REPORT

WESTON & SAMPSON ENGINEERS, INC.

712 Brook Street, Suite 103
Rocky Hill, CT 06067
tel: 860.513.1473

September 2021
Connecticut Department of Energy and Environmental Protection
Statewide PFAS Initiative

Sampling Plan – Per- and Polyfluoroalkyl

Substances at Publicly Owned Treatment
Works – REVISION 2
Project Name: PFAS at POTWs Title: Sampling Plan – PFAS at POTWs
Project Location: Connecticut Revision Number:2
Revision Date: 09/02/2021 Page 1 of 21

SECTION I – TITLE AND APPROVAL PAGE

Document Title: Sampling Plan – Per- and Polyfluoroalkyl Substances at Publicly Owned Treatment
Works
Prepared For: Connecticut Department of Energy and Environmental Protection
Prepared By: John Zbell and Loren McGrath, Weston & Sampson Engineers, Inc.
Address/Telephone Number: 712 Brook Street, Suite 103, Rocky Hill, CT 06067 (860) 513-1473
Date: 09/02/2021

Weston & Sampson Project Manager:

Signature
John G. Zbell 09/02/2021
Printed Name/Date

Weston & Sampson Project QA Officer:

Signature
Loren A. McGrath 09/02/2021
Printed Name/Date

CTDEEP Project Manager: on behalf of

Signature
Rowland Denny 9/8/2021
Printed Name/Date

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Project Location: Connecticut Revision Number:2
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TABLE OF CONTENTS Page

SECTION I – TITLE AND APPROVAL PAGE ...................................................................................... 1

SECTION II – INTRODUCTION ......................................................................................................... 4

SECTION III – PROJECT ORGANIZATION AND RESPONSIBILITY ................................................... 5

Organization and Responsibility ...................................................................................................5

SECTION IV – PROBLEM DEFINITION ............................................................................................. 7

SECTION V – PROJECT DESCRIPTION/PROJECT TIMELINE .......................................................... 8

Project Description .......................................................................................................................8
Data Quality Objectives ..............................................................................................................10
Applicable Standards .................................................................................................................10
Project Timeline ..........................................................................................................................10

SECTION VI – SAMPLING DESIGN................................................................................................. 11

SECTION VII – SAMPLING AND ANALYTICAL METHOD REQUIREMENTS.................................... 12

SECTION VIII – METHOD AND SOP REFERENCE TABLE .............................................................. 13

SECTION IX – ANALYTICAL SENSITIVITY, PRECISION, AND ACCURACY ..................................... 14

SECTION X – FIELD DATA EVALUATION ....................................................................................... 15

Verification and Validation Requirements....................................................................................15

SECTION XI – LABORATORY DATA EVALUATION ......................................................................... 16

Laboratory Data Review Process ................................................................................................16
Data Review Process ..................................................................................................................16
Review Laboratory Report Narratives..........................................................................................17
Verification Summary Report ......................................................................................................17

SECTION XII – DATA USABILITY..................................................................................................... 18

Precision .....................................................................................................................................18
Accuracy .....................................................................................................................................18
Representativeness ....................................................................................................................19
Completeness ............................................................................................................................20
Comparability .............................................................................................................................20
Data Sensitivity ...........................................................................................................................20
Usability Summary ......................................................................................................................20

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Project Name: PFAS at POTWs Title: Sampling Plan – PFAS at POTWs
Project Location: Connecticut Revision Number:2
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LIST OF TABLES

TABLE 1 ...................................................................................................... POTWs to be Sampled

LIST OF APPENDICES

Appendix A .............................................................................................. CTDEEP Letters to POTWs

Appendix B .......................................................................................................... Sampling Schedule
Appendix C ....................................................................................................... Field Sampling SOPs
Appendix D .............................................................................................................. Laboratory SOPs
Appendix E ............................................................................................... Analytical Sensitivity Table

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Project Name: PFAS at POTWs Title: Sampling Plan – PFAS at POTWs
Project Location: Connecticut Revision Number:2
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SECTION II – INTRODUCTION
Weston & Sampson has prepared this Sampling Plan on behalf of the Connecticut Department of Energy
and Environmental Protection (CTDEEP) for sampling and laboratory analysis of environmental media
to evaluate the presence of per- and polyfluoroalkyl substances (PFAS) at publicly owned treatment
works (POTWs) throughout Connecticut. This project is being completed with funding provided by the
CTDEEP under a statewide PFAS Initiative.
Background
The CTDEEP is conducting an investigation to identify the presence of PFAS at POTWs geographically
distributed throughout the State. This project is focused on evaluating the presence of PFAS in POTW
influent, effluent, and sludge at 35 POTW locations. In addition, 4 biosolid incinerators will be sampled
from the input sludge and incinerator scrubber water. Finally, receiving surface water bodies and select
species of fish tissue within these water bodies at 10 locations co-located near sampled POTWs will
also be sampled. The results will be used by the CTDEEP to evaluate the general distribution of PFAS
at POTWs and the water bodies into which they discharge.

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Project Name: PFAS at POTWs Title: Sampling Plan – PFAS at POTWs
Project Location: Connecticut Revision Number:2
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SECTION III – PROJECT ORGANIZATION AND RESPONSIBILITY

Organization and Responsibility

CTDEEP

Rowland Denny
Project Manager

Weston & Sampson

Project Management
John Figurelli, PG, LEP
Principal-In-Charge Technical Review

John Zbell, PG, LEP Steven LaRosa

Project Manager

POTW/Surface Water Sampling Biota Sampling

William Sirotnak Anchor, QEA
Megan Shanahan
Richard Manandhar
Robert Colantonio

Analytical Laboratory Data QA/QC GIS/Data Management

Pace Analytical Services, LLC Loren McGrath Daniel Shinnick, GISP
East Longmeadow, MA

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Project Name: PFAS at POTWs Title: Sampling Plan – PFAS at POTWs
Project Location: Connecticut Revision Number:2
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The following briefly describes project responsibilities for personnel involved in this Sampling Plan:
 CTDEEP – Responsible for initial coordination with POTWs, contract and project management,
document review, approval, and project changes.
Rowland Denny phone: 860-424-3749 email: [email protected]
Connecticut Department of Energy and Environmental Protection
79 Elm Street
Hartford, CT 06106-5127

 Weston & Sampson:

o Project Management – Responsible for ensuring the project schedule and scope adheres to this
Sampling Plan. Responsible for regularly informing the CTDEEP of work progress and for
ensuring that project quality control and quality assurance elements adhere to this Sampling
Plan.
o Field Sampling – Responsible for ensuring the project fieldwork is performed in accordance with
the Sampling Plan and coordinating field investigations with Anchor, QEA.
John Zbell phone: 959-777-5822 email: [email protected]
Steven LaRosa phone: 802-244-5051x6007 email: [email protected]

Weston & Sampson Engineers, Inc.

712 Brook Street, Suite 103
Rocky Hill, CT 06067

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Project Name: PFAS at POTWs Title: Sampling Plan – PFAS at POTWs
Project Location: Connecticut Revision Number:2
Revision Date: 09/02/2021 Page 7 of 21

SECTION IV – PROBLEM DEFINITION

In July 2019, the Governor of Connecticut established the Interagency PFAS Task Force to protect
Connecticut residents and the environment from the harmful effects of PFAS. PFAS are a large group
of human-made chemicals that have been used for decades due to their resistance to heat, oil, stains,
grease, and water. PFAS have been documented in surface water, groundwater, soils, food, indoor dust,
plants, invertebrates, fish, and mammals. These compounds are stable, persistent, and considered
highly mobile.
In November 2019, the Governor released the “PFAS Action Plan”, which was prepared by the Task
Force. The Plan includes a number of actions items and steps to evaluate and address PFAS in
Connecticut. The responsibility for implementing the Plan has transitioned to the CTDEEP, Connecticut
Department of Public Health, and other state agencies.
As part of their efforts, the CTDEEP is presently undertaking the development of an interagency
geographic information system (GIS) to document sources of PFAS contamination and evaluate the
potential exposure to nearby sensitive receptors. The CTDEEP is also interested in identifying likely
sources of PFAS contamination. The scope of work addressed under this Sampling Plan, will specifically
focus on the assessment of PFAS associated with POTWs. These facilities are believed to be impacted
by discharges of PFAS to the sanitary sewer system from industrial and commercial processes.

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Project Name: PFAS at POTWs Title: Sampling Plan – PFAS at POTWs
Project Location: Connecticut Revision Number:2
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SECTION V – PROJECT DESCRIPTION/PROJECT TIMELINE

Project Description
The CTDEEP has identified 35 POTWs where they plan to sample and analyze environmental media to
evaluate the occurrence of PFAS. In addition, surface water and fish tissue sampling are planned at 10
of the POTWs. The locations of sample collection are summarized on the attached Table 1.
Task 1: Kick-Off Meeting and Sampling Plan Preparation:
On June 11, 2021, staff from Weston & Sampson, Anchor QEA and the CTDEEP held a virtual project
kick-off meeting to review the proposed scope of work and schedule.
Weston & Sampson has prepared this Sampling Plan for review and approval by the CTDEEP.
Task 2: Coordinate Access with POTWs:
On March 10 and May 28, 2021, the CTDEEP provided letters to the Superintendents of the 35 POTWs,
which had been selected for sampling. The letters, which are attached as Appendix A, outlined the
Statewide PFAS initiative, the scope for the proposed sampling and requested access to the facility
under Connecticut General Statues Section 22a-6(a)(6).
Upon approval of this Sampling Plan, Weston & Sampson will contact each of the POTWs to reiterate
the sampling scope and coordinate dates and times to conduct the sampling.
Task 3: Field Sampling:
Prior to the visiting each POTW, our interactive data collection system, iDataCollect, will be populated
with contact information, addresses, and additional information for each POTW. The iDataCollect
system integrates with Geographic Information System and EnvrioData8 platforms, which will be used
to create databases and maps of the results.
Influent, effluent and sludge samples will be collected at each of the 35 POTWs during two seasonal
sampling events: one in the summer and the other in the winter. At each of the 4 facilities that have
active sludge treatment, influent sludge and scrubber water samples will also be collected. Samples
will be collected at locations already established by the POTW operators. If a sample location has not
previously been established, we will work in concert with the POTW operator to determine an appropriate
sample location. If the optimal sampling location is not located within a regularly occupied space and/or
may pose a health and safety risk, in consultation with our in-house safety administrators we will develop
an acceptable method for sampling the location. A digital tablet will be used to collect real-time site-
specific sample information (date, time, location, visual observations, etc.). Photographs will be taken
at each sampling location to document the condition of the sample and facilitate the collection of
samples from the same location during subsequent sampling events.
An attempt will be made to schedule sampling of the POTWs, within a 2-week time timeframe for both
the summer and winter sampling events. If possible, sampling of multiple POTWs will be completed on
the same day.
Surface water sampling and fish collection will be conducted at the ten locations listed on Table 1.
Surface water sampling will be conducted by Weston & Sampson and fish collection will be conducted
by Anchor QEA. Sampling locations will extend from the POTW outfalls to 1.5 kilometers (km)
downstream. Sampling may also extend upstream of the outfalls but will not extend beyond barriers to
fish passage or into major confluence areas. Attempts will be made to avoid fish collection in areas of
other potential substantial PFAS discharge. Distances will be measured with a Differential Global

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Positioning System (DGPS) and start and end points of the sampling transects will be recorded on the
DGPS. Detailed logs of land use on each bank within the sampling reach will be kept noting direct
discharges and facilities with the potential to be using PFAS. Prior to initiating fish collection, a data
collection permit will be obtained from CTDEEP.
Fish samples will be collected by electrofishing. The method of electrofishing will be dependent on
water depth and may include the use of a backpack unit for shallow water, tote barge for wadeable
water up to waist deep and a vessel for deeper water (i.e., the Connecticut River sites).
Electrofishing will occur over the entirety of the site (from the outfall downstream 1.5 km, bank to bank),
targeting areas of preferred habitat which may include pools, riffles and varying structure. Sampling will
generally be conducted starting upstream and moving downstream to aid in netting the fish as the
current will keep them in front of the netting crew.
Target species are smallmouth bass (Micropterus dolomieu) and white sucker (Catostomus
commersonii). These species were selected to include a predatory gamefish species (smallmouth bass)
residing in the top trophic level and a bottom dwelling species (white sucker). Substitute species may
be considered based on availability. The order of preference for alternate gamefish species are
largemouth bass, then yellow perch. The order of preference for alternate bottom fish is catfish,
common carp and then fallfish. Stocked species such as trout will not be targeted due to spending less
time in the waterbodies and limiting the amount of bioaccumulation over time. Species substitutions
and/or size range modifications will be discussed between the field crew and project managers and will
need CTDEEP approval before a final determination is made.
At each of the ten locations, three composite samples of gamefish consisting of five fish of each species,
will be collected for a total of 30 samples. Composites of gamefish will consist of legal-size fish of similar
size (CTDEEP proposed that fish should be within +/- 1 inch in length). If necessary, we will collect
composites of more than one species to achieve the 3 composite samples per location (e.g., 2
composites of smallmouth bass and 1 composite of perch) depending upon species abundance.
Three composites of bottom feeding fish will also be collected at each location consisting of similar size
fish (within +/- 1 inch) for a total of 30 samples. Each composite will be made up of the same species
If necessary, we will collect composites of more than one species to achieve the 3 composite samples
per location (e.g., 2 composites of white sucker and 1 composite of carp) depending upon species
abundance.
If the target number of fish is not reached at a location, composite samples may consist of a minimum
of two fish.
Based on our experience, achieving the proposed size requirement of within 1 inch in length for all fish
within a composite may be difficult. Any field modification to target size range will approved by CTDEEP
as a field modification, e.g., to allow retained fish to be within 75% of the length of the composite batch,
which is a standard approach.
Field processing of fish will be limited to avoid cross-contamination. Stunned fish will be netted and
placed in HDPE buckets to recover. Non-target fish and target fish with deformities/abnormalities will
be released upon recovery. Target species will be placed in a resealable plastic bag to measure length
and inspect for abnormalities. Each bag will be labelled and then placed in a second bag prior to storage
on wet ice for transport to Weston & Sampson’s Rocky Hill, Connecticut office for pickup by a laboratory
courier. In the event the fish need to remain at the office overnight they will be placed in the freezer.

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Upon receipt, the laboratory will freeze the fish samples prior to shipping to the processing and analytical
laboratory.
Fish will be processed in the analytical laboratory by descaling and filleting prior to homogenization as
composite samples. The composite samples will utilize the left-side fillets. It has been assumed that
sufficient tissue mass will be available from a single side fillet from 3-5 fish for each composite. The
right-side fillets will be utilized if more sample mass is required. If sufficient mass is present from the
left-side fillets, the right-side fillets will be archived for future use, if necessary (e.g., QA/QC issues with
original sample analysis). Following completion of data validation, unused samples can be archived at
the laboratory for 6 months or shipped to CTDEEP for archiving.
Michigan Department of Environmental Quality (MDEQ) has developed a reference guide summarizing
procedures and protocols for the sampling of tissue for PFAS which will be adhered to for this sampling
event. Further information regarding these procedures is presented in Section VIII.
Task 4: Laboratory Analysis:
All samples will be submitted to Con-Test Analytical Laboratory, a Pace Analytical Laboratory, for
analysis. The POTW influent, effluent, incinerator scrubber water and surface water samples will be
analyzed in accordance with Con-Test SOP 454, an in-house isotope dilution method for analyzing PFAS
in non-drinking water samples that complies with Department of Defense Quality Systems Manual 5.3
B-15. Sludge samples will be analyzed by Contest SOP 466. Fish tissue samples will be analyzed by
SOP MIN4-0178. All three methods report up to at least 34 PFAS, including the 18 PFAS specified in
EPA Method 537. A summary of the analytes and detection limits by media are included in Appendix
E. The results will be reported to Weston & Sampson as an electronic data deliverable (EDD), EPA Level
IV data package, within 28 days of sample receipt. Electronic data deliverables will also be formatted
for upload to the EPA Water Quality Exchange database.

Data Quality Objectives

The primary data quality objective (DQO) for this project is to provide information regarding overall PFAS
loading at POTWs, the distribution of PFAs in the POTW treatment train, contribution of PFAS to receiving
surface water bodies, and biota within those surface water bodies.

Applicable Standards
There are currently no applicable standards for PFAS in the media being sampled as part of this project.

Project Timeline
Upon approval of this sampling plan, Weston & Sampson anticipates commencing the first of two
rounds of sampling in Late August/early September 2021. The second round of sampling would be
completed in February/March 2022. The schedule outlining project tasks, major milestones,
anticipated dates, and development of deliverables is included in Appendix B.

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SECTION VI – SAMPLING DESIGN

The detailed scope of work addressing sample collection and analysis is provided below.

Sampling - Weston & Sampson will coordinate with the facility operators to collect the designated media
samples at the locations shown in Table 1 attached. The table below summarizes the total number of
samples to be collected of each media.

Number Sample Total # of

Sample Location Sample Type
Frequency Samples
Influent 2 rounds 70

Effluent 35 2 rounds 70
POTW Sludge 2 rounds 70

Incinerator Sludge 2 rounds 8

4
Scrubber Water 2 rounds 8

Upstream 1 round 10

Downstream 1 round 10
Surface Water 10
Predator 3/location 30
Fish Tissue
Bottom Feeder 3/location 30

The sampling locations will be dependent on each individual facility design and available sampling
ports. Weston& Sampson will utilize the same sampling locations as the facility uses for influent, effluent,
and sludge as applicable. Samples of the various media will be collected in accordance with the Field
Sampling Standard Operating Procedures (SOPs) provided in Appendix C. Additional information
regarding sampling methods is presented in Section VII.

Sample Analyses - Each of the samples will be shipped to Con-Test Analytical under chain-of-custody
in sealed coolers. The analytical methods used for PFAS quantification will be dependent upon the
media being analyzed.

POTW influent, POTW effluent, POTW scrubber water and surface water samples will be analyzed using
a proprietary method following SOP 454, which includes the use of solid phase extraction and internal
isotope dilution. Solids/sludges will be analyzed using a proprietary method following SOP 466 Rev 7
using internal isotope dilution. Fish tissue samples will be analyzed via MIN4-0178, which includes the
use of internal isotope dilution. All three methods comply with Department of Defense Quality Systems
Manual 5.3 B-15 and are included in Appendix D. Additional information regarding analytical methods
is presented in Sections VII and VIII.

The list of PFAS to be reported by each method of analysis and the limits of detection and quantification
are included in the table in Appendix E.

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SECTION VII – SAMPLING AND ANALYTICAL METHOD REQUIREMENTS

The parameters and number of samples associated with each analysis for the different types of media are presented in the following table.

Sampling Analytical Maximum

Parameter Media Type Sample # Analytical SOP Containers Preservation
SOP*** Method Holding Time
Extracted within
148 samples + (2) 250-mL high-density
14 days and
8 Blanks + 14 SOP-25 polyethylene (HDPE) or Cool to ≤ 10°C but
PFAS Aqueous* DoD QSM 5.3 See Section VIII extracts
Field Duplicates SOP-26 polypropylene (PP) containers not frozen
analyzed within
Total=170 with unlined plastic screw caps
28 days
Extracted within
78 samples + (1) 250-mL high-density
SOP-1 14 days and
7 Field polyethylene (HDPE) or Cool to ≤ 10°C but
PFAS Solid** SOP-10 DoD QSM 5.3 See Section VIII extracts
Duplicates polypropylene (PP) containers not frozen
SOP-26 analyzed within
Total=85 with unlined plastic screw caps
28 days
Extracted within
20 samples + (2) 250-mL high-density
14 days and
Surface 2 Field SOP-13 polyethylene (HDPE) or Cool to ≤ 10°C but
PFAS DoD QSM 5.3 See Section VIII extracts
Water Duplicates SOP-26 polypropylene (PP) containers not frozen
analyzed within
Total=22 with unlined plastic screw caps
28 days
60 samples +
2 Field Cool to ≤ 6°C to
See Lab Resealable plastic bags
PFAS Fish Tissue Duplicates DoD QSM 5.3 See Section VIII lab for One year
SOP (double)
(second fillet) homogenization****
Total=62
* Aqueous samples include influent, effluent and scrubber water
**Solid samples include sludges and incinerator sludges
***Sampling SOPs can be found in Appendix C
****Samples will be frozen at processing laboratory

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SECTION VIII – METHOD AND SOP REFERENCE TABLE

The following table includes the laboratory SOP references, included in Appendix D.

Matrix Analytical Method Reference: Testing Facility Laboratory Analytical SOPs:

Determination of Selected Perfluorinated Alkyl Substances (PFAS) SOP ID 454 PFAS Water Isotope Dilution
Aqueous by Solid Phase Extraction and Liquid Chromatography/Tandem Con-Test Revision #7
Mass Spectrometry Isotope Dilution (LC/MS/MS) 06/30/21

Determination of Selected Perfluorinated Alkyl Substances (PFAS) SOP ID 466 PFAS Water Isotope Dilution
Soil/Solid Samples by Liquid Chromatography/Tandem Mass
Solid Con-Test Revision #7
Spectrometry Isotope Dilution (LC/MS/MS)
06/30/21

USEPA National Guidance for Assessing Chemical Contaminant ENV-SOP-GBAY-0129 Sample Homogenization,
Tissue Data for Use in Fish Advisories, Volume 1: Fish Sampling and Compositing and Sub-Sampling
Analysis – 3rd Edition
Con-Test Revision #3
(Preparation)
02/09/21
Department of Defense Department of Energy Consolidated
Quality Systems Manual (QSM) for Environmental Laboratories, ENV-SOP-MIN4-0178 Determination of Selected 36
Tissue Version 5.3, Appendix B, Table B-15, 06/19
Per- and Polyfluoroalkyl Substances (PFAS) by
Con-Test
(Analysis)
LC/MS/MS (Isotope Dilution)
DoD Guidance for PFAS Analysis in Biota. 04/20

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SECTION IX – ANALYTICAL SENSITIVITY, PRECISION, AND ACCURACY

The method detection limits and reporting limits related to the analytical testing methods are provided
in Appendix E. All reporting limits have been reviewed for accuracy as of the date of this Sampling Plan.

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SECTION X – FIELD DATA EVALUATION

Verification and Validation Requirements

This section provides a discussion of the type and extent of quality control evaluation that will be
completed in conjunction with the analytical data collected at the POTWs. Results of this evaluation will
be used to provide verification for reported sample concentrations.
Verification of Sampling Procedures
The following criteria will be used to evaluate the field sampling data:
• Documentation of field equipment calibration activities;
• reviewing data for technical credibility vs. the sample site setting;
• auditing field sample data records and chain-of-custody;
• auditing of sample handling and preservation procedures; and
• holding times for sample analyses.
Sampling procedures will be evaluated by the Field Lead and/or the Project Manager as appropriate.
The results of the evaluation will be included in reports prepared after each of the seasonal sampling
rounds and resulting impacts to the data will be discussed.
Data Verification and Validation
Data evaluation reports, in general accordance with EPA Region 1 Functional Guidelines, will be
submitted with each of the seasonal reports. Deviations from the standard evaluation process, along
with justification for why the changes were made will be described in the reports.

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SECTION XI – LABORATORY DATA EVALUATION

Laboratory Data Review Process

Every laboratory report will include a certification pertaining to the analytical procedures, and associated
QC criteria and performance standards for all data included in the reports. This Analytical Report
Certification Form includes a statement attesting to the validity of the results, a dated signature of a
qualified laboratory representative and a set of “yes” or “no” answers pertaining to questions regarding
compliance with method-specific QC criteria, performance standards, analytes reported and
consistency with COC documentation. Any question answered with a “no” should be fully explained in
the project narrative, with additional information and/or documentation attached as necessary.

Data Review Process

When Weston & Sampson receives the laboratory data package, Weston & Sampson will verify the data
is complete, correct, and contractually compliant. If Weston & Sampson finds that data are missing or
incorrect and the discrepancies are not already noted in the laboratory’s case narrative for that data
package, Weston & Sampson will request missing data and an explanation from the laboratory. The
EPA “New England Environmental Data Review Program Guidance”, dated April 22, 2013, will be used
as guidance for the data evaluation.
Tier I Evaluation
Data review evaluation will include a review of tabulated quality control results and comparison against
EPA Region I validation limits and/or project specific criteria to identify bias or other interferences that
could affect the quality of sample results. Specific quality control components to be evaluated include
the following:
 Data completeness check
 Holding times
 Sample preservation
 Blank results. The 5x and 10x rules (EPA, 1996) will be used to qualify sample concentrations
that have detections in associated blanks
 Surrogate recoveries
 Matrix spike and matrix spike duplicate results
 Field duplicates
 Laboratory control sample results
Tier II Evaluation
If, during data evaluation, significant problems with data are encountered, then a Tier II validation
process may be performed. Specific quality control components to be evaluated in the Tier II review
include everything from the Tier 1 review plus the following:
 Initial and continuing calibration results
 Internal standard results
 GC/MS tuning results
 Serial dilution results

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Review Laboratory Report Narratives

If any QC issues are documented in the Laboratory Report Narrative, which are not included in the data
package, then Weston & Sampson, during the data validation process, will contact the laboratory and
obtain copies of relevant necessary QC information. Surrogate recovery, laboratory control sample,
duplicate, field blank, trip blank and temperature blanks checks are part of the data validation process
that Weston & Sampson will complete and are thoroughly reviewed in the Tier II-type data
validation/usability assessment process. Data limitations and bias will be identified within the report
analytical summary tables.

Verification Summary Report

Weston & Sampson will include a data verification section in the text of the seasonal summary reports
that summarizes overall data quality observations and conclusions and explains any limitations of the
data.

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SECTION XII – DATA USABILITY

Weston & Sampson will review the data in general accordance with the “U.S. Environmental Protection
Agency Guidance on Quality Assurance Project Plans CI0 2106-G-05 QAPP”. Weston & Sampson will
evaluate the data quality objectives (DQOs) in general accordance with the PARCCS (precision,
accuracy, representativeness, completeness, comparability and sensitivity) parameters outlined in the
above-referenced guidance document.
Data Qualifiers
Based on validation results, qualifiers will be added to reported analyte concentrations to indicate
uncertainty or potential bias or interferences. Specific data qualifiers which will be applied to organic
sample concentration include the following:
 U - The analyte was not detected above the practical quantitation limit.
 J - The analyte was detected but the associated reported concentration is approximate and is
considered estimated.
 R - The reported analyte concentration is rejected due to serious deficiencies with associated
quality control results. The presence or absence of the analyte cannot be confirmed.
 UJ - The analyte was not detected above the PQL. However, due to quality control results that
did not meet acceptance criteria, the quantitation limit is uncertain and may not accurately
represent the actual limit.
Weston & Sampson will evaluate the data to determine if it meets the PARCCS (precision, accuracy,
representativeness, completeness, comparability and sensitivity) parameters outlined in the above-
referenced guidance document. The PARCCS criteria are described below.

Precision
Precision is a measure of mutual agreement among individual measurements of the same property and
is generally expressed as the reproducibility of the analytical result between the initial sample and the
field duplicate sample as expressed by the relative percent difference (% RPD). Weston & Sampson will
include field duplicate results in the report tables. Duplicate sample results will be listed adjacent to the
existing sample results so that a direct analyte by analyte concentration comparison can be made.
Weston & Sampson will calculate the RPD for each duplicate sample result by calculating:
| (sample concentration) – (duplicate concentration) | x 100 = RPD (%)
(simple average of sample and duplicate concentration)
The acceptable RPD values for POTW influent, effluent, sludges and fish tissue will be <50% and for
surface water will be <30%. Any duplicate results that are above these RPD limits will be flagged on
the data tables.

Accuracy
Accuracy is the degree of measurement with an accepted reference or true value. The difference
between the measurement and the true value is usually expressed as a percentage ratio. Weston &

westonandsampson.com
18
Project Name: PFAS at POTWs Title: Sampling Plan – PFAS at POTWs
Project Location: Connecticut Revision Number: 1
Revision Date: 08/23/2021 Page 19 of 21

Sampson will evaluate accuracy by reviewing the following: laboratory control sample (LCS) results,
surrogate results, matrix spike/matrix spike duplicate (MS/MSD) results, calibration QC results, and field
and laboratory blank results. Poor accuracy may be the result of laboratory error, field error, or the natural
sample matrix. In the data usability reports, Weston & Sampson will evaluate the cause of bias/accuracy
problems. For environmental samples, poor accuracy can be due to interferences present in the natural
sample.
Continuing calibration accuracy checks are assessed by comparing the true value against the reported
concentration. The percent difference between the results is calculated as follows:
Accuracy may be expressed as a percent difference (%D) calculated by the following equation:

%D = (Vt - Vm)/Vt x 100

Where:
Vt = the true or real value expected.
Vm = the measured or observed value.
The degree of accuracy demonstrated for laboratory control and matrix spike samples is expressed as
a percent recovery. The percent recovery indicates the amount of known concentration of an analyte
that has been detected by the associated instrumentation. The percent recovery (%R) is calculated as
follows:
%R = (SSR – SR)/SA x 100
Where:
SSR = the spiked sample result.
SR = the unspiked sample result.
SA = the value of the spike added.

The objective for accuracy of laboratory determinations is to demonstrate that the analytical
instrumentation provides consistent measurements, which are within EPA and statistically derived
method-specific accuracy criteria. Laboratory data quality objectives for accuracy as measured by
“%Recovery” are provided in the laboratory SOPs (see Appendix D).

Representativeness
Representativeness expresses the degree to which data accurately and precisely represent a
characteristic of a population, parameter variation, or environmental condition. Weston & Sampson will
review data qualitatively for representativeness. Sample representativeness will be reviewed qualitatively
through the review of the proper use of the specified sampling SOPs, through internal field sampling
audits, and through the use of field QC samples including field duplicates and trip blanks. Any
unexpected, unusual, or anomalous results will be mentioned in the report text.

westonandsampson.com
19
Project Name: PFAS at POTWs Title: Sampling Plan – PFAS at POTWs
Project Location: Connecticut Revision Number: 1
Revision Date: 08/23/2021 Page 20 of 21

Completeness
Completeness is a measure of the amount of valid data obtained from a measurement system relative
to the amount expected under normal conditions. Completeness is expressed as a percentage and is
determines as follows:
Completeness (A%) = # of valid values reported for a parameter x 100
# of samples collected for analysis for that particular parameter
A% = Acceptance Percentage
The project completeness goal is 85-90% percent of the samples being successfully analyzed. If less
than 85 percent completeness is obtained, further sampling may need to be performed. Incomplete
sampling from critical areas may result in the need for further sampling, even if the overall 85-90%
completeness goal is met.

Comparability
Comparability refers to the ability to generate data for each parameter that is both comparable between
sampling locations and over time. Comparability is a qualitative parameter that expresses the
confidence with which data sets can be compared to one another. Comparable data allows for the
ability to combine analytical results (e.g., current data with historic data) acquired from the various media
to be sampled at each of the POTWs. Comparability relies upon precision and accuracy within the
individual data sets to be acceptable to promote confidence in the data sets.

Data Sensitivity
Sensitivity is a measure of whether the laboratory method was sufficient to report detected contaminants
at concentrations at or below the applicable guidance criteria. Prior to laboratory analysis, anticipated
laboratory reporting limits will be compared to the most recent and applicable state and EPA guidance
levels and procedures will be adjusted as needed to ensure that reporting limits are at or below the
required standards. The applicable reporting limits will likely vary based on site conditions, sensitive
receptors, etc.
Any non-detected data result that has a quantitation limit above the applicable state and EPA guidance
concentration will be flagged on the data tables. For non-detected chemical results for which the project
quantitation limit (QL) was not achieved, the risk assessor may determine that the use of one-half of the
reported QL is sufficient as a conservative approach in risk calculations.

Usability Summary
Any or all of the following considerations for precision, accuracy, and completeness may be evaluated
to determine if DQOs have been met. To meet these requirements, quality control criteria are provided
in the standard laboratory methodologies. These criteria include the use of field duplicates and matrix
spike samples to assess precision, matrix spikes, laboratory control samples and calibration results to
assess accuracy; blank samples to determine representativeness; field duplicates to assess
comparability. The amount (percentage) of valid data obtained from validation will be used to determine
completeness. The results of the data usability evaluation will be included in the seasonal reports, and
impacts and/or limitations on the use of the data will be discussed.

westonandsampson.com
20
Project Name: PFAS at POTWs Title: Sampling Plan – PFAS at POTWs
Project Location: Connecticut Revision Number: 1
Revision Date: 08/23/2021 Page 21 of 21

Weston & Sampson will include a data usability section in each report’s text that summarizes overall
data usability and answers whether the data is usable overall for report findings and conclusions.
Data usability will be performed by the project staff with support from the Project QA Officer and
confirmed by the Project Manager.

