, if they are contractually compliant, and the extent to which they meet the objectives of the NCCA 2020 Great Lakes human health fish tissue studies. Every laboratory data package submitted for these studies (2020 GLHHFFTS and ORD-Duluth 2020 Great Lakes special study) will be subjected to data verification by qualified CSRA staff who have been trained in procedures for verifying data and who are familiar with the laboratory methods used to analyze the samples. This includes all of the mercury, PFAS, PCB congener, Aroclor, and fatty acid data generated under this QAPP and any subsequent QAPP revisions. The verification process is designed to identify and correct data deficiencies as early as possible in order to maximize the amount of usable data generated during the studies. The CSRA Project Leader will verify the summary level results for these analytical data, determine if they meet the project objectives in this QAPP, and report the verification findings to OST. D1.3 Data Validation Data validation is the process of evaluating the quality of the results relative to their intended use. Data need not be "perfect" to be usable for a particular project, and the validation process is designed to identify data quality issues uncovered during the verification process that may affect the intended use. One goal of validation is to answer the "So what?" question with regard to any data quality issues. CSRA data review staff will validate all of the mercury, PFAS, PCB congener, Aroclor, and fatty acid analysis results to be generated under this QAPP and any subsequent QAPP revisions. D2. Verification and Validation Methods D2.1 Verification Methods In the first stage of the data verification process, the CSRA data review chemists will perform a "Data Completeness Check" in which all elements in each laboratory submission will be evaluated to verify that results for all specified samples are provided, that data are reported in the ------- NCCA 2020 Great Lakes Fish Fillet Sample Analysis QAPP Revision 4 Date: November 19, 2021 Page 42 of 45 correct format, and that all relevant information, such as preparation and analysis records, are included in the data package. Corrective action procedures will be initiated if deficiencies are noted. The second stage of the verification process will focus on an "Instrument Performance Check" in which the CSRA data review chemists will verify that calibrations, calibration verifications, standards, and calibration blanks were analyzed at the appropriate frequency and met method or study performance specifications. If errors are noted at this stage, corrective action procedures will be initiated immediately. Stage three of the verification process will focus on a "Laboratory Performance Check" in which the CSRA data review chemists will verify 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 the laboratory control samples, method blanks, matrix spike samples and/or reference samples, where applicable. Corrective action procedures will be initiated with the laboratories to resolve any deficiencies identified. In stage four of the verification process, the CSRA data review chemists will perform a "Method/Matrix Performance Check" to discern whether any QC failures are a result of laboratory performance or difficulties with the method or sample matrix. Data evaluated in this stage may include matrix spike and reference sample results. The CSRA data review chemists also will verify that proper sample dilutions were performed and that necessary sample cleanup steps were taken. If problems are encountered, the CSRA data review chemists will immediately implement corrective actions. D2.2 Validation Methods CSRA data review chemists will perform a data quality and usability assessment in which the overall quality of data is evaluated against the performance criteria (see Section B5 for a description of performance criteria). This assessment will strive to maximize use of data gathered in this study based on performance criteria established for these 2020 Great Lakes human health fish tissue studies. This will be accomplished by evaluating the overall quality of a particular data set rather than focusing on individual QC failures. Results of this assessment will be documented in project-specific QA reports developed after all of the results have been evaluated, and before they are used in any final decision making. During this assessment, data qualifier flags are applied to project results to identify any results that did not meet the method- or project-specific requirements; CSRA data review chemists still may also apply additional qualifiers that indicate an assessment of the impact of the problem. For example, individual sample results are often qualified based on the presence of the analyte in a method blank associated with samples prepared together (e.g., extracted or digested in the same batch). While it is important to identify any result associated with the presence of the analyte in the blank, the relative significance of the potential for sample contamination will be assessed using commonly accepted "rules." In instances where the amount of the analyte found in the