\\wse03.local\WSE\Projects\CT\CT DEEP\PFAS WWTF Sampling\Sampling Plan\Text_PFAS WWTF_ POTW_Sampling Plan.docx

westonandsampson.com
21
 TABLE 1
CTDEEP STATEWIDE PFAS INITIATIVE
SAMPLING OF PUBLICLY OWNED TREATMENT WORKS

POTWs TO BE SAMPLED

PLANT Sample Types per Location

Incinerator Input Incinerator
Influent Effluent Sludge Surface Water Fish Tissue
Sludge Scrubber Water
BEACON FALLS WPCF     
BRIDGEPORT WEST WPCF   
BRIDGEPORT WPCF   
BRISTOL WPCF     
CHESHIRE WPCF   
DANBURY WPCF   
EAST HADDAM WPCF   
FARMINGTON WPCF     
GEORGETOWN WPCF   
GRASS ISLAND WPCF   
HARTFORD MDC WPCF       
HERITAGE VILLAGE WATER CO   
LEDYARD WPCF   
MATTABASSETT DISTRICT WPCF
    
MERIDEN WPCF   
MONTVILLE WPCF   
NAUGATUCK WPCF     
NEW HAVEN WPCF     
NEW LONDON WPCF   
NEWTOWN WPCF   
NORWALK WPCF   
SALISBURY WPCF   
SHARON WPCF   
SOMERS WPCF     
STAFFORD WPCF   
THOMASTON WPCF   
THOMPSON WPCF   
TORRINGTON WPCF   
UNIVERSITY OF CONNECTICUT WPCF
  
VERNON WPCF     
WALLINGFORD WPCF     
WATERBURY WPCF     
WINDHAM WPCF   
WINDSOR POQUONOCK WPCF
    
WINSTED WPCF     

\\wse03.local\WSE\Projects\CT\CT DEEP\PFAS WWTF Sampling\Sampling Plan\WPCF PFAS Testing List - Alpha w emails for distribution
 APPENDIX A

westonandsampson.com
79 Elm Street • Hartford, CT 06106-5127 www.ct.gov/deep Affirmative Action/Equal Opportunity Employer

March 10, 2021

RE: Statewide PFAS Initiative

Superintendent:

PFAS are a family of man-made chemicals that contain a high content of fluorine atoms, which gives them many useful
properties, including the abilities to repel water, prevent staining, and increase heat resistance. As such, PFAS are used in
thousands of industrial processes and consumer products, including coatings for fabrics and nonstick cookware, grease-
resistant food packaging materials (e.g., microwave popcorn bags), and firefighting foam used to put out flammable liquid
fires. Once released to the environment, PFAS are persistent and do not readily biodegrade or break down and have serious
adverse impacts on human health and the environment, even at low levels. Currently, neither the U.S. Environmental
Protection Agency (EPA) nor the Connecticut Department of Energy and Environmental Protection (DEEP) has a regulatory
water quality standard for PFAS. DEEP has undertaken a statewide initiative to study PFAS, including the levels present in
wastewater.

As part of the Statewide PFAS Initiative the DEEP will be working with a contractor to conduct sampling of domestic
wastewater treatment plant (POTW) influent, effluent, sludge and/or scrubber water (if applicable) for PFAS compounds
from 34 POTWs around the state.

Your facility has been selected for this effort and a separate letter will be sent to inform you who the commissioner’s agent
is once selected. You will then be contacted by the commissioner’s authorized agent to schedule a date for sample
collection.

There is no cost to your facility for this effort nor for the testing. When the report is finalized an electronic copy will be
provided to you.

Please provide access for this sampling effort as directed by CGS 22a-6(a)(6). Under this statute the commissioner may
undertake any studies, inquiries, surveys or analyses she may deem relevant, through the personnel of the department or in
cooperation with any public or private agency, to accomplish the functions, powers and duties of the commissioner.

Thank you for your cooperation in our effort to determine the prevalence of PFAS in our state.

If you have any questions or comments please contact Rowland C. Denny of my staff at 860-424-3749 or
[email protected].

Sincerely,

Jennifer Perry, P.E., Director

Water Planning and Management Division
Water Planning and Land Reuse Bureau
79 Elm Street • Hartford, CT 06106-5127 www.ct.gov/deep Affirmative Action/Equal Opportunity Employer

May 28, 2021

RE: Statewide PFAS Initiative

Superintendent:

As a follow up to my letter to you dated March 8, 2021, the Department of Energy and Environmental Protection
(DEEP) has selected Weston & Sampson Engineers to conduct sampling of publicly owned treatment works
(POTW) influent, effluent, sludge and scrubber water (if applicable) for PFAS compounds from 34 POTWs
around the state.

As part of the Statewide PFAS Initiative, your facility has been selected for this effort. You will be contacted by
Weston & Sampson Engineers, the commissioner’s authorized agent, to schedule a date(s) for sample collection.

There is no cost to your facility for this effort, including but not limited to, testing and analysis costs. When the
report is finalized, an electronic copy will be provided to you.

Please provide access to the necessary sampling locations for this sampling effort as directed by Connecticut
General Statutes Section 22a-6(a)(6). Under this statute, the commissioner may undertake any studies, inquiries,
surveys or analyses she may deem relevant, through the personnel of the department or in cooperation with any
public or private agency, to accomplish the functions, powers and duties of the commissioner.

The intent of this effort is to assist the DEEP in directing our efforts to evaluate the introduction of PFAS into
sanitary sewers and determine the extent and degree of PFAS at POTWs at various points in the treatment train.

Thank you for your cooperation in our effort to determine the prevalence of PFAS in our state.

If you have any questions or comments please contact Rowland C. Denny of my staff at 860-424-3749 or
[email protected].

Sincerely,

Jennifer Perry, P.E., Director

Water Planning and Management Division
Water Protection and Land Reuse Bureau
 APPENDIX B

westonandsampson.com
 CTDEEP STATEWIDE PFAS INTIATIVE
SAMPLING OF PUBLICLY OWNED TREATMENT WORKS

Project Schedule

2021 2022
ACTIVITY
8 9 10 11 12 1 2 3 4 5 6
CTDEEP Review and Comment on Sampling
Plan
Finalize Sampling Plan

Communication/Scheduling with POTWs

Sampling of POTWs (24 locations with no
surface water or fish tissue sampling)
Sampling of POTWs (10 locations with surface
water and fish tissue sampling)
Laboratory Analysis (up to 28 days)

QA/QC and Data Tabulation and Review

Prepare Draft Report

CTDEEP Review and Comment on Draft Report

Finalize Report

Communication/scheduling with POTWs

Round 2 Sampling of POTWs (24 locations with
no surface water or fish tissue sampling)
Laboratory Analysis (up to 14 days)

QA/QC and Data Tabulation and Review

Draft Report Generation

CTDEEP Review and Comment on Draft Report

Finalize Report
 APPENDIX C

westonandsampson.com
 SOP 24 – Composite Sludge Sampling
Environmental, Geotechnical, and Energy Division
Page 1 of 2

SOP 1: Composite Sludge Sampling

Purpose: A Composite Sludge Sample is a mixture of one or more individual “grab
samples” from several discrete locations in a stockpile and/or discrete
depths within a tank or lagoon.
Applicable for: Compositing sludge samples while in the field for laboratory analysis of
metals, polychlorinated biphenyls (PCBs), or other suitable analyte.
Not applicable for: Sludge being collected for volatile organic compounds (VOCs) analysis.

PREREQUISITES MATERIALS NEEDED

None, unless otherwise noted.  Sludge collection/measuring device (i.e. stainless steel
trowel, pre-cleaned/ disposable scoop/bucket auger with
“T” handle, “Sludge-Judge”);
 Stainless steel mixing bowl or inert plastic sheeting;
 Pre-cleaned mixing spoon/device (i.e. wooden tongue
depressor);
 Pre-cleaned laboratory container; and
 Personal protection equipment: at a minimum protective
gloves and eye protection.

PROCEDURE

1) Determine the number of “grab” locations to be sampled. This will be dependent on the
volume, solids content and storage method of the sludge. Generally, at least 4 “grab”
locations should be sampled. The intent is to collect sufficient “grabs” from locations likely
representative of the entire sludge mass.
2) Select locations and/or depths from which sludge “grab” samples are desired. Tank samples
can be selected from several access points at depths of 1/3 to 2/3 of the tank depth. Lagoon
bottom samples should be collected from locations both near and distal to the lagoon inlet.
3) Prepare a sampling sketch showing the general location of each “grab” sample. Label each
“grab” location uniquely.
4) Collect a known volume of equal amount of sludge “grab” sample from each location using a
collection/measuring device (decontaminated HDPE bottle, sludge-judge, scoop).
5) Empty each sludge “grab” sample into the homogenization container or onto a mixing surface.
6) Thoroughly mix or “composite” sludge using an inert mixing device.
7) Using a pre-cleaned, stainless steel scoop, plastic spoon, or trowel, remove composite soil
sample and place required volume into designated pre-cleaned laboratory container for
analysis.
8) Label sampling container with a waterproof label using a waterproof ink pen with: unique
name, project name, W&S project number, date, time of collection, sampler initials,
preservative utilized and required analyses.

Revision 0 SOP Owner: W&S

Implementation Date: 08/20/19 Approval: SJL
Last Update Date: 08/20/19 Page 1 of 2
 SOP 24 – Composite Sludge Sampling
Environmental, Geotechnical, and Energy Division
Page 2 of 2

9) Transfer sampling label information to the laboratories preferred Chain of Custody.

10) Place sample container into a cooler/box and prepare for transportation to the laboratory
(ice, vermiculite, bubble wrap, etc.).

REFERENCES

1. MassDEP. 1991. Standard References for Monitoring Wells (MassDEP Policy #WSC-310-91).

2. USACE, 2001, Requirements for the Preparation of Sampling and Analysis Plans (USACE EM
200-1-3). February 1

Revision 0 SOP Owner: W&S

Implementation Date: 08/20/19 Approval: SJL
Last Update Date: 08/20/19 Page 2 of 2
 SOP 10 – Sampling Soil with a Scoop or Hand Auger
Environmental, Geotechnical, and Energy Division
Page 1 of 4

SOP 10: Sampling Soil with a Scoop or Hand Auger

Purpose: This procedure covers the method and equipment used to collect surface
and near-surface samples of soils and physically similar materials using
a scoop or hand auger.
Applicable for: Collecting samples of contaminated soils and similar materials primarily
near the surface. Subsurface samples can be obtained by first removing
higher layers using a shovel or other suitable equipment and collecting
the sample with the scoop.
Not applicable for: Water samples.

PREREQUISITES MATERIALS NEEDED

✓ Stainless steel hand auger;
Driller to contact utility clearance
Phone (811) at least 72 hours and ✓ Ruler or Measuring tape;
no more than 10 days prior to any ✓ Decontamination equipment;
drilling activities and AFTER pre- ✓ Sampling Plan;
marking is performed.
✓ Soil collection/measuring device (e.g. Encore or
Terracore syringe or pre-cleaned stainless steel trowel,
Additional notification of disposable scoop);
applicable local municipalities ✓ Pre-cleaned, pre-preserved laboratory containers;
and/or private utilities where
✓ Sample container label; and
required.
✓ Personal Protective Equipment: at a minimum
protective gloves and eye protection
Refer to SOP 5: Pre-Marking
Boring Locations if needed.

PROCEDURE
Non VOC Analysis
1. Samples should be collected in accordance with an appropriate work plan.
2. Remove the top layers of material down to the required sample depth using a shovel or other
suitable equipment. A shovel or other suitable equipment can be used for the initial removal
of overburden material. This equipment should be manufactured from material that is
compatible with the soil or waste to be sampled.
3. Measure to the depth at which the sample will be collected with a ruler or tape measure.
Record this information in a field log book.
4. Remove the thin layer of material that was in contact with the overburden removal equipment
and discard it using a clean scoop. The project scope will define if the scoop may or may not
be reused to collect the actual sample.

Revision 2 SOP Owner: W&S

Implementation Date: 11/21/2013 Approval: SJL
Last Update Date: 03/28/2019 Page 1 of 4
 SOP 10 – Sampling Soil with a Scoop or Hand Auger
Environmental, Geotechnical, and Energy Division
Page 2 of 4

5. Collect a suitable volume of sample with the scoop* (the same scoop can be used to collect
multiple scoopfuls to obtain sufficient volume to fill the container). Use a new (or
decontaminated) scoop for each sample. Transfer the sample into the suitable container.
Samples should be contained in plastic, glass, or other non-reactive certified-clean containers.
6. Close the sample container and label with a waterproof label using a waterproof ink pen with:
unique name, project name, W&S project number, date, time of collection, sampler initials,
preservative utilized and required analyses.
7. Place sample container into a cooler/box and prepare for transportation to the laboratory (ice,
vermiculite, bubble wrap, etc.).
8. Complete the field log book with all relevant information and observations about the sample
location and chain-of-custody form.
9. Decontaminate the reusable equipment in accordance with the protocol specified in the
project scope and refer to SOP 8: Decontaminating Equipment.

VOC Analysis
This method is suitable for collecting soil samples for volatile organic compounds (VOC) for high
and low level detection limits via EPA Method 5035. In most cases samples are being collected
as a part of an initial subsurface investigation and general VOC concentrations are unknown.
Therefore, samples should be collected for both “low concentration” and “high concentration”
analysis methods. By collecting samples for both analysis methods you are assured results will
be reported with appropriate detection limits. The procedure below collects sufficient samples for
both analysis methods.
1. Organize sampling containers near sampling device.
a. Low Concentration – 40 ml vial with 5ml water and 1g sodium bisulfate
b. High Concentration – 40 ml vial with 10 mls methanol
c. Moisture Content – 40 ml vial with no preservative
2. Select desired sampling interval(s) within sampling device.
3. Remove the thin layer of material that was in contact with the overburden removal equipment
and discard it using a clean scoop.
4. Using an appropriate sample collection device (preferably Encore or Terracore syringe or
clean stainless steel scoop), collect approximately 5g of sample as soon as possible after the
surface of the soil or other solid material has been exposed to the atmosphere: generally within
a few minutes at most.
5. Transfer sample from syringe/scoop to pre-preserved sample container. Clean thread and
secure cap. Repeat Step 3 for each pre-preserved container. Fill remaining unpreserved
container nearly full and secure cap.
6. Label each container with a waterproof label using a waterproof ink pen with: unique name,
project name, W&S project number, date, time of collection, sampler initials, preservative
utilized and required analyses.

Revision 2 SOP Owner: W&S

Implementation Date: 11/21/2013 Approval: SJL
Last Update Date: 03/28/2019 Page 2 of 4
 SOP 10 – Sampling Soil with a Scoop or Hand Auger
Environmental, Geotechnical, and Energy Division
Page 3 of 4

7. Place sample containers into a cooler/box and prepare for transportation to the laboratory (ice,
vermiculite, bubble wrap, etc.).
8. Complete the field log book with all relevant information and observations about the sample
location and chain-of-custody form.
9. Complete the field log book and chain-of-custody form.
Decontaminate the reusable equipment in accordance with the protocol specified in the project
scope and refer to SOP 8: Decontaminating Equipment.
REFERENCES
1. USEPA. 2005. Region 9 – Technical Guidelines for Accurately Determining Volatile Organic
Compound (VOC) Concentrations in Soil and Solid Matrices (EPA-R9QA/05.2). December.
2. USACE. 2001. Requirements for the Preparation of Sampling and Analysis Plans (USACE EM 200-
1-3). February 1.
3. USACE. 1998. Sample Collection and Preparation Strategies for Volatile Organic Compounds in
Solids (USACE 1998b).

Revision 2 SOP Owner: W&S

Implementation Date: 11/21/2013 Approval: SJL
Last Update Date: 03/28/2019 Page 3 of 4
 SOP 10 – Sampling Soil with a Scoop or Hand Auger
Environmental, Geotechnical, and Energy Division
Page 4 of 4

Revision 2 SOP Owner: W&S

Implementation Date: 11/21/2013 Approval: SJL
Last Update Date: 03/28/2019 Page 4 of 4
 SOP 13 – Surface Water Sampling
Environmental, Geotechnical, and Energy Division
Page 1 of 2

SOP 13: Surface Water Sampling

Purpose: Surface water sampling is the process of collecting surface water samples
from a body of water for laboratory analysis.
Applicable for: Surface water sampling while in the field for laboratory analysis of
polychlorinated biphenyls (PCBs), volatile organic compounds (VOCs),
metals or other suitable analyte.
Not applicable for: Soil samples and groundwater samples.

PREREQUISITES MATERIALS NEEDED

✓ Sampling Plan
None, unless otherwise noted by
✓ Selected Sampling Equipment: Disposable Bailers,
Project Engineer.
Peristaltic Pump, Kemmerer or Von Dorn sampler.
✓ Nitrile disposable gloves
✓ Paper Towels
✓ Pre-cleaned sample containers, preserved as appropriate
✓ Field notebook
✓ Personal protective equipment (i.e., hard hat, waterproof
boots, waders, PFD, etc.)
✓ Cooler and ice

PROCEDURE

1) Select locations from which surface water grab samples are desired using Site
Sampling Plan. Select the appropriate sampling method for each sample location and
data needs.
2) Initiate sampling at the downstream location and proceed up stream to each sampling
location.
3) Collect Sample:
Dipping Method
A sample may be collected directly into the sample container when the surface water
source is accessible by wading or other means. The sampler should face upstream if
there is a current and collect the sample without disturbing the bottom sediment. The
surface water sample should always be collected prior to the collection of a sediment
sample at the same location. The sampler should be careful not to displace the
preservative from a pre-preserved sample container, such as the 40-ml VOC.
Bailer Method
Submerge the bailer into the water column until the bailer is filled with water. Carefully
transfer the water from the bailer into the pre-cleaned laboratory sample container,
making sure that the bailer does not come in contact with the sample containers.

Revision 2 SOP Owner: W&S

Implementation Date: 11/21/2013 Approval: SJL
Last Update Date: 03/27/2019 Page 1 of 2
 SOP 13 – Surface Water Sampling
Environmental, Geotechnical, and Energy Division
Page 2 of 2

Peristaltic Pump Method

The peristaltic pump can be used to collect a water sample from any depth if the pump
is located at or near the surface water elevation. There is no suction limit for these
applications. The use of a metal conduit to which the tubing is attached, allows for the
collection of a vertical sample (to about a 25-foot depth) which is representative of the
water column. The tubing intake is positioned in the water column at the desired depth
by means of the conduit. Using this method, discrete samples may be collected by
positioning the tubing intake at one depth or a vertical composite may be collected by
moving the tubing intake at a constant rate vertically up and down the water column
over the interval to be composited. New tubing is to be used at each sampling
location.
Kemmerer/Von Dorn Method
The Kemmerer sampler is a brass cylinder with rubber stoppers that leave the ends of
the sampler open while being lowered in a vertical position, thus allowing free passage
of water through the cylinder. The Van Dorn sampler is plastic and is lowered in a
horizontal position. In each case, a messenger is sent down a rope when the sampler
is at the designated depth, to cause the stoppers to close the cylinder, which is then
raised. Water is removed through a valve to fill respective sample containers. With a
rubber tube attached to the valve, dissolved oxygen sample bottles can be properly
filled by allowing an overflow of the water being collected. With multiple depth
samples, care should be taken not to disturb the bottom sediment, thus biasing the
sample. When metals and organic compounds parameters are of concern, then a
double-check valve, stainless steel bailer or Kemmerer sampler should be used to
collect the sample. The sampler must be decontaminated after each sample is
collected.
4) Transfer surface water sample to appropriate sample containers. For volatile organic
compounds (VOC) analysis ensure as little agitation or disturbance as possible.
5) Close the sample container and complete and attach the sample label.
6) Record all relevant information and observations about the sample location.
7) Complete the field log book and chain-of-custody form.
8) Decontaminate the reusable equipment in accordance with the protocol specified in
the project scope and refer to SOP 8: Decontaminating Equipment.

REFERENCES
1. USEPA. 2016. Science and Ecosystem Support Division Operating Procedure – Surface Water
Sampling (EPA-SESDPROC-201-R4). Revision 4, Effective December 16

Revision 2 SOP Owner: W&S

Implementation Date: 11/21/2013 Approval: SJL
Last Update Date: 03/27/2019 Page 2 of 2
 SOP 26: PFAS Related Sampling Considerations
Page 1 of 2

SOP 26: Per-and Polyfluorinated Alkyl Substance Related Sampling

Purpose: Guidelines to be applied IN ADDITION to environmental media sampling
and laboratory analysis for the presence of per- and polyfluorinated alkyl
substances (PFAS).
Applicable for: PFAS sampling while in the field for laboratory analysis via EPA Method 537
Ver. 1.1, 537.1, 533; DoD QSM 5.2; or lab proprietary PFAS methods.
Not applicable for: N/A

PREREQUISITES MATERIALS NEEDED

Media specific SOP, soil,  Sampling Plan
groundwater, surface water etc.  Sampling Materials (spoons, bailers, etc)
 Laboratory provided bottles and blanks
 Field notebook
 Paper towels
 Sampling Sheet and/or iPad with appropriate
iDataCollectSM Form
 Nitrile gloves
 Cooler and ice

PROCEDURE
PFAS are frequently present in the clothing, sampling materials and PPE used for routine
environmental media sampling. PFAS presence should be considered ubiquitous in all consumer
materials we use, unless specifically noted as “PFAS free”. Therefore, extra care must be taken
when collecting samples for PFAS analyses. The following provides guidance to be utilized to
minimize the risk of inadvertently contaminating media samples with PFAS. Adherence to
“standard”/”good practice” sampling protocols will greatly reduce the risk of inadvertently
contaminating samples. These additional measures should be taken to further reduce potential
contamination of samples.
1) Sampler must assure that no potential PFAS containing materials are utilized during
sampling. No materials containing Teflon, Goretex, or other waterproofing can be
utilized while sampling. LDPE products should be avoided (tubing, tarps, spoons,
bailers) in favor of HDPE. Aluminum foil should also be avoided as it may have a thin
layer of coating with PFAS.
2) Extra care to assure clothing, storage containers, and sampling equipment do not
contain potential PFAS must be taken. Clothing should not be “weather proof”, “water
proof” or contain Teflon. Natural fiber clothing that has been laundered without fabric
softener is preferred. If rain gear must be worn, field blanks should be collected at
each sample location to determine if PFAS impacts occur during sampling.
3) Use of cosmetics, insect repellant, deodorant, perfumes, and sun block should be
minimized if possible. PFAS free products may exist and should be utilized when
available. The following products are acceptable:

Revision 1 SOP Owner: W&S

Implementation Date: 07/23/20 Approval: SJL
Last Update Date: 07/23/20 Page 1 of 2
 SOP 26: PFAS Related Sampling Considerations
Page 2 of 2

• Sunscreens - Alba Organics Natural Sunscreen, Yes To Cucumbers, Aubrey

Organics, Jason Natural Sun Block, Kiss my face, and baby sunscreens that are
“free” or “natural”
• Insect Repellents - Jason Natural Quit Bugging Me, Repel Lemon Eucalyptus Insect
repellant, Herbal Armor, California Baby Natural Bug Spray, BabyGanics
• Sunscreen and insect repellant – Avon Skin So Soft Bug Guard Plus – SPF 30 Lotion
4) Decontamination solutions and rinse waters must also be assured to be PFAS free. If
deemed necessary samples of the decontamination materials may be needed to
confirm lack of PFAS.
5) Record all relevant information and observations about the sample location including
an inventory of potential PFAS containing items in the area nearby the sampling point,
if any exist. Avoid “Rite-in-the-Rain” notebooks and waterproof paper, sharpies and
pencils. Ballpoint pen and loose leaf paper is preferred.
6) Place bottles in the shipping container, add ice and packing materials sufficient to
keep samples cool and protected from damage during shipping. DO NOT UTILIZE
CHEMICAL ICE PACKS.

References: ITRC PFAS Fact Sheets, 2019

Revision 1 SOP Owner: W&S

Implementation Date: 07/23/20 Approval: SJL
Last Update Date: 07/23/20 Page 2 of 2
 APPENDIX D

westonandsampson.com
 ENV-SOP-GBAY-0129, Rev 03

Document Information
Document Number: ENV-SOP-GBAY-0129 Revision: 03

Document Title: Sample Homogenization, Compositing and Sub-Sampling

Department(s):
Other

Date Information

Effective Date: 09 Feb 2021

Notes

Document Notes:

All Dates and Times are listed in: Central Time Zone

1 of 19
 ENV-SOP-GBAY-0129, Rev 03

Signature Manifest
Document Number: ENV-SOP-GBAY-0129 Revision: 03
Title: Sample Homogenization, Compositing and Sub-Sampling
All dates and times are in Central Time Zone.

ENV-SOP-GBAY-0129-Rev.03 Sample Homogenization, Compositing and Sub-Sampling

QM Approval

Name/Signature Title Date Meaning/Reason

Kate Verbeten (007119) Manager - Quality 08 Feb 2021, 07:26:11 PM Approved

Management Approval

Name/Signature Title Date Meaning/Reason

Nils Melberg (007142) General Manager 2 09 Feb 2021, 08:41:28 AM Approved
Christopher Haase (007121) Manager 09 Feb 2021, 01:28:41 PM Approved

2 of 19
 ENV-SOP-GBAY-0129, Rev 03

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Sample Homogenization, Compositing and Sub-Sampling
TEST METHOD NA
ISSUER: Pace ENV – Green Bay Quality – GBAY
COPYRIGHT © 2002 - 2021 Pace Analytical Services, LLC

1.0 SCOPE AND APPLICATION

This standard operating procedure (SOP) describes the laboratory procedure for homogenizing soil,
liquid, and biota samples to obtain a representative sample aliquot used for analysis or composite.
This procedure is restricted to use by, or under the supervision of, technicians experienced in the
preparation of samples.

2.0 SUMMARY OF METHOD

Solid, liquid, or biological samples are thoroughly mixed or blended to ensure that any aliquots taken
are representative of the sample as a whole. The samples are mixed in either their original containers
or, in the event that original container does not allow for adequate mixing, transferred to an inert
container for thorough homogenization.
Necropsy and/or filleting of whole-body animals may be performed to isolate the individual organs or
portions of the specimen to be homogenized and utilized for analysis.
This SOP involves instruction to chop, grind, and blend plant materials, biological tissue, sediment and
synthetic materials into a homogenized sample compatible with preparation of semi volatile organic
extracts and metals digestates.
Analysts must make reasonable judgments when sub-sampling materials in order to obtain a
homogenous, representative aliquot of the material. Because of the nature of environmental samples,
the analyst may have to treat samples on a case-by-case basis ensuring that all aspects of the
analytical method are performed. In the event that the analyst cannot reasonably determine what
constitutes a representative sample, the project manager or supervisor must be involved so that the
client can help ensure an appropriate representative aliquot is utilized.

3.0 INTERFERENCES
Metallic Devices – Samples to be analyzed for metal constituents must not be homogenized using any
metallic mixing devices or containers as it may result in contamination of the sample with a variety of
metals. Use only glass, plastic or ceramic materials when working with these sample types. This may
not be applicable for tissue samples since metallic devices (blenders, etc.) may be necessary for
grinding and chopping prior to sample homogenization.
Plastic Devices – Samples to be analyzed for organic constituents must not be homogenized using
any plastic mixing devices or containers as it may result in both positive and negative interferences.
Use only glass and ceramic devices when working with these sample types. Metal instruments may
also be used if analysis for metals is not required from the same sample
Solvents, reagents, glassware, and other sample processing hardware may yield discrete artifacts
and/or elevated baselines causing misinterpretation of the analytical results. All these materials must
be free from interferences under the conditions of the analysis, demonstrated by performing method
blanks.

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released, or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

3 of 19
 ENV-SOP-GBAY-0129, Rev 03

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Sample Homogenization, Compositing and Sub-Sampling
TEST METHOD NA
ISSUER: Pace ENV – Green Bay Quality – GBAY
COPYRIGHT © 2002 - 2021 Pace Analytical Services, LLC

4.0 DEFINITIONS
Refer to the Laboratory Quality Manual for a glossary of common lab terms and definitions.
Biota- the flora or fauna of a region.
Composite- combining the typical or essential characteristics of individuals, making up a group.
Fillet- to cut an edible portion of fish. This may or may not contain the ribcage and belly flap and is
dependent upon the regulatory, scientific, and data quality objectives for the project.
Head- the upper or anterior division of the animal body that contains the brain, the chief sense organs,
and the mouth.

5.0 HEALTH AND SAFETY

The toxicity or carcinogenicity of each chemical material used in the laboratory has not been fully
established. Each chemical should be regarded as a potential health hazard and exposure to these
compounds should be as low as reasonably achievable.
The laboratory maintains documentation of hazard assessments and OSHA regulations regarding the
safe handling of the chemicals specified in each method. Safety data sheets for all hazardous
chemicals are available to all personnel. Employees must abide by the health, safety and
environmental (HSE) policies and procedures specified in this SOP and in the Pace Chemical Hygiene
/ Safety Manual.
Personal protective equipment (PPE) such as safety glasses, gloves, and a laboratory coat must be
worn in designated areas and while handling samples and chemical materials to protect against
physical contact with samples that contain potentially hazardous chemicals and exposure to chemical
materials used in the procedure. Hearing protection should be worn when a blender is in operation, or
during sediment processing when applicable.
Liquid Nitrogen presents additional hazards and may cause cryogenic burns or displace oxygen and
cause rapid suffocation. Use in a well-ventilated area with additional PPE designed for handing these
materials.
Contact your supervisor or local HSE coordinator with questions or concerns regarding safety protocol
or safe handling procedures for this procedure.

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released, or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

4 of 19
 ENV-SOP-GBAY-0129, Rev 03

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Sample Homogenization, Compositing and Sub-Sampling
TEST METHOD NA
ISSUER: Pace ENV – Green Bay Quality – GBAY
COPYRIGHT © 2002 - 2021 Pace Analytical Services, LLC

6.0 SAMPLE COLLECTION, PRESERVATION, HOLDING TIME, AND STORAGE

The laboratory does not perform sample collection for this procedure. Samples should be collected in
accordance with a sampling plan and procedures appropriate to achieve the regulatory, scientific, and
data quality objectives for the project. The laboratory will record any nonconformance to these
requirements in the laboratory’s sample receipt record.
Details concerning sample shipping, preservation and storage can be found in the applicable extraction
and/or analytical SOPs, or as specifically discussed below. Samples must be stored separately from
all standards and reagents. Where possible, samples for trace analysis should be segregated from
highly contaminated samples to avoid cross contamination. Food or drink products must always be
kept away from samples and never stored in the same area with samples.
Biota samples must be kept frozen at ≤-10˚C in their original sample containers until the
homogenization process occurs.
Small rodents must undergo a special procedure to destroy any Hantavirus which may be present.
Refer to the Pace SOP: ENV-SOP-GBAY-0130 Small Rodent Handling and Homogenization (most
recent revision or replacement) for details.
After homogenization, biota samples are kept frozen at ≤-10˚C in glass jars sized closest to the
prepared homogenate sample volume. Individual jars of samples are grouped together as appropriate
and stored in a labeled cardboard box within the freezer.
Sediment samples which will be air-dried are received at ≤6°C. After the dry and grind procedure is
completed the samples are retained at room temperature.
Synthetic materials may be kept at room temperature.

7.0 EQUIPMENT AND SUPPLIES

7.1 Equipment
Table 7.1: General Laboratory Homogenization Equipment and Supplies
Supply Vendor* Model / ID* Catalog #* Description
Spatula Fisher Stainless Steel S50822 -
Spatula Fisher Cooper Surgical Inc 11080 Plastic scrapers
Capable of weighing Analytical or top
Analytical Balance Varied NA
nearest sensitivity loading
Sample Mixing Metal, plastic, glass or Dependent upon
Varied NA
Containers ceramic sample composition
*Or equivalent

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released, or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

5 of 19
 ENV-SOP-GBAY-0129, Rev 03

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Sample Homogenization, Compositing and Sub-Sampling
TEST METHOD NA
ISSUER: Pace ENV – Green Bay Quality – GBAY
COPYRIGHT © 2002 - 2021 Pace Analytical Services, LLC

Table 7.2: Biota Homogenization Equipment and Supplies

Supply Vendor* Model / ID* Catalog # Description
Spatula Fisher Stainless Steel S50822 -
Spoons Fisher Spoonulet 14-375-2 Lab spoon
Cutting Board Cooking Supply HDPE Stainless Steel NA Inert
Stainless Steel or
Knives Sharp Hi Carbon NA Inert
Titanium
Meat Cleaver Sharp Hi Carbon Lamson NA Stainless Steel
Mallet Stanley 2lb Mallet NA Plastic Face, 2-3lb
Robot Coupe Robot Coupe R2UB NA Serial# 2471016003J-11
Meat Grinder Hobart E-222 / Cast Iron NA Serial# 9243-0011-02990
Bell Housing Berkle Cast Iron NA For Meat Grinder
Stainless steel blade and
blender cup (may use
Blender Waring Industrial Grade NA glass)
Scaler NA NA NA Stainless Steel
Aluminum Foil NA Heavy Duty NA NA
Pliers NA Stainless Steel NA NA
Analytical Balance Mettler Toledo PE-16 NA Capable 15000±0.1g
Analytical Balance Sargent Welch 400DR NA Capable of 150±0.001g
Analytical Balance A&D GH200 NA Capable of 50±0.0001g
Vial C&G 40mL NA Amber Glass
2oz CG
Wide-mouth 4oz AG Clear or Amber Glass with
Container C&G or QEC 9oz AG Various Teflon lined cap
Foam ear plugs or
noise reducing
Ear plugs or Earmuffs Howard Leight earmuffs NA Moldex
Paper Towels NA 11x8.8inch NA Georgia Pacific
*Or equivalent

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released, or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

6 of 19
 ENV-SOP-GBAY-0129, Rev 03

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Sample Homogenization, Compositing and Sub-Sampling
TEST METHOD NA
ISSUER: Pace ENV – Green Bay Quality – GBAY
COPYRIGHT © 2002 - 2021 Pace Analytical Services, LLC

Table 7.3: Sediment Homogenization Equipment and Supplies

Supply Vendor* Model / ID Catalog # Description

Spatula Fisher Stainless Steel S50822 -
HDPE Stainless
Cutting Board Cooking Supply NA Inert
Steel
Mallet Stanley 2lb Mallet NA Plastic Face, 2-3lb
Rolling Pin Cooking Supply Marble NA NA
Aluminum Foil NA Heavy Duty NA Reynolds
Scissors Varies Stainless Steel NA NA
Pliers NA Stainless Steel NA NA
Weigh Boat Big Science Large 80060 Plastic
Analytical Balance Sargent Welch Capable 0.001g NA NA
Vial C&G 40mL NA Amber Glass
Quart size
Plastic Bags NA Gallon size NA Ziploc®
Foam ear plugs or
noise reducing
Ear plugs or Earmuffs Howard Leight earmuffs NA Moldex
Dust Mask Modex NA 4240-01-255-7856 NA
Paper Towels NA 11x8.8inch NA Georgia Pacific
*Or equivalent

Table 7.4 Computer Hardware and Software

Hardware/Software* Description* Make/ModelVersion*

Computers Laptop or desktop Various
Horizon Data reporting software (LIMS) See master list for current version.
Electronic Prep Log Electronic bench sheet software See master list for current version.
*Or equivalent

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released, or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

7 of 19
 ENV-SOP-GBAY-0129, Rev 03

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Sample Homogenization, Compositing and Sub-Sampling
TEST METHOD NA
ISSUER: Pace ENV – Green Bay Quality – GBAY
COPYRIGHT © 2002 - 2021 Pace Analytical Services, LLC

8.0 REAGENTS AND STANDARDS

8.1 Reagents
Table 8.1: Biota Reagents and Standards
Requirements/
Reagent Concentration/ Description Expiration Date
Vendor/ Item #*
Manufacturer’s recommended
Liquid expiration date or 5 years from
Nitrogen To homogenize biota matrices. 11700 receipt, whichever is sooner.
Canned / matrix for SVOA Starkist in water or None assigned until blended, then
Tuna parameters as required Equivalent given a 1-year expiration date.
Ground / matrix for metals and
Chicken wetchem parameters as required NA
DI Water Type II ASTM US Filter 18Ω NA
Alconox Cleaning Solution Fisher / 50-212-165 NA
Cleaning Solution / Commercial Clorox or
Bleach Grade equivalent NA
Dry Ice For shipping purposes NA NA
*Or Equivalent
Table 8.2: Sediment Homogenization Reagents and Standards
Requirements/
Reagent Concentration/ Description Expiration Date
Vendor/ Item #*
DI Water Type II ASTM US Filter 18Ω NA
Manufacturer’s recommended
expiration date or 2 years
from receipt, whichever is
Methanol Pesticide Grade Fisher / 141.40 sooner.
*Or equivalent

9.0 PROCEDURE
9.1 Analytical Balance Calibration
9.1.1 Annual Calibration – The balance must be calibrated at least annually by an outside agency
and checked daily before each use using Class 1 or 2 weights. Refer to Pace ENV-SOP-
GBAY-0115 Support Equipment (current revision or replacement).
9.1.2 Daily Calibration Check
9.1.2.1 Clean the balance and surrounding area prior to starting the daily calibration check.
9.1.2.2 Check the sight level on the balance. If it needs adjusting, level the balance.
9.1.2.3 The weight set ID indicated in the logbook is used as the primary set. If an
alternate weight set ID is used, that ID must be recorded in the comment section
of the balance calibration logbook for that day.

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released, or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

8 of 19
 ENV-SOP-GBAY-0129, Rev 03

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Sample Homogenization, Compositing and Sub-Sampling
TEST METHOD NA
ISSUER: Pace ENV – Green Bay Quality – GBAY
COPYRIGHT © 2002 - 2021 Pace Analytical Services, LLC

9.1.2.4 Tare the balance before weighing the NIST certified weights.
9.1.2.5 Use forceps or other means to lift each weight (Do not touch the weights with
fingertips as the residue may artificially adjust the true value of the weights).
Record the date of the calibration check, the true value of the weight, and the actual
measured weight in the logbook. Repeat this procedure for the other certified
weights. If calibration weights differ from the certified weights by more than
specified in the balance calibration logbook, corrective action must be taken (see
Section 9.1.3).
9.1.3 Corrective Action
9.1.3.1 Clean the balance and balance pan. Check the sight level on the balance and
adjust if necessary. Re-tare and reweigh all the certified weights.
9.1.3.2 The internal calibration function (if available) of the balance may be used as a
means of corrective action.
9.1.3.3 Utilize the internal calibration function and diagnostics. Refer to instrument
manual.
9.1.3.4 Contact the QA office for assistance if the balance does not meet the calibration
tolerances.
9.1.3.5 If the above action does not correct the problem, the balance should be taken out
of service and appropriately labeled to avoid improper usage. A service technician
should be contacted.
9.1.3.6 Record any corrective action. Initial and date all entries in the logbook.
9.2 Soil/Solid Sample Homogenization and Sub-sampling
9.2.1 Soil samples that are collected in regulated domestic areas or that are of foreign origin must
be handled in accordance with the Pace SOP: ENV-SOP-GBAY-0121, Regulated Soil
Handling (current revision or replacement).
9.2.2 An analyst examines the sample as received by the laboratory. If standing water is noted on
top of a soil or sediment sample, the water is incorporated into the sample. Any foreign
material, not subject to analysis, must be removed and discarded. This material may include
sticks, leaves, rocks, etc. that are not or cannot be analyzed for that particular procedure.
Any questions related to the nature of the sample should be directed to the Project Manager
(PM). The PM will work with the client to help determine what should be considered foreign
material and how to appropriately qualify the final report, if necessary.
9.2.3 The remaining sample must be homogenized until the point of an even consistency
throughout the entire sample. The easiest option, for non-volatile samples, is to mix the
sample thoroughly in the original container. The sample is considered thoroughly mixed once
all layers, colors, and inconsistencies have been evenly distributed throughout the container.
If additional mixing is still required to get an even consistency, proceed to the next step.
9.2.4 If mixing is difficult in the original container, the analyst should transfer the entire sample into
a larger, clean container and attempt homogenization there. After mixing, return the
homogenized sample to its original container. If homogenization within the container was
successful, this step may be omitted.

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released, or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

9 of 19
 ENV-SOP-GBAY-0129, Rev 03

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Sample Homogenization, Compositing and Sub-Sampling
TEST METHOD NA
ISSUER: Pace ENV – Green Bay Quality – GBAY
COPYRIGHT © 2002 - 2021 Pace Analytical Services, LLC

9.2.5 After mixing thoroughly, the analyst must take out a representative aliquot to use for
preparation or analysis. The analyst must not add or remove minute quantities of sample
from the representative aliquot in an attempt to target a specific final weight. If an analyst
weighs out slightly more than the required weight of sample, they have not adversely
impacted the reporting limit for that sample. If applicable, the analyst should also weigh out
portions of the sample for duplicate or spike analysis at this time so that all aliquots are
approximately equal to each other. When taking aliquots for QC samples, they must be taken
one after the other from the container. A larger amount of sample may not be removed from
the container, mixed further, and then subdivided.
9.2.6 For volatile soil samples, the analyst must attempt homogenization as quickly as possible to
minimize the loss of volatile analytes. This may limit the amount of mixing that can be done
on the sample. The analyst can quickly scrape off the top layer of soil, core down to the
center of the jar, and take an aliquot to acquire a sufficiently representative sample. It should
also be noted that for sample containers that are received for multiple analyses, the volatiles
department must take their aliquot first to minimize loss of volatiles and minimize the chance
of contamination from other departments. Then the sample can be processed for percent
moisture, organic extractions, etc.
9.2.7 Other options for soil homogenization may be employed if requested by a particular client.
When employing a client-requested method, there must be clear documentation in the
preparatory and analytical logbooks stating the method used.
9.3 Water/Liquid Homogenization and Sub-sampling
9.3.1 Water samples are homogenized by shaking the sample bottle prior to pouring an aliquot.
This applies to non-volatile analytical methods only. If a sample contains distinct layers –
either liquid or solid – the department supervisor must be consulted to determine the most
appropriate means of homogenizing and splitting the sample.
9.4 Biota (Biological Tissue and Plant) Homogenization:
9.4.1 Clean the work area by wiping the surfaces with a damp cloth. Follow procedure outlined in
SOP: ENV-SOP-GBAY-0143 Labware Cleaning Procedures (current revision or
replacement) to prepare utensils and grinders for use.
9.4.2 Depending on the sample matrix and specific instructions provided by the customer, the
method for ensuring homogeneity may vary. Necropsy and/or filleting of whole-body animals
may be performed to isolate the individual organs or portions of the specimen to be
homogenized and utilized for analysis. The project manager must be contacted for
clarification prior to thawing the samples if there are any questions.
9.4.3 Select a set of samples for processing. Depending on the size of the specimen, remove the
samples from the freezer to allow the specimen to partially thaw. Large specimens typically
need to thaw overnight at room temperature. Small specimens require a shorter amount of
time and may be placed in a refrigerator overnight or thawed at room temperature for 2-3
hours during the day of processing. It is important to make sure that each specimen is not
touching another specimen during the thawing process.
9.4.4 Record the date that the specimens are removed from the freezer to thaw in the Electronic
Biota Homogenization Log.

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released, or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

10 of 19
 ENV-SOP-GBAY-0129, Rev 03

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Sample Homogenization, Compositing and Sub-Sampling
TEST METHOD NA
ISSUER: Pace ENV – Green Bay Quality – GBAY
COPYRIGHT © 2002 - 2021 Pace Analytical Services, LLC

9.4.5 Pre-label the appropriate sized sample jars with the LIMS numbers. Samples should be
placed in the appropriate sized container dependent upon the sample mass received (40mL,
2oz, 4oz, 9oz). Transport the clean, dry utensils and pre-labeled jars to the countertop work
area.
9.4.6 If the client requires an equipment blank to be processed with the samples, the same
equipment which is used to process the samples must be Deionized Water (DI) rinsed prior
to sample processing. Multiple equipment blanks may be processed with each batch. The
equipment blank must be logged into the LIMs system to report with the sample data.
9.4.7 Once the specimen is adequately thawed, processing may begin. Compare the label on the
specimen with the pre-labeled jar to verify errors have not occurred.
9.4.8 Small fish, such as minnows, are usually collected as composites and will represent a single
composite sample. Large whole fish that require compositing are chopped into cubes and
put through the meat grinder together (refer to 9.4.10) and aliquots of the ground tissue are
blended with liquid nitrogen (refer to 9.4.11 through 9.4.12).
9.4.9 If the specimen requires filleting prior to homogenization, thaw the fish to the point that it can
be cut into with a sharp clean knife. Skinning or scaling may be necessary prior to filleting
the fish.
9.4.9.1 Skinning: Catfish, bullheads, and other fish may need to be skinned prior to
removing fillets. With a sharp knife slice the skin front to back along the dorsal
side of fish. Make another incision from top to bottom just behind the gills. Hold
the fish head with one hand and grasp an edge of the skin just behind the gill with
pliers. Peel the skin back toward the tail. The skin may also be removed by placing
the fillet with skin attached, skin side down on the cutting board. Remove the fillet
by running knife along the skin between the skin and fillet.
9.4.9.2 Scaling: If scales are to be removed prior to filleting, lay the fish flat on a cutting
board. Grasp the fish with one hand and with the other hand use a scaler to scrape
the scales off the fish. Work the scaler from the tail toward the head. Rinse the
scales and slime from fish prior to filleting.
9.4.9.3 Filleting: Begin with an incision just behind the gills, cutting through the fish from
back to belly. Next, make a clean cut along the dorsal ridge towards the tail. Be
careful not to cut into the gut cavity. After cutting through to the tail, separate the
fillet from the rib cage, peeling the fillet from the carcass with the non-cutting hand.
Pick out any bones. (Rib cage may be removed with the fillet at the client’s
request).
9.4.10 Chop large whole-body specimens, plant material, or fillets into 2-3 inch cubes using a sharp
knife and mallet. Smaller samples of limited quantity must be finely ground using the blender
in step 9.4.11.
9.4.11 Grind the cubes in a large commercial meat grinder to coarse texture. Repeat the procedure
a minimum of two times to ensure proper texture.
9.4.12 Transfer the course ground sample to a stainless-steel bowl containing liquid nitrogen. Place
the frozen sample in a blender cup and blend the frozen tissue to a powder consistency.
9.4.13 Transfer the blended sample into the pre-labeled jars.

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released, or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

11 of 19
 ENV-SOP-GBAY-0129, Rev 03

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Sample Homogenization, Compositing and Sub-Sampling
TEST METHOD NA
ISSUER: Pace ENV – Green Bay Quality – GBAY
COPYRIGHT © 2002 - 2021 Pace Analytical Services, LLC

9.4.14 Samples such as eggs, insects, and small individual organs (liver, brain) may be stirred
vigorously with a metal spatula in an appropriate sized container without liquid nitrogen to
avoid loss of sample. The technician documents that the sample was prepared in the
container on the prep worksheet or notebook.
9.4.15 Clean the work area and the utensils in accordance with the procedure outlined in SOP: ENV-
SOP-GBAY-0143 Labware Cleaning Procedures (current revision or replacement) between
samples.
9.4.16 Periodically, canned tuna or chicken is homogenized using this procedure for use as a quality
control matrix within the laboratory. Each analysis may require a different quality control
matrix, See Section 11 for additional information.
9.4.17 Sample integrity must be maintained throughout the digestion and analytical processes,
therefore samples which were blended with liquid nitrogen should be kept within a freezer at
≤-10°C up until the weighing process..
9.5 Sediment Homogenization Procedure (Air Dry-Grind Procedure):
9.5.1 Clean the work area by wiping the surfaces with Methanol. Follow procedure outlined in SOP:
ENV-SOP-GBAY-0143 Labware Cleaning Procedure (current revision or replacement) to
prepare utensils and grinding equipment for use.
9.5.2 Pre-label the appropriate aluminum sample drying tray with 2 removable labels containing
the sample LIMS numbers. Double bagged client samples should be placed behind the
aluminum sample drying tray. The secondary technician must verify the correct bag is placed
behind the correct drying tray.
9.5.3 Remove the interior sample bag and place on top of the exterior client bag. Rip along both
side seams of the interior bag. Place the opened bag inside out slightly above the drying tray
(the sample should fall from the bag into the tray). Ensure no contact occurs between the
outside of the bag and the drying tray. The technician may need to manually transfer the
sediment from the bag to the tray with their gloved hand. Once transferred, the sample is
spread evenly throughout the bottom of the drying tray.
9.5.3.1 An aliquot of the wet sample must be taken in order to complete the dry weight
analysis. Please see SOP: ENV-SOP-GBAY-0004 Measurement of Percent
Moisture in Soils and Solids (current revision or replacement).
9.5.4 The interior sample bag is then placed bag into the exterior client bag. Both bags are then
placed into a 2-gallon plastic bag labeled with the batch workorder number.
9.5.5 Prior to placing samples on the cart for transfer, clean the surface of the sample cart by
sweeping it to remove visible particulates. Then wipe the surfaces clean with Methanol.
9.5.6 Place all drying trays on the clean sample cart and deliver to the drying room. Use Methanol
to wipe the sample racks prior to placing the drying trays in the drying room and record the
drying room temperature, which should not exceed 100 degrees Fahrenheit. Adjust the
temperature as needed and log any changes in the temperature logbook.
9.5.7 Samples should dry for a minimum of 8 hours or until moisture content is less than or equal
to 10%. Once the sediment is adequately dried, processing may begin.
9.5.8 Clean the work area and all processing equipment with Methanol following SOP: ENV-SOP-
GBAY-0143 Labware Cleaning Procedure (current revision or replacement).

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released, or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

12 of 19
 ENV-SOP-GBAY-0129, Rev 03

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Sample Homogenization, Compositing and Sub-Sampling
TEST METHOD NA
ISSUER: Pace ENV – Green Bay Quality – GBAY
COPYRIGHT © 2002 - 2021 Pace Analytical Services, LLC

9.5.9 Obtain the appropriate number of plastic bags. For each sample, separately transfer the
labels on the drying tray to the plastic bag. Immediately transfer the dried sample to the
plastic bag by sliding the drying tray into the plastic bag and transferring the dried sediment.
9.5.10 Using a rubber mallet and/or a rolling pin, pulverize the sample until the sample is free flowing
in nature.
9.5.11 Transfer the labels on the processing bag to a new pre-labeled plastic bag. Immediately
transfer the ground/homogenized sample to the new bag, shifting all sediment to one side of
the bag, and cutting one corner off with a methanol cleaned scissors.
9.5.11.1 An aliquot of the air-dried sample must be taken in order to complete the air-dry
dry weight analysis, please see SOP: ENV-SOP-GBAY-0004 Measurement of
Percent Moisture in Soils and Solids (current revision or replacement).
9.5.12 Clean the work area and all processing equipment with Methanol following SOP: ENV-SOP-
GBAY-0143 Labware Cleaning Procedure (current revision or replacement), between
samples.
9.5.13 Dried/grind samples are stored at room temperature in individually labeled boxes by Work
order.
9.6 Sample Compositing Procedure for Homogenized Sediment:
9.6.1 Clean the work area by wiping the surfaces with Methanol. Follow procedure outlined in SOP:
ENV-SOP-GBAY-0143 Labware Cleaning Procedure (current revision or replacement) to
prepare the work surface for use.
9.6.2 Pre-label the appropriate sample composite container with the LIMS numbers for the
composite sample with one permanent label and one removable label.
9.6.3 Set out all samples that will be used to create the composite sample.
9.6.4 Transfer the Pace work order label from the “parent” sample to the “child” composite sample
container prior to the sample aliquot being taken. Weigh 20g of the first sample that will be
used to create the composite sample into a large weigh boat. Record the mass of the sample
in the composite logbook. (Please see Appendix II).
9.6.5 Immediately after the sample aliquot is measured into the large weigh boat, transfer the 20g
aliquot to the pre-labeled plastic Ziploc® bag for the composite sample.
9.6.6 Repeat steps 9.6.4 through 9.6.5 using a new large weigh boat each time, until all sample
aliquots have been sub-sampled and placed into the composite sample container.
9.6.7 Thoroughly mix the sample in the container to create a homogenized sample.
9.6.8 Record the composite date and time in the composite logbook. The time should be the end
time of the compositing process.
9.6.9 An aliquot of the air-dried sample must be taken in order to complete the air-dry dry weight
analysis. Please see SOP: ENV-SOP-GBAY-0004 Measurement of Percent Moisture in Soils
and Solids (current revision or replacement).
9.6.10 Clean the work area and all processing equipment with Methanol following SOP: ENV-SOP-
GBAY-0143 Labware Cleaning Procedure (current revision or replacement), between
samples.

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released, or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

13 of 19
 ENV-SOP-GBAY-0129, Rev 03

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Sample Homogenization, Compositing and Sub-Sampling
TEST METHOD NA
ISSUER: Pace ENV – Green Bay Quality – GBAY
COPYRIGHT © 2002 - 2021 Pace Analytical Services, LLC

9.7 Synthetic Material Homogenization:

9.7.1 With Methanol, clean the countertop inside of the prep hood.
9.7.2 Randomly arrange synthetic material into 4 separate quadrants labeled 1,2,3,4 or A, B, C, D.
A dust mask and gloves are required to be worn.
9.7.3 Randomly draw a number or letter that corresponds to a quadrant from the sample square.
Diagram and log the selection into the fluff prep logbook.
9.7.4 Breakdown all of the material into “penny” sized pieces by chopping, clipping, cutting or
tearing the material within the selected square. The sample pieces should be small enough
to fit into an extraction thimble.
9.7.5 Place the prepped pieces into a sample bag labeled with the sample number. Return any
unused material to its original bag.
9.7.6 Clean the benchtop and all of the utensils used with methanol. Replace your gloves before
proceeding to the next sample.

10.0 DATA ANALYSIS AND CALCULATIONS

Not applicable to this SOP.

11.0 QUALITY CONTROL AND METHOD PERFORMANCE

11.1 Tuna is prepared as outlined in Section 9 to be utilized as the Method Blank (MB) and Laboratory
Control Spike (LCS) matrix for organic analysis by EPA 8081A/B, 8082, 8082A, and 8270C-SIM.
The tuna is an analyte free biota matrix.
11.2 Chicken is prepared as outlined in Section 9. The Chicken Blank (CB) must be prepared for every
biota batch analyzed for metals analysis by EPA 6020/A, EPA 7471B and EPA 245.6. The CB will
contain detectable amounts of elements such as K, Ca, Na, Mg, and P etc., and is used to ensure
acceptable performance of the laboratory control spike. The chicken is also used as the matrix
modifier for the Laboratory Control Spike (LCS) matrix.
11.3 Air drying of sediment samples is a minimum of 8 hours or until moisture content is ≤10%.

12.0 DATA REVIEW AND CORRECTIVE ACTION

12.1 Data Review
Pace’s data review process includes a series of checks performed at different stages of the
analytical process by different people to ensure that SOPs were followed, the analytical record is
complete and properly documented, proper corrective actions were taken for QC failure and other
nonconformance(s), and that test results are reported with proper qualification.
The review steps and checks that occur as employees complete tasks and review their own work
is called primary review.

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released, or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

14 of 19
 ENV-SOP-GBAY-0129, Rev 03

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Sample Homogenization, Compositing and Sub-Sampling
TEST METHOD NA
ISSUER: Pace ENV – Green Bay Quality – GBAY
COPYRIGHT © 2002 - 2021 Pace Analytical Services, LLC

All data and results are also reviewed by an experienced peer or supervisor. Secondary review
is performed to verify SOPs were followed, that calibration, instrument performance, and QC
criteria were met and/or proper corrective actions were taken, qualitative ID and quantitative
measurement is accurate, all manual integrations are justified and documented in accordance with
the Pace ENV’s SOP for manual integration, calculations are correct, the analytical record is
complete and traceable, and that results are properly qualified.
A third-level review, called a completeness check, is performed by reporting or project
management staff to verify the data report is not missing information and project specifications
were met.
Refer to laboratory SOP ENV-SOP-GBAY-0120 Data Review and Final Report Process (current
revision or replacement) for specific instructions and requirements for each step of the data review
process.
12.2 Corrective Action
Corrective action is expected any time QC or sample results are not within acceptance criteria. If
corrective action is not taken or was not successful, the decision/outcome must be documented
in the analytical record. The primary analyst has primary responsibility for taking corrective action
when QA/QC criteria are not met. Secondary data reviewers must verify that appropriate action
was taken and/or that results reported with QC failure are properly qualified. Please see Section
9 for support equipment corrective actions.
12.3 Trouble Shooting
Not applicable to this SOP.

13.0 POLLUTION PREVENTION AND WASTE MANAGEMENT

Pace proactively seeks ways to minimize waste generated during our work processes. Some
examples of pollution prevention include but are not limited to: reduced solvent extraction, solvent
capture, use of reusable cycletainers for solvent management, and real-time purchasing. Additional
information can be found in the Pace’s Chemical Hygiene Plan / Safety Manual (current revision or
replacement).
The EPA requires that laboratory waste management practice to be conducted consistent with all
applicable federal and state laws and regulations. Excess reagents, samples and method process
wastes must be characterized and disposed of in an acceptable manner in accordance with Pace’s
SOP ENV-SOP-GBAY-0125 Waste Handling and Management (current revision or replacement).
Regulated soil samples are to be handled in accordance with Pace SOP: ENV-SOP-GBAY-0121,
Regulated Soil Handling (current revision or replacement).

14.0 MODIFICATIONS
A modification is a change to a reference test method made by the laboratory. For example, changes
in stoichiometry, technology, quantitation ions, reagent or solvent volumes, reducing digestion or
extraction times, instrument runtimes, etc. are all examples of modifications. Refer to Pace ENV
corporate SOP ENV-SOP-CORQ-0011 Method Validation and Instrument Verification (current revision
or replacement) for the conditions under which the procedures in test method SOPs may be modified
and for the procedure and document requirements.
Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released, or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

15 of 19
 ENV-SOP-GBAY-0129, Rev 03

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Sample Homogenization, Compositing and Sub-Sampling
TEST METHOD NA
ISSUER: Pace ENV – Green Bay Quality – GBAY
COPYRIGHT © 2002 - 2021 Pace Analytical Services, LLC

15.0 RESPONSIBILITIES
Employees that perform any step of this procedure must have a completed Read and Acknowledgment
Statement for this version of the SOP in their training record. In addition, prior to unsupervised
(independent) work on any client sample, analysts that complete dry weight analysis must have
successful initial demonstration of capability (IDOC) and must successfully demonstrate on-going
proficiency on an annual basis. Successful means the initial and on-going DOC met criteria,
documentation of the DOC is complete, and the DOC record is in the employee’s training file. Refer to
laboratory SOP ENV-SOP-GBAY-0094 Orientation and Training Procedures (current revision or
replacement) for more information.
Pace supervisors/managers are responsible for training employees on the procedures in this SOP and
monitoring the implementation of this SOP in their work area.

16.0 ATTACHMENTS
16.1 Attachment I: Sediment Dry-Grind Tracking Logbook
16.2 Attachment II: Fox River Sediment Composite Logbook

17.0 REFERENCES
17.1 Pace Analytical Services, LLC – Green Bay, WI Quality Assurance Manual- current version.
17.2 TNI Standard, Management and Technical Requirements for Laboratories Performing
Environmental Analyses, EL-VI-2016-Rev.2.1.
17.3 Department of Defense (DoD) Quality Systems Manual - current version.
17.4CERCLA Quality Assurance Manual; October 1989.

17.5 USEPA National Guidance for Assessing Chemical Contaminant Data for Use in Fish
Advisories, Volume I: Fish Sampling and Analysis-Third Edition
17.6 Pace Analytical SOP ENV-SOP-GBAY-0130 Small Rodent Handling and Homogenization
(current revision or replacement).
17.7 Pace Analytical SOP: ENV-SOP-GBAY-0143 Labware Cleaning Procedures, (current revision
or replacement).
17.8 Pace Analytical SOP: ENV-SOP-GBAY-0121 Regulated Soil Handling, (current revision or
replacement).

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released, or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

16 of 19
 ENV-SOP-GBAY-0129, Rev 03

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Sample Homogenization, Compositing and Sub-Sampling
TEST METHOD NA
ISSUER: Pace ENV – Green Bay Quality – GBAY
COPYRIGHT © 2002 - 2021 Pace Analytical Services, LLC

18.0 REVISION HISTORY

This Version: ENV-SOP-GBAY-0129-Rev.03
Section Description of Change
7.4 Added computer hardware and software.
12.3 Added Trouble shooting
Attachment Updated to current document.
I, II

This document supersedes the following document(s):

Document Number Title Version
ENV-SOP-GBAY- Sample Homogenization, Compositing and Sub-Sampling 02
0129
ENV-SOP-GBAY- Sample Homogenization, Compositing and Sub-Sampling 01
0129
ENV-SOP-GBAY- Biological Tissue, Plant, Sediment, and Synthetic Material 00
0129 Preparation
ENV-SOP-GBAY- Sample Homogenization and Sub-Sampling 01
0110

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released, or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

17 of 19
 ENV-SOP-GBAY-0129, Rev 03

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Sample Homogenization, Compositing and Sub-Sampling
TEST METHOD NA
ISSUER: Pace ENV – Green Bay Quality – GBAY
COPYRIGHT © 2002 - 2021 Pace Analytical Services, LLC

Attachment I: Sediment Dry-Grind Tracking Logbook

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released, or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

18 of 19
 ENV-SOP-GBAY-0129, Rev 03

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Sample Homogenization, Compositing and Sub-Sampling
TEST METHOD NA
ISSUER: Pace ENV – Green Bay Quality – GBAY
COPYRIGHT © 2002 - 2021 Pace Analytical Services, LLC

Attachment II: Fox River Sediment Composite Logbook

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released, or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

19 of 19
 ENV-SOP-MIN4-0178, Rev 01

Document Information
Document Number: ENV-SOP-MIN4-0178 Revision: 01

Document Title: Determination of Selected 36 Per- and Polyfluoroalkyl Substances (PFAS) by

LC/MS/MS (Isotope Dilution)

Department(s):
Dioxin

Date Information

Effective Date: 31 Dec 2020

Notes

Document Notes:

All Dates and Times are listed in: Central Time Zone

1 of 58
 ENV-SOP-MIN4-0178, Rev 01

Signature Manifest
Document Number: ENV-SOP-MIN4-0178 Revision: 01
Title: Determination of Selected 36 Per- and Polyfluoroalkyl Substances (PFAS) by LC/MS/MS (Isotope
Dilution)
All dates and times are in Central Time Zone.

ENV-SOP-MIN4-0178

QM Approval

Name/Signature Title Date Meaning/Reason

Janielle Ward (007319) Manager - Quality 30 Dec 2020, 07:54:27 PM Approved

Management Approval

Name/Signature Title Date Meaning/Reason

Adam Haugerud (005828) General Manager 2 23 Dec 2020, 03:18:11 PM Approved
Krista Carlson (004514) Project Manager 1 23 Dec 2020, 03:42:42 PM Approved
Keith Sturgeon (003603) Manager 28 Dec 2020, 10:26:58 AM Approved

2 of 58
 ENV-SOP-MIN4-0178, Rev 01

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Determination of Selected 36 Per- and Polyfluoroalkyl Substances (PFAS) by
LC/MS/MS
TEST METHOD: Isotope Dilution
ISSUER: Pace ENV – Minneapolis – MIN4
COPYRIGHT © 2020 Pace Analytical Services, LLC

1.0 SCOPE AND APPLICATION

The purpose of this Standard Operating Procedure (SOP) is to document the procedure used for the
identification and simultaneous measurement of per- and polyfluoroalkyl substances (PFAS) in non-
potable waters, leachate, solid (e.g. soil, sediment, and wipe), and tissue matrices using LC/MS/MS
technology based on Table B-15 of the Department of Defense Quality Systems Manual Version 5.3,
Appendix B (DoD QSM 5.3), and the Wisconsin PFAS Aqueous (Non-Potable Water) and Non-Aqueous
Matrices Method Expectations.
1.1 Target Analyte List and Limit of Quantitation (LOQ)
The target analytes and the normal LOQ that can be achieved with this procedure are provided in
Table 1, Appendix A.
LOQ are established in accordance with Pace policy and SOPs for method validation and for the
determination of detection limits (DL) and quantitation limits (LOQ). DL and LOQ are routinely
verified and updated when needed. The current LOQ for each target analyte that can be
determined by this SOP as of the effective date of this SOP is provided in Table 1, Appendix A.
The reporting limit (RL) is the value to which analytes are reported as detected or not detected in
the final report. When the RL is less than the lower limit of quantitation (LLOQ), all detects and
non-detects at the RL are qualitative. The LLOQ is the lowest point of the calibration curve used
for each target analyte.
DL, LOQ, and RL are always adjusted to account for actual amounts used and for dilution.