method blank has very limited potential to affect the field sample result, an additional data qualifier will be applied to that field sample result to indicate that the result was not affected by the observed blank contamination. Similar assessments made for other data quality concerns ------- NCCA 2020 Great Lakes Fish Fillet Sample Analysis QAPP Revision 4 Date: November 19, 2021 Page 43 of 45 may result in the application of additional flags that reconcile the observed data quality concerns with the user requirements and warn the end user of any limitations to the results (i.e., potential low or high bias, blank contamination, etc.). All of the data qualifiers will be included in the data file along with summary level comments that explain the implication in relatively plain English. Where data quality concerns suggest that no valid result was produced for a given analyte, the result for the analyte will be flagged for exclusion in the project-specific databases, and the comments will provide the rationale for the exclusion. The final report of fish tissue study results generated from each database and provided to EPA will not include such invalid results, although the records marked for exclusion will be retained in the database for transparency. As noted earlier, the overall verification and validation process is designed to maximize the amount of usable data for each fish tissue study, so flagging results for exclusion in each final fish tissue study database is intended as a last resort. D3. Reconciliation with User Requirements The QC results for the analyses of the homogenized fish fillet tissue samples for mercury, PFAS, PCB congeners, Aroclors, and fatty acids will be assessed against the QC acceptance criteria for those respective analyses. CSRA will track laboratory performance, notify the OST Project Manager of any issues, initiate corrective actions, and track progress by each sample analysis laboratory. ------- NCCA 2020 Great Lakes Fish Fillet Sample Analysis QAPP Revision 4 Date: November 19, 2021 Page 44 of 45 REFERENCES ALS. 2020. ALS-Environmental Kelso Facility Quality Assurance Manual. ALSKL-QAM. Revision 28.0. October 21, 2020. Clarkson. 2021. Quality Assurance Project Plan and Program Level Quality Documentation Plan - The Great Lakes Fish Monitoring and Surveillance Program GLFMSP: Building on Success. Version 2. Clarkson University Centre for Air Resources Engineering and Science. October 2, 2021. Eurofins-TestAmerica. 2020. Quality Assurance Manual, PT-QA-M-001. Revision No. 6. April 22, 2020. SGS-AXYS. 2020. Quality Assurance/Quality Control (QA/QC) Policies and Procedures Manual, QDO-001. Revision No. 32. June 11, 2020. USEPA. 1994. Method 3541, Revision 0. Automated Soxhlet Extraction. September 1994. Third Edition of the Test Methods for Evaluating Solid Waste, Physical/Chemical Methods, EPA publication SW-846. U.S. Environmental Protection Agency, Office of Land and Emergency Management, Washington, DC. USEPA. 1994. Method 3640A, Revision 1. Gel-Permeation Cleanup. September 1994. Third Edition of the Test Methods for Evaluating Solid Waste, Physical/Chemical Methods, EPA publication SW-846. U.S. Environmental Protection Agency, Office of Land and Emergency Management, Washington, DC. USEPA. 1995. Good Automated Laboratory Practices. EPA Manual 2185. U.S. Environmental Protection Agency, Office of Information Resources Management, Research Triangle Park, NC, August 1995. USEPA. 1996. Method 3660, Revision 0. Sulfur Cleanup. December 1996. Third Edition of the Test Methods for Evaluating Solid Waste, Physical/Chemical Methods, EPA publication SW-846. U.S. Environmental Protection Agency, Office of Land and Emergency Management, Washington, DC. USEPA. 1996. Method 3665A, Revision 1. Sulfuric Acid/Permanganate Cleanup. December 1996. Third Edition of the Test Methods for Evaluating Solid Waste, Physical/Chemical Methods, EPA publication SW-846. U.S. Environmental Protection Agency, Office of Land and Emergency Management, Washington, DC. USEPA. 2001a. EPA Requirements for Quality Assurance Project Plans. EPAQA/R-5. EPA/240/B-01/003. U.S. Environmental Protection Agency, Office of Environmental Information, Washington, DC. USEPA. 2001b. Appendix to Method 1631, Total Mercury in Tissue, Sludge, Sediment, and Soil by Acid Digestion and BrCl Oxidation. EPA No. EPA-821-R-01-013. January 2001. U.S. Environmental Protection Agency, Office of Water, Washington, DC. ------- NCCA 2020 Great Lakes Fish Fillet Sample Analysis QAPP Revision 4 Date: November 19, 2021 Page 45 of 45 USEPA. 2002. Method 1631, Revision E: Mercury in Water by Oxidation, Purge and Trap, and Cold Vapor Atomic Fluorescence Spectrometry. EPA No. EPA-821-R-02-019. August 2002. U.S. Environmental Protection Agency, Office of Water, Washington, DC. USEPA. 2007. Method 8082A, Revision 1. Polychlorinated Biphenyls (PCBs) by Gas Chromatography. February 2007. Third Edition of the Test Methods for Evaluating Solid Waste, Physical/Chemical Methods, EPA publication SW-846. U.S. Environmental Protection Agency, Office of Land and Emergency Management, Washington, DC. USEPA. 