2.0 SUMMARY OF METHOD

A 250-mL water sample is fortified with a known quantity of isotope dilution extracted internal standards
(EIS) and then passed through a solid phase extraction (SPE) cartridge (e.g., StrataTM PFAS,
WAX/GCB sorbent, weak anion exchange mixed-mode) to extract the method analytes and EIS. The
analytes and EIS are eluted from the cartridge with a small amount of ammonia/methanol solution. The
method for the analysis of PFAS in solid materials extracts 5 g of material with a total of 9-mL aliquot
of 0.2% ammonia/methanol. The extract is treated with 50 mg ENVI-Carb™ and filtered prior to nitrogen
concentration. For tissue samples, 2 g of material is extracted with 7 mL of 1% ammonia acetonitrile for
16 hours. The extract is treated with ENVI-Carb™ and filtered prior to SPE cleanup. The water or solid
sample extract is concentrated to ~0.8 mL while the tissue extract is concentrated to ~0.1 mL with
nitrogen and spiked with Injection Internal Standards (IIS) and then brought to 1 mL with 96:4% (vol/vol)
methanol:H2O solution prior to LC/MS/MS analysis. A 3-µL injection is made into a Liquid
Chromatography (LC) System equipped with a C18 column that is interfaced to a tandem mass
spectrometer (MS/MS). The concentration of each analyte is determined by using the isotope dilution
and internal standard techniques, depending on target analyte. EIS is added to all calibration standards,
field samples, blanks and QC samples to monitor the extraction efficiency of the method analytes.

3.0 INTERFERENCES
3.1 All glassware must be meticulously cleaned. Wash glassware with non-phosphate alkaline
detergent and deionized (DI) water, rinse with DI water and reagent water, followed by a methanol
rinse. Non-volumetric glassware can be heated in a muffle furnace at 400 °C for 2 h or solvent

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

3 of 58
 ENV-SOP-MIN4-0178, Rev 01

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Determination of Selected 36 Per- and Polyfluoroalkyl Substances (PFAS) by
LC/MS/MS
TEST METHOD: Isotope Dilution
ISSUER: Pace ENV – Minneapolis – MIN4
COPYRIGHT © 2020 Pace Analytical Services, LLC

rinsed. Volumetric glassware should be solvent rinsed and not be heated in an oven above 120 °C.
Store clean glassware inverted or capped. Do not cover with aluminum foil because PFAS can
be potentially transferred from the aluminum foil to the glassware.
NOTE: PFAS standards, extracts and samples should not come in contact with any glass
containers or pipettes as these analytes can potentially adsorb to glass surfaces. PFAS analytes,
EIS and IIS commercially purchased in glass ampoules are acceptable; however, all subsequent
transfers or dilutions performed by the analyst must be prepared and stored in polypropylene or
equivalent containers
3.2 Method interferences may be caused by contaminants in solvents, reagents (including reagent
water), sample bottles and caps, and other sample processing hardware that lead to discrete
artifacts and/or elevated baselines in the chromatograms. The method analytes in this method can
also be found in many common laboratory supplies and equipment, such as PTFE
(polytetrafluoroethylene) products, LC solvent lines, methanol, aluminum foil, SPE sample transfer
lines, etc. All items such as these must be routinely demonstrated to be free from interferences
(less than 1/2 the RL) under the conditions of the analysis by analyzing method blanks. Subtracting
blank values from sample results is not permitted.
3.3 Matrix interferences may be caused by contaminants that are co-extracted from the sample. The
extent of matrix interferences will vary considerably from source to source, depending upon the
nature of the water. Humic and/or fulvic material can be co-extracted during SPE and high levels
can cause enhancement and/or suppression in the electrospray ionization source or low recoveries
on the SPE sorbent. Total organic carbon (TOC) is a good indicator of humic content of the sample.
Under the LC conditions used during method development, matrix effects due to TOC were not
observed.
3.4 SPE cartridges can be a source of interferences. The analysis of field and method blanks can
provide important information regarding the presence or absence of such interferences. Brands and
lots of SPE devices should be tested to ensure that contamination does not preclude analyte
identification and quantitation.

4.0 DEFINITIONS
Refer to the Laboratory Quality Manual for a glossary of common lab terms and definitions.
4.1 Confirmation Ion – One of the product ions used to help qualitatively confirm presence of the
analytes. The product ion chosen is typically one of the remaining ions with high sensitivity and
minimum interferences after the quantitation ion has been chosen. Not all precursor ions provide
confirmation ions.
4.2 Extraction Internal Dilution standards (EIS) – Isotopically labeled internal standards that
undergo the same extraction and analysis as the other analytes in the sample. The EIS are added
to the sample at the beginning of the procedure before extraction, centrifugation, filtering, or phase
separation. Ideally, there are exact isotopically labeled analogs of the native analytes so that
identical behavior can be assumed. The recoveries of these standards are used to adjust the native
analyte results.
4.3 Internal Standard Quantitation – measurement of native analytes using an alternate analog
isotope (one that has the same chemical behavior and is close in retention time to the native

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

4 of 58
 ENV-SOP-MIN4-0178, Rev 01

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Determination of Selected 36 Per- and Polyfluoroalkyl Substances (PFAS) by
LC/MS/MS
TEST METHOD: Isotope Dilution
ISSUER: Pace ENV – Minneapolis – MIN4
COPYRIGHT © 2020 Pace Analytical Services, LLC

analyte), thus providing a close approximation of matrix effects and losses that can occur during
the preparation and analysis. The native analyte concentration is adjusted for the recovery of the
alternate analog isotope. An alternate analog isotope is typically used when an exact analog isotope
is not available.
4.4 Isotope Dilution Quantification – measurement of native analytes using an exact analog isotope
of the native analyte. The native analyte concentration is adjusted for the recovery of the exact
analog isotope that has been included in the preparatory and analytical procedure.
4.5 Precursor Ion – For the purpose of this method, the precursor ion is the deprotonated molecule
([M-H]-) of the method analyte. In MS/MS, the precursor ion is mass selected and fragmented by
collisionally activated dissociation to produce distinctive product ions of smaller m/z.
4.6 Product Ion – For the purpose of this method, a product ion is one of the fragment ions produced
in MS/MS by collisional activated dissociation of the precursor ion.
4.7 Primary Dilution Standard (PDS) solution – A solution containing the analytes prepared in the
laboratory from stock standard solutions and diluted as needed to prepare calibration solutions and
other needed analyte solutions.
4.8 Preparation Batch – A group of up to 20 field samples (not including QC samples) extracted
together by the same person(s) during a workday (24 hours) using the same lot of SPE devices,
solvents, surrogate, internal standard and fortifying solutions. Required QC samples include MB,
LCS, MS, and MSD.
4.9 Quantitation Ion – One of the product ions used to quantitate analyte concentrations. The product
ion chosen is typically one of high sensitivity and minimum interference

5.0 HEALTH AND SAFETY

The toxicity or carcinogenicity of each chemical material used in the laboratory has not been fully
established. Each chemical should be regarded as a potential health hazard and exposure to these
compounds should be as low as reasonably achievable.
The laboratory maintains documentation of hazard assessments and OSHA regulations regarding the
safe handling of the chemicals specified in each method. Safety data sheets for all hazardous chemicals
are available to all personnel. Employees must abide by the health, safety and environmental (HSE)
policies and procedures specified in this SOP and in the Pace Chemical Hygiene / Safety Manual.
Personal protective equipment (PPE) such as safety glasses, gloves, and a laboratory coat must be
worn in designated areas and while handling samples and chemical materials to protect against
physical contact with samples that contain potentially hazardous chemicals and exposure to chemical
materials used in the procedure.
Concentrated corrosives present additional hazards and are damaging to skin and mucus membranes.
Use these acids in a fume hood whenever possible with additional PPE designed for handing these
materials. If eye or skin contact occurs, flush with large volumes of water. When working with acids,
always add acid to water to prevent violent reactions. Any processes that emit large volumes of solvents
(evaporation/concentration processes) must be in a hood or apparatus that prevents employee
exposure.

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

5 of 58
 ENV-SOP-MIN4-0178, Rev 01

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Determination of Selected 36 Per- and Polyfluoroalkyl Substances (PFAS) by
LC/MS/MS
TEST METHOD: Isotope Dilution
ISSUER: Pace ENV – Minneapolis – MIN4
COPYRIGHT © 2020 Pace Analytical Services, LLC

Contact your supervisor or local HSE coordinator with questions or concerns regarding safety protocol
or safe handling procedures for this procedure.

6.0 SAMPLE COLLECTION, PRESERVATION, HOLDING TIME, AND STORAGE

Samples should be collected in accordance with a sampling plan and procedures appropriate to achieve
the regulatory, scientific, and data quality objectives for the project.
The laboratory does not perform sample collection or field measurements for this test method. To
assure sample collection and field checks and treatment are performed in accordance with applicable
regulations. Pace project managers will inform the client of these requirements at the time of request
for analytical services when the request for testing is received prior to sample collection. If samples
were already collected, the laboratory will record any nonconformance to these requirements in the
laboratory’s sample receipt record when sufficient information about sample collection is provided with
the samples.
The laboratory will provide containers for the collection of samples upon client request for analytical
services. Bottle kits are prepared in accordance with laboratory SOP ENV-SOP-MIN4-009 Bottle
Preparation (current version or equivalent replacement).
Requirements for container type, preservation, and field quality control (QC) for the common list of test
methods offered by Pace are included in the laboratory’s quality manual.
General Requirements
Routine Minimum
Matrix Preservation Holding Time
Container Sample Amount
Aqueous 250 mL HDPE bottle 250 mL Thermal: <6 °C but Collection to Prep: 28 Days
fitted with polyethylene >0 °C Prep to Analysis: 28 Days
screw-cap lid
Chemical: NA Extract stored at 0-6oC
Solid 250 mL HDPE bottle 5g Thermal: <6 °C but Collection to Prep: 28 Days
fitted with polyethylene >0 °C Prep to Analysis: 28 Days
screw-cap lid
Chemical: NA Extract stored at 0-6oC
Tissue 250 mL HDPE bottle 2g Thermal: Frozen Collection to Prep: 1 year
fitted with Chemical: NA Prep to Analysis: 30 Days
polyethylene screw- Extract stored at 0-6oC
cap lid

6.1 Sample Collection

6.1.1 The sample handler must wash their hands before sampling and wear nitrile gloves while
filling and sealing the sample bottles. PFAS contamination during sampling can occur from
a number of common sources, such as food packaging and certain foods and beverages.
Proper hand washing and wearing nitrile gloves will aid in minimizing this type of accidental
contamination of the samples.
6.1.2 Fill sample bottles. Do not fill aqueous sample containers completely.
6.1.3 Matrix spike (MS) and matrix spike duplicate (MSD) sample. Analysis of a MS is requested
in each extraction batch and is used to determine that the sample matrix does not adversely
affect method accuracy.

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

6 of 58
 ENV-SOP-MIN4-0178, Rev 01

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Determination of Selected 36 Per- and Polyfluoroalkyl Substances (PFAS) by
LC/MS/MS
TEST METHOD: Isotope Dilution
ISSUER: Pace ENV – Minneapolis – MIN4
COPYRIGHT © 2020 Pace Analytical Services, LLC

6.1.4 Field Duplicates. Collect one per sampling event for each sampling site
6.2 Sample Shipment –Aqueous and solid samples must be chilled during shipment and must not
exceed 6 °C after collection. Ship tissue samples frozen.
6.3 Sample Receipt and Storage - Thermal preservation is checked and recorded on receipt in the
laboratory in accordance with laboratory SOP ENV-SOP-MIN4-0008 Sample Management (current
version or equivalent replacement).
6.3.1 Aqueous and solid sample temperature must be confirmed to be at or below 6 °C when the
samples are received at the laboratory.
6.3.2 Samples stored in the laboratory must be held at or below 6 °C until extraction but should
not be frozen.
6.3.3 Tissue samples received frozen can be documented as “frozen” at sample receipt. Store
tissue samples at less than or equal to -10oC at the laboratory.

7.0 EQUIPMENT AND SUPPLIES

7.1 Equipment
7.1.1 Brand names and catalogue numbers represent materials in use at the time of this revision.
Due to potential adsorption of analytes onto glass, polypropylene containers were used for
all standard, sample and extraction preparations. Other materials which meet the QC
requirements may be substituted.
Table 7.1.1 - Equipment and Supplies (Including Computer Software)
Vendor/Item
Supply Description
#/Description
SPE cartridge Phenomenex StrataTM PFAS, WAX/GCB, Phenomenex, Cat# CS0-
200 mg/50 mg 6 mL 9207 or equivalent
Extraction manifold An automatic/robotic sample preparation Supelco Cat# 57030 and
system designed for use with SPE 57275 or equivalent
cartridges
Analytical column Gemini® 100 × 3 mm 3 µm C18 reverse Phenomenex Cat# 00D-
phase LC column 4439-Y0 or equivalent
HPLC 1100/1290 infinity series/NexeraXR Agilent/Shimadzu
MS API 4000/5500 quadrupole Sciex
Analyst® Data acquisition software Version 1.6.3
TM
Multiquant Data processing software Version 3.0.2
Avalon Data reporting software See master list for current
version
Nitrogen N-EVAPTM 112 nitrogen evaporator Oasys Heating system
evaporator equivalent nitrogen evaporator/heated (Berlin, MA, USA)
waterbath capable of heating 25-60oC
Balance Electronic, capable of weighing to 0.001 g NA
or equivalent.

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

7 of 58
 ENV-SOP-MIN4-0178, Rev 01

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Determination of Selected 36 Per- and Polyfluoroalkyl Substances (PFAS) by
LC/MS/MS
TEST METHOD: Isotope Dilution
ISSUER: Pace ENV – Minneapolis – MIN4
COPYRIGHT © 2020 Pace Analytical Services, LLC

Vendor/Item
Supply Description
#/Description
Syringe pump Model # NE-300 or equivalent system New Era Pump Systems,
capable of delivering variable flow rates. Inc
Ultrasonicator Branson ultrasonicator Branson Model 8510
Sample container High density polyethylene (HDPE) or C&C Container, Cat#
polypropylene, 250 mL, wide mouth, with 183277
screw top
Centrifuge tube and 15-mL and 50-mL conical polypropylene BD Falcon, P/N 352096
cap tubes with polypropylene screw caps for and P/N352070
collection and storage of the extracts
Polypropylene 4-mL narrow-mouth polypropylene bottles Thermo Cat# 2006-9125
bottles
Polypropylene 15-mLnarrow-mouth polypropylene bottles Thermo Cat# 2002-9050
bottles
Autosampler vials Polypropylene 0.3-mL autosampler vials Phenomenex Cat# AR0-
with polypropylene caps 9995-12-C
Adjustable auto- Ranges 10-100 µL, 100-1000 µL, and NA
pipettors 1000-5000 µL.
Laboratory or aspirator vacuum system
ENVI-Carb Supelclean™ ENVI-Carb™ SPE Bulk Sigma Aldrich, Cat# 57210-
Packing U.
Vacuum extraction A manual vacuum manifold with Visiprep Supelco Cat# 57250-U and
manifold volume sampler (Supelco Cat# 57030 and 57275 or equivalent
57275 or equivalent) for extraction, or an
automatic/robotic sample preparation
system designed for use with SPE
cartridges, may be used if all QC
requirements are met.

7.1.2 Liquid Chromatography (LC)/ Tandem Mass Spectrometer (MS/MS)

7.1.2.1 LC System – Liquid chromatography (LC) system with binary pump, autosampler,
column heater. All solvent lines were replaced with (polyether ether ketone) PEEK
tubing. PFAS isolator column (Phenomenex Luna ® 30 × 3 mm 5 µm C18 reverse
phase LC column, Cat# 00A-4252-Y0) and stainless steel tubing installed
between the mixing chamber and injection port. Other equivalent automated LC
system capable of reproducibly injecting up to 5-µL aliquots and performing binary
linear gradients at a constant flow rate near the flow rate used for development of
this method (0.5 mL/min) may be used.
7.1.2.2 LC/MS/MS – The LC/MS/MS must be capable of negative ion electrospray
ionization (ESI) near the suggested LC flow rate of 0.6 mL/min. The system must
be capable of performing MS/MS to produce unique product ions for the method
analytes within specified retention time segments. A minimum of 10 scans across
the chromatographic peak is required to ensure adequate precision.

8.0 REAGENTS AND STANDARDS

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

8 of 58
 ENV-SOP-MIN4-0178, Rev 01

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Determination of Selected 36 Per- and Polyfluoroalkyl Substances (PFAS) by
LC/MS/MS
TEST METHOD: Isotope Dilution
ISSUER: Pace ENV – Minneapolis – MIN4
COPYRIGHT © 2020 Pace Analytical Services, LLC

8.1 Gases, Reagents, and Solvents – LC/MS grade or equivalent is used (Fisher equivalent is
Optima). Other grades may be used, provided it is first determined that the reagent is of sufficiently
high purity to permit its use without lessening the quality of the determination. Fisher solvents are
preferred for mobile phases (water, methanol, acetonitrile) as the one liter bottles can be directly
loaded on the instrument removing a transfer step with the inherent low level contamination.

Reagent/Standard Concentration/ Description Requirements/Vendor/Item #

Ammonium Acetate Optima LC/MS grade, demonstrated to be Fisher or equivalent
(NH4C2H3O2, CAS# 631-61-8) free of analytes and interferences
Acetonitrile Optima HPLC grade Fisher or equivalent
(CH3CN, CAS# 75-05-8) Cat# A955-4
Ammonium Hydroxide Certified ACS Plus grade demonstrated to be Fisher or equivalent
(NH4OH, 28-30% in water) free of analytes and interferences
Ammonia/Methanol Solution Optima HPLC grade, w = 0.2% mass fraction. Fisher or equivalent
Mix 0.72 mL of 28-30% ammonia solution
with 99.28 mL of methanol
Glacial Acetic Acid HPLC grade. Demonstrated to be free of VWR Analytical
(C2H3CO2H, CAS# 64-19-7) analytes and interferences Cat# BDH20108
Sodium hydroxide Certified ACS. High purity demonstrated to be Fisher, or equivalent
(NaOH, 1310-73-2) free of analytes and interferences
Methanol Optima HPLC grade, demonstrated to be free Fisher, or equivalent
(CH3OH, CAS# 67-56-1) of analytes and interferences
Reagent water Optima HPLC grade, demonstrated to be free Fisher or equivalent
(H2O, CAS# 7732-18-5) of analytes and interferences Cat# W7-4
Aqueous Mobile Phase To prepare 1 L, add 1.54 g ammonium Fisher or equivalent
(20 mM Ammonium acetate) acetate to 1 L of reagent water.
Acetate Buffer Mix 0.5 mL of acetic acid with 349.5 mL of Fisher or Equivalent
(25 mM, pH 4) water. Dissolve 0.116 g of ammonium acetate
in 60 mL of water. Mix 200 mL of the diluted
acetic acid with 50 mL of the ammonium
acetate solution
PPG Tuning Solutions Instrument tuning compound. Using Sciex
standards chemical kit with low/high P/N 4406127
concentration is recommend, however
solution can be prepared from a neat
material.
Ottawa Sand To prepare method blank, LCSs, for the EMD or equivalent
extraction of soil samples Cat# SX0075-3
Nitrogen Nitrogen aids in aerosol generation of the ESI Ultra-High Purity or equivalent
(N2) liquid spray and is used as collision gas in
some MS/MS instruments. The nitrogen (Ultra
High Purity or equivalent) used should meet
or exceed instrument manufacturer’s
specifications
Canola Oil Canola oil, or equivalent, for Oil quality control Local grocery store
sample matrix,
Lake Michigan Fish Tissue Standard reference materials (SRM) for NIST, 1947
tissue analysis

8.2 Stock Standards – non-neat standards purchased from vendors that are used for the preparation
of working standards. Standards containing both branched and linear isomers must be used when
commercially available. If not available, the total response of the analyte must be integrated, (i.e.

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

9 of 58
 ENV-SOP-MIN4-0178, Rev 01

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Determination of Selected 36 Per- and Polyfluoroalkyl Substances (PFAS) by
LC/MS/MS
TEST METHOD: Isotope Dilution
ISSUER: Pace ENV – Minneapolis – MIN4
COPYRIGHT © 2020 Pace Analytical Services, LLC

accounting for peaks that are identified as linear and branched isomers) and quantitated using a
calibration curve which includes the linear isomer only for that analyte, i.e. PFOA.
If no expiration date is assigned by the vendor, expiration date is 1 year from the date of receipt.
For open stock standards, the expiration is date is 1 year for the open date.
PFBS, PFPeS, PFHxS, PFHpS, PFOS, PFNS, PFDS, 4:2FTS, 6:2FTS, 8:2FTS, DONA, 9Cl-
PF3ONS and 11Cl-PF3OUdS are not available as the acid form, but rather as their corresponding
salts, such as Na+ and K+. These salts are acceptable for use as stock standards as long as the
weight is corrected for the salt content according to the equation below:

= ×

Where: MW (acid) = the molecular weight of PFAS

MW (salt) = the molecular weight of purchased salt
NOTE: All standards purchased are greater than or equal to 98% purity, therefore the weight can
be used without correction to calculate the concentration of stock standards. Primary stock
standards are stored at ≤ 4 ± 2 oC. Stock solution is brought to room temperature before using.
PFAS may be purchased in glass ampoules however all further solutions and storage is in
polypropylene or equivalent containers.
Table 8.2 – Description and Concentration of Each Analyte in Stock Solution
Analyte/Concentration Used to prepare
Wellington Laboratories PFAC-30PAR (µg/mL)
PFBA 1
PFPeA 1
PFHxA 1
PFHpA 1
PFOA 1
PFNA 1
PFDA 1
PFUdA 1 Cal curve and spiking
PFDoA 1 solution.
PFTrDA 1
PFTeDA 1
PFOSA 1
N-EtFOSAA* 1
N-MeFOSAA* 1
HFPO-DA 1
DONA 0.945
PFBS 0.887

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

10 of 58
 ENV-SOP-MIN4-0178, Rev 01

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Determination of Selected 36 Per- and Polyfluoroalkyl Substances (PFAS) by
LC/MS/MS
TEST METHOD: Isotope Dilution
ISSUER: Pace ENV – Minneapolis – MIN4
COPYRIGHT © 2020 Pace Analytical Services, LLC

Analyte/Concentration Used to prepare

PFPeS 0.941
PFHxS* 0.914
PFHpS 0.953
PFOS* 0.928
PFNS 0.962
PFDS 0.965
4:2FTS 0.937
6:2FTS 0.951
8:2FTS 0.960
9Cl-PF3ONS 0.933
11Cl-PF3OUdS 0.943
Wellington Laboratories PFAC-8Native (µg/mL)
10:2FTS 48.2
N-MeFOSA 50
N-EtFOSA 50
N-MeFOSE 50
N-EtFOSE 50
PFDoS 48.4
PFHxDA 50
PFODA 50
Wellington Laboratories PFAC-24PAR (µg/mL)
PFBA 2
PFPeA 2
PFHxA 2
PFHpA 2
PFOA 2
PFNA 2
PFDA 2 Use for the ICV
PFUdA 2
PFDoA 2
PFTrDA 2
PFTeDA 2
PFOSA 2
N-EtFOSAA 2
N-MeFOSAA 2
PFBS 1.77

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

11 of 58
 ENV-SOP-MIN4-0178, Rev 01

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Determination of Selected 36 Per- and Polyfluoroalkyl Substances (PFAS) by
LC/MS/MS
TEST METHOD: Isotope Dilution
ISSUER: Pace ENV – Minneapolis – MIN4
COPYRIGHT © 2020 Pace Analytical Services, LLC

Analyte/Concentration Used to prepare

PFPeS 1.88
PFHxS* 1.82
PFHpS 1.90
PFOS* 1.86
PFNS 1.92
PFDS 1.93
4:2FTS 1.87
6:2FTS 1.90
8:2FTS 1.92
Wellington Laboratories PFAC-12Native (µg/mL)
10:2FTS 48.2
HFPO-DA 50
DONA 47.25
N-MeFOSA 50
N-EtFOSA 50
Use for the ICV
N-MeFOSE 50
N-EtFOSE 50
9Cl-PF3ONS 46.6
11Cl-PF3OUdS 47.1
PFDoS 48.4
PFHxDA 50
PFODA 50
Wellington Laboratories MPFAC-24ES (µg/mL)
13C4_PFBA 1
13C5_PFPeA 1
13C3_PFBS 0.929
13C2_4:2FTS 0.935
13C5_PFHxA 1
13C4_PFHpA 1 Isotopically labelled Extracted Internal
13C3_PFHxS 0.946 Standards
13C2_6:2FTS 0.949
13C8_PFOA 1
13C9_PFNA 1
13C8_PFOS 0.957
13C2_8:2FTS 0.958
13C6_PFDA 1

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

12 of 58
 ENV-SOP-MIN4-0178, Rev 01

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Determination of Selected 36 Per- and Polyfluoroalkyl Substances (PFAS) by
LC/MS/MS
TEST METHOD: Isotope Dilution
ISSUER: Pace ENV – Minneapolis – MIN4
COPYRIGHT © 2020 Pace Analytical Services, LLC

Analyte/Concentration Used to prepare

d3-MeFOSAA 1
13C8_FOSA 1
d5-EtFOSAA 1
13C7_PFUdA 1
13C2_PFDoA 1
13C2_PFTeDA 1
Wellington Laboratories MPFAC-6ES (µg/mL)
13C3_HFPO-DA 50
13C2_PFHxDA 50
Isotopically labelled Extracted Internal
d7-N-MeFOSE 50
Standards
d9-N-EtFOSE 50
d3-N-MeFOSA 50
d5-N-EtFOSA 50
Wellington Laboratories MPFAC-Injection Internal
Standards (µg/mL)
13C2_PFHxA 50
Isotopically labelled Injection Internal
13C4_PFOA 50 Standards (IIS)
13C2_PFDA 50
13C4_PFOS 50
Wellington Laboratories T-PFOA (µg/mL) Qualitative Standard for PFOA (branch
T-PFOA* 50 isomer of PFOA)
Note(s):
Asterisked (*) analytes indicate the presence of both linear and branch isomers.
See Appendix A for additional information about acronyms variations.

9.0 PROCEDURE
9.1 Equipment Preparation
9.1.1 Instrument
9.1.1.1 Routine Instrument Operating Conditions
Table 9.1.1.1 – LC-MS/MS Operating Conditions
Syringe Size 100 µL
Sample Loop vol. 40 µL
Injector
Injection Volume: 3 µL
Needle Wash 1 100% Methanol
Pump Flow rate 400 µL/min

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

13 of 58
 ENV-SOP-MIN4-0178, Rev 01

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Determination of Selected 36 Per- and Polyfluoroalkyl Substances (PFAS) by
LC/MS/MS
TEST METHOD: Isotope Dilution
ISSUER: Pace ENV – Minneapolis – MIN4
COPYRIGHT © 2020 Pace Analytical Services, LLC

Flow method Gradient

Mobile Phase 1 20 mM Ammonium Acetate H2O
Mobile Phase 2 LCMS Acetonitrile
Time % Mobile Phase 1 % Mobile Phase 2
Initial 90 10
0.5 90 10
8.0 20 80
Gradient Program 10.0 20 80
10.1 5 95
12.0 5 95
12.1 90 10
15.0 90 10
Type: Phenomenex Gemini® (or equivalents)
Part Number: 00D-4439-Y0
Running temp 40 oC
Column
Length: 100 mm
Diameter: 3 mm
Particle Size 3.0 µm
Collision Gas 10 psi
Curtain Gas 25 psi
Ion Source Gas 1 40 psi
Ion Source Gas 2 50 psi
Nominal IonSpray Voltage -4500 v
Tune Temperature 450 oC
Values
ESI polarity Negative
Declustering Potential
Collision Energy Optimized for each analyte
(See Appendix C for reference)
Collision Cell Exit
Potential

9.1.2 Routine Instrument Maintenance

9.1.2.1 Routine instrument maintenance is critical to achieve optimum method sensitivity.
All laboratory materials must be determined to be free of contamination to ensure
potential background interferences are minimized.
9.1.2.2 Please refer to the instrument manual for maintenance procedures performed by
the lab.

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

14 of 58
 ENV-SOP-MIN4-0178, Rev 01

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Determination of Selected 36 Per- and Polyfluoroalkyl Substances (PFAS) by
LC/MS/MS
TEST METHOD: Isotope Dilution
ISSUER: Pace ENV – Minneapolis – MIN4
COPYRIGHT © 2020 Pace Analytical Services, LLC

9.1.2.3 All maintenance activities are listed in maintenance logs that are assigned to each
separate instrument.
9.1.2.4 LC Maintenance – LC system components, as well as the mobile phase
constituents, contain many of the method analytes in this method. Thus, these
PFAS will build up on the head of the LC column during mobile phase
equilibration.
9.1.2.5 Column Equilibrate – To minimize the background PFAS peaks and to keep
background levels constant, the time the LC column sits at initial conditions must
be kept constant and as short as possible (while ensuring reproducible retention
times).
9.1.2.6 Column Flush – In addition, prior to daily use, flush the column with 95%
methanol for at least 15 min before initiating a sequence. It may be necessary on
some systems to flush other LC components such as wash syringes, sample
needles or any other system components before daily use.
9.1.3 MS Maintenance – Please refer to the instrument manual for maintenance procedures
performed by the lab. Common maintenance procedures are listed below
9.1.3.1 Source Cleaning – Clean the ion source parts which include curtain plate, orifice
plate or skimmer, Q0, and etc. with reagent water and methanol. Tuning or
optimizing the instrument followed the instrument manual. Refer to the Operating
Instruction – Tune and Calibrate, the ion source operator guide, or the Analyst®
software Help system.
9.1.3.2 Pump Oil – Check pump oil level and color periodically. Add or change pump oil
when necessary followed the manual instruction.
9.1.4 Troubleshooting
9.1.4.1 Any deviations from the norm encountered while conducting this analysis must be
noted and brought to the attention of the section supervisor. This section contains
basic information for troubleshooting basic system issues. Certain activities may
be carried out by the Agilent and AB SCIEX trained Qualified Maintenance Person
(QMP) in the laboratory. For advanced troubleshooting, contact field service
agents of the instruments.
9.1.4.2 LC Troubleshooting – Please refer to the instrument manual for troubleshooting
procedures performed by the lab. Common LC issues are listed below.
9.1.4.2.1 Pressure Issue – Large pressure variation could cause by the
presence of air bubble in the system, blockage of the system, column
contamination, system leaking, and etc. High pressure issue could be
solved through system solvent purge, column rinse, clean or change
of column inlet frit, injection valve, needle seat, and etc. Low pressure
issue could usually be fixed by tighten or replace the capillary
connection or other parts such as pump seals.
9.1.4.2.2 Peak Shape Issue – Split peaks, peak tailing, poor efficiency, and
inconsistent response are usually associated with issues like column
contamination, partially plugged frit, column void, injection solvent

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

15 of 58
 ENV-SOP-MIN4-0178, Rev 01

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Determination of Selected 36 Per- and Polyfluoroalkyl Substances (PFAS) by
LC/MS/MS
TEST METHOD: Isotope Dilution
ISSUER: Pace ENV – Minneapolis – MIN4
COPYRIGHT © 2020 Pace Analytical Services, LLC

effects, or sample overload effects. Rinsing or changing the column,

preparing fresh mobile solvent, reducing sample injection volume
could usually
9.1.4.2.3 Retention Time – Deviation of retention time from originally values
could cause by column aging or contamination, insufficient system
equilibration, mobile phase variation, change in column temperature,
or other instrument issues. Cleaning the HPLC system and column
could Ammonium acetate is volatile and may cause the shift of
retention time over certain period of time. Prepare fresh mobile phase
solvent when necessary.
9.1.4.2.4 Background Contamination – After multiple injections or long period
of operation, background interference may accumulate at the gradient
proportional valve, needle seat, or other instrument parts. Rinsing and
cleaning corresponding parts with reagent water, methanol or stronger
solvents to remove the interference.
9.1.4.2.5 MS Troubleshooting – Please refer to the instrument manual for
troubleshooting procedures performed by the lab. Common MS issues
are listed below:
9.1.4.2.5.1 Sensitivity Loss – The possible causes for intensity decrease
could be contamination of TurboV ion spray, or the instrument
requires tuning and optimization. Clean the ion source
including curtain plate, orifice plate or skimmer, Q0. Tune or
optimize the instrument following the instrument manual.
Refer to the Operating Instruction – Tune and Calibrate, the
ion source operator guide, or the Analyst ® software Help
system.
9.1.4.2.5.2 Low Vacuum Pressure – Low pump oil level could cause the
vacuum pressure issue. Check the pump oil level and add oil
if necessary.
9.2 Initial Calibration
9.2.1 Calibration Design
9.2.1.1 Demonstration and documentation of acceptable initial calibration is required
before any samples are analyzed. After the initial calibration is successful, a
continuing calibration verification (CCV) is required at the beginning and end of
each period in which analyses are performed, and after every tenth field sample.
Samples must be bracketed with CCV’s passing for all criteria, or the samples
should be re-analyzed (with the single exception mentioned in Appendix B).
9.2.1.2 ESI-MS/MS Tune – Tuning should occur at least every six months using PPGs
for tuning. Tune the system when ICAL won't pass, when the peak shape is
significantly off (indicating an MS problem), when major maintenance is
performed, or instrument is moved.
9.2.1.3 The instrument tuning with PPGs has its own manufacturing criteria- see the
documentation. After PPGs passes, calibration must be verified to be +/- 0.5 amu

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

16 of 58
 ENV-SOP-MIN4-0178, Rev 01

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Determination of Selected 36 Per- and Polyfluoroalkyl Substances (PFAS) by
LC/MS/MS
TEST METHOD: Isotope Dilution
ISSUER: Pace ENV – Minneapolis – MIN4
COPYRIGHT © 2020 Pace Analytical Services, LLC

of true values by acquiring a full scan continuum mass spectrum of a PFAS stock
standard.
9.2.1.4 Mass calibration range must bracket the ion masses of interest.
9.2.1.5 When done, run the Compound Optimization or Manual Tuning under the Tune
and Calibrate tab to optimize response and peak shape.
9.2.1.6 Prepare a set of six CAL standards (Table 9.2.1.20A as an example). Analyze
each standard level with the same acquisition method used to analyze samples,
changes to retention times or other analytical parameters are saved as part of the
local method generated with each analytical sequence, these parameters can be
adjusted mid-sequence so long as they are applied to all data.
9.2.1.7 The LC/MS/MS system is calibrated using the isotope dilution and internal
standard technique. Use the LC/MS/MS data system software to generate a linear
regression calibration curve for each of the relevant analytes. This curve may be
concentration weighted, if necessary.
9.2.1.8 A calibration meets criteria when the recovery for each calibration point reads
back at ±30% for all calibration points.
9.2.1.8.1 For Wisconsin samples, re-quantitated concentrations for all target
compounds at all concentration levels must be within the range 70-
130% of their actual concentrations, except for the lowest calibration
concentration level, which must be within the range of 50-150% of
actual concentration.
9.2.1.9 Provided a minimum of five calibration points are still being used, a point at the
top or bottom of the calibration curve may be dropped to achieve recovery
requirements across the remaining points. Dropping high concentration points
lowers the PQL of the calibration and may require that more dilutions are
performed. Dropping low calibration points can potentially elevate the RL for this
sequence.
9.2.1.10 An ICV (prepared from a second source standard or by different analyst) is run
with every initial calibration curve (ICAL). The acceptance criteria are ±30% of the
true value.
9.2.1.11 Additional calibration procedures (where applicable) can be found in ENV-POL-
CORQ-0005 Acceptable Calibration Practices for Instrument Testing (or
equivalent replacement).
9.2.1.12 537 Mix – Mix 40 mL of Optima grade Water with 960 mL of Optima grade
Methanol. Expires 1 year from prep.
9.2.1.13 PFAC_EIS – PFAC (Extracted Internal Standards) (0.05 µg/mL, 25 EIS)
9.2.1.13.1 Dissolve 40 µL of each MPFAC-6ES standard in 537 Mix. Dilute to 2
mL.
9.2.1.13.2 Dissolve each 1 mL of MPFAC-24ES and 10.3.2.1 mix solution in 537
Mix. Dilute to 20 mL. Added 100 µL to each field sample, standard,
blanks and QC samples prior to extraction.

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

17 of 58
 ENV-SOP-MIN4-0178, Rev 01

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Determination of Selected 36 Per- and Polyfluoroalkyl Substances (PFAS) by
LC/MS/MS
TEST METHOD: Isotope Dilution
ISSUER: Pace ENV – Minneapolis – MIN4
COPYRIGHT © 2020 Pace Analytical Services, LLC

9.2.1.13.3 Used for curve and sample prep. Store at room temperature. Expires
180 days from prep.
9.2.1.14 PFAC_IIS – PFAC (Injection Internal Standards) (0.05 µg/mL)
9.2.1.14.1 Dissolve 50 µL of each MPFAC-Injection Internal Standards in 537
Mix. Dilute to 50 mL. Added 100 µL to each field sample, standard,
blanks and QC samples just prior to analysis.
9.2.1.14.2 Used for curve prep and sample extraction. Store at room
temperature. Expires 180 days from prep.
9.2.1.15 PFAC_Native Spike Solution
9.2.1.15.1 Dissolve 100 µL of each PFAC-8Native in 537 Mix. Dilute to 5 mL.
9.2.1.15.2 PFAC_Native Spike 1 (~0.2 µg/mL) – Dissolve 1 mL of PFAC-30PAR
and 1 mL of 9.2.1.16.1 mix solution to 537 Mix. Dilute to 5 mL.
9.2.1.15.3 PFAC_Native Spike 2 (~0.050 µg/mL) – Dissolve 2.5 mL of
PFAC_Native Spike 1 to 537 Mix. Dilute to 10 mL.
9.2.1.15.4 Used for curve prep and sample extraction. Store at room
temperature. Expires 180 days from prep.
9.2.1.16 PFAC_ICV Spike Solution
9.2.1.16.1 Dissolve each of 100 µL PFAC-12Native in 537 Mix. Dilute to 2.5 mL.
9.2.1.16.2 PFAC_ICV Spike 1 (~0.2 µg/mL) – Dissolve 1 mL of PFAC-24PAR
and 1 mL of 9.2.1.17.1 mix solution to 537 Mix. Dilute to 10 mL.
9.2.1.16.3 PFAC_ICV Spike 2 (~0.050 µg/mL) – Dissolve 2.5 mL of PFAC_ICV
Spike 1 to 537 Mix. Dilute to 10 mL.
9.2.1.16.4 Used for ICV prep. Store at room temperature. Expires 180 days from
prep.
9.2.1.17 Isomer Check PDS – Isomer check Qualitative primary standard Spike
9.2.1.17.1 Dissolve 40 µL of T-PFOA stock solution in 537 Mix, dilute to 1 mL.
Expires 180 days from prep.
9.2.1.17.2 PFOA qualitative dilution standard spike – Dissolve 50 µL PFOA
qualitative primary standard spike in 537 Mix, dilute to 2 mL. Expires
180 days from prep.
9.2.1.18 Calibration Curve – Different volumes of PFAC_Native Spike solutions at various
concentrations are added to 1 mL 537 Mix (Table 9.2.1.20A). A known amount of
EIS is added into each calibration point. The corresponding concentration in 1 mL
final solvent is shown in Table 9.2.1.20B.
9.2.1.19 Table 9.2.1.20A – Example Calibration Curve
Extracted Extracted
Calibration Native std. Native std. Injection IS Injection IS
IS Soln IS Soln
Standard Soln added Soln conc. Soln added Soln conc.
added conc.
Point (µL) (µg/mL) (µL) (µg/mL)
(µL) (µg/mL)
CS-1 10 0.05 100 0.05 100 0.05

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

18 of 58
 ENV-SOP-MIN4-0178, Rev 01

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Determination of Selected 36 Per- and Polyfluoroalkyl Substances (PFAS) by
LC/MS/MS
TEST METHOD: Isotope Dilution
ISSUER: Pace ENV – Minneapolis – MIN4
COPYRIGHT © 2020 Pace Analytical Services, LLC

Extracted Extracted
Calibration Native std. Native std. Injection IS Injection IS
IS Soln IS Soln
Standard Soln added Soln conc. Soln added Soln conc.
added conc.
Point (µL) (µg/mL) (µL) (µg/mL)
(µL) (µg/mL)
CS-2 20 0.05 100 0.05 100 0.05
CS-3 40 0.05 100 0.05 100 0.05
CS-4 100 0.05 100 0.05 100 0.05
CS-5 200 0.05 100 0.05 100 0.05
CS-6 100 0.20 100 0.05 100 0.05
CS-7 250 0.20 100 0.05 100 0.05
CS-8 500 0.20 100 0.05 100 0.05
ICV 100 0.05 100 0.05 100 0.05
T-PFOA
(Qualitative 100 0.05 100 0.05 100 0.05
Calibration)

Table 9.2.1.20B – Concentrations of Each Analyte in 1 mL Final Solvent (ng/L)

Analyte CS-1 CS-2 CS-3 CS-4 CS-5 CS-6 CS-7 CS-8 ICV
PFBA 0.50 1.00 2.00 5.00 10.00 20.00 50.00 100.00 5.00
PFPeA 0.50 1.00 2.00 5.00 10.00 20.00 50.00 100.00 5.00
PFHxA 0.50 1.00 2.00 5.00 10.00 20.00 50.00 100.00 5.00
PFHpA 0.50 1.00 2.00 5.00 10.00 20.00 50.00 100.00 5.00
PFOA 0.50 1.00 2.00 5.00 10.00 20.00 50.00 100.00 5.00
PFNA 0.50 1.00 2.00 5.00 10.00 20.00 50.00 100.00 5.00
PFDA 0.50 1.00 2.00 5.00 10.00 20.00 50.00 100.00 5.00
PFUdA 0.50 1.00 2.00 5.00 10.00 20.00 50.00 100.00 5.00
PFDoA 0.50 1.00 2.00 5.00 10.00 20.00 50.00 100.00 5.00
PFTrDA 0.50 1.00 2.00 5.00 10.00 20.00 50.00 100.00 5.00
PFTeDA 0.50 1.00 2.00 5.00 10.00 20.00 50.00 100.00 5.00
PFOSA 0.50 1.00 2.00 5.00 10.00 20.00 50.00 100.00 5.00
N-EtFOSAA 0.50 1.00 2.00 5.00 10.00 20.00 50.00 100.00 5.00
N-MeFOSAA 0.50 1.00 2.00 5.00 10.00 20.00 50.00 100.00 5.00
PFBS 0.44 0.89 1.77 4.43 8.85 17.70 44.25 88.50 4.43
PFPeS 0.47 0.94 1.88 4.70 9.40 18.80 47.00 94.00 4.70
PFHxS* 0.46 0.91 1.82 4.55 9.10 18.20 45.50 91.00 4.55
PFHpS 0.48 0.95 1.90 4.75 9.50 19.00 47.50 95.00 4.75
PFOS* 0.47 0.93 1.86 4.65 9.30 18.60 46.50 93.00 4.65
PFNS 0.48 0.96 1.92 4.80 9.60 19.20 48.00 96.00 4.80
PFDS 0.48 0.97 1.93 4.83 9.65 19.30 48.25 96.50 4.83

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

19 of 58
 ENV-SOP-MIN4-0178, Rev 01

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Determination of Selected 36 Per- and Polyfluoroalkyl Substances (PFAS) by
LC/MS/MS
TEST METHOD: Isotope Dilution
ISSUER: Pace ENV – Minneapolis – MIN4
COPYRIGHT © 2020 Pace Analytical Services, LLC

Analyte CS-1 CS-2 CS-3 CS-4 CS-5 CS-6 CS-7 CS-8 ICV
4:2FTS 0.47 0.94 1.87 4.68 9.35 18.70 46.75 93.50 4.68
6:2FTS 0.48 0.95 1.90 4.75 9.50 19.00 47.50 95.00 4.75
8:2FTS 0.48 0.96 1.92 4.80 9.60 19.20 48.00 96.00 4.80
10:2FTS 0.48 0.97 1.93 4.83 9.65 19.30 48.25 96.50 4.83
HFPO-DA 0.50 1.00 2.00 5.00 10.00 20.00 50.00 100.00 5.00
DONA 0.47 0.95 1.89 4.73 9.45 18.90 47.25 94.50 4.73
N-MeFOSA 0.50 1.00 2.00 5.00 10.00 20.00 50.00 100.00 5.00
N-EtFOSA 0.50 1.00 2.00 5.00 10.00 20.00 50.00 100.00 5.00
N-MeFOSE 0.50 1.00 2.00 5.00 10.00 20.00 50.00 100.00 5.00
N-EtFOSE 0.50 1.00 2.00 5.00 10.00 20.00 50.00 100.00 5.00
9Cl-PF3ONS 0.47 0.93 1.86 4.66 9.32 18.64 46.60 93.20 4.66
11Cl-PF3OUdS 0.47 0.94 1.88 4.71 9.42 18.84 47.10 94.20 4.71
PFDoS 0.48 0.97 1.94 4.84 9.68 19.36 48.40 96.80 4.84
PFHxDA 0.50 1.00 2.00 5.00 10.00 20.00 50.00 100.00 5.00
PFODA 0.50 1.00 2.00 5.00 10.00 20.00 50.00 100.00 5.00

9.2.1.20 Store at room temperature. Expires 180 days from prep.

NOTE: Stock standards (Section 8.2) were stored at ≤ 4± 2 °C. Primary dilution
standards were stored at room temperature to prevent adsorption of the method
analytes onto the container surfaces that may occur when refrigerated. Storing
the standards at room temperature will also minimize daily imprecision due to the
potential of inadequate room temperature stabilization.
9.2.2 Calibration Sequence
9.2.2.1 ESI-MS/MS Tune – Tuning should occur at least every six months using PPGs
for tuning. Tune the system when ICAL won't pass, when the peak shape is
significantly off (indicating an MS problem), when major maintenance is
performed, or instrument is moved.
9.2.2.1.1 Load a 500 µL syringe filled with PPGs tuning solution in the syringe
pump and connect it directly to the probe. Hold syringe pump power
button for a few seconds to purge the line. Use the SCIEX Analyst®
1.6.3 software to adjust the parameters under the Tune and Calibrate
tab to a relative signal maxima for peaks 44.998, 585.385, 933.636,
1223.845, 1572.097, 1863.306, 2037.431, 2211.557 in negative mode
and 59.050, 175.133, 616.464, 906.673, 1254.925, 1545.134,
2010.469, 2242.637 in positive mode.
9.2.2.1.2 Mass assignment of tuning standard within 0.5 amu of true value.
9.2.2.1.3 When done, run the Compound Optimization or Manual Tuning under
the Tune and Calibrate tab to optimize response and peak shape.

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

20 of 58
 ENV-SOP-MIN4-0178, Rev 01

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Determination of Selected 36 Per- and Polyfluoroalkyl Substances (PFAS) by
LC/MS/MS
TEST METHOD: Isotope Dilution
ISSUER: Pace ENV – Minneapolis – MIN4
COPYRIGHT © 2020 Pace Analytical Services, LLC

9.2.2.2 Optimize the precursor ion and product ion for each target analyte by infusing a
standard mix from calibration curve to MS. The MS parameters (voltages,
temperatures, gas flows, etc.) and the MS/MS parameters (collision energy,
declustering potential, collision cell exit potential, etc.) are determined to achieve
optimal analyte responses.
NOTE: There have been reports that not all product ions in the linear PFOS are
produced in all branched PFOS isomers. (This phenomenon may exist for many
of the PFAS.) Thus, to reduce PFOS, PFBS and PFHxS bias, it is required that
the precursor m/z → m/z 80 transition be used as the quantitation transition. Some
MS/MS instruments may not be able to scan a product ion with such a wide mass
difference from the precursor ion; therefore, if the MS/MS cannot measure the
precursor m/z → m/z 80 transition they may not be used for this method if PFOS,
PFBS, or PFHxS analysis is to be conducted.
9.2.2.3 Establish LC operating parameters that optimize resolution and peak shape.
9.2.2.4 Inject a mid-level CAL standard under optimized LC/MS/MS conditions to ensure
that each target analyte is observed in its retention time window and that there
are at least 10 scans across the peak for optimum precision.
9.2.2.5 Prepare a set of at least five calibration point standards for linear fit (Table
9.2.1.20A as an example). Analyze each standard level with the same acquisition
method used to analyze samples, changes to retention times or other analytical
parameters are saved as part of the local method generated with each analytical
sequence, these parameters can be adjusted mid-sequence so long as they are
applied to all data. Use the LC/MS/MS data system software to generate a linear
regression curve for each of the relevant analytes. This curve may be
concentration weighted, if necessary. Forcing zero is not allowed for this analysis.
9.2.2.5.1 Calibration points at the top or the bottom of the curve may be dropped
to achieve recovery requirements across the remaining points
provided the minimum number of calibration points are still being used
based on the curve fit. Dropping high concentration points lowers the
upper QL of the calibration and may require that more dilutions are
performed. Dropping low calibration points may elevate the reporting
limit for samples associated with this calibration. The RL must be met
without exception.
9.2.2.6 Analyte quantification uses the isotope dilution technique for the analytes having
commercially available isotopically labeled analogs. The internal standard
technique is used when a labeled analog is not commercially available for the
target analyte. Details in analytes quantification refer to Section 10.5.
9.2.3 ICAL Evaluation
9.2.3.1 Calibration factors have RSD that is ≤ 20% for all analytes
9.2.3.2 Linear regressions have a coefficient of determination that is r2 ≥0.99 and a
minimum of five non-zero concentration standards is used.
9.2.3.3 Do not force linear regression through zero.

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

21 of 58
 ENV-SOP-MIN4-0178, Rev 01

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Determination of Selected 36 Per- and Polyfluoroalkyl Substances (PFAS) by
LC/MS/MS
TEST METHOD: Isotope Dilution
ISSUER: Pace ENV – Minneapolis – MIN4
COPYRIGHT © 2020 Pace Analytical Services, LLC

9.2.3.4 For each calibration standard, reprocess the target (native) analyte against the
chosen calibration function. The reprocessed recoveries are expected to be within
±30% of true value. For some data uses, the lowest concentration standard
reprocessed recoveries are expected to be within ±50% of true value.
9.2.3.5 The lowest concentration ICAL standard must be ≤ reporting level (RL).
9.2.3.6 S/N Ratio: ≥ 10: 1 for all quantification ions and S/N Ratio of ≥ 3:1 for confirmation
ions
9.2.3.7 Demonstration and documentation of acceptable initial calibration is required
before any samples are analyzed. After the initial calibration is successful, a CCV
is required at the beginning and end of each period in which analyses are
performed, and after every tenth field sample.
9.2.4 Relative Standard Error (RSE) – Percent error between the calculated and expected
amounts of an analyte should be ≤ 30% for all standards. For some data uses, ≤ 50% may
be acceptable for the lowest calibration point.
9.2.5 Initial Calibration Verification
9.2.5.1 Initial Calibration Verification (ICV) – analyze an ICV sample from a source
different from the source of the CAL standards with each new ICAL before sample
analysis. If a second vendor is not available, then a different lot of the standard
should be used. The ICV should be prepared and analyzed just like a CCV.
Acceptance criteria for the ICV are identical to the CCV; the calculated amount
for each analyte must be ± 30% of the expected value. If measured analyte
concentrations are not of acceptable accuracy, check the entire analytical
procedure to locate and correct the problem.
9.2.6 Continuing Calibration Verification
9.2.6.1 CCVs are run at the beginning, end, and bracketing every 10 field samples.
Blanks, rinses, and spiked QC (LCS/LCSD/MS/MSD) are not considered field
samples, and so can be run in addition to 10 field samples in a CCV window.
9.2.6.2 The opening CCV for any batch must be below or at the RL (CS-1), all further
CCVs cycle between mid and high level calibration point.
9.2.6.3 Calculate the concentration of each analyte in the CCV. The calculated amount
for each analyte must be within ± 30% of the true value. Determine that the
absolute areas of the quantitation ions of the EIS and IIS are within ±50% from
the mid-point measured during initial calibration. On days when ICAL is not
performed, the peak areas must be within ±50% of the peak area measured in
daily initial CCV. If any of the EIS and IIS areas has changed by more than these
amounts, adjustments must be made to restore system.
9.2.6.3.1 For Wisconsin samples, the calculated amount for each analyte must
be within ± 30% of the true value except for the lowest ICAL point, for
which the calculated amount for each analyte must be within ± 50% of
the true value.

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

22 of 58
 ENV-SOP-MIN4-0178, Rev 01

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Determination of Selected 36 Per- and Polyfluoroalkyl Substances (PFAS) by
LC/MS/MS
TEST METHOD: Isotope Dilution
ISSUER: Pace ENV – Minneapolis – MIN4
COPYRIGHT © 2020 Pace Analytical Services, LLC

9.2.6.4 If the CCV fails high for a particular analyte, and the field sample shows no
detection for that analyte, samples may be reported without re-analysis.
9.2.7 Laboratory Control Sample/Laboratory Control Sample Duplicate (LCS/LCSD)
9.2.7.1 A LCS is required with each extraction batch. See DoD acceptance criteria for
LCS targets in Appendix E for aqueous and solid matrices. If the LCS results do
not meet the criteria listed in Appendix E for method analytes, then all data for the
problem analyte(s) must be considered invalid for all samples in the extraction
batch. For target analytes not included in the DoD Limits for batch control Table
B-15 per DoD QSM 5.3, , limits of 70-140% recovery will be used as acceptance
criteria. For tissue batches, the recoveries are expected to be within 60-140%.
9.2.7.1.1 For Wisconsin samples, the recoveries are expected to be within 60-
135%, except for the low range (1 – 2x RL) where the recoveries are
expected to be within 50-150% in aqueous and solid batches. For
tissue batches, spike the LCS at midrange. For tissue batches, the
recoveries are expected to be within 60-135% with the following
exceptions: for PFHxDA, PFODA, and NMeFOSA, the recoveries are
expected to be within 50-135%; for PFDS, PFDoS, and 4:2 FTS, the
recoveries are expected to be within 40-135%.
9.2.8 EIS Recovery
9.2.8.1 The EIS is fortified into all samples, CCVs, MBs, LCSs, MSs, MSDs, and FD prior
to extraction. It is also added to the CAL standards. The EIS is a means of
assessing method performance from extraction to final chromatographic
measurement.
9.2.8.2 A minimal signal to noise ratio of 10:1 is expected for each EIS. Do not report
results with a qualifier if this minimum is not achieved.
9.2.8.3 EIS recovery must be in ±50% of the mid-point ICAL when the day the ICAL was
performed. When EIS recovery from a sample, blank, or CCV failed the criteria,
check 1) calculations to locate possible errors, 2) standard solutions for
degradation, 3) contamination, and 4) instrument performance. Correct the
problem and re-analyze the extract.
9.2.8.3.1 For Wisconsin samples, all EIS compounds must recover within the
range 25-150%, except 13C8-PFOSA, d3-MeFOSA, d5-EtFOSA, d7-
MeFOSE, and d9-EtFOSE, which must recover within the range 10-
150%. Recovery will be based on area counts.
9.2.8.4 If the EIS recoveries in a chromatographic run do not meet these criteria, inject a
second aliquot of that extract from a new capped auto-sampler vial.
9.2.8.5 If the reinjected aliquot produces an acceptable EIS recoveries, report results for
that aliquot.
9.2.8.6 If recoveries are acceptable for QC samples, but not field samples, the field
samples must be re-prepped and reanalyzed (greater dilution may be needed). If
recoveries are unacceptable for QC samples, correct problem, and reanalyze all
associated failed field samples.

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

23 of 58
 ENV-SOP-MIN4-0178, Rev 01

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Determination of Selected 36 Per- and Polyfluoroalkyl Substances (PFAS) by
LC/MS/MS
TEST METHOD: Isotope Dilution
ISSUER: Pace ENV – Minneapolis – MIN4
COPYRIGHT © 2020 Pace Analytical Services, LLC

9.2.8.7 Apply Q-flag and discuss in the Case Narrative only if reanalysis confirms failures
in exactly the same manner.
9.2.8.8 If the extract re-analysis meets the EIS recovery criterion, report only data for the
re-analyzed extract.
9.2.8.9 If the extract re-analysis fails the criterion, the analyst should check the calibration
by injecting the last CAL standard that passed. If the CAL standard fails the
criteria, re-calibration is in order. If the CAL standard is acceptable, extraction of
the sample should be repeated provided the sample is still within the holding time.
If the re-extracted sample also fails the recovery criterion, report all data for that
sample as suspect recovery to inform the data user that the results are suspect
due to EIS recovery. Alternatively, collect a new sample and re-analyze.
9.2.9 IIS Recovery
9.2.9.1 The IIS is fortified into all samples, CCVs, MBs, LCSs, MSs, MSDs, and FD prior
to extraction. It is also added to the CAL standards. The IIS is a means of
assessing instrument performance.
9.2.9.2 A minimal signal to noise ratio of 10:1 is expected for each IIS. Do not report
results with a qualifier if this minimum is not achieved.
9.2.9.3 IIS recovery must be in ±50% of the mid-point ICAL when the day the ICAL was
performed. When IIS recovery from a sample, blank, or CCV is failed the criteria,
check 1) calculations to locate possible errors, 2) standard solutions for
degradation, 3) contamination, and 4) instrument performance. Correct the
problem and re-analyze the extract.
9.2.9.4 If the EIS recoveries in a chromatographic run do not meet these criteria, inject a
second aliquot of that extract from a new capped auto-sampler vial.
9.2.9.5 If the reinjected aliquot produces an acceptable IIS recovery, report results for
that aliquot.
9.2.9.6 If recoveries are acceptable for QC samples, but not field samples, the field
samples must be re-prepped and reanalyzed (greater dilution may be needed). If
recoveries are unacceptable for QC samples, correct problem, and reanalyze all
associated failed field samples.
9.2.9.7 If the extract re-analysis meets the IIS recovery criterion, report only data for the
re-analyzed extract.
9.2.10 Additional calibration procedures (where applicable) can be found in ENV-POL-CORQ-0005
Acceptable Calibration Practices for Instrument Testing (or equivalent replacement).
9.3 Sample Preparation
9.3.1 Some of the PFAS adsorb to surfaces, including polypropylene. Therefore, the aqueous
sample bottles must be rinsed with the elution solvent. The bottle rinse is passed through
the cartridge to elute the method analytes and is then collected.

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

24 of 58
 ENV-SOP-MIN4-0178, Rev 01

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Determination of Selected 36 Per- and Polyfluoroalkyl Substances (PFAS) by
LC/MS/MS
TEST METHOD: Isotope Dilution
ISSUER: Pace ENV – Minneapolis – MIN4
COPYRIGHT © 2020 Pace Analytical Services, LLC

NOTE: The SPE cartridges and sample bottles described in this section are designed as
single use items and should be discarded after use. They may not be refurbished for reuse
in subsequent analyses.
9.3.2 Solid Sample Preparation
9.3.2.1 Homogenize the entire solid sample received in the sample container in which it
was collected in by stirring the solid with a clean spatula or other implement.
9.3.2.2 5 g of solid sample is weighed in a tared 50-mL polypropylene bottle
9.3.2.3 Add a 100 µL aliquot of the PFAC_EIS to all field and QC samples at the very
beginning of the procedure.
9.3.2.4 QC samples for each batch include a MB, LCS and MS/MSD which are extracted
along with each prep batch.
9.3.2.4.1 MB is required for each prep batch. Each batch contains a LCS and a
pair of MS/MSD. LCS/MS/MSD spike at concentrations ≥ LOQ and ≤
the mid-level calibration concentration. If insufficient sample is
available for a pair of MS/MSD, an MS, Dup, and LCSD at the same
level of LCS may be used.
9.3.2.4.2 The LCS/LCSD/MS/MSD is spiked with 20 µL of the PFAC_Native
Spike 2.
9.3.2.5 5 mL of 0.2% ammonia/methanol is added to all samples and QC, bottles are
sealed and put on an ultrasonicator for 20 minutes and then shake for one hour.
9.3.2.6 Centrifuge the samples and QC for 5 minutes after shake.
9.3.2.7 Decant the supernatant layer in a 50-mL polypropylene bottle with 50 mg of ENVI-
Carb powder.
9.3.2.8 Repeat sections 9.3.2.5 with 4 mL of 0.2% ammonia/methanol and centrifuge. All
supernatant are collected and combined.
9.3.2.9 The combined supernatant is shaken for one hour and then centrifuge for 5
minutes after shake.
9.3.2.10 Clean the filter with 10 mL of 1% of ammonia/acetonitrile.
9.3.2.11 Condition the pre-cleaned filter with 10 mL methanol. Pass the combined
supernatant through the filter. Rinse the filter with additional 1 mL 0.2%
ammonia/methanol. Collect the filtrate and turn on the vacuum for 10 minutes.
9.3.3 Aqueous Sample Preparation
9.3.3.1 Sample volume is determined gravimetrically. The full sample bottle is weighed
and the empty bottle is weighted after extraction. The sample volume is the
difference between the full and empty bottle weights. Sample density is assumed
at 1 g/mL. When the sample has significant solids, the laboratory should account
for the weight or volume displaced by the solid in the initial sample volume
determination and include this information in the report.

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

25 of 58
 ENV-SOP-MIN4-0178, Rev 01

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Determination of Selected 36 Per- and Polyfluoroalkyl Substances (PFAS) by
LC/MS/MS
TEST METHOD: Isotope Dilution
ISSUER: Pace ENV – Minneapolis – MIN4
COPYRIGHT © 2020 Pace Analytical Services, LLC

9.3.3.2 pH is taken using strips in the lab. This is accomplished via the use of common
laboratory grade pH strips (Whatman Indicator Paper pH 0-14 Type CF Cat. No.
2613-991). Adjust the pH to ~5 with acetic acid or 10 mM sodium hydroxide
solution when necessary.
9.3.3.3 Add a 100 µL aliquot of the PFAC_EIS to all field and QC samples at the very
beginning of the procedure, before extraction, centrifuging, filtering or phase
separation takes place. Cap and invert and mix.
9.3.3.4 Ideally, whole samples will pass through the cartridge as received. If particulates
in the sample is greater than one percent, centrifuge the sample and take the
liquid phase through the SPE after spiking the PFAC_EIS.
9.3.3.5 QC samples for each batch include a MB, LCS and MS/MSD which are extracted
along with each prep batch.
9.3.3.5.1 MB is required for each prep batch.
9.3.3.5.2 Each batch contains a LCS and a pair of MS/MSD. LCS/MS/MSD
spike at concentrations ≥ LOQ and ≤ the mid-level calibration
concentration. If insufficient sample is available for a pair of MS/MSD,
an MS, Dup, and LCSD at the same level of LCS may be used.
9.3.3.5.3 The LCS/LCSD/MS/MSD is spiked with 20 µL of the PFAC_Native
Spike 2.
9.3.3.6 Proceed with SPE procedure in section 9.3.6.
9.3.4 Tissue Sample Preparation
9.3.4.1 Homogenization will be performed on the entire tissue sample in accordance with
SOP ENV-SOP-GBAY-0129 Sample Homogenization, Compositing and Sub-
Sampling (or equivalent replacement) by the Pace Green Bay laboratory.
9.3.4.2 2 g of tissue sample is weighed in a tared 50-mL HDPE bottle.
9.3.4.3 Add a 100 µL aliquot of the PFAC_EIS to all field and QC samples (canola oil and
SRM) at the very beginning of the procedure.
9.3.4.4 QC samples for each batch include a MB, LCS, MS/MSD, and SRM which are
extracted along with each prep batch.
9.3.4.4.1 MB is required for each prep batch. Each batch contains a LCS and a
pair of MS/MSD. LCS/MS/MSD spike at concentrations at
concentrations ≥ LOQ and ≤ the mid-level calibration concentration. If
insufficient sample is available for a pair of MS/MSD, an MS, Dup, and
LCSD at the same level of LCS may be used.
9.3.4.4.2 The LCS/LCSD/MS/MSD is spiked with 40 µL of the PFAC_Native
Spike 2.
9.3.4.5 7 mL of 1% ammonia/acetonitrile is added to all samples and QC, bottles are
sealed and put on a shaker for 16 hours.
9.3.4.6 Centrifuge the samples and QC for 5 minutes after shake.