2009. Method 537, Determination of Selected Perfluorinated Alkyl Acids in Drinking Water by Solid Phase Extraction and Liquid Chromatography/Tandem Mass Spectrometry (LC/MS/MS), Revision 1.1. EPA/600/R-08/092. September 2009. US Environmental Protection Agency, National Exposure Research Laboratory, Office of Research and Development, Cincinnati, OH. USEPA. 2010. Method 1668C, Chlorinated Biphenyl Congeners in Water, Soil, Sediment, Biosolids, and Tissue by HRGC/HRMS, April 2010. U.S. Environmental Protection Agency, Office of Water, Washington, DC. USEPA. 2020a. National Coastal Condition Assessment 2020 Quality Assurance Project Plan. Version 1.1. April 2020. U.S. Environmental Protection Agency, Office of Wetlands, Oceans, and Watersheds, Washington, DC. USEPA. 2020b. National Coastal Condition Assessment 2020 Field Operations Manual. Version 1.1. April 2020. U.S. Environmental Protection Agency, Office of Water, Washington, DC. USEPA. 2020c. Quality Assurance Project Plan for Preparation for National Coastal Condition Assessment (NCCA) 2020 Great Lakes Human Health Fish Sample Preparation. July 2020. U.S. Environmental Protection Agency, Office of Science and Technology, Washington, DC. Vista. 2020. Vista Analytical Laboratory Quality Manual. Revision 30. December 10, 2020. ------- 2020 NCCA Fish Fillet Sample Analysis QAPP Revision 4 Date: November 19, 2021 Appendix A Target Lists of NCCA 2020 Great Lakes Human Health Whole Fish Sampling Locations Target Lists of NCCA 2020 Great Lakes Human Health Whole Fish Sampling Locations A-l ------- 2020 NCCA Fish Fillet Sample Analysis QAPP Revision 4 Date: November 19, 2021 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 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 Fluron MI NGL20 MI-10020 43.661172 -83.813745 Lake Fluron MI NGL20 MI-10021 43.246154 -82.464205 Lake Fluron MI NGL20 MI-10022 45.750362 -84.563951 Lake Fluron MI NGL20 MI-10023 44.839345 -83.242500 Lake Fluron MI NGL20 MI-10024 43.691878 -83.607161 Lake Fluron MI NGL20 MI-10025 45.963069 -84.714304 Lake Fluron MI NGL20 MI-10026 45.378104 -83.647971 Lake Fluron MI NGL20 MI-10027 44.005051 -83.227579 Lake Fluron MI NGL20 MI-10028 45.939652 -84.669392 Lake Fluron MI NGL20 MI-10029 45.006597 -83.359057 Lake Fluron MI NGL20 MI-10030 43.879908 -83.436643 Lake Fluron MI NGL20 MI-10031 44.013500 -82.767393 Lake Fluron MI NGL20 MI-10032 45.960709 -84.419150 Lake Fluron MI NGL20 MI-10033 45.186507 -83.333887 Lake Fluron MI NGL20 MI-10034 44.262727 -83.475720 Lake Fluron MI NGL20 MI-1003 5 45.365339 -83.558984 Lake Fluron MI NGL20 MI-10036 43.933393 -83.375104 Lake Fluron MI NGL20 MI-10037 44.058141 -82.890460 Lake Fluron MI NGL20 MI-1003 8 44.975283 -83.442236 Target Lists of NCCA 2020 Great Lakes Human Health Whole Fish Sampling Locations A-2 ------- 2020 NCCA Fish Fillet Sample Analysis QAPP Revision 4 Date: November 19, 2021 2020 Great Lakes Human Health Fish Fillet Tissue Study Nearshore Target Sampling Locations (226)1 Lake State Site ID Latitude Longitude Lake Fluron MI NGL20 MI-10039 43.987738 -83.643622 Lake Fluron MI NGL20 MI-10040 43.762320 -82.615251 Lake Fluron MI NGL20 MI-10041 45.700899 -84.357468 Lake Fluron MI NGL20 MI-10042 44.380129 -83.305156 Lake Fluron MI NGL20 MI-10043 43.998848 -82.682268 Lake Fluron MI NGL20 MI-10044 45.637261 -84.224843 Lake Fluron MI NGL20 MI-10045 44.220559 -83.436314 Lake Fluron MI NGL20 MI-10046 44.560749 -83.269783 Lake Fluron MI NGL20 MI-10047 43.325180 -82.522957 Lake Fluron MI NGL20 MI-10048 45.931044 -84.370602 Lake Fluron MI NGL20 MI-10049 43.747200 -83.482180 Lake Fluron MI NGL20 MI-10050 43.069218 -82.418883 Lake Fluron MI NGL20 MI-10051 45.510818 -84.053375 Lake Fluron MI NGL20 MI-10052 44.995087 -83.402484 Lake Fluron MI NGL20 MI-10053 43.763117 -83.450645 Lake Fluron MI NGL20 MI-10054 43.866042 -83.384851 Lake Fluron MI NGL20 MI-1005 5 45.399225 -83.640746 Lake Fluron MI NGL20 MI-10056 44.881911 -83.263937 Lake Fluron MI NGL20 MI-10057 43.672811 -83.905286 Lake Fluron MI NGL20 MI-1005 8 44.082561 -82.990724 Lake Fluron MI NGL20 MI-10059 44.563966 -83.305080 Lake Fluron MI NGL20 MI-10060 43.915173 -83.840608 Lake Fluron MI NGL20 MI-10061 43.857134 -82.597289 Lake Fluron MI NGL20 MI-10062 44.062607 -83.575569 Lake Fluron MI NGL20 MI-10063 45.079141 -83.300411 Lake Fluron 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 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 Target Lists of NCCA 2020 Great Lakes Human Health Whole Fish Sampling Locations A-3 ------- 2020 NCCA Fish Fillet Sample Analysis QAPP Revision 4 Date: November 19, 2021 2020 Great Lakes Human Health Fish Fillet Tissue Study Nearshore Target Sampling Locations (226)1 Lake State Site ID Latitude Longitude 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-10035 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-10038 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-10055 43.985167 -76.060517 Lake Ontario NY NGL20 NY-10056 43.822188 -76.317569 Lake Ontario NY NGL20 NY-10057 43.340606 -77.703826 Lake Ontario NY NGL20 NY-10058 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 Target Lists of NCCA 2020 Great Lakes Human Health Whole Fish Sampling Locations A-4 ------- 2020 NCCA Fish Fillet