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

26 of 58
 ENV-SOP-MIN4-0178, Rev 01

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Determination of Selected 36 Per- and Polyfluoroalkyl Substances (PFAS) by
LC/MS/MS
TEST METHOD: Isotope Dilution
ISSUER: Pace ENV – Minneapolis – MIN4
COPYRIGHT © 2020 Pace Analytical Services, LLC

9.3.4.7 Decant the supernatant layer in a 50-mL polypropylene bottle with 100 mg of
ENVI-Carb powder and shake for 1 hour and then centrifuge for 5 minutes after
shake
9.3.4.8 Clean the 250 mg ENVI-Carb cartridge with 10 mL of 1% ammonia/acetonitrile.
9.3.4.9 Condition the pre-cleaned cartridge with 10 mL methanol. Pass the supernatant
through the cartridge. Rinse the filter with additional 1 mL 1%
ammonia/acetonitrile.
9.3.4.10 Collect the filtrate and dilute the filtrate with 125 mL H2O and adjust pH to ~5.
Proceed with SPE procedure in 9.3.5.
9.3.5 Cartridge SPE Procedure
9.3.5.1 Cartridge Clean-up – Rinse each cartridge with 20 mL of 1%
ammonia/acetonitrile solution.
9.3.5.2 Cartridge Conditioning – Do NOT allow cartridge packing material to go dry
during any of the conditioning steps. Condition each cartridge with 10 mL of 0.2%
ammonia/methanol solution following with 10 mL of methanol. Next, rinse each
cartridge with 10 mL of reagent water following with 10 mL of acetate buffer,
without allowing the water to drop below the top edge of the packing. If the
cartridge goes dry during the conditioning phase, the conditioning must be started
over. Turn on the vacuum, and begin adding sample to the cartridge through the
attached plastic sample transfer reservoir.
9.3.5.3 Sample Extraction – Adjust the vacuum so that the approximate flow rate is 6-
10 mL/min. Do not allow the cartridge to go dry before all the sample has passed
through.
9.3.5.4 Sample Bottle and Cartridge Rinse – Rinse the sample bottles with two 5-mL
aliquots of reagent water, then draw each aliquot through the plastic sample
transfer reservoir and the cartridges. Draw air through the cartridge for 25 min at
high vacuum (10-15 in. Hg).
NOTE: If transfer tubes are used in place of the sample transfer tubes to pass the
samples through the cartridges, these reservoirs must be treated like the empty
plastic sample transfer reservoirs. After the entire sample has passed through the
cartridge, the tubes must be rinsed to waste with reagent water.
9.3.5.5 Sample Bottle and Cartridge Elution – Turn off and release the vacuum. Lift the
extraction manifold top and insert a rack with collection tubes into the extraction
tank to collect the extracts as they are eluted from the cartridges. Rinse the
sample bottles with 3 mL of 0.2% ammonia/methanol twice and elute the analytes
from the cartridges by pulling the additional 3 mL of 0.2% ammonia/methanol
through the sample plastic reservoirs and the cartridges. Turn the vacuum on for
20 minutes between each elution. The elution solvent used to rinse the sample
bottles must be swirled down the sides of the reservoirs while eluting the cartridge
to ensure that any method analytes on the surface of the reservoirs are transferred
to the extract.

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

27 of 58
 ENV-SOP-MIN4-0178, Rev 01

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Determination of Selected 36 Per- and Polyfluoroalkyl Substances (PFAS) by
LC/MS/MS
TEST METHOD: Isotope Dilution
ISSUER: Pace ENV – Minneapolis – MIN4
COPYRIGHT © 2020 Pace Analytical Services, LLC

NOTE: After centrifuging, it is expected that the solid phase remains in the bottom
of the container when rinsing the container with elution solvent. If rinsing the
container disrupts the solid phase significantly, the container can be centrifuged
again before removing the solvent for use during the elution step.
9.3.6 Extract Concentration – Concentrate the extract to approximately 0.8 mL for water and
solid extract and approximate 0.2 mL for tissue extract under a gentle stream of nitrogen
without a heated water bath. Add 100 µL of PFAC_IIS and fill the sample vial to 1 mL mark
with 537 Mix. Then vortex for 5-10 seconds. Transfer a ~100 µL to a 300 µL polypropylene
autosampler vial with a plastic pipette. The remaining extract is stored at 0-6 °C.
9.4 Analysis
9.4.1 Establish operating conditions equivalent to those summarized in Appendix C. Instrument
conditions and columns should be optimized prior to the initiation of the IDOC.
9.4.2 Establish an appropriate retention time window for each analyte. This should be based on
measurements of actual retention time variation for each method analyte in Calibration
(CAL) standard solutions analyzed on the LC over the course of time. A value of plus or
minus three times the standard deviation of the retention time obtained for each method
analyte while establishing the initial calibration and completing the IDC can be used to
calculate a suggested window size. However, the experience of the analyst should weigh
heavily on the determination of the appropriate retention window size.
9.4.3 Retention Time (RT) acceptance – RT of each analyte and EIS analyte must fall within 0.4
minutes (±0.2 minutes) of the predicted retention times from the daily CCV or, on days when
ICAL is performed, from the midpoint standard of the ICAL. Analytes must elute within 0.1
minute of the associated EIS. This criterion applies only to analyte and labeled analog pairs.
9.4.4 Calibrate the system by either the analysis of a calibration curve or by confirming the initial
calibration is still valid by analyzing a CCV. If establishing an initial calibration, complete the
IDC.
9.4.5 Begin analyzing field samples, including QC samples, at their appropriate frequency by
injecting 3 µL of final sample extractant, under the same conditions used to analyze the
ICAL standards.
9.4.6 At the conclusion of data acquisition, use the same software that was used in the calibration
procedure to identify peaks of interest in predetermined retention time windows. Use the
data system software to examine the ion abundances of the peaks in the chromatogram.
Identify an analyte by comparison of its retention time with that of the corresponding method
analyte peak in a reference standard. Comparison of the MS/MS mass spectra is not
particularly useful given the limited ±0.5 amu mass range around a single product ion for
each method analyte.
9.4.7 Dilution – When the concentrations of target analytes exceed the highest concentration of
ICAL, dilution analyses are required.
9.4.7.1 An appropriate dilution should be in the upper half of the calibration range, or
close to the CCV. The diluted extract must maintain the same methanol/water
ratio as the original extract

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

28 of 58
 ENV-SOP-MIN4-0178, Rev 01

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Determination of Selected 36 Per- and Polyfluoroalkyl Substances (PFAS) by
LC/MS/MS
TEST METHOD: Isotope Dilution
ISSUER: Pace ENV – Minneapolis – MIN4
COPYRIGHT © 2020 Pace Analytical Services, LLC

9.4.7.2 If an analyte concentration exceeds the range of the initial calibration curve, the
extract is diluted with 537 Mix. Re-inject the diluted extract. Incorporate the dilution
factor into the final concentration calculations. Acceptable injection internal
standard (IIS) performance is determined from the undiluted sample extract. The
resulting data is documented as a dilution, with an increased LOQ.
9.4.8 In validating this method, concentrations were calculated by measuring the product ions
listed in Appendix C. Two transitions and the ion transition ratio per analyte shall be
monitored and documented with the exception of PFBA and PFPeA. In order to avoid biasing
results high due to known interferences for some transitions, the following transitions must
be used for the quantification of the following analytes: PFOA: 413 —› 369, PFOS: 499 —›
80, PFHxS: 399 —› 80, PFBS: 299 —› 80, 4:2FTS: 327 —› 307, 6:2FTS: 427 —› 407
8:2FTS: 527 —› 507, N-EtFOSAA: 584 —› 419, N-MeFOSAA: 570 —› 419. If these
transitions are not used, the reason must be technically justified and documented (e.g.,
alternate transition was used due to observed interferences).
9.4.9 Calculate analyte concentrations using the multipoint calibration established in Section
9.2.1.20. Do not use daily calibration verification data to quantitate analytes in samples.
Adjust final analyte concentrations to reflect the actual sample volume.
9.4.10 Prior to reporting the data, the chromatogram is reviewed for any incorrect peak identification
or poor integration. Modify if necessary.
9.4.11 Calculations must utilize all available digits of precision, but final reported concentrations
are rounded to an appropriate number of significant figures (one digit of uncertainty),
typically two, and not more than three significant figures.
9.4.12 For native analytes, the Signal to Noise (S/N) ratio should be ≥3:1 for both quantitation and
confirmation ions. If S/N is not achieved, the analyte would be reported as not detected.
9.4.13 Ion Ratios – For analytes with two ion transitions (quantitation and confirmation) are
analyzed, the area ratio between the confirmation and quantitation transitions shall be
monitored and documented. The ion ratio for all analytes in each injection should be within
± 50% of the mid ICAL ion ratio for the same analyte in the ICAL. On days ICAL is not
performed, the ion ratio should be within ±50% of the initial CCV standard.
9.4.14 Report results in acid form.
9.4.15 Perform a moisture analysis on solid samples (on a subsample different than that used for
extraction) and adjust the final concentration of solid sample for the percent moisture.
9.4.16 DoD acceptance criteria for LCS and MS target analytes are listed in Appendix E. If the LCS
results do not meet the criteria listed then all data for the problem analyte(s) must be
considered invalid for all samples in the extraction batch. For target analytes not included in
the DoD Limits for batch control table in Table B-15 per DoD QSM 5.3, limits of 70-140%
recovery for water and soil, 60-140% recovery for tissue will be used as acceptance criteria.
9.5 Analytical Sequence
9.5.1 Example analytical sequence
Sequence
Instrument Blank (ICB)

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

29 of 58
 ENV-SOP-MIN4-0178, Rev 01

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Determination of Selected 36 Per- and Polyfluoroalkyl Substances (PFAS) by
LC/MS/MS
TEST METHOD: Isotope Dilution
ISSUER: Pace ENV – Minneapolis – MIN4
COPYRIGHT © 2020 Pace Analytical Services, LLC

Initial Calibration (ICAL)

Instrument Blank (ICB)
Initial Calibration Verification (ICV)
Continuing Calibration Curve (CCV)
Continuing Calibration Blank (CCB)
Qualitative Standard for PFOA (T-PFOA)
Method Blank (MB)
Laboratory Control Spike/Dup (LCS/LCSD)
Matrix Spike/Dup (MS/MSD)
Field Samples
Continuing Calibration Curve (CCV)
Continuing Calibration Blank (CCB)
Field Samples
Continuing Calibration Curve (CCV)
Continuing Calibration Blank (CCB)

10.0 DATA ANALYSIS AND CALCULATIONS

10.1 Qualitative Identification
10.1.1 Manual Integration
Manual changes to automated integration is called manual integration. Manual integration
is sometimes necessary to correct inaccurate automated integrations but must never be
used to meet QC criteria or to substitute for proper instrument maintenance and/or method
set-up. To assure that all manual integrations are performed consistently and are ethically
justified, all manual integrations must be performed, reviewed, and recorded in accordance
with corporate SOP ENV-SOP-CORQ-0006, Manual Integration.
10.2 Quantitative Identification
10.2.1 Complete chromatographic resolution is not necessary for accurate and precise
measurements of analyte concentrations using MS/MS. In validating this method,
concentrations were calculated by measuring the product ions listed in Appendix C. Other
ions may be selected at the discretion of the analyst.
10.2.2 Prior to reporting the data, the chromatogram should be reviewed for any incorrect peak
identification or poor integration.
10.2.3 For PFHxS, PFOS, N-MeFOSAA and N-EtFOSAA, all the chromatographic peaks observed
in the standard must be integrated and the areas summed. Chromatographic peaks in all
Field Samples and QC samples must be integrated in the same way as the CAL standard
for analytes with quantitative standards containing the branched and linear isomers.
10.2.4 For PFOA, identify the branched isomers by analyzing a qualitative standard (T-PFOA) that
includes both linear and branched isomers and compare retention times and tandem mass
spectrometry transitions. Quantitate Field Samples and QC samples by integrating the total
response (i.e., accounting for peaks that are identified as linear and branched isomers) and
relying on the initial calibration with a linear isomer quantitative PFOA standard. This
qualitative PFOA standard is not used for quantitation. This branched isomer identification
check must be repeated any time changes occur that affect the analyte retention times.

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

30 of 58
 ENV-SOP-MIN4-0178, Rev 01

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Determination of Selected 36 Per- and Polyfluoroalkyl Substances (PFAS) by
LC/MS/MS
TEST METHOD: Isotope Dilution
ISSUER: Pace ENV – Minneapolis – MIN4
COPYRIGHT © 2020 Pace Analytical Services, LLC

10.2.5 Peaks that are consistent with branched isomers have been observed with other target
analytes, in particular PFOA. Quantitate of PFOA by integrating the total response (i.e.
accounting for peaks that are identified as linear and branched isomers) and relying on the
initial calibration with the linear-isomer quantitative standard is acceptable.
10.2.6 All analytes are quantified using the isotope dilution or internal standard technique.
10.2.7 The native analytes are quantified by comparison of their responses to the mass-labelled
internal standards. Relative response factors are calculated from analyses of standard
mixtures containing native analytes at six concentration levels, and the concentration
remains at a constant level for each internal standard. The target analytes response factors
are calculated by comparing the response from the native ion mass monitored to the
response from the ion mass of the corresponding isotopically labelled internal standard (See
Appendix D for reference).
10.3 Calculations
See the laboratory SOP ENV-SOP-MIN4-0171 Laboratory Calculations (or equivalent replacement)
for equations for common calculations.
10.3.1 Linear Calibration Using Average Response Factors
For each target analyte, calculate the response factor of each calibration level as follows:
Equation 1
RFi = (AaQs)/AsQa
Where, RF = Response factor
Aa = Sum of integrated areas for analyte
Qs = Quantity of labeled standard
As = Sum of integrated areas for labeled standard
Qa = Quantity of analyte
10.3.2 The levels of native analytes in the samples are quantified using the following
equations:
Equation 2
C = (AnQis)/Ais×W×RF
Where, RF = Response factor
An = Sum of integrated areas for target isomer
Qis = Quantity of labeled internal standard added to the sample
Ais = Sum of integrated areas for labeled internal standard
W = Sample amount

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

31 of 58
 ENV-SOP-MIN4-0178, Rev 01

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Determination of Selected 36 Per- and Polyfluoroalkyl Substances (PFAS) by
LC/MS/MS
TEST METHOD: Isotope Dilution
ISSUER: Pace ENV – Minneapolis – MIN4
COPYRIGHT © 2020 Pace Analytical Services, LLC

C = Concentration of target isomer

Equation 3
∑
Average Response Factor = =( )
Where, n = Number of calibration levels
RFi = Response factor for the ith level
10.3.3 The relative standard deviation (RSD) is calculated as follows:
Equation 4

(%) = × 100%

Where SD is the standard deviation of the average RF, which is calculated as

follows:

∑ ( − )
=
−1

10.3.4 Linear regression fit: y = mx + b

Equation 5
Ax/Ais = m(Cx/Cis) + b
Where, Ax = Response area for analyte
Ais = Response area for the internal standard
Cx = Analyte concentration of calibration standard
Cis = Internal standard concentration
m = Slope
b = y-intercept

10.3.5 The levels of native analytes in the samples are quantified using the following
equation:
Equation 6
Csx = (Ax/Ais -b)*Cis/m
Where, Csx = Unknown sample analyte concentration

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

32 of 58
 ENV-SOP-MIN4-0178, Rev 01

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Determination of Selected 36 Per- and Polyfluoroalkyl Substances (PFAS) by
LC/MS/MS
TEST METHOD: Isotope Dilution
ISSUER: Pace ENV – Minneapolis – MIN4
COPYRIGHT © 2020 Pace Analytical Services, LLC

Ax = Response area for analyte

Ais = Response area for the internal standard
Cis = Internal standard concentration
m = Slope
b = y-intercept
10.3.6 For Wisconsin samples, report sample results and all quality control blank results to the MDL
and include the RL for each result reported. Quality results reported between the MDL and
RL are estimated concentrations and reported with a J-flag.
Example 1
Laboratory Report Result as
MDL = 0.6 0.6
MRL = 2.0 2.0
Sample Result = 0.4 < 0.6

Example 2 (Estimated Concentration)

Laboratory Report Result as
MDL = 0.6 0.6
MRL = 2.0 2.0
Sample Result = 0.8 0.8 J

11.0 QUALITY CONTROL AND METHOD PERFORMANCE

11.1 Quality Control
The following QC samples are prepared and analyzed with each batch of samples. Refer to
Appendix B for acceptance criteria and required corrective action.

QC Item Frequency
Method Blank (MB) 1 per batch of 20 or fewer samples. If batch exceeds, 20
samples, every 20.
Laboratory Control Sample (LCS) 1 per batch of 20 or fewer samples. If batch exceeds, 20
samples, every 20.
Laboratory Control Sample Duplicate As needed
(LCSD)
Matrix Spike (MS) 1 per batch of 20 or fewer samples. If batch exceeds, 20
samples, every 20.
Matrix Spike Duplicate (MSD) 1 per batch of 20 or fewer samples. If batch exceeds, 20
samples, every 20.
Field Duplicate 1 per batch of 20 or fewer samples. If batch exceeds, 20
samples, every 20.

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

33 of 58
 ENV-SOP-MIN4-0178, Rev 01

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Determination of Selected 36 Per- and Polyfluoroalkyl Substances (PFAS) by
LC/MS/MS
TEST METHOD: Isotope Dilution
ISSUER: Pace ENV – Minneapolis – MIN4
COPYRIGHT © 2020 Pace Analytical Services, LLC

QC Item Frequency
Extraction Internal Standard All samples and QC
Injection Internal Standard All samples and QC
Standard Reference Material 1 per batch of 20 or fewer tissue samples. If batch
exceeds, 20 samples, every 20.

11.2 Instrument QC
The following Instrument QC checks are performed. Refer to Appendix B for acceptance criteria
and required corrective action.

QC Item Frequency
Tune Every 6 months or when ICAL won't pass, the peak shape is
significantly off, major maintenance is performed, or instrument is
moved.
Initial Calibration At instrument set up, after CCV failure
Initial Calibration Verification Once per calibration at mid-level of ICAL
Initial Calibration Blank One following the highest standard analyzed and prior to ICV
Continuing Calibration Verification At the beginning, end, and bracketing every 10 field samples
Continuing Calibration Blank 1 after each CCV
RT Window RT of each analyte and EIS analyte must fall within 0.4 minutes
(±0.2 minutes) of the predicted retention times from the daily
CCV or, on days when ICAL is performed, from the midpoint
standard of the ICAL.
Relative Retention Time Analytes must elute within 0.1 minute of the associated EIS. This
criterion applies only to analyte and labeled analog pairs

11.3 Method Performance

11.3.1 Method Validation
11.3.1.1 Detection Limits
Detection limits (DL) and limits of quantitation (LOQ) are established at initial
method setup and verified on an on-going basis thereafter. Refer to Pace ENV
corporate SOP ENV-SOP-CORQ-0011 Method Validation and Instrument
Verification.
11.4 Analyst Qualifications and Training
11.4.1 Employees that perform any step of this procedure must have a completed Read and
Acknowledgment Statement for this version of the SOP in their training record. In addition,
prior to unsupervised (independent) work on any client sample, analysts that prepare or
analyze samples must have successful initial demonstration of capability (IDOC) and must
successfully demonstrate on-going proficiency on an annual basis. Successful means the
initial and on-going DOC met criteria, documentation of the DOC is complete, and the DOC
record is in the employee’s training file. Refer to laboratory SOP ENV-SOP-MIN4-0165
Orientation and Training Procedures (or equivalent replacement) for more information.

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

34 of 58
 ENV-SOP-MIN4-0178, Rev 01

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Determination of Selected 36 Per- and Polyfluoroalkyl Substances (PFAS) by
LC/MS/MS
TEST METHOD: Isotope Dilution
ISSUER: Pace ENV – Minneapolis – MIN4
COPYRIGHT © 2020 Pace Analytical Services, LLC

11.4.2 For each analyte, the mean accuracy is true value ±30%. The RSD must be less than 20%.
If any target analyte fails to meet this criterion, the source of the problem must be corrected
and the test repeated
11.4.2.1 For Wisconsin samples, the mean accuracy is true value ±35%. The RSD must
be less than or equal to 30%. If any target analyte fails to meet this criterion, the
source of the problem must be corrected and the test repeated

12.0 DATA REVIEW AND CORRECTIVE ACTION

12.1 Data Review
Pace’s data review process includes a series of checks performed at different stages of the
analytical process by different people to ensure that SOPs were followed, the analytical record is
complete and properly documented, proper corrective actions were taken for QC failure and other
nonconformance(s), and that test results are reported with proper qualification.
The review steps and checks that occur as employee’s complete tasks and review their own work
is called primary review.
All data and results are also reviewed by an experienced peer or supervisor. Secondary review is
performed to verify SOPs were followed, that calibration, instrument performance, and QC criteria
were met and/or proper corrective actions were taken, qualitative ID and quantitative measurement
is accurate, all manual integrations are justified and documented in accordance with the Pace
ENV’s SOP for manual integration, calculations are correct, the analytical record is complete and
traceable, and that results are properly qualified.
A third-level review, called a completeness check, is performed by reporting or project management
staff to verify the data report is not missing information and project specifications were met.
Refer to laboratory SOP ENV-SOP-MIN4-0092 Data Review Process (or equivalent replacement)
for specific instructions and requirements for each step of the data review process.
12.2 Corrective Action
Corrective action is expected any time QC or sample results are not within acceptance criteria. If
corrective action is not taken or was not successful, the decision/outcome must be documented in
the analytical record. The primary analyst has primary responsibility for taking corrective action
when QA/QC criteria are not met. Secondary data reviewers must v erify that appropriate action
was taken and/or that results reported with QC failure are properly qualified.
Corrective action is also required when carryover is suspected and when results are over range.
Samples analyzed after a high concentration sample must be checked for carryover and reanalyzed
if carryover is suspected. Carryover is usually indicated by low concentration detects of the analyte
in successive samples analyzed after the high concentration sample.
Sample results at concentrations above the upper limit of quantitation must be diluted and
reanalyzed. The result in the diluted samples should be within the upper half of the calibration
range. Results less than the mid-range of the calibration indicate the sample was over diluted and
analysis should be repeated with a lower level of dilution. If dilution is not performed, any result

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

35 of 58
 ENV-SOP-MIN4-0178, Rev 01

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Determination of Selected 36 Per- and Polyfluoroalkyl Substances (PFAS) by
LC/MS/MS
TEST METHOD: Isotope Dilution
ISSUER: Pace ENV – Minneapolis – MIN4
COPYRIGHT © 2020 Pace Analytical Services, LLC

reported above the upper range is considered a qualitative measurement and must be qualified as
an estimated value.
Refer to Appendix B for a complete summary of QC, acceptance criteria, and recommended
corrective actions for QC associated with this test method.

13.0 POLLUTION PREVENTION AND WASTE M ANAGEMENT

Pace proactively seeks ways to minimize waste generated during our work processes. Some examples
of pollution prevention include but are not limited to: reduced solvent extraction, solvent capture, use of
reusable cycletainers for solvent management, and real-time purchasing.
The EPA requires that laboratory waste management practice to be conducted consistent with all
applicable federal and state laws and regulations. Excess reagents, samples and method process
wastes must be characterized and disposed of in an acceptable manner in accordance with Pace’s
Chemical Hygiene Plan / Safety Manual.

14.0 MODIFICATIONS
A modification is a change to a reference test method made by the laboratory. For example, changes
in stoichiometry, technology, quantitation ions, reagent or solvent volumes, reducing digestion or
extraction times, instrument runtimes, etc. are all examples of modifications. Refer to Pace ENV
corporate SOP ENV-SOP-CORQ-0011 Method Validation and Instrument Verification for the conditions
under which the procedures in test method SOPs may be modified and for the procedure and document
requirements.

15.0 RESPONSIBILITIES
Pace ENV employees that perform any part this procedure in their work activities must have a signed
Read and Acknowledgement Statement in their training file for this version of the SOP. The employee
is responsible for following the procedures in this SOP and handling temporary departures from this
SOP in accordance with Pace’s policy for temporary departure.
Pace supervisors/managers are responsible for training employees on the procedures in this SOP and
monitoring the implementation of this SOP in their work area.

16.0 ATTACHMENTS
Appendix A – Target Analyte List and Routine LOQ
Appendix B – QC Summary
Appendix C – Typical MS/MS Method Conditions
Appendix D – PFAS Analyte and Recommended Extracted Internal Standard Used for Quantification
Appendix E –PFAS by LCMSMS Compliant with DoD QSM Batch Control Limits
Appendix F – DoD QSM 5.3, Appendix B, Table B-15 - Per- and Polyfluoroalkyl Substances (PFAS)
Using Liquid Chromatography Tandem Mass Spectrometry (LC/MS/MS) With Isotope Dilution or
Internal Standard Quantification in Matrices Other Than Drinking Water

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

36 of 58
 ENV-SOP-MIN4-0178, Rev 01

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Determination of Selected 36 Per- and Polyfluoroalkyl Substances (PFAS) by
LC/MS/MS
TEST METHOD: Isotope Dilution
ISSUER: Pace ENV – Minneapolis – MIN4
COPYRIGHT © 2020 Pace Analytical Services, LLC

17.0 REFERENCES
Department of Defense Department of Energy Consolidated Quality Systems Manual (QSM) for
Environmental Laboratories, Version 5.3, Appendix B, Table B-15, June 2019.
Wisconsin Department of Natural Resources, Wisconsin PFAS Aqueous (Non-Potable Water) and Non-
Aqueous Matrices Method Expectations, EA-19-0001, December, 2019.
DoD Guidance for PFAS Analysis in Biota. April, 2020.
USEPA, Method 537.1, Version 1.0 “Determination of selected per- and polyfluorinated alkyl
substances in drinking water by solid phase extraction and liquid chromatography/tandem mass
spectrometry (LC/MS/MS)”; November 2018.
USEPA, Technical Advisory, “Technical Advisory - Laboratory Analysis of Drinking Water Samples for
Perfluorooctanoic Acid (PFOA) Using EPA Method 537 Rev. 1.1”; September 2016.
JT Baker, Application Technical Support Group, Endothall extraction using BAKERBOND Speedisk
SAX, PN-8058-06, 2006.
Pace Quality Assurance Manual- most current version.
TNI Standard, Management and Technical Requirements for Laboratories Performing Environmental
Analyses, EL-V1-2009.
TNI Standard, Management and Technical Requirements for Laboratories Performing Environmental
Analyses, EL-VI-2016-Rev.2.1.
USEPA, “Manual for the Certification of Laboratories Analyzing Drinking Water”; Fifth Edition, January
2005.
USEPA, “Supplement 1to the Fifth Edition of the Manual for the Certification of Laboratories Analyzing
Drinking Water”; June 2008.
40 CFR Appendix B to Part 136, Definition and Procedure for the Determination of the Method Detection
Limit - Rev 2, August 28, 2017.

18.0 REVISION HISTORY

This Version:
Section Description of Change
All Updated sections with "Wisconsin" reference and removed verbiage following, in regard
to "compliance" or "non-compliance samples", etc. Updated short-hand reference
throughout SOP to “DoD QSM 5.3”—reference of Table B-15 where applicable.
Header Updated test method reference.
6.0 Added back some verbiage to section 6.0 that are part of the SOP template. Deleted
6.1.1. Split 6.2 into two sections, 6.3 for receipt and storage.
Table Added “Used to Prepare” entry for “Wellington Laboratories MPFAC-6ES” rows. Updated
8.2 note below table verbiage.
10.0 Fixed/added missing numbering issues. Reworded 10.3.6.

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

37 of 58
 ENV-SOP-MIN4-0178, Rev 01

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Determination of Selected 36 Per- and Polyfluoroalkyl Substances (PFAS) by
LC/MS/MS
TEST METHOD: Isotope Dilution
ISSUER: Pace ENV – Minneapolis – MIN4
COPYRIGHT © 2020 Pace Analytical Services, LLC

16.0 Appendix A – added back verbiage from template. Appendix F – updated format and
added missing info (First 11 sections/QC Check info from DoD Table B-15 were missing).

This document supersedes the following document(s):

Document Number Title Version
ENV-MIN4-SOP-0178 Determination of Selected 36 Per- and Polyfluoroalkyl 00
Substances (PFAS) by LC/MS/MS

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

38 of 58
 ENV-SOP-MIN4-0178, Rev 01

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Determination of Selected 36 Per- and Polyfluoroalkyl Substances (PFAS) by
LC/MS/MS
TEST METHOD: Isotope Dilution
ISSUER: Pace ENV – Minneapolis – MIN4
COPYRIGHT © 2020 Pace Analytical Services, LLC

Appendix A: Target Analyte List and Routine LOQ1

Table 1: Routine Analyte List and Limits of Quantitation (LOQ)1

LOQ
Analyte Acronym(s)3 CAS# Water Solid Tissue
(ng/L) (ng/kg) (ng/kg)
Perfluorobutanoic acid PFBA 375-22-4 2 100 250
Perfluoropentanoic acid PFPeA 2706-90-3 2 100 250
HFPO-DA2
Hexafluoropropylene oxide dimer acid 13252-13-6 2 100 250
PFPrOPrA
Perfluorohexanoic acid PFHxA 307-24-4 2 100 250
Perfluoroheptanoic acid PFHpA 375-85-9 2 100 250
Perfluorooctanoic acid PFOA 335-67-1 2 100 250
Perfluorononanoic acid PFNA 375-95-1 2 100 250
PFOSAm
Perfluorooctanesulfonamide PFOSA 754-91-6 2 100 250
FOSA
MeFOSA2
N-methylperfluorooctane sulfonamide N-MeFOSA 31506-32-8 2 100 250
NMeFOSA
Perfluorodecanoic acid PFDA 335-76-2 2 100 250
EtFOSAm 2
N-ethylperfluorooctane sulfonamide N-EtFOSA 4151-50-2 2 100 250
NEtFOSA
PFUnDA
Perfluoroundecanoic acid PFUnA 2058-94-8 2 100 250
PFUdA
N-methyl NMeFOSAA
2355-31-9 2 100 250
perfluorooctanesulfonamidoacetic acid N-MeFOSAA
N-ethyl NEtFOSAA
2991-50-6 2 100 250
perfluorooctanesulfonamidoacetic acid N-EtFOSAA
PFDOA
Perfluorododecanoic acid PFDoA 307-55-1 2 100 250
PFDoDA
MeFOSE2
N-methylperfluorooctane
N-MeFOSE 24448-09-7 2 100 250
sulfonamidoethanol
NMeFOSE
EtFOSE2
N-ethylperfluorooctane
N-EtFOSE 1691-99-2 2 100 250
sulfonamidoethanol
NEtFOSE
PFTrDA
Perfluorotridecanoic acid PFTriA 72629-94-8 2 100 250
PFTrA

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

39 of 58
 ENV-SOP-MIN4-0178, Rev 01

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Determination of Selected 36 Per- and Polyfluoroalkyl Substances (PFAS) by
LC/MS/MS
TEST METHOD: Isotope Dilution
ISSUER: Pace ENV – Minneapolis – MIN4
COPYRIGHT © 2020 Pace Analytical Services, LLC

LOQ
Analyte Acronym(s)3 CAS# Water Solid Tissue
(ng/L) (ng/kg) (ng/kg)
PFTDA
PFTeDA
Perfluorotetradecanoic acid 376-06-7 2 100 250
PFTA
PFTeA
Perfluorohexadecanoic acid PFHxDA2 67905-19-5 2 100 250
2
Perfluorooctandecanoic acid PFODA 16517-11-6 2 100 250
Perfluorobutanesulfonic acid PFBS 375-73-5 1.77 88.5 221.3
Perfluoropentanesulfonic acid PFPeS 2706-91-4 1.88 94 235
Perfluorohexanesulfonic acid PFHxS 355-46-4 1.82 91 227.5
Perfluoroheptanesulfonic acid PFHpS 375-92-8 1.90 95 237.5
Perfluorooctanesulfonic acid PFOS 1763-23-1 1.85 92.5 231.3
Perfluorononanesulfonic acid PFNS 68259-12-1 1.92 96 240
Perfluorodecanesulfonic acid PFDS 335-77-3 1.93 96.5 241.3
PFDoS2
Perfluorododecanesulfonic acid 79780-39-5 1.94 97 242.5
PFDoDS
4:2 FTS
4:2 Fluorotelomer sulfonic acid 4:2 FTSA 757124-72-4 1.87 93.5 233.8
4:2FTS
6:2 FTS
6:2 Fluorotelomer sulfonic acid 6:2 FTSA 27619-97-2 1.90 95 237.5
6:2FTS
8:2 FTS
8:2 Fluorotelomer sulfonic acid 8:2 FTSA 39108-34-4 1.93 96.5 241.3
8:2FTS
10:2 FTS2
10:2 Fluorotelomer sulfonic acid 120226-60-0 1.93 96.5 241.3
10:2 FTSA
DONA2
4,8-Dioxa-3H-perfluorononanoic acid 919005-14-4 1.89 94.5 236.3
ADONA
9-Chlorohexadecafluoro-3-oxanonane- 9Cl-PF3ONS2
756426-58-1 1.86 93 232.5
1-sulfonic acid F-53B Major
11Cl-
11-Chloroeicosafluoro-3-oxaundecane-
PF3OUdS2 763051-92-9 1.88 94 235
1-sulfonic acid
F-53B Minor
1
Values in place as of effective date of this SOP. LOQs are subject to change. For the most up to date
LOQ, refer to the LIMS or contact the laboratory.
2
DoD currently does not have guidance for the analyte in Table B-15 per DoD QSM 5.3, Appendix B, as of
June 2019.
3
All possible acronym variations are listed as the acronym used and/or referenced may vary depending on
the State data is being reported to.

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

40 of 58
 ENV-SOP-MIN4-0178, Rev 01

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Determination of Selected 36 Per- and Polyfluoroalkyl Substances (PFAS) by
LC/MS/MS
TEST METHOD: Isotope Dilution
ISSUER: Pace ENV – Minneapolis – MIN4
COPYRIGHT © 2020 Pace Analytical Services, LLC

Appendix B: QC Summary

Acceptance Criteria
QC Item Frequency Wisconsin Table B-15 per Corrective Action Qualification
Guidance DoD QSM 5.3
ICAL At instrument Lowest ICAL ± All points ±30% Identify and correct source None.
set up, after 50% of true value. of problem, repeat Do not
CCV failure Other points ± proceed with
30% For any curve fit analysis
other than
For any curve fit Average RF
other than (RSD), curve
Average RF must also pass
(RSD), curve RSE test at the
must also pass low and
RSE test at the midpoint
low and midpoint calibration
calibration standard.
standard.
Curve Whenever Must meet one of Must meet one Identify and correct source None. Do not
Refitting ICAL is curve fit options of curve fit of problem, repeat proceed with
performed presented in options analysis
Section 10.0. presented in
Section 10.0.
ICV 1 after each True value ± 30% True value ± Identify source of problem, Qualify
ICAL 30% re-analyze. If repeat analytes with
failure, repeat ICAL. ICV out of
Analysis may proceed if it criteria.
can be demonstrated that
the ICV exceedance has no
impact on analytical
measurements. For
example, the ICV %R is
high, CCV is within criteria,
and the analyte is not
detected in sample(s).
RT Once per Position is set Position is set NA NA
Window ICAL and at using the mid- using the mid-
Position the beginning point of the ICAL point of the
(Daily) of the on the day ICAL is ICAL on the day
analytical performed; ICAL is
window. otherwise mid- performed;
point of CCV is otherwise mid-
used point of CCV is
used
RT At method Window is ±0.2 Window is ±0.2 Correct problem and NA
Window set-up and minutes the daily minutes the reanalyze samples
Study after major CCV or, on days daily CCV or, on
instrument when ICAL is days when ICAL
maintenance performed, from is performed,

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

41 of 58
 ENV-SOP-MIN4-0178, Rev 01

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Determination of Selected 36 Per- and Polyfluoroalkyl Substances (PFAS) by
LC/MS/MS
TEST METHOD: Isotope Dilution
ISSUER: Pace ENV – Minneapolis – MIN4
COPYRIGHT © 2020 Pace Analytical Services, LLC

Acceptance Criteria
QC Item Frequency Wisconsin Table B-15 per Corrective Action Qualification
Guidance DoD QSM 5.3
the midpoint from the
standard of the midpoint
ICAL. standard of the
Analytes must ICAL.
elute within 0.1 Analytes must
minute of the elute within 0.1
associated EIS minute of the
associated EIS
Continuing Daily, before CCV at low level: True value Perform necessary Qualify
Calibration sample True value ±50% ±30% maintenance and analytes with
Curve analysis, after Other CCV: True demonstrate stability by CCV out of
(CCV) every 10 field value ±30% analyzing an initial criteria.
samples, and calibration before resuming
at end of sample analysis. Samples
analytical between passing CCV and
window. failing CCV should be re-
analyzed.
Extracted Every field Must meet criteria Must meet If the CCV fails high for a Qualify
Internal sample, specified in criteria specified particular analyte, and the outages and
Standards standard and Section 9.2.8 in Section 9.2.8 field sample is non-detect explain in case
(EIS) QC sample for that analyte, samples narrative.
may be reported without re-
analysis.
Injection Every field Must meet criteria Must meet Troubleshoot instrument Qualify
Internal sample, specified in criteria specified performance. Reanalyze outages and
Standards standard and Section 9.2.9 in Section 9.2.9 samples. explain in case
(IIS) QC sample narrative.
Method 1 per batch Analytes <1/2 the Analytes <1/2 1) If sample ND, report Qualify
Blank RL or 1/10th the the RL or 1/10th sample without outages and
(MB) amount measured the amount qualification. explain in case
in any sample measured in 2) If sample result >10x MB narrative
any sample or detects and sample cannot
1/10th the be reanalyzed, report
regulatory limit, sample with appropriate
whichever is qualifier indicating blank
greater contamination.
3) If sample result <10x
MB detects, report sample
with appropriate qualifier to
indicate an estimated
value. Client must be
alerted to give authorization
to report this data.
4) Analyte detection or
failure of internal standard
fails entire batch.

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

42 of 58
 ENV-SOP-MIN4-0178, Rev 01

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Determination of Selected 36 Per- and Polyfluoroalkyl Substances (PFAS) by
LC/MS/MS
TEST METHOD: Isotope Dilution
ISSUER: Pace ENV – Minneapolis – MIN4
COPYRIGHT © 2020 Pace Analytical Services, LLC

Acceptance Criteria
QC Item Frequency Wisconsin Table B-15 per Corrective Action Qualification
Guidance DoD QSM 5.3
LCS/LCSD 1 spiked at a Reanalyze and/or re- Qualify
concentration True Value ≤ 1-2x See Appendix E prepare batch of samples outages and
≥ LOQ and ≤ RL with new LCS. explain in case
the mid-level 50-150% RPD ≤ 30% If LCS rec > QC limits and narrative
calibration True Value > 2x these compounds are non-
concentration RL detect in the associated
60-135% samples, the sample data
may be reported with
appropriate data qualifiers.
RPD ≤ 30%
If these criteria are not met,
where extra samples are
available, a re-extract is
analyzed or else data is
narrated.
MS/MSD 1 pair/batch True Value ≤ 1-2x See Appendix E Failures are flagged but do Qualify
spiked at a RL 50-150% not prevent reporting data if outages and
concentration True Value > 2x RPD ≤ 30% MB and LCS meet criteria. explain in case
≥ LOQ and ≤ RL 60-135% narrative
the mid-level
calibration RPD ≤ 30%
concentration
Field 1 per batch. NA NA If these criteria are not met, Qualify
Duplicate A MSD may results are labeled suspect outages and
be due to matrix effects explain in case
(FD)
substituted narrative
for a sample
duplicate if
sample is
insufficient
Tune Every six See section 9.2.1 See section Refer to manufacture NA
Standard month, when for reference 9.2.1 for criteria
ICAL won't reference
pass, when
the peak
shape is
significantly
off (indicating
an MS
problem),
when major
maintenance
is performed,
or instrument
is moved
Instrument 1 following < ½ RL < ½ RL If acceptance criteria are Flagging is
Blank the highest not met after the highest only
(ICB) standard calibration standard, appropriate in
analyzed calibration must be cases when

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

43 of 58
 ENV-SOP-MIN4-0178, Rev 01

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Determination of Selected 36 Per- and Polyfluoroalkyl Substances (PFAS) by
LC/MS/MS
TEST METHOD: Isotope Dilution
ISSUER: Pace ENV – Minneapolis – MIN4
COPYRIGHT © 2020 Pace Analytical Services, LLC

Acceptance Criteria
QC Item Frequency Wisconsin Table B-15 per Corrective Action Qualification
Guidance DoD QSM 5.3
performed using a lower the sample
concentration for the cannot be
highest standard until reanalyzed
acceptance criteria is met. and when
there is no
more sample
left.
CCB 1 following < ½ RL NA If acceptance criteria are Flagging is
the CCV and not met after the CCV. only
prior to Clean the system and appropriate in
sample prepare new CCV if cases when
analysis needed. the sample
cannot be
reanalyzed
and when
there is no
more sample
left.

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

44 of 58
 ENV-SOP-MIN4-0178, Rev 01

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Determination of Selected 36 Per- and Polyfluoroalkyl Substances (PFAS) by
LC/MS/MS
TEST METHOD: Isotope Dilution
ISSUER: Pace ENV – Minneapolis – MIN4
COPYRIGHT © 2020 Pace Analytical Services, LLC

Appendix C: Typical MS/MS Method Conditions

Precursor Ion Product Ion RT Declustering Collision Collision Cell
Analyte
(m/z) (m/z) (min) Potential (v) Energy (v) Exit Potential (v)
PFBA 212.9 169 2.43 -45 -12 -11
PFPeA 262.9 219 3.33 -25 -12 -11
HFPO-DA 285 169 4.3 -70 -12 -11
HFPO-DA_2 285 185 4.29 -70 -24 -13
PFBS 298.9 80 4.27 -65 -58 -9
PFBS_2 298.9 99 4.27 -65 -40 -9
PFHxA 313 269 4.03 -25 -12 -19
PFHxA_2 313 119 4.02 -25 -28 -19
4:2FTS 327 307 3.75 -65 -28 -13
4:2FTS_2 327 81 3.75 -65 -56 -11
PFPeS 349 80 4.94 -45 -68 -9
PFPeS_2 349 99 4.94 -65 -40 -9
PFHpA 363 319 4.63 -50 -14 -15
PFHpA_2 363 169 4.63 -50 -24 -11
DONA 377 251 4.84 -50 -16 -11
DONA_2 377 85 4.84 -50 -36 -11
PFHxS 399 80 5.52 -55 -84 -9
PFHxS_2 399 99 5.52 -55 -68 -11
PFOA 413 369 5.17 -55 -14 -17
PFOA_2 413 169 5.17 -55 -24 -9
6:2FTS 427 407 4.89 -65 -32 -17
6:2FTS_2 427 81 4.89 -65 -68 -7
PFHpS 449 80 6.07 -105 -92 -9
PFHpS_2 449 99 6.07 -80 -80 -13
PFNA 463 419 5.7 -70 -16 -15
PFNA_2 463 169 5.7 -70 -26 -11
PFOSA 498 78 7.47 -130 -90 -11
PFOS 499 80 6.58 -65 -112 -9
PFOS_2 499 99 6.58 -65 -90 -11
N-MeFOSA 512 169 8.76 -55 -36 -11
N-MeFOSA_2 512 218.9 8.75 -60 -34 -19
PFDA 513 469 6.21 -80 -16 -19
PFDA_2 513 169 6.21 -80 -28 -13
N-EtFOSA 526 169 9.23 -40 -36 -13
N-EtFOSA_2 526 219.15 9.23 -15 -34 -9

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

45 of 58
 ENV-SOP-MIN4-0178, Rev 01

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Determination of Selected 36 Per- and Polyfluoroalkyl Substances (PFAS) by
LC/MS/MS
TEST METHOD: Isotope Dilution
ISSUER: Pace ENV – Minneapolis – MIN4
COPYRIGHT © 2020 Pace Analytical Services, LLC

Precursor Ion Product Ion RT Declustering Collision Collision Cell

Analyte
(m/z) (m/z) (min) Potential (v) Energy (v) Exit Potential (v)
8:2FTS 527 507 5.92 -70 -38 -21
8:2FTS_2 527 81 5.92 -70 -92 -9
9Cl-PF3ONS 530.9 351 6.97 -75 -36 -15
9Cl-PF3ONS_2 530.9 83 6.97 -75 -70 -11
PFNS 549 80 7.08 -65 -118 -13
PFNS_2 549 99 7.08 -85 -96 -11
PFUdA 563 519 6.72 -30 -18 -21
PFUdA_2 563 169 6.71 -30 -32 -11
N-MeFOSAA 570 419 6.11 -125 -28 -33
N-MeFOSAA_2 570 483 6.11 -125 -22 -33
N-EtFOSAA 584 419 6.34 -125 -28 -33
N-EtFOSAA_2 584 526 6.33 -125 -28 -33
PFDS 599 80 7.57 -85 -122 -11
PFDS_2 599 99 7.57 -85 -100 -11
PFDoA 613 569 7.21 -25 -18 -23
PFDoA_2 613 169 7.21 -25 -34 -11
N-MeFOSE 616 59 8.58 -20 -76 -5
10:2FTS 627 607 6.92 -50 -44 -25
10:2FTS_2 627 81 6.91 -50 -108 -9
N-EtFOSE 630 59 9.03 -20 -58 -27
11Cl-PF3OUdS 630.9 451 7.94 -90 -40 -19
11Cl-PF3OUdS_2 630.9 99 7.94 -90 -92 -5
PFTrDA 663 619 7.69 -75 -20 -25
PFTrDA_2 663 169 7.69 -75 -34 -9
PFDoS 699 80 8.46 -30 -134 -9
PFDoS_2 699 99 8.46 -20 -132 -11
PFTeDA 713 669 8.17 -85 -20 -27
PFTeDA_2 713 169 8.17 -85 -36 -13
PFHxDA 813 769 9.23 -30 -22 -33
PFHxDA_2 813 169 9.23 -30 -38 -9
PFODA 913 869 9.72 -5 -22 -35
PFODA_2 913 169 9.72 -5 -42 -11
Note(s):
Analyte_2 Ions used for confirmation purposes.

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

46 of 58
 ENV-SOP-MIN4-0178, Rev 01

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Determination of Selected 36 Per- and Polyfluoroalkyl Substances (PFAS) by
LC/MS/MS
TEST METHOD: Isotope Dilution
ISSUER: Pace ENV – Minneapolis – MIN4
COPYRIGHT © 2020 Pace Analytical Services, LLC

Appendix D: PFAS Analyte and Recommended Extracted Internal Standard Used

for Quantification
Analyte EIS Name
PFBA 13C4_PFBA
PFPeA 13C5_PFPeA
HFPO-DA 13C3_HFPO-DA
PFBS 13C3_PFBS
PFHxA 13C5_PFHxA
4:2FTS 13C2_4:2FTS
PFPeS 13C3_PFHxS
PFHpA 13C4_PFHpA
DONA 13C8_PFOA
PFHxS 13C3_PFHxS
PFOA 13C8_PFOA
6:2FTS 13C2_6:2FTS
PFHpS 13C3_PFOS
PFNA 13C9_PFNA
PFOSA 13C8_PFOSA
PFOS 13C8_PFOS
N-MeFOSA d3-N-MeFOSA
PFDA 13C6_PFDA
N-EtFOSA d5-N-EtFOSA
8:2FTS 13C2_8:2FTS
9Cl-PF3ONS 13C8_PFOS
PFNS 13C8_PFOS
PFUdA 13C7_PFUdA
N-MeFOSAA d3-MeFOSAA
N-EtFOSAA d5-EtFOSAA
PFDS 13C8_PFOS
PFDoA 13C2_PFDoA
N-MeFOSE d7-N-MeFOSE
10:2FTS 13C2_8:2FTS
N-EtFOSE d9-N-EtFOSE
11Cl-PF3OUdS 13C8_PFOS
PFTrDA 13C2_PFDoA
PFDoS 13C8_PFOS
PFTeDA 13C2_PFTeDA
PFHxDA 13C2_PFHxDA
PFODA 13C2_PFHxDA

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
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47 of 58
 ENV-SOP-MIN4-0178, Rev 01

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Determination of Selected 36 Per- and Polyfluoroalkyl Substances (PFAS) by
LC/MS/MS
TEST METHOD: Isotope Dilution
ISSUER: Pace ENV – Minneapolis – MIN4
COPYRIGHT © 2020 Pace Analytical Services, LLC

Appendix E: PFAS by LCMSMS Compliant with DoD QSM Batch Control Limits
Aqueous Tissue
Solid Matrix
Matrix Matrix3
Analyte Lower Upper Lower Upper Lower Upper
CAS#
Acronym Control Control Control Control Control Control
Limit Limit Limit Limit Limit Limit
(%REC) (%REC) (%REC) (%REC) (%REC) (%REC)
2991-50-6 N-EtFOSAA 61 135 61 139 60 140
2355-31-9 N-MeFOSAA 65 136 63 144 60 140
757124-72-4 4:2 FTS 63 143 62 145 60 140
27619-97-2 6:2 FTS 64 140 64 140 60 140
39108-34-4 8:2 FTS 67 138 65 137 60 140
375-73-5 PFBS 72 130 72 128 60 140
375-22-4 PFBA 73 129 71 135 60 140
335-77-3 PFDS 53 142 59 134 60 140
335-76-2 PFDA 71 129 69 133 60 140
307-55-1 PFDoA 72 134 69 135 60 140
375-92-8 PFHpS 69 134 70 132 60 140
375-85-9 PFHpA 72 130 71 131 60 140
355-46-4 PFHxS 68 131 67 130 60 140
307-24-4 PFHxA 72 129 70 132 60 140
68259-12-1 PFNS 69 127 69 125 60 140
375-95-1 PFNA 69 130 72 129 60 140
754-91-6 PFOSA 67 137 67 137 60 140
1763-23-1 PFOS 65 140 68 136 60 140
335-67-1 PFOA 71 133 69 133 60 140
2706-91-4 PFPeS 71 127 73 123 60 140
2706-90-3 PFPeA 72 129 69 132 60 140
376-06-7 PFTeDA 71 132 69 133 60 140
72629-94-8 PFTrDA 65 144 66 139 60 140
2058-94-8 PFUdA 69 133 64 136 60 140
31506-32-8 N-MeFOSA1 68 141 70 140 60 140
4151-50-2 N-EtFOSA2 70 140 70 140 60 140
120226-60-0 10:2FTS2 70 140 70 140 60 140
13252-13-6 HFPO-DA2 70 140 70 140 NA NA
919005-14-4 DONA1 70 140 70 140 60 140
756426-58-1 9Cl-PF3ONS2 70 140 70 140 60 140
763051-92-9 11Cl-PF3OUDS2 70 140 70 140 60 140

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
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48 of 58
 ENV-SOP-MIN4-0178, Rev 01

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Determination of Selected 36 Per- and Polyfluoroalkyl Substances (PFAS) by
LC/MS/MS
TEST METHOD: Isotope Dilution
ISSUER: Pace ENV – Minneapolis – MIN4
COPYRIGHT © 2020 Pace Analytical Services, LLC

Aqueous Tissue
Solid Matrix
Matrix Matrix3
Analyte Lower Upper Lower Upper Lower Upper
CAS#
Acronym Control Control Control Control Control Control
Limit Limit Limit Limit Limit Limit
(%REC) (%REC) (%REC) (%REC) (%REC) (%REC)
24448-09-7 N-MeFOSE2 70 140 70 140 60 140
1691-99-2 N-EtFOSE2 70 140 70 140 60 140
67905-19-5 PFHxDA2 70 140 70 140 60 140
16517-11-6 PFODA2 70 140 70 140 60 140
79780-39-5 PFDoS2 70 140 70 140 60 140

1
DoD currently does not have guidance for the analyte in solid matrix in Table B-15 per DoD QSM 5.3,
Appendix B, as of June 2019.
2
DoD currently does not have guidance for the analyte in both aqueous and solid matrix in Table B-15 per
DoD QSM 5.3, Appendix B, as of June 2019.
3
DoD currently does not have guidance for the analyte in tissue matrix in Table B-15 per DoD QSM 5.3,
Appendix B, as of June 2019.

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

49 of 58
 ENV-SOP-MIN4-0178, Rev 01

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Determination of Selected 36 Per- and Polyfluoroalkyl Substances (PFAS) by
LC/MS/MS
TEST METHOD: Isotope Dilution
ISSUER: Pace ENV – Minneapolis – MIN4
COPYRIGHT © 2020 Pace Analytical Services, LLC

Appendix F: DoD QSM 5.3, Appendix B, Table B-15 - Per- and Polyfluoroalkyl
Substances (PFAS) Using Liquid Chromatography Tandem Mass Spectrometry
(LC/MS/MS) With Isotope Dilution or Internal Standard Quantification in Matrices
Other Than Drinking Water

Minimum Flagging
QC Check Acceptance Criteria Corrective Action Comments
Frequency Criteria
Aqueous Sample Each sample Solid Phase Extraction NA. NA. Samples with >1%
Preparation and associated (SPE) must be used solids may require
batch QC unless samples are centrifugation prior to
samples. known to contain high SPE extraction.
PFAS concentrations Pre-screening of
(e.g., Aqueous Film separate aliquots of
Forming Foam (AFFF) aqueous samples is
formulations). Inline SPE recommended.
is acceptable.

Entire sample plus bottle

rinsate must be extracted
using SPE.

Known high PFAS

concentration samples
require serial dilution be
performed in duplicate.

Documented project
approval is needed for
samples prepared by
serial dilution as opposed
to SPE.
Solid Sample Each sample Entire sample received NA. NA. NA.
Preparation and associated by the laboratory must be
batch QC homogenized prior to
samples. subsampling.
Biota Sample Each sample Sample prepared as L NA. NA.
Preparation and associated defined by the project
batch QC (e.g., whole fish versus
samples. filleted fish).
AFFF and AFFF Each sample Each field sample must NA. NA. Adsorption onto bottle
Mixture Samples and associated be prepared in duplicate is negligible
Preparation batch QC (equivalent to matrix compared to sample
samples. duplicate). concentration so
subsampling is
Serial dilutions must be allowed.
performed to achieve the
lowest LOQ possible for Multiple dilutions will
each analyte. most likely have to be
reported in order to
achieve the lowest

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

50 of 58
 ENV-SOP-MIN4-0178, Rev 01

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Determination of Selected 36 Per- and Polyfluoroalkyl Substances (PFAS) by
LC/MS/MS
TEST METHOD: Isotope Dilution
ISSUER: Pace ENV – Minneapolis – MIN4
COPYRIGHT © 2020 Pace Analytical Services, LLC

Minimum Flagging
QC Check Acceptance Criteria Corrective Action Comments
Frequency Criteria
LOQ possible for
each analyte.
Sample Cleanup Each sample ENVI-CarbTM or NA. Flagging is not Cleanup should
Procedure and associated equivalent must be used appropriate. reduce bias from
batch QC on each sample and matrix
samples. batch QC sample. interferences.

Not applicable to
AFFF and AFFF
Mixture
Samples.
Mass Calibration Instrument must Calibrate the mass scale If the mass calibration Flagging is not Problem must be
have a valid of the MS with calibration fails, then recalibrate. appropriate. corrected. No
mass calibration compounds and If it fails again, consult samples may be
prior to any procedures described by manufacturer analyzed under a
sample analysis. the manufacturer. instructions on failing mass
corrective calibration.
Mass calibration Mass calibration range maintenance.
is verified after must bracket the ion The mass calibration
each mass masses of interest. The is updated on an as-
calibration, prior most recent mass needed basis (e.g.,
calibration must be used QC failures, ion
to initial
for every acquisition in an masses fall outside of
calibration analytical run. the ±0.5 amu of the
(ICAL). true value, major
Mass calibration must be instrument
verified to be ±0.5 amu of maintenance is
the true value, by performed, or the
acquiring a full scan instrument is moved).
continuum mass
spectrum of a PFAS
stock standard.
Mass Spectral Each analyte, A minimum of 10 spectra NA. Flagging is not NA.
Acquisition Rate Extracted scans are acquired appropriate.
Internal across each
Standard (EIS) chromatographic peak.
Analyte.
Calibration, All analytes. Standards containing NA. Flagging is not Standards containing
Calibration both branched and linear appropriate. both branched and
Verification, and isomers must be used linear isomers are to
Spiking when commercially be used during
Standards available. method validation and
when reestablishing
PFAS method analytes retention times, to
may consist of both ensure the total
branched and linear response is
isomers, but quantitative quantitated for that
standards that contain
analyte.
the linear and branched
isomers do not exist for
all method analytes. Technical grade

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released or the SOP is made obsolete.
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51 of 58
 ENV-SOP-MIN4-0178, Rev 01

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Determination of Selected 36 Per- and Polyfluoroalkyl Substances (PFAS) by
LC/MS/MS
TEST METHOD: Isotope Dilution
ISSUER: Pace ENV – Minneapolis – MIN4
COPYRIGHT © 2020 Pace Analytical Services, LLC

Minimum Flagging
QC Check Acceptance Criteria Corrective Action Comments
Frequency Criteria
standards cannot be
For PFAS that do not used for quantitative
have a quantitative analysis.
branched and linear
standard, identify the
branched isomers by
analyzing a qualitative
standard that includes
both linear and branched
isomers and determine
retention times,
transitions and transition
ion ratios. Quantitate
samples by integrating
the total response (i.e.,
accounting for peaks that
are identified as linear
and branched isomers)
and relying on the initial
calibration that uses the
linear isomer quantitative
standard.
Sample PFAS All analytes The chemical derivation NA. PFAS identified, For example: Ion
Identification detected in a of the ion transitions with Ion ratios Ratio = (quant ion
sample. must be documented. A that fail abundance/confirm
minimum of two ion acceptance ion abundance)
transitions (Precursor → criteria, must be
quant ion and precursor flagged. Calculate the average
→ confirmation ion) and ratio (A) and standard
the ion transitions ratio Any quantitation deviation (SD) using
per analyte are required ion peak that the ICAL standards.
for confirmation. does not meet
Exception is made for An acceptance range
the maximization of ratio could be
analytes where two criteria shall be
transitions do not exist within A ±3SD for
included in the
(PFBA and PFPeA). confirmation of
summed
integration and detection.
Documentation of the the resulting
primary and confirmation data flagged as
transitions and the ion “estimated,
ratio is required. biased high”.

In-house acceptance
criteria for evaluation of
ion ratios must be used
and must not exceed 50-
150%.

Signal to Noise Ratio

(S/N) must be ≥ 10 for all
ions used for
quantification and must

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Uncontrolled copies are not tracked or replaced when new versions are released or the SOP is made obsolete.
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52 of 58
 ENV-SOP-MIN4-0178, Rev 01

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Determination of Selected 36 Per- and Polyfluoroalkyl Substances (PFAS) by
LC/MS/MS
TEST METHOD: Isotope Dilution
ISSUER: Pace ENV – Minneapolis – MIN4
COPYRIGHT © 2020 Pace Analytical Services, LLC

Minimum Flagging
QC Check Acceptance Criteria Corrective Action Comments
Frequency Criteria
be ≥ 3 for all ions used
for confirmation.

Quant ion and

confirmation ion must be
present and must
maximize simultaneously
(±2 seconds).
Ion Transitions Every field In order to avoid biasing NA. Flagging is not NA.
(Precursor-> sample, results high due to known appropriate
Product) standard, blank, interferences for some
and QC sample. transitions, the following
transitions must be used
for the quantification of
the following analytes:

PFOA: 413 → 369

PFOS: 499 → 80
PFHxS: 399 → 80
PFBS: 299 → 80
4:2 FTS: 327 → 307
6:2 FTS: 427 → 407
8:2 FTS: 527 → 507
NEtFOSAA: 584 → 419
NMeFOSAA: 570 → 419

If these transitions are

not used, the reason
must be technically
justified and documented
(e.g., alternate transition
was used due to
observed interferences).
Initial Calibration At instrument The isotopically labeled Correct problem, then Flagging is not No samples shall be
(ICAL) set-up and after analog of an analyte repeat ICAL. appropriate. analyzed until ICAL
ICV or CCV (Extracted Internal has passed.
failure, prior to Standard Analyte) must
sample analysis. be used for quantitation if External Calibration is
commercially available not allowed for any
(Isotope Dilution analyte.
Quantitation).
Calibration can be
Commercial PFAS linear (minimum of 5
standards available as standards) or
salts, are acceptable, quadratic (minimum of
providing the measured 6 standards);
mass is corrected to the weighting is allowed.
neutral acid
concentration. Results
shall be reported as the
neutral acid with

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

53 of 58
 ENV-SOP-MIN4-0178, Rev 01

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Determination of Selected 36 Per- and Polyfluoroalkyl Substances (PFAS) by
LC/MS/MS
TEST METHOD: Isotope Dilution
ISSUER: Pace ENV – Minneapolis – MIN4
COPYRIGHT © 2020 Pace Analytical Services, LLC

Minimum Flagging
QC Check Acceptance Criteria Corrective Action Comments
Frequency Criteria
appropriate CAS number.

If a labeled analog is not

commercially available,
the Extracted Internal
Standard Analyte with
the closest retention time
or chemical similarity to
the analyte must be used
for quantitation. (Internal
Standard Quantitation)

Analytes must be within

70-130% of their true
value for each calibration
standard.

ICAL must meet one of

the two options below:

Option 1: The RSD of the

RFs for all analytes must
be ≤20%.

Option 2: Linear or non-

linear calibrations must
have r2 ≥ 0.99 for each
analyte.
Retention Time Once per ICAL Position shall be set NA. NA. Calculated for each
window position and at the using the midpoint analyte and EIS.
establishment beginning of the standard of the ICAL
analytical curve when ICAL is
sequence. performed. On days
when ICAL is not
performed, the initial
CCV is used.
Retention Time Every field RT of each analyte and Correct problem and NA. Calculated for each
(RT) window sample, EIS analyte must fall reanalyze samples. analyte and EIS.
width standard, blank, within 0.4 minutes of the
and QC sample. predicted retention times
from the daily calibration
verification or on days
when ICAL is performed,
from the midpoint
standard of the ICAL.

Analytes must elute

within 0.1 minutes of the
associated EIS. This
criterion applies only to
analyte and labeled
analog pairs.

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

54 of 58
 ENV-SOP-MIN4-0178, Rev 01

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Determination of Selected 36 Per- and Polyfluoroalkyl Substances (PFAS) by
LC/MS/MS
TEST METHOD: Isotope Dilution
ISSUER: Pace ENV – Minneapolis – MIN4
COPYRIGHT © 2020 Pace Analytical Services, LLC

Minimum Flagging
QC Check Acceptance Criteria Corrective Action Comments
Frequency Criteria
Instrument Prior to analysis Analyte concentrations Correct problem, Flagging is not No samples shall be
Sensitivity Check and at least once must be at LOQ; rerun ISC. If problem appropriate. analyzed until ISC
(ISC) every 12 hours. concentrations must be persists, repeat ICAL. has met acceptance
within ±30% of their true criteria.
values.
ISC can serve as the
initial daily CCV.
Initial Calibration Once after each Analyte concentrations Correct problem, Flagging is not No samples shall be
Verification (ICV) ICAL, analysis of must be within ±30% of rerun ICV. If problem appropriate. analyzed until
a second source their true value. persists, repeat ICAL. calibration has been
standard prior to verified.
sample analysis.
Continuing Prior to sample Concentration of analytes Immediately analyze If reanalysis Results may not be
Calibration analysis, after must range from the LOQ two additional cannot be reported without valid
Verification every 10 field to the mid-level consecutive performed, data CCVs.
(CCV) samples, and at calibration concentration. CCVs. If both pass, must be qualified
the end of the samples may be and explained in Instrument Sensitivity
analytical Analyte concentrations reported without the Case Check (ISC) can
sequence. must be within ±30% of reanalysis. If either Narrative. serve as a bracketing
fails, or if two
their true value. CCV.
consecutive CCVs Apply Q-flag to
cannot be run, all results for the
perform corrective specific
action(s) and repeat analyte(s) in all
CCV and all samples since
associated samples the last
since last successful acceptable
calibration
CCV.
verification.

Alternately,
recalibrate if
necessary; then
reanalyze all
associated samples
since the last
acceptable CCV.
Instrument Immediately Concentration of each If acceptance criteria Flagging is only No samples shall be
Blanks following the analyte must be ≤ ½ the are not met after the appropriate in analyzed until
highest standard LOQ. highest calibration cases when instrument
analyzed and standard, calibration the sample blank has met
daily prior to Instrument Blank must must be performed cannot be acceptance criteria.
sample analysis. contain EIS to enable using a lower reanalyzed and
quantitation of concentration for the when there is no Note: Successful
contamination. highest standard until more sample analysis following the
acceptance criteria is left. highest standard
met. analyzed determines
the highest
concentration that
If sample carryover does not
concentrations occur.
exceed the highest

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

55 of 58
 ENV-SOP-MIN4-0178, Rev 01

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Determination of Selected 36 Per- and Polyfluoroalkyl Substances (PFAS) by
LC/MS/MS
TEST METHOD: Isotope Dilution
ISSUER: Pace ENV – Minneapolis – MIN4
COPYRIGHT © 2020 Pace Analytical Services, LLC

Minimum Flagging
QC Check Acceptance Criteria Corrective Action Comments
Frequency Criteria
allowed standard and When the highest
the sample(s) standard analyzed is
following exceed this not part of the
calibration curve, it
acceptance criteria
cannot be used to
(>1/2 LOQ), they extend out the
must be reanalyzed. calibration range, it is
used only to
document a higher
concentration at
which carry over still
does not occur.
Extracted Every field Added to solid sample Correct problem. If Apply Q-flag and Failing analytes shall
Internal sample, prior to extraction. Added required, re-extract discuss in the be thoroughly
Standard (EIS) standard, blank, to aqueous samples, into and reanalyze Case Narrative documented in
Analytes and QC the original container, associated field and only if reanalysis the Case Narrative.
sample. prior to extraction. QC samples. confirms failures
in exactly the EIS should be 96%
For aqueous samples If recoveries are same manner. (or greater) purity.
prepared by serial acceptable for QC When the impurity
dilution instead of SPE, samples, but not field
consists of the
added to final dilution of samples, the field
samples must be re- unlabeled analyte, the
samples prior to analysis.
extracted and EIS can result in a
analyzed (greater background artifact in
Extracted Internal
dilution may be every sample,
Standard Analyte
needed). standard and blank, if
recoveries must be within
50% to 150% of ICAL the EIS is fortified at
Samples may be re- excessive
midpoint standard area
extracted and concentrations.
or area measured in the
analyzed outside of
initial CCV on days when
hold times, as
an ICAL is not
necessary for
performed.
corrective action
associated with QC
failure.
Method Blank One per No analytes detected > Correct problem. If If reanalysis Results may not be
(MB) preparatory required, re-extract cannot be reported without a
batch. ½ LOQ or > 1/10th the and reanalyze MB performed, data valid MB.
amount measured in any and all QC samples must be
sample or 1/10th the and field samples qualified and Flagging is only
regulatory limit, processed with the explained in the appropriate in cases
whichever is greater. contaminated blank. Case Narrative. where the samples
cannot be reanalyzed.
Samples may be re- Apply B-flag to
extracted and all results for the
analyzed outside of
specific
hold times, as
necessary for analyte(s) in all
corrective action samples in the
associated with QC associated
failure. preparatory
batch.