Sample Analysis QAPP Revision 4 Date: November 19, 2021 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-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 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-5 ------- 2020 NCCA Fish Fillet Sample Analysis QAPP Revision 4 Date: November 19, 2021 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 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-6 ------- 2020 NCCA Fish Fillet Sample Analysis QAPP Revision 4 Date: November 19, 2021 Appendix B 2020 NCCA Detection and Quantitation Limits for Great Lakes Human Health Fish Fillet Tissue Analyses 2020 NCCA Detection and Quantitation Limits for Great Lakes Human Health Fish Fillet Tissue Analyses B-l ------- 2020 NCCA Fish Fillet Sample Analysis QAPP Revision 4 Date: November 19, 2021 Method Detection Limits (MDLs) and Minimum Levels (MLs) for 2020 NCCA Great Lakes Fish Fillet Tissue Target Analytes Mercury MDL and ML (based on a 0.5-g sample) MDL1 (ng/g) ML (ng/g) 0.2 1 1 The MDL is based on the EPA procedure described at 40 CFR 136, Appendix B, Revision 2, from August 2017. The PFAS analytes to be determined in this project are listed in the table below, along with their common abbreviations. The method detection and quantitation limits (also referred to as minimum levels) were provided by the laboratory as part of its bid submission. PFAS MDLs and MLs (based on a 2-g tissue sample and a 250-mL rinsate sample) Name Abbreviation Tissue Samples (ng/g) Rinsate Samples (ng/L) MDL1 ML MDL1 ML Perfluorobutanoic acid PFBA 0.593 0.8 0.661 6.4 Perfluoropentanoic acid PFPeA 0.083 0.4 0.392 3.2 Perfluorohexanoic acid PFHxA 0.096 0.2 0.636 1.6 Perfluoroheptanoic acid PFHpA 0.088 0.2 0.443 1.6 Perfluorooctanoic acid PFOA 0.086 0.2 0.604 1.6 Perfluorononanoic acid PFNA 0.160 0.2 0.442 1.6 Perfluorodecanoic acid PFDA 0.124 0.2 0.666 1.6 Perfluoroundecanoic acid PFUnA 0.152 0.2 0.527 1.6 Perfluorododecanoic acid PFDoA 0.130 0.2 0.758 1.6 Perfluorotridecanoic acid PFTrDA 0.086 0.2 0.476 1.6 Perfluorotetradecanoic acid PFTeDA 0.185 0.2 0.527 1.6 Perfluorobutanesulfonic acid PFBS 0.070 0.2 0.491 1.6 Perfluoropentanesulfonic acid PFPeS 0.032 0.2 0.407 1.6 Perfluorohexanesulfonic acid PFHxS 0.083 0.2 0.434 1.6 Perfluoroheptanesulfonic acid PFHpS 0.043 0.2 0.274 1.6 Perfluorooctanesulfonic acid PFOS 0.294 0.3 0.654 1.6 Perfluorononanesulfonic acid PFNS 0.114 0.2 0.606 1.6 Perfluorodecanesulfonic acid PFDS 0.101 0.2 0.668 1.6 Perfluorododecanesulfonic acid PFDoS 0.177 0.2 0.358 1.6 1H, 1H, 2H, 2H-perfluorohexane sulfonic acid 4:2 FTS 0.740 0.8 4.561 6.4 1H, 1H, 2H, 2H-perfluorooctane sulfonic acid 6:2 FTS 1.149 1.3 7.946 8.7 1H, 1H, 2H, 2H-perfluorodecane sulfonic acid 8:2 FTS 0.373 0.8 3.132 6.4 2H, 2H, 3H, 3H-perfluorohexanoic acid 3:3 FTC A 0.247 0.8 1.441 6.4 2H, 2H, 3H, 3H-perfluorooctanoic acid 5:3 FTC A 1.537 5.0 10.133 40.0 2H, 2H, 3H, 3H-perfluorodecanoic acid 7:3 FTC A 0.845 5.0 11.885 40.0 Perfluorooctanesulfonamide PFOSA 0.094 0.2 0.454 1.6 N-Methylperfluorooctanesulfonamide N-MeFOSA 0.161 0.2 0.392 1.8 N-Ethylperfluorooctanesulfonamide N-EtFOSA 0.169 0.5 1.169 4.0 2020 NCCA Detection and Quantitation Limits for Great Lakes Human Health Fish Fillet Tissue Analyses B-2 ------- 2020 NCCA Fish Fillet Sample Analysis QAPP Revision 4 Date: November 19, 2021 PFAS MDLs and MLs (based on a 2-g tissue sample and a 250-mL rinsate sample) Name Abbreviation Tissue Samples (ng/g) Rinsate Samples (ng/L) MDL1 ML MDL1 ML N-Methylperfluoro-l-octanesulfonamidoacetic acid N-MeFOSAA 0.930 0.2 0.650 0.0 N-Ethylperfluoro-1 -octanesulfonamidoacetic acid N-EtFOSAA 0.138 0.2 2.040 12.0 N-Methylperfluoro-l-octanesulfonamidoethanol N-MeFOSE 9.977 11.0 2.383 16.0 N-Ethylperfluoro-1 -octanesulfonamidoethanol N-EtFOSE 1.500 1.7 2.043 12.0 2,3,3,3-Tetrafluoro-2-(l,l,2,2,3,3,3- heptafluoropropoxypropionic acid HFPO-DA 0.161 0.8 0.811 6.1 4,8-dioxa-3H-perfluorononanoic acid ADONA 0.082 0.8 1.558 6.4 Perfluoro-3,6-dioxaheptanoic acid NFDHA 0.294 0.4 2.768 3.2 Perfluoro-3-methoxypropanoic acid PFMPA 0.070 0.4 0.354 3.2 Perfluoro-4-methoxybutanoic acid PFMBA 0.069 0.2 0.233 1.6 9-chlorohexadecafluoro-3 -oxanonane-1 -sulfonic acid 9C1-PF30NS 0.152 0.8 1.742 6.4 11 -chloroeicosafluoro-3 -oxaundecane-1 -sulfonic acid llCl-PF30UdS 0.312 0.8 1.639 6.4 Perfluoro(2-ethoxyethane)sulfonic acid PFEESA 0.045 0.2 0.274 1.6 1 The MDL is based on the EPA procedure described at 40 CFR 136, Appendix B, Revision 2, from August 2017. PFAS analytes in NARS fish tissue studies from 2008 to 2015 PFAS analytes new for 2018-19 NRSA Fish Tissue Study PFAS analytes new to this study (2020 GLHHFFTS) The PCB congeners to be determined in this project are listed in the table below. The method detection and quantitation limits (also referred to as minimum levels) were provided by the laboratory as part of its bid submission. PCB MDLs and MLs in pg/g (in elution order, based on a 10-g sample) Analyte MDL1 ML PCB-1 0.14 1 PCB-2 0.13 1 PCB-3 0.22 1 PCB-4/10 0.29 1 PCB-5/8 0.39 1 PCB-6 0.21 1 PCB-7/9 0.21 1 PCB-11 1.13 1 PCB-12/13 0.24 1 PCB-14 0.14 1 PCB-15 0.11 1 PCB-16/32 0.34 1 PCB-17 0.19 1 PCB-18 0.30 1 PCB-19 0.28 1 PCB-20/21/33 0.74 PCB-22 0.21 1 PCB-23 0.20 