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

56 of 58
 ENV-SOP-MIN4-0178, Rev 01

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Determination of Selected 36 Per- and Polyfluoroalkyl Substances (PFAS) by
LC/MS/MS
TEST METHOD: Isotope Dilution
ISSUER: Pace ENV – Minneapolis – MIN4
COPYRIGHT © 2020 Pace Analytical Services, LLC

Minimum Flagging
QC Check Acceptance Criteria Corrective Action Comments
Frequency Criteria
Examine the project-
specific requirements.
Contact the client as
to additional
measures to be
taken.
Laboratory One per Blank spiked with all Correct problem, then If reanalysis Results may not be
Control preparatory analytes at a re- extract and cannot be reported without a
Sample (LCS) batch. concentration ≥ LOQ and reanalyze the performed, data valid LCS.
≤ the mid-level calibration LCS and all samples must be
concentration. in the associated qualified and Flagging is only
preparatory explained in the appropriate in cases
A laboratory must use batch for failed Case Narrative. where the samples
the DoD/DOE QSM analytes if sufficient cannot be reanalyzed.
Appendix C Limits for sample material Apply Q-flag to
batch control if project is available. specific
limits are not specified. If
analyte(s) in all
the analyte(s) are not
Samples may be re- samples in the
listed, use in-house LCS
extracted and associated
limits if project limits are analyzed outside of preparatory
not specified. hold times, as batch.
necessary for
corrective action
associated with QC
failure.

Examine the project-

specific requirements.
Contact the client as
to additional
measures to be taken.
Matrix Spike One per Sample spiked with all Examine the project- For the specific For matrix evaluation
(MS) preparatory analytes at a specific requirements. analyte(s) only. If MS results are
batch. concentration ≥ LOQ and Contact the client as in the parent outside the limits, the
≤ the mid-level calibration to additional sample, apply data shall be
Not required for concentration. measures to be J-flag if evaluated to
aqueous taken. acceptance determine the
samples A laboratory must use criteria are not source(s) of
prepared by the DoD/DOE QSM met and explain difference (i.e., matrix
serial dilution Appendix C Limits for in the Case effect or analytical
instead of SPE. batch control if project Narrative. error).
limits are not specified.

If the analyte(s) are not

listed, use in-house LCS
limits if project limits are
not specified.
Matrix Spike For MSD: One For MSD: Sample spiked Examine the project- For the specific The data shall be
Duplicate (MSD) per preparatory with all analytes at a specific requirements. analyte(s) evaluated to
or Matrix batch. concentration ≥ LOQ and determine
Duplicate

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

57 of 58
 ENV-SOP-MIN4-0178, Rev 01

TEST METHOD STANDARD OPERATING PROCEDURE

TITLE: Determination of Selected 36 Per- and Polyfluoroalkyl Substances (PFAS) by
LC/MS/MS
TEST METHOD: Isotope Dilution
ISSUER: Pace ENV – Minneapolis – MIN4
COPYRIGHT © 2020 Pace Analytical Services, LLC

Minimum Flagging
QC Check Acceptance Criteria Corrective Action Comments
Frequency Criteria
(MD) ≤ the mid-level Contact the client as in the parent the source of
For MD: Each calibration concentration. to additional sample, apply difference.
aqueous sample measures to be J-flag if
prepared by A laboratory must use taken. acceptance For Sample/MD: RPD
serial dilution the DoD/DOE QSM If the analyte(s) are criteria are not criteria only apply to
instead of SPE. Appendix C Limits for not listed, use in- met and explain analytes whose
batch control if project in the Case concentration in the
house LCS limits if
limits are not specified. sample is ≥LOQ.
project limits are not Narrative.
specified.
If the analyte(s) are not The MD is a second
listed, use in-house LCS aliquot of the field
limits if project limits are sample that has been
not specified. prepared by serial
dilution.
RPD ≤ 30% (between
MS and MSD or sample
and MD).
Post Spike Only applies to Spike all analytes When analyte Flagging is not When analyte
Sample aqueous reported as <LOQ into concentrations are appropriate. concentrations are
samples the dilution that the result calculated as <LOQ, calculated as <LOQ,
prepared by for that analyte is and the spike results may not be
serial dilution reported from. The spike recovery does not reported without
instead of SPE must be at the LOQ meet the acceptance acceptable post spike
that have concentration to be criteria, the sample, recoveries.
reported value of reported for this sample sample duplicate, and
<LOQ for as <LOQ. post spike sample
analyte(s). must be reanalyzed at
When analyte consecutively higher
concentrations are dilutions until the
calculated as <LOQ, the criteria is met.
post spike for that
analyte must recover
within 70-130% of its true
value.

Any printed copy of this SOP and all copies of this SOP outside of Pace are uncontrolled copies.
Uncontrolled copies are not tracked or replaced when new versions are released or the SOP is made obsolete.
Users of the SOP should verify the copy in possession is the current version of the SOP before use.

58 of 58
CONTEST, A PACE ANALYTICAL LABORATORY SOP 454 PFAS Water Isotope Dilution Author: BLH
39 Spruce Street Doc 454. Revision No. 7
East Longmeadow, MA 01028-0591 Effective Date: 06/30/2021
Page 1 of 37

Determination of Selected Per- and Polyfluorinated Alkyl

Substances (PFAS) by Solid Phase Extraction & Isotope
Dilution by Liquid Chromatography/Tandem Mass
Spectrometry (LC/MS/MS)

Approved:

_________________________ _________________________

Tod Kopyscinski Katherine Allen

Laboratory Director QA Officer

Revision Number: 7

DRAFT
 CONTEST, A PACE ANALYTICAL LABORATORY SOP 454 PFAS Water Isotope Dilution Author: BLH
39 Spruce Street Doc 454. Revision No. 7
East Longmeadow, MA 01028-0591 Effective Date: 06/30/2021
Page 2 of 37

Change Record

Revision Date Responsible Description of Change

Person
0 06/01/2017 Brianna Original
McLaughlin
1 10/10/2017 Brianna Changes to Recovery Allowances for specific Compounds; Changes to LC
McLaughlin method conditions
2 11/13/2018 Katherine Updates from Sept 2018 NH audit: “Effective” date added to header, Sec 3.4 (should
Allen/KAF replaced with must), Sec 7.3.4 (expansion of Isotope Dilution Technique-note added),
Sec 7.3.5 (surrogates should change to surrogates must), Sec 7.5.2 (Blank Spike
Duplicate deleted), Sec 7.5.3 (addition of DI water rinse after methanol), Sec 7.6.1.1
(blank extraction replaced with blank subtraction), Sec 7.7.3 (delete to prove system is
contamination free), deletion of section 8.1.11 (IDOC), Sec 12.0 (addition of
calculations), and Sec 5.7(updated volumes), Sec 5.8 (changed water to methanol),
Appendix B(added pH adjustment method). Addition of section 7.3.8 for explanation of
isotope dilution technique.
3 03/17/2020 Brianna Update to make entire new procedure: Entire SOP edited.
Mclaughlin/James
Constantino
4 07/01/2020 Brianna Update to make entire new procedure: Entire SOP edited and CAS number
Henriquez/James added.
Constantino
5 08/17/2020 Brianna Henriquez Updates from NH 2020 Audit: Section 7.3.5 should to shall, 7.3.7 & 7.3.8
updates to current procedure and clarification. Section 7.5.11 grammatical
error fixed. Section 12.5 removed. Section 8.2.4 – MS/MSD spiked at mid-
level added, Edit to Appendix C to including 40CFR reference, Addition to
section 7.3.4 to include verbiage of using branched and linear isomers, and
Sec 12.0 (removal of calc not needed).

6 11/18/2020 Katherine Allen Update to holding time in Section 3.5 – update from 14 days to 28 days to be
the same as EPA 533.

7 06/30/21 Brianna Henriquez Updates to Procedure: Overall: References to Con-Test changed to Pace. Section 1.1:
added specific instruments (6470 and 6495). Section 4: Updated column PNs. Removed
envi-carb cartridge that isn’t used in this method. Section 5: 20MM mobile phase
changed to 5MM, mobile phase expiration changed. Section 7: Sampling instructions
changed. Removed section on separate internal standard. Section 7.1: Sampling
instructions changed. Section 7.2.6: Prepared and in-use standards stored at room
temperature. Added 24PAR and certain individual analyte stocks for separate spike
sourcing. Standard prep and calibration tables were changed. CCVs changed to only be
run at the mid-level (level 4) after an opening low-level. Section 7.3: Updated
calibration method, technique, and requirements (removal of IS/surrogate now
functioning as both surrogate and IS). Added section 7.3.5.1. Section. Section 7.4.1:
Requirement added of IBL and low-level CCV every 12 hours. Section 7.5.1: pH range
changed from ± 0.5 pH units to ± 1.0 pH units. Section 7.5.4: Surrogate and spike
step combined, and are now always the same amount. Sec 7.5: changed Dup and MS to
MS and MSD and specified balance, Section 7.5.5: Added volume by weight step, and
special instructions for sediment-heavy samples (Appendix F). Section 7.5.11: Added
cartridge soaking step before solvent collection. Section 7.5.14: Rest of new volume by
weight step. Section 7.6.2: N-Me-FOSAA was written incorrectly and changed. Section
7.6.2.1: TPFOA branched check section added. Section 7.7.2: Instrument method
conditions changed. Cell accelerator voltage moved to this section. Section 8.2.4:
MS/MSD spike levels held constant. 8.2.7: Updated how isotope dilution technique is
calculated. Section 8.2.9: Section reworded and changed to reflect proper CCV criteria.
Appendices: Removed Appendices A and C, changing other appendices letters.
Appendix A: Now has transitions and conditions for both instruments, and qualifier
transitions. Added Appendix F with instructions on extracting samples with heavy
sediment.
 CONTEST, A PACE ANALYTICAL LABORATORY SOP 454 PFAS Water Isotope Dilution Author: BLH
39 Spruce Street Doc 454. Revision No. 7
East Longmeadow, MA 01028-0591 Effective Date: 06/30/2021
Page 3 of 37

Distribution/Training List

See Employee Training Record File for signed training statements for trained users.

1.0 SUMMARY, SCOPE, AND APPLICATION

1.1 This method is used to analyze water samples for selected per- and polyfluorinated alkyl acids
(PFAAs). A water sample of approximately 250mLs (preserved with Trizma if chlorinated source) is
fortified with spikes and surrogates and extracted via Solid Phase Extraction (SPE). The sample is then
concentrated to near dryness and subsequently brought up to a final volume of 1mL. All samples are
analyzed using a Triple Quad LC/MS 6470 or 6495 (LC/MS/MS) system. Target analytes are identified by
comparing mass spectra and retention times to reference spectra and retention times of calibration
standards. Analytes are quantitated using the isotope dilution technique explained in the initial calibration
section. The following compounds can be identified by this method:

Analyte Acronym Standard RL Cas Number

(ng/L)
11- 11‐chloroeicosafluoro‐3‐oxanone‐1‐ 11Cl-PF3OUdS 2.0 763051-92-9
sulfonic acid
9 -Chlorohexadecafluoro-3- 9Cl-PF3ONS 2.0 756426-58-1
oxapentane-1-sulfonic acid
4,8-Dioxa-3H-perfluorononanoic acid ADONA 2.0 919005-14-4
Hexafluoropropylene oxide dimer acid HFPO-DA 2.0 13252-13-6
Perfluoro‐3,6‐dioxaheptanoic acid NFDHA 2.0 151772-58-6
Perfluorobutanoic acid PFBA 2.0 375-22-4
Perfluorobutanesulfonic acid PFBS 2.0 375-73-5
8:2 Fluorotelomer sulfonic acid 8:2FTS 2.0 39108-34-4
Perfluorodecanoic acid PFDA 2.0 335-76-2
Perfluorododecanoic acid PFDoA 2.0 307-55-1
Perfluoro(2-ethoxyethane)sulfonic acid PFEESA 2.0 113507-82-7
Perfluoroheptanesulfonic acid PFHpS 2.0 375-92-8
Perfluoroheptanoic acid PFHpA 2.0 375-85-9
4:2 Fluorotelomer sulfonic acid 4:2FTS 2.0 757124-72-4
Perfluorohexanesulfonic acid PFHxS 2.0 355-46-4
Perfluorohexanoic acid PFHxA 2.0 307-24-4
Perfluoro-3-methoxypropanoic acid PFMPA 2.0 377-73-1
Perfluoro-4-methoxybutanoic acid PFMBA 2.0 863090-89-5
Perfluorononanoic acid PFNA 2.0 375-95-1
6:2 Fluorotelomer sulfonic acid 6:2FTS 2.0 27619-97-2
 CONTEST, A PACE ANALYTICAL LABORATORY SOP 454 PFAS Water Isotope Dilution Author: BLH
39 Spruce Street Doc 454. Revision No. 7
East Longmeadow, MA 01028-0591 Effective Date: 06/30/2021
Page 4 of 37

Perfluorooctanesulfonic acid PFOS 2.0 1763-23-1

Perfluorooctanoic acid PFOA 2.0 335-67-1
Perfluoropentanoic acid PFPeA 2.0 2706-90-3
Perfluoropentanesulfonic acid PFPeS 2.0 2706-91-4
Perfluoroundecanoic acid PFUnA 2.0 2058-94-8
N-ethyl perfluorooctanesulfonamidoacetic NEtFOSAA 2.0 2991-50-6
acid
N‐methyl NMeFOSAA 2.0 2355-31-9
perfluorooctanesulfonamidoacetic acid
Perfluoro‐1‐butanesulfonamide FBSA 2.0 30334-69-1

Perfluoro‐1‐hexanesulfonamide FHxSA 2.0 41997-13-1

Perfluorotetradecanoic acid PFTA 2.0 376-06-7

Perfluorotridecanoic acid PFTrDA 2.0 72629-94-8

Perfluorooctanesulfonamide FOSA 2.0 754-91-6

Perfluorononanesulfonic acid PFNS 2.0 68259-12-1

Perfluorodecanesulfonic acid PFDS 2.0 335-77-3

2.0 INTERFERENCES
2.1 Standards and samples should not come into contact with glass other than standards purchased in
glass ampules. PFAS commonly adsorb to the surface and could result in recovery discrepancies.

2.2 Matrix interferences may be caused by co-extracted contaminants present in the sample.
2.3 Method interferences may be caused by contaminants in solvents, reagents, and other sample
processing hardware.
2.3.1 Other common lab supplies that are associated with PFAAs and should be avoided where possible:
aluminum foil, permanent marker, and PTFE.
2.3.1.1 To eliminate any residual PTFE from the Agilent LC, an inline filter column has been installed to
reduce any background contamination prior to sample introduction into the system. See Equipment and
Supplies Section 4.0.
2.3.2 Organic contaminants can pose a threat of interference due to the high quantities of de-
chlorinating agent added to samples.
2.3.3 Contamination levels should be monitored and all blanks should be free from interferences (less
than 1/2 the MRL) in all Laboratory Reagent Blanks (LRB).
2.3.4 Blank subtraction is not permitted in this method.
2.3.5 There is a possibility of matrix effects due to co-extracted organic material. When high levels of
TOC are present, this can affect the ionization of 4:2 FTS considerably
CONTEST, A PACE ANALYTICAL LABORATORY SOP 454 PFAS Water Isotope Dilution Author: BLH
39 Spruce Street Doc 454. Revision No. 7
East Longmeadow, MA 01028-0591 Effective Date: 06/30/2021
Page 5 of 37

3.0 SAMPLE PRESERVATION/STORAGE/HOLDING TIME

3.1 Samples should be collected in a wide mouth 250-mL polypropylene bottle fitted with a
polypropylene screw cap.
3.2 Prior to shipment to the field for sampling, Trizma will be added for any chlorinated sources or
drinking water samples.
3.2.1 It is important that no amount of preservative is spilled from the container or overflowed during
sampling.
3.3 Samples cannot exceed 10°C during the first 48 hours following sample collection. Samples must
be received at or below 10°C OR have ice remaining in the cooler.
3.4 Prior to extraction, samples must be stored at or below 6°C and cannot be frozen.
3.5 Samples collected must be extracted within 28 days. Extracted samples must be run within 28
days of extraction and remain stored at room temperature.

4.0 EQUIPMENT & SUPPLIES

4.1 Triple Quad LC/MS System
4.2 Inline Delay column
4.2.1 Agilent Zorbax Eclipse Plus C18 3.0x50mm 1.8-micron P.N. PFCDELAY or
equivalent
4.3 Analytical Column
4.3.1 Agilent Zorbax Eclipse Plus C18 3.0x50mm 1.8-micron P.N. 959757-02 or
equivalent
4.4 Auto-pipettors: 0-10uL, 10-100uL, 100-1000uL
4.5 Polypropylene pipet tip: 0-10uL, 10-100uL, 100-1000uL
4.6 Polypropylene transfer pipets
4.7 Polypropylene graduated cylinder: 10mL, 50mL, 100mL, 1000mL
4.8 Vials: 2ml polypropylene vials
4.9 Caps: 11mm polypropylene snap caps
4.10 Sample containers: 250ml wide mouth polypropylene containers
4.11 Polypropylene centrifuge tubes
4.12 SPE System
4.12.1 Supelco Visiprep manifold or equivalent
4.12.2 Non-PTFE SPE Reservoirs and/or sample transfer tubing with adapters
4.12.3 Phenomenex Pre-Stacked WAX/GCB Cartridge or equivalent
4.12.3.1 Each lot should be checked to be free of contamination prior to usage for any field samples or
batch QC. This can be done in the form of an additional LRB prior to usage for extraction.
4.12.4 Vacuum pump capable of reaching up to 20” Hg
4.12.5 13L Safety coated Pyrex waste collection container
4.12.6 Polypropylene tubing for vacuum pump and manifold
4.13 N-Evap concentrator system
CONTEST, A PACE ANALYTICAL LABORATORY SOP 454 PFAS Water Isotope Dilution Author: BLH
39 Spruce Street Doc 454. Revision No. 7
East Longmeadow, MA 01028-0591 Effective Date: 06/30/2021
Page 6 of 37

4.14 Balance: Analytical, capable of accurately weighing 0.0001g

4.15 Vortexer
4.16 Polypropylene inserts

5.0 REAGENTS & STANDARDS

5.1 Reagent Water : interferent free

5.2 Methanol : LC/MS Grade
5.3 Nitrogen : Ultra high purity
5.4 Ammonium Acetate : LC/MS Grade
5.5 Stock Standard Solutions: Purchased as either certified solutions or neat standards.
5.6 Surrogate, Internal Standard, and ESI-L Low concentration tuning mix purchased as certified
solutions
5.7 5 mM Ammonium Acetate reagent water: Prepared by adding 1.54 grams of Ammonium Acetate
to 1000mL of reagent water and mixing until solids are into solution.
5.8 1 g/L Ammonium Acetate reagent water.
5.9 96:4 Methanol:Water- Made fresh every 2 days.
5.10 Ammonium hydroxide (56.6% w/w)
5.11 0.1 M Sodium Phosphate Monobasic
5.12 0.1 M Sodium Phosphate Dibasic
5.13 0.1 M Phosphate Buffer: Prepared by mixing 500 mL of dibasic sodium phosphate with 275 mL
of monobasic sodium phosphate. Verify solution pH of about 7.0.
5.14 Agilent ESI-L Low Concentration Tuning Mix
5.15 Sodium Hydroxide
5.16 Elution Solvent: 1% ammonium hydroxide in methanol (made fresh daily)

6.0 SAFETY
See Material Safety Data Sheets (MSDS) and Pace Chemical Hygiene Plan.

7.0 PROCEDURE
7.1 Sampling
7.1.1 Samples are to be collected in lab-provided plastic containers. Guidance for sampling is obtained
through Pace corporate website or preferably local project authorities/municipalities.
7.2 Surrogate/Spike/Isotope Performance Standard Preparation
7.2.1 All standards must be documented in Element and have Certificate of Analysis forms attached
electronically. All information should be documented and each standard should be given an Element Standard
ID#.
7.2.2 Standards may be received in purchased glass ampoules but any transfer or dilution must be stored
in polypropylene vials with Non-PTFE caps.
CONTEST, A PACE ANALYTICAL LABORATORY SOP 454 PFAS Water Isotope Dilution Author: BLH
39 Spruce Street Doc 454. Revision No. 7
East Longmeadow, MA 01028-0591 Effective Date: 06/30/2021
Page 7 of 37

7.2.3 All standards purchased from Wellington come pre-treated with sodium hydroxide for compound
stability. If making standards from solid, standards must be stored under basic condition to prevent
esterification of fluorinated carboxylic acids. See calculation 1 in section 12.1.
7.2.4 PFAS Surrogate Preparation
7.2.4.1 All purchased surrogate stock standards are to be stored until expiration date provided by
manufacturer at 4°C.
Compound Abbreviation PDS, ng/mL
Perfluoro‐n‐[1,2,3,4‐ MPFBA 1000
13C4]butanoic acid
Perfluoro‐n‐[1,2,3,4,5‐ M5PFPeA 1000
13C5]pentanoic acid
Sodium perfluoro‐1‐[2,3,4‐ M3PFBS 929
13C3]butanesulfonate
Sodium 1H,1H,2H,2H‐ M2‐4:2FTS 935
perfluoro‐1‐[1,2‐13C2]hexane
sulfonate
Perfluoro‐n‐[1,2,3,4,6‐ M5PFHxA 1000
13C5]hexanoic acid
Perfluoro‐n‐[1,2,3,4‐ M4PFHpA 1000
13C4]heptanoic acid
Sodium perfluoro‐1‐[1,2,3‐ M3PFHxS 946
13C3]hexanesulfonate
Sodium 1H,1H,2H,2H‐ M2‐6:2FTS 949
perfluoro‐1‐[1,2‐13C2]‐octane
sulfonate
Perfluoro‐n‐[13C8]octanoic M8PFOA 1000
acid
Perfluoro‐n‐[13C9]nonanoic M9PFNA 1000
acid
Sodium perfluoro‐ M8PFOS 957
[13C8]octanesulfonate
Sodium 1H,1H,2H,2H‐ M2‐8:2FTS 958
perfluoro‐1‐[1,2‐13C2]‐decane
sulfonate
Perfluoro‐n‐[1,2,3,4,5,6‐ M6PFDA 1000
13C6]decanoic acid
Perfluoro‐n‐[1,2,3,4,5,6,7‐ M7PFUnA 1000
13C7]undecanoic acid
2,3,3,3‐Tetrafluoro‐2‐ M3HFPO‐DA 1000
(1,1,2,2,3,3,3‐
heptafluoropropoxy‐13C3‐
propanoic acid
Perfluoro‐n‐[1,2‐ M2PFDoA 1000
13C2]dodecanoic acid
Perfluoro‐n‐[12‐ M2PFTA 1000
13C2]tetradecanoic acid
CONTEST, A PACE ANALYTICAL LABORATORY SOP 454 PFAS Water Isotope Dilution Author: BLH
39 Spruce Street Doc 454. Revision No. 7
East Longmeadow, MA 01028-0591 Effective Date: 06/30/2021
Page 8 of 37

Perfluoro‐1‐ M8FOSA 1000

[13C8]octanesulfonamidoaceti
c acid
N‐methyl‐d3‐perfluoro‐1‐ d3‐N‐MeFOSAA 1000
octansulfonamidoacetic acid
N‐ethyl‐d5‐perfluoro‐1‐ d5‐N‐EtFOSAA 1000
octansulfonamidoacetic acid
Ordered as MPFAC-24ES from Wellington Labs

2,3,3,3‐Tetrafluoro‐2‐ M3HFPO‐DA 1000

(1,1,2,2,3,3,3‐
heptafluoropropoxy‐13C3‐
propanoic acid
Ordered as M3HFPO-DA from Wellington Labs
7.2.6 Analyte Primary Dilution Standard Preparation
7.2.6.1 All purchased PFAA spike standard stock standards are to be stored until expiration date provided
by manufacturer at 4°C.
7.2.6.2 Prepared and in use PFAA stock standard solutions should be stored at room temperature and
vortexed prior to usage. These standards will expire 2 months after preparation date or manufacturer’s
expiration date, whichever comes first.
Analyte Standards
Compound Vendor Concentration of Standard
(ng/ mL)
PFAC-30PAR Wellington 1000*
PFAC-24PAR Wellington 2000*
PFEESA Wellington 50000*
NFDHA Wellington 50000
PFMPA Wellington 50000
PFMBA Wellington 50000
HPFO-DA Wellington 50000
NaDONA Wellington 50000*
9ClPF3ONS Wellington 50000*
11ClPF3OUdS Wellington 50000*
FBSA Wellington 50000
FHxSA Wellington 50000
*Individual analyte concentration may vary due to amount of anion present in solution. All
calculations must use the anion concentration, not the salt concentration. See Calculation
2 in section 12.2.
7.2.6.3 PFEESA, NFDHA, PFMPA, PFMBA, HPFO-DA, NaDONA, 9ClPF3ONS, 11ClPF3OUdS,
FBSA, and FHxSA are not included in the 24PAR mixture and are mixed into a separate
“supplemental” 1000 ng/ mL stock. See below table for mixture.
7.2.6.4 The Supplemental Stock is used to make a 500ppb spike dilution with 24PAR so that a different
lot is used for spiking. The calibration stock is made using 30PAR and directly adding
CONTEST, A PACE ANALYTICAL LABORATORY SOP 454 PFAS Water Isotope Dilution Author: BLH
39 Spruce Street Doc 454. Revision No. 7
East Longmeadow, MA 01028-0591 Effective Date: 06/30/2021
Page 9 of 37

PFEESA, NFDHA, PFMPA, and PFMBA, which are missing from that mixture. See below table
for prep instructions:
Stock Dilution Prep Table
Volume of Volume of Final Final
Compound/Standard Mixtures Methanol Volume Concentrati
(µL) on
(µL) (µL)
(ng/mL)
Supplemental 100µL – HPFO-DA 4000µL 5000µL 1000
Stock
100µL – NaDONA
100µL – 9ClPF3ONS
100µL – 11ClPF3OUdS
100µL – PFEESA
100µL – PFMBA
100µL – PFMPA
100µL – NFDHA
100µL – FBSA
100µL – FHxSA
500 ng/ mL 1250µL – PFAC24PAR 1250µL 5000µL 500
Spike
2500µL – Supplemental Stock
100 ng/ mL 250µL – PFAC24PAR 4250µL 5000µL 100
Spike
500µL – Supplemental Stock
100 ng/ mL 500µL – PFAC30PAR 4460µL 5000µL 100
Cal Stock 10µL – PFEESA
10µL – PFMBA
10µL – PFMPA
10µL – NFDHA
7.2.6.5 The calibration is prepared as follows using the stock dilutions prepared above.
Calibration Table
Volume Volume Volume Volume of Volume of Volume of Final Final
Surrogate Stock M3HFPODA 96:4 Volume
100ppb 30PAR Supplementa (µL) Surrogate Stock Methanol: Concentration
l Water (µL) (µL)
Stock Stock (ng/mL)
Standard (µL) Standard Stock
(µL) Standard
(µL)

12.5 0 0 25 25 4937.5 5000 0.25*

25 0 0 25 25 4925 5000 0.5*

50 0 0 25 25 4900 5000 1.0*

125 0 0 25 25 4825 5000 2.5*

250 0 0 25 25 4700 5000 5.0*

CONTEST, A PACE ANALYTICAL LABORATORY SOP 454 PFAS Water Isotope Dilution Author: BLH
39 Spruce Street Doc 454. Revision No. 7
East Longmeadow, MA 01028-0591 Effective Date: 06/30/2021
Page 10 of 37

0 50 50 25 25 4850 5000 10.0*

0 125 125 25 25 4700 5000 25.0*

*Individual analyte concentration may vary due to amount of anion present in solution. All
calculations must use the anion concentration, not the salt concentration. See Calculation 2 in section
12.2.
7.2.6.6 Continuing calibration verification (CCVs) standards are made at the mid-level, identically to the
4th calibration level above. The ICV/QCS is made similarly, just like the 5th calibration level above, except
instead of calibration stock 100 ppb Spike is used as shown in the table below:
QCS/ICV Preparation Table
Standard Volume Standards Volume of Final Final
Name 96:4 Volume
Concentration
Methanol:Wa
(µL)
ter (µL) (ng/mL)
QCS/ICV 25µL- PFAC-24ES 4700µL 5000µL 5.0
25µL- M3HPFO-DA
Surrogate Dilution
250µL- 100ppb Spike

7.3 Initial Calibration Criteria

7.3.1 All analytes must first be product ion optimized with the LC/MS/MS system using the
MassHunter Optimizer program to determine optimal fragmentor and collision cell energy for applicable
ions. This optimization should be done using a high-level standard for each analyte and using all of the LC
parameters used in the analytical method. A level 4 standard must then be run to identify all retention time
windows for all compounds of interest (See Section 7.7.2). A minimum of 10 spectra scans are acquired
across each chromatographic peak. See Appendix A for further analyte MS/MS conditions.
7.3.2 Prior to initial calibrations, and when the instrument is having difficulty passing regular
calibrations, a mass calibration will be performed using Agilent ESI-L Low Concentration Tuning Mix.
The MassHunter program has an autotune feature that performs the mass calibration using this mix and
verifies it in a report.
7.3.3 A calibration is to be run when continuing calibration checks or surrogates do not pass QC criteria.
A calibration should also be performed when any hardware is changed or major instrument maintenance is
performed.
7.3.4 The initial calibration must contain a minimum of five points with the lowest point being at or
below the MRL. A minimum of six points is required for a quadratic calibration. The total of the branched
and linear isomers must be used for calibration for the following target analytes: PFOS, PFHxS,
NEtFOSAA, NMeFOSAA.
7.3.5 The LC/MS/MS system is to be calibrated using the isotope dilution technique. Therefore, isotope
dilution analogues are added at a constant concentration in all standards prior to injection. Either linear or
quadratic regression can be used, but it must always be forced through zero and can be concentration
weighted. Forcing through zero allows for more sensitivity to detect background contamination within the
system. The calibration shall be done using the same LC conditions as the samples (See Section 7.7.2).
Because the isotope dilution analogues are added in equal concentrations, calibrate for them using an
average response factor. Not all analytes have an exact mass-labeled analogue, in which case the closest
analogue is used (either by chemical properties or retention time). See Appendix A for the list of isotope
analogues and corresponding target analytes.
CONTEST, A PACE ANALYTICAL LABORATORY SOP 454 PFAS Water Isotope Dilution Author: BLH
39 Spruce Street Doc 454. Revision No. 7
East Longmeadow, MA 01028-0591 Effective Date: 06/30/2021
Page 11 of 37

7.3.5.1 The isotope dilution technique utilizes extracted compounds to serve as a traditional
internal standard. In this case, the extracted analogues must pass criteria listed in section 8.2.6. The
analogue is then used as the internal standard compound for associated target analytes.
7.3.6 Calibration levels for linear or non-linear analyte targets must have a r2 ≥ 0.99 for each analyte and
the recovery for each analyte must be within 70-130% of the true value. Surrogate and internal standards
must have an RSD of the RFs for all analytes of ≤20%.
7.3.7 A quality control standard (QCS) will serve as an initial calibration verification (ICV) and be run
following initial calibration and all subsequent calibrations. The ICV shall be prepared from a separate
dilution of a different stock standard. This sample must be run following a calibration or quarterly,
whichever comes first. The accepted values for the ICV are 70-130% of the true value for each analyte.
7.3.8 If any instrumentation or analytical setpoints are changed to the instrument calibration, an initial
demonstration of capability (IDOC) for the procedure and instrumentation shall be performed. See
Appendix E.

7.4 Continuing Calibration

7.4.1 An instrument blank (IBL) and a low-level instrument sensitivity check (ISC) must be run at the
MRL before any other injections and once every 12 hours. The results must be between 70-130% of the
true value for all analytes. The IBL should have no hits greater than ½ the MRL. The ISC can serve as your
initial CCV for the day.
7.4.2 Prior to samples analysis