1 2020 NCCA Detection and Quantitation Limits for Great Lakes Human Health Fish Fillet Tissue Analyses B-3 ------- 2020 NCCA Fish Fillet Sample Analysis QAPP Revision 4 Date: November 19, 2021 PCB MDLs and MLs in pg/g (in elution order, based on a 10-g sample) Analyte MDL1 ML PCB-24/27 0.39 1 PCB-25 0.18 1 PCB-26 0.25 1 PCB-28 0.36 1 PCB-29 0.21 1 PCB-30 0.20 1 PCB-31 0.41 1 PCB-34 0.17 1 PCB-35 0.17 1 PCB-36 0.14 1 PCB-37 0.15 1 PCB-38 0.14 1 PCB-39 0.18 1 PCB-40 0.24 1 PCB-41/64/71/72 0.62 PCB-42/59 0.40 1 PCB-43/49 0.28 1 PCB-44 0.17 1 PCB-45 0.28 1 PCB-46 0.19 1 PCB-47 0.64 1 PCB-48/75 0.21 1 PCB-50 0.25 1 PCB-51 0.20 1 PCB-52/69 0.42 1 PCB-53 0.31 1 PCB-54 0.15 1 PCB-55 0.24 1 PCB-56/60 0.25 1 PCB-57 0.08 1 PCB-58 0.10 1 PCB-61/70 0.24 1 PCB-62 0.26 1 PCB-63 0.18 1 PCB-65 0.29 1 PCB-66/76 0.30 1 PCB-67 0.29 1 PCB-68 0.16 1 PCB-73 0.16 1 PCB-74 0.28 1 PCB-77 0.14 1 PCB-78 0.13 1 PCB-79 0.12 1 PCB-80 0.19 1 PCB-81 0.13 1 PCB-82 0.32 1 2020 NCCA Detection and Quantitation Limits for Great Lakes Human Health Fish Fillet Tissue Analyses B-4 ------- 2020 NCCA Fish Fillet Sample Analysis QAPP Revision 4 Date: November 19, 2021 PCB MDLs and MLs in pg/g (in elution order, based on a 10-g sample) Analyte MDL1 ML PCB-83 0.29 1 PCB-84/92 0.40 1 PCB-85/116 0.46 1 PCB-86 0.43 1 PCB-87/117/125 0.52 PCB-88/91 0.66 1 PCB-89 0.22 1 PCB-90/101 0.32 1 PCB-93 0.52 1 PCB-94 0.26 1 PCB-95/98/102 0.59 PCB-96 0.32 1 PCB-97 0.25 1 PCB-99 0.29 1 PCB-100 0.17 1 PCB-103 0.24 1 PCB-104 0.17 1 PCB-105 0.16 1 PCB-106/118 0.41 1 PCB-107/109 0.31 1 PCB-108/112 0.46 1 PCB-110 0.37 1 PCB-111/115 0.31 1 PCB-113 0.24 1 PCB-114 0.24 1 PCB-119 0.16 1 PCB-120 0.18 1 PCB-121 0.41 1 PCB-122 0.21 1 PCB-123 0.15 1 PCB-124 0.24 1 PCB-126 0.16 1 PCB-127 0.13 1 PCB-128/162 0.33 1 PCB-129 0.13 1 PCB-130 0.23 1 PCB-131/133 0.32 1 PCB-132/161 0.29 1 PCB-134/143 0.39 1 PCB-135 0.30 1 PCB-136 0.17 1 PCB-137 0.22 1 PCB-138/163/164 0.61 PCB-139/149 0.72 1 PCB-140 0.31 1 PCB-141 0.22 1 2020 NCCA Detection and Quantitation Limits for Great Lakes Human Health Fish Fillet Tissue Analyses B-5 ------- 2020 NCCA Fish Fillet Sample Analysis QAPP Revision 4 Date: November 19, 2021 PCB MDLs and MLs in pg/g (in elution order, based on a 10-g sample) Analyte MDL1 ML PCB-142 0.22 1 PCB-144 0.18 1 PCB-145 0.35 1 PCB-146/165 0.31 1 PCB-147 0.17 1 PCB-148 0.43 1 PCB-150 0.25 1 PCB-151 0.16 1 PCB-152 0.21 1 PCB-153 0.37 1 PCB-154 0.35 1 PCB-155 0.34 1 PCB-156 0.14 1 PCB-157 0.12 1 PCB-15 8/160 0.32 1 PCB-159 0.10 1 PCB-166 0.14 1 PCB-167 0.17 1 PCB-168 0.14 1 PCB-169 0.22 1 PCB-170 0.37 1 PCB-171 0.35 1 PCB-172 0.26 1 PCB-173 0.20 1 PCB-174 0.29 1 PCB-175 0.16 1 PCB-176 0.14 1 PCB-177 0.21 1 PCB-178 0.26 1 PCB-179 0.22 1 PCB-180 0.49 1 PCB-181 0.23 1 PCB-182/187 0.36 1 PCB-183 0.18 1 PCB-184 0.16 1 PCB-185 0.18 1 PCB-186 0.18 1 PCB-188 0.17 1 PCB-189 0.22 1 PCB-190 0.29 1 PCB-191 0.16 1 PCB-192 0.26 1 PCB-193 0.27 1 PCB-194 0.15 1 PCB-195 0.22 1 PCB-196/203 0.55 1 2020 NCCA Detection and Quantitation Limits for Great Lakes Human Health Fish Fillet Tissue Analyses B-6 ------- 2020 NCCA Fish Fillet Sample Analysis QAPP Revision 4 Date: November 19, 2021 PCB MDLs and MLs in pg/g (in elution order, based on a 10-g sample) Analyte MDL1 ML PCB-197 0.24 1 PCB-198 0.37 1 PCB-199 0.42 1 PCB-200 0.35 1 PCB-201 0.21 1 PCB-202 0.22 1 PCB-204 0.23 1 PCB-205 0.16 1 PCB-206 0.23 1 PCB-207 0.21 1 PCB-208 0.19 1 PCB-209 0.27 1 1 The Vista Analytical Laboratory MDLs are based on Revision 2 of the MDL procedure published in August 2017. The Aroclors to be determined in this project are listed in the table below. The method detection and quantitation limits (also referred to as minimum levels) were provided by the laboratory as part of its bid submission. Aroclor MDLs and MLs in ng/g (based on a 10-g sample) Analyte MDL1 ML2 Aroclor 1016 0.5568 2.5 Aroclor 1221 0.8868 2.5 Aroclor 1232 0.6063 2.5 Aroclor 1242 0.3642 2.5 Aroclor 1248 0.6057 2.5 Aroclor 1254 0.7476 2.5 Aroclor 1260 0.7437 2.5 Aroclor 1268 0.8778 2.5 1 The Eurofins-TestAmerica MDLs are based on Revision 2 of the MDL procedure published in August 2017. 2 The Eurofins-TestAmerica ML values are based on the concentration of the lowest calibration standard analyzed. 2020 NCCA Detection and Quantitation Limits for Great Lakes Human Health Fish Fillet Tissue Analyses B-7 ------- 2020 NCCA Fish Fillet Sample Analysis QAPP Revision 4 Date: November 19, 2021 The fatty acids to be determined in this project are listed in the table below, along with the method detection and quantitation limits that were provided by the laboratory as part of its bid submission. The quantitation limits (QLs) in the table below are three times the MDL and rounded to two decimal places. Fatty Acid MDLs and QLs in jig/g (based on a 0.5-g sample) Fatty Acid Lipid Number Abbreviation* MDL QL Decanoic acid C10:0 3.00 9.00 Undecanoic acid C11:0 3.00 9.00 Dodecanoic acid C12:0 3.00 9.00 Tridecanoic acid C13:0 0.09 0.27 Myristic acid C14:0 1.89 5.67 Myristoleic acid C14:l 0.18 0.54 Pentadecanoic acid C15:0 0.36 1.08 cw-10-Pentadecenoic acid CI 5:1 0.09 0.27 Palmitic acid C16:0 6.72 20.16 cw-7-Hexadecenoic acid C16:l 7.14 21.42 Heptadecanoic acid C17:0 0.33 0.99 ccis-10-Heptadecenoic acid C17:l 0.09 0.27 Stearic acid C18:0 0.69 2.07 Elaidic acid C18:ln9 trans 0.30 0.90 Oleic acid C18:ln9 cis 10.6 31.8 cw-vaccenic acid C18:lw7 2.26 6.78 Linolelaidic acid C18:2 trans 0.93 2.79 Linoleic acid C18:2n6 cis 1.60 4.80 gamma-Linolenic acid C18:3n6 0.34 1.02 Arachidic acid C20:0 0.09 0.27 Linolenic acid C18:3n3 3.50 10.50 Eicosenoic acid C20:ln9 3.50 10.50 Octadecatetraenoic acid C18:4n3 3.50 10.50 Heneicosanoic acid C21:0 0.12 0.36 Eicosadienoic acid C20:2 0.16 0.48 Dihomo-gararaa-linolenic acid C20:3n6 0.32 0.96 Behenic acid C22:0 0.09 0.27 Eicosatrienoic acid C20:3n3 2.04 6.12 Arachidonic acid C20:4n6 1.67 5.01 Cetoleic acid C22:lnll 0.14 0.42 Erucic acid C22:ln9 0.14 0.42 Tricosanoic acid C23:0 0.15 0.45 13,16-Docosadienoic acid C22:2 1.84 5.52 Eicosapentaenoic acid C20:5n3 0.20 0.60 Lignoceric acid C24:0 0.09 0.27 Nervonic acid C24:ln9 0.63 1.89 Docosapentaenoic acid C22:5n3 2.49 7.47 Docosahexaenoic acid C22:6n3 2.49 7.47 * Lipid numbers take the form C:DnX, where C is the number of carbon atoms in the fatty acid and D is the number of double bonds. Where applicable, the fatty acid double bond location is identified by nX, where X is the carbon number of the first double bond relative to the terminal alkyl end of the fatty acid. 2020 NCCA Detection and Quantitation Limits for Great Lakes Human Health Fish Fillet Tissue Analyses B-8 ------- 2020 NCCA Fish Fillet Sample Analysis QAPP Revision 4 Date: November 19, 2021 Appendix C 2020 NCCA Quality Control (QC) Acceptance Criteria for PFAS Analyses of Great Lakes Fish Fillet Tissue Samples and QC Rinsate Samples 2020 NCCA Quality Control (QC) Acceptance Criteria for PFAS Analyses of Tissue and Rinsate Samples C-l ------- 2020 NCCA Fish Fillet Sample Analysis QAPP Revision 4 Date: November 19, 2021 Calibration Verification (CCV), LCS, and Labeled Compound Recovery QC Acceptance Criteria for PFAS Analyses LCS Recovery (%) Labeled Compound Recovery in Samples (%) Analyte CCV (%) Tissues Rinsates Tissues Rinsates Target Analytes PFBA 70 - 130 70 - 130 PFPeA 70 - 130 70 - 130 PFHxA 70 - 130 70 - 130 PFHpA 70 - 130 70 - 130 PFOA 70 - 130 70 - 130 PFNA 70 - 130 70 - 130 PFDA 60 - 130 70 - 130 PFUnA 70 - 140 70 - 130 PFDoA 70 - 130 70 - 130 PFTrDA 70 - 130 70 - 130 PFTeA 70 - 130 70 - 130 PFBS 60 - 130 70 - 130 PFPeS 70 - 130 70 - 130 PFHxS 70 - 130 70 - 130 PFHpS 70 - 130 70 - 130 PFOS 70 - 140 70 - 130 PFNS 60 - 150 70 - 130 PFDS 40 - 150 70 - 130 PFDoS 70 - 140 60 - 130 4:2 FTS 70 -130 40 - 150 70 - 130 NA NA 6:2 FTS 70 - 130 70 - 130 8:2 FTS 70 - 170 70 - 130 3:3 FTCA 70 - 130 65 - 130 5:3 FTCS 70 - 180 70 - 130 7:3 FTCA 70 - 130 70 - 130 PFOSA 70 - 130 70 - 130 N-MeFOSA 50 - 140 70 - 130 N-EtFOSA 70 - 130 70 - 130 N-MeFOSAA 60 - 160 70 - 130 N-EtFOSAA 60 - 160 70 - 130 N-MeFOSE 70 - 150 70 - 130 N-EtFOSE 70 - 130 70 - 130 HFPO-DA 70 - 130 70 - 130 ADONA 70 - 130 70 - 130 NFDHA 60 - 130 65 - 140 PFMPA 70 - 130 70 - 130 PFMBA 70 - 130 70 - 130 9C1-PF30NS 70 - 130 70 - 130 11C1-PF30UDS 60 - 130 70 - 130 PFEESA 70 - 130 70 - 130 2020 NCCA Quality Control (QC) Acceptance Criteria for PFAS Analyses of Tissue and Rinsate Samples C-2 ------- 2020 NCCA Fish Fillet Sample Analysis QAPP Revision 4 Date: November 19, 2021 LCS Recovery (%) Labeled Compound Recovery in Samples (%) Analyte CCV (%) Tissues Rinsates Tissues Rinsates Labeled Compounds 13c4-pfba 50 - 150 50 - 150 50 - 150 50 - 150 13C5-PFPeA 50 - 150 50 - 150 50 - 150 50 - 150 13C5-PFHxA 50 - 150 50 - 150 50 - 150 50 - 150 13C4-PFHpA 50 - 150 50 - 150 50 - 150 50 - 150 13c6-pfoa 50 -150 50 - 150 50 - 150 50 - 150 50 - 150 13c9-pfna 50 - 150 50 - 150 50 - 150 50 - 150 13c6-pfda 50 - 180 50 - 150 50 - 180 50 - 150 13C7-PFUnA 50 - 150 50 - 150 50 - 150 50 - 150 13C2-PFDoA 50 - 150 50 - 150 50 - 150 50 - 150 13C2-PFTeA 50 - 150 50 - 150 50 - 150 50 - 150 13c3-pfbs 50 - 150 50 - 150 50 - 150 50 - 150 13C3-PFHxS 50 - 150 50 - 150 50 - 150 50 - 150 13c8-pfos 50 - 150 50 - 150 50 - 150 50 - 150 13C2-4:2 FTS 50 - 220 50 - 150 50 - 220 50 - 150 13C2-6:2 FTS 50 - 180 50 - 150 50 - 180 50 - 150 13C2-8:2 FTS 50 - 300 50 - 150 50 - 300 50 - 150 13c8-pfosa 50 - 150 50 - 150 50 - 150 50 - 150 50 - 150 D3-N-MeFOSA 5 - 150 30-150 20 - 200 30-150 Ds-N-EtFOSA 10 - 150 20 - 150 20 - 200 20 - 150 D3-N-MeFOSAA 50 - 180 50 - 150 20 - 200 50 - 150 Ds-N-EtFOSAA 50 - 250 50 - 150 20 - 200 50 - 150 D-N-McFOSE 2- 150 30-150 20 - 200 30-150 Dg-N-EtFOSE 2- 150 30-150 20 - 200 30-150 13C3-HFPO-DA 50 - 150 50 - 150 50 - 150 50 - 150 2020 NCCA Quality Control (QC) Acceptance Criteria for PFAS Analyses of Tissue and Rinsate Samples C-3 ------- 2020 NCCA Fish Fillet Sample Analysis QAPP Revision 4 Date: November 19, 2021 Appendix D 2020 NCCA Quality Control (QC) Acceptance Criteria for PCB Congener Analysis of Great Lakes Fish Fillet Tissue Samples 2020 NCCA Quality Control (QC) Acceptance Criteria for PCB Congener Analyses of Tissue Samples D-l ------- 2020 NCCA Fish Fillet Sample Analysis QAPP Revision 4 Date: November 19, 2021 Calibration Verification Limits (%), Laboratory Control Sample Recovery Limits (%), and Labeled Compound Recovery Limits (%) for PCB Congener Analyses QC Acceptance Criteria for VER1, OPR2, and Labeled Compounds3 in Samples Congener Name Congener Number VER (%) OPR Recovery (%) Labeled Compound Recovery in Samples (%) 2-MonoCB 1 75-125 60-135 3-MonoCB 2 75-125 60-135 4-MonoCB 3 75-125 60-135 2,2'-DiCB/2,6-DiCB 4/10 75-125 60-135 2,3-DiCB/2,4'-DiCB 5/8 75-125 60-135 2,3'-DiCB 6 75-125 60-135 2,4 -DiCB/2,5 -DiCB 7/9 75-125 60-135 3,3'-DiCB 11 75-125 60-135 3,4-DiCB/3,4'-DiCB 12/13 75-125 60-135 3,5-DiCB 14 75-125 60-135 4,4'-DiCB 15 75-125 60-135 2,2',3-TrCB/2,4',6-TrCB 16/32 75-125 60-135 2,2',4-TrCB 17 75-125 60-135 2,2',5-TrCB 18 75-125 60-135 2,2',6-TrCB 19 75-125 60-135 2,3,3'-TrCB/2,3,4-TrCB/2',3,4-TrCB 20/21/33 75-125 60-135 2,3,4'-TrCB 22 75-125 60-135 2,3,5-TrCB 23 75-125 60-135 2,3,6-TrCB/2,3',6-TrCB 24/27 75-125 60-135 2,3',4-TrCB 25 75-125 60-135 2,3',5-TrCB 26 75-125 60-135 2,4,4'-TrCB 28 75-125 60-135 2,4,5-TrCB 29 75-125 60-135 2,4,6-TrCB 30 75-125 60-135 2,4',5-TrCB 31 75-125 60-135 2,3',5'-TrCB 34 75-125 60-135 3,3',4-TrCB 35 75-125 60-135 NA 3,3',5-TrCB 36 75-125 60-135 3,4,4'-TrCB 37 75-125 60-135 3,4,5-TrCB 38 75-125 60-135 3,4',5-TrCB 39 75-125 60-135 2,2',3,3'-TeCB 40 75-125 60-135 2,2',3,4-TeCB/2,3,4',6-TeCB/2,3',4',6-TeCB/2,3',5,5'-TeCB 41/64/71/72 75-125 60-135 2,2',3,4'-TeCB/2,3,3',6-TeCB 42/59 75-125 60-135 2,2',3,5-TeCB/2,2',4,5'-TeCB 43/49 75-125 60-135 2,2',3,5'-TeCB 44 75-125 60-135 2,2',3,6-TeCB 45 75-125 60-135 2,2',3,6'-TeCB 46 75-125 60-135 2,2',4,4'-TeCB 47 75-125 60-135 2,2' ,4,5 -T eCB/2,4,4',6-TeCB 48/75 75-125 60-135 2,2',4,6-TeCB 50 75-125 60-135 2,2',4,6'-TeCB 51 75-125 60-135 2,2',5,5'-TeCB/2,3',4,6-TeCB 52/69 75-125 60-135 2,2',5,6'-TeCB 53 75-125 60-135 2,2',6,6'-TeCB 54 75-125 60-135 2,3,3',4-TeCB 55 75-125 60-135 2,3,3',4'-TeCB/2,3,4,4'-TeCB 56/60 75-125 60-135 2,3,3',5-TeCB 57 75-125 60-135 2,3,3',5'-TeCB 58 75-125 60-135 2,3,4,5-TeCB/2,3',4',5-TeCB 61/70 75-125 60-135 2,3,4,6-TeCB 62 75-125 60-135 2,3,4',5-TeCB 63 75-125 60-135 2,3,5,6-TeCB 65 75-125 60-135 2020 NCCA Quality Control (QC) Acceptance Criteria for PCB Congener Analyses of Tissue Samples D-2 ------- 2020 NCCA Fish Fillet Sample Analysis QAPP Revision 4 Date: November 19, 2021 QC Acceptance Criteria for VER1, OPR2, and Labeled Compounds3 in Samples Congener Name Congener Number VER (%) OPR Recovery (%) Labeled Compound Recovery in Samples (%) 2,3',4,5-TeCB 67 75-125 60-135 2,3',4,5'-TeCB 68 75-125 60-135 2,3',4',5-TeCB 70 75-125 60-135 2,3',5',6-TeCB 73 75-125 60-135 2,4,4',5-TeCB 74 75-125 60-135 2',3,4,5-TeCB/2,3',4,4'-TeCB 76/66 75-125 60-135 3,3',4,5-TeCB 77 75-125 60-135 3,3',4,5'-TeCB 78 75-125 60-135 3,3',5,5'-TeCB 79 75-125 60-135 3,4,4',5-TeCB 80 75-125 60-135 2,2',3,3',4-PeCB 81 75-125 60-135 2,2',3,3',5-PeCB 82 75-125 60-135 2,2',3,3',5-PeCB 83 75-125 60-135 2,2',3,3',6-PeCB/2,2',3,5,5'-PeCB 84/92 75-125 60-135 2,2',3,4,4'-PeCB/2,3,4,5,6-PeCB 85/116 75-125 60-135 2,2',3,4,5-PeCB 86 75-125 60-135 2,2',3,4,5'-PeCB/2,3,4',5,6-PeCB/2',3,4,5,6'-PeCB 87/117/125 75-125 60-135 2,2' ,3,4,6-PeCB/2,2' ,3,4' ,6-PeCB 88/91 75-125 60-135 2,2',3,4,6'-PeCB 89 75-125 60-135 2,2',3,4',5-PeCB/2,2',4,5,5'-PeCB 90/101 75-125 60-135 2,2',3,5,6-PeCB 93 75-125 60-135 2,2',3,5,6'-PeCB 94 75-125 60-135 2,2',3,5',6-PeCB/2,2',3',4,6-PeCB/2,2',4,5,6'-PeCB 95/98/102 75-125 60-135 2,2',3,6,6'-PeCB 96 75-125 60-135 2,2',3,4',5-PeCB 97 75-125 60-135 2,2',4,4',5-PeCB 99 75-125 60-135 2,2',4,4',6-PeCB 100 75-125 60-135 2,2',4,5',6-PeCB 103 75-125 60-135 2,2',4,4,6'-PeCB 104 75-125 60-135 NA 2,3,3',4,4'-PeCB 105 75-125 60-135 2,3',4,4',5-PeCB/2,3,3',4,5-PeCB 118/106 75-125 60-135 2,3,3',4',5-PeCB/2,3,3',4,6-PeCB 107/109 75-125 60-135 2,3,3',4,5'-PeCB/2,3,3',5,6-PeCB 108/112 75-125 60-135 2,3,3',4',6-PeCB 110 75-125 60-135 2,3,3',5,5'-PeCB/2,3,4,4',6-PeCB 111/115 75-125 60-135 2,3,3',5',6-PeCB 113 75-125 60-135 2,3,4,4',5-PeCB 114 75-125 60-135 2,3',4,4',6-PeCB 119 75-125 60-135 2,3',4,5,5'-PeCB 120 75-125 60-135 2,3',4,5',6-PeCB 121 75-125 60-135 2,3,3',4',5'-PeCB 122 75-125 60-135 2,3',4,4',5'-PeCB 123 75-125 60-135 2,3',4',5,5'-PeCB 124 75-125 60-135 3,3'4,4',5-PeCB 126 75-125 60-135 3,3',4,5,5'-PeCB 127 75-125 60-135 2,2',3,3',4,4'-HxCB/2,3,3',4',5,5'-HxCB 128/162 75-125 60-135 2,2',3,3',4,5-HxCB 129 75-125 60-135 2,2',3,3',4,5'-HxCB 130 75-125 60-135 2,2',3,3',4,6-HxCB 131 75-125 60-135 2,2',3,3',4,6'-HxCB/2,3,3',4,5',6-HxCB 132/161 75-125 60-135 2,2',3,3',5,5'-HxCB/2,2',3,4,5,6-HxCB 133/142 75-125 60-135 2,2',3,3',5,6-HxCB/2,2',3,4,5,6'-HxCB 134/143 75-125 60-135 2,2',3,3',5,6'-HxCB 135 75-125 60-135 2,2',3,3',6,6'-HxCB 136 75-125 60-135 2,2',3,4,4',5-HxCB 137 75-125 60-135 2,2',3,4,4',5'-HxCB/2,3,3',4',5,6-HxCB/2,3,3',4',5',6-HxCB 138/163/164 75-125 60-135 2,2',3,4,4',6-HxCB/2,2',3,4',5',6-HxCB 139/149 75-125 60-135 2020 NCCA Quality Control (QC) Acceptance Criteria for PCB Congener Analyses of Tissue Samples D-3 ------- 2020 NCCA Fish Fillet Sample Analysis QAPP Revision 4 Date: November 19, 2021 QC Acceptance Criteria for VER1, OPR2, and Labeled Compounds3 in Samples Congener Name Congener Number VER (%) OPR Recovery (%) Labeled Compound Recovery in Samples (%) 2,2',3,4,4',6'-HxCB 140 75-125 60-135 2,2',3,4,5,5'-HxCB 141 75-125 60-135 2,2',3,4,5',6-HxCB 144 75-125 60-135 2,2',3,4,6,6'-HxCB 145 75-125 60-135 2,2',3,4',5,5'-HxCB/2,3,3',5,5',6-HxCB 146/165 75-125 60-135 2,2' ,3,4' ,5,6-HxCB 147 75-125 60-135 2,2',3,4',5,6'-HxCB 148 75-125 60-135 2,2',3,4',6,6'-HxCB 150 75-125 60-135 2,2' ,3,5,5' ,6-HxCB 151 75-125 60-135 2,2',3,5,6,6'-HxCB 152 75-125 60-135 2,2',4,4',5,5'-HxCB 153 75-125 60-135 2,2',4,4',5,6'-HxCB 154 75-125 60-135 2,2' ,4,4' ,6,6' -HxCB 155 75-125 60-135 2,3,3',4,4',5-HxCB 156 75-125 60-135 2,3,3',4,4',5'-HxCB 157 75-125 60-135 2,3,3',4,4',6-HxCB/2,3,3',4,5,6-HxCB 158/160 75-125 60-135 2,3,3',4,5,5'-HxCB 159 75-125 60-135 2,3,4,4',5,6-HxCB 166 75-125 60-135 2,3',4,4',5,5'-HxCB 167 75-125 60-135 2,3',4,4',5',6-HxCB 168 75-125 60-135 3,3',4,4',5,5'-HxCB 169 75-125 60-135 2,2',3,3',4,4',5-HpCB 170 75-125 60-135 2,2',3,3',4,4',6-HpCB 171 75-125 60-135 2,2',3,3',4,5,5'-HpCB 172 75-125 60-135 2,2',3,3',4,5,6-HpCB 173 75-125 60-135 2,2',3,3',4,5,6'-HpCB 174 75-125 60-135 2,2',3,3',4,5',6-HpCB 175 75-125 60-135 2,2',3,3'4,6,6'-HpCB 176 75-125 60-135 2,2',3,3',4',5,6-HpCB 177 75-125 60-135 NA 2,2',3,3',5,5',6-HpCB 178 75-125 60-135 2,2',3,3',5,6,6'-HpCB 179 75-125 60-135 2,2',3,4,4',5,5'-HpCB 180 75-125 60-135 2,2',3,4,4',5,6-HpCB 181 75-125 60-135 2,2',3,4,4',5,6'-HpCB/2,2',3,4',5,5',6-HpCB 182/187 75-125 60-135 2,2',3,4,4',5',6-HpCB 183 75-125 60-135 2,2',3,4,4',6,6'-HpCB 184 75-125 60-135 2,2',3,4,5,5',6-HpCB 185 75-125 60-135 2,2',3,4,5,6,6'-HpCB 186 75-125 60-135 2,2',3,4',5,6,6'-HpCB 188 75-125 60-135 2,3,3',4,4',5,5'-HpCB 189 75-125 60-135 2,3,3',4,4',5,6-HpCB 190 75-125 60-135 2,3,3',4,4',5',6-HpCB 191 75-125 60-135 2,3,3',4,5,5',6-HpCB 192 75-125 60-135 2,3,3',4',5,5',6-HpCB 193 75-125 60-135 2,2',3,3',4,4',5,5'-OcCB 194 75-125 60-135 2,2',3,3',4,4',5,6-OcCB 195 75-125 60-135 2,2',3,3',4,4',5,6'-OcCB/2,2',3,4,4',5,5',6-OcCB 196/203 75-125 60-135 2,2',3,3',4,4',6,6'-OcCB 197 75-125 60-135 2,2',3,3',4,5,5',6-OcCB 198 75-125 60-135 2,2',3,3',4,5,5',6'-OcCB 199 75-125 60-135 2,2',3,3',4,5,6,6'-OcCB 200 75-125 60-135 2,2',3,3',4,5',6,6'-OcCB 201 75-125 60-135 2,2',3,3',5,5',6,6'-OcCB 202 75-125 60-135 2,2',3,4,4',5,6,6'-OcCB 204 75-125 60-135 2,3,3',4,4',5,5',6-OcCB 205 75-125 60-135 2,2',3,3',4,4',5,5',6-NoCB 206 75-125 60-135 2,2',3,3',4,4',5,6,6'-NoCB 207 75-125 60-135 2020 NCCA Quality Control (QC) Acceptance Criteria for PCB Congener Analyses of Tissue Samples D-4 ------- 2020 NCCA Fish Fillet Sample Analysis QAPP Revision 4 Date: November 19, 2021 QC Acceptance Criteria for VER1, OPR2, and Labeled Compounds3 in Samples OPR Labeled Compound Congener Name Congener Number VER (%) Recovery (%) Recovery in Samples (%) 2,2',3,3',4,5,5',6,6'-NoCB 208 75-125 60-135 NA DeCB 209 75-125 60-135 Labeled Compounds 13Ci2-2-MonoCB 1L 50-145 15-145 5-145 13Ci2-4-MonoCB 3L 50-145 15-145 5-145 13Ci2-2,2'-DiCB 4L 50-145 15-145 5-145 13Ci2-2,5-DiCB 9L 50-145 15-145 5-145 13Ci2-3,3'-DiCB 11L 50-145 15-145 5-145 13Ci2-2,2',6-TrCB 19L 50-145 15-145 5-145 13Ci2-2,4,4'-TrCB 28L 50-145 15-145 5-145 13Ci2-2,4',6-TrCB 32L 50-145 15-145 5-145 13Ci2-3,4,4'-TrCB 37L 50-145 15-145 5-145 13Ci2-2,2',4,4'-TeCB 47L 50-145 15-145 5-145 13Ci2-2,2',5,5'-TeCB 52L 50-145 15-145 5-145 13Ci2-2,2',6,6'-TeCB 54L 50-145 15-145 5-145 13Ci2-2,3',4',5-TeCB 70L 30-135 15-145 10-145 13Ci2-3,3',4,4'-TeCB 77L 50-145 40-145 10-145 13Ci2-3,4,4',5-TeCB 80L 50-145 40-145 10-145 13Ci2-3,3',4,4'-TeCB 81L 50-145 40-145 10-145 13C12-2,2', 3,5' ,6-PeCB 95L 50-145 40-145 10-145 13Ci2-2,2',3,4',5-PeCB 97L 50-145 40-145 10-145 13C12-2,2' ,4,5,5' -PeCB 101L 50-145 40-145 10-145 13C12-2,2' ,4,6,6' -PeCB 104L 50-145 40-145 10-145 13C12-2,3,3' ,4,4' -PeCB 105L 50-145 40-145 10-145 13Ci2-2,3,4,4',5-PeCB 114L 50-145 40-145 10-145 13C12-2,3' ,4,4' ,5 -PeCB 118L 50-145 40-145 10-145 13Ci2-2',3,4,4',5-PeCB 123L 50-145 40-145 10-145 13Ci2-3,3',4,4',5-PeCB 126L 50-145 40-145 10-145 13Ci2-3,3',4,5,5'-PeCB 127L 50-145 40-145 10-145 13Ci2-2,2',3,4,4',5'-HxCB 138L 50-145 40-145 10-145 13Ci2-2,2',3,4,5,5'-HxCB 141L 50-145 40-145 10-145 13Ci2-2,2',4,4',5,5'-HxCB 153L 50-145 40-145 10-145 13Ci2-2,2',4,4',6,6'-HxCB 155L 50-145 40-145 10-145 13Ci2-2,3,3',4,4',5-HxCB 156L 50-145 40-145 10-145 13Ci2-2,3,3',4,4',5'-HxCB 157L 50-145 40-145 10-145 13C i 2-2,3,3' ,4,5,5' -HxCB 159L 50-145 40-145 10-145 13Ci2-2,3',4,4',5,5'-HxCB 167L 50-145 40-145 10-145 13Ci2-3,3',4,4',5,5'-HxCB 169L 50-145 40-145 10-145 13Ci2-2,2',3,3',4,4',5-HpCB 170L 50-145 40-145 10-145 13Ci2-2,2',3,4,4',5,5'-HpCB 180L 50-145 40-145 10-145 13Ci2-2,2',3,4',5,6,6'-HpCB 188L 50-145 40-145 10-145 13Ci2-2,3,3',4,4',5,5'-HpCB 189L 50-145 40-145 10-145 13Ci2-2,2',3,3',4,4',5,5'-OcCB 194L 50-145 40-145 10-145 13Ci2-2,2',3,3',5,5',6,6'-OcCB 202L 50-145 40-145 10-145 13Ci2-2,2',3,3',4,4',5,5',6-NoCB 206L 50-145 40-145 10-145 13Ci2-2,2',3,3',4,5,5',6,6'-NoCB 208L 50-145 40-145 10-145 13Ci2-DeCB 209L 50-145 40-145 10-145 Cleanup Standards 13Ci2-3,3',4,5'-TeCB 79L 50-145 40-145 10-145 13Ci2-2,2'3,3'5,5'6-HpCB 178L 50-145 40-145 10-145 'VER = Calibration verification 2OPR = Ongoing precision and recovery 3The suffix "L" in a congener number indicates an isotopically labeled compound. 2020 NCCA Quality Control (QC) Acceptance Criteria for PCB Congener Analyses of Tissue Samples D-5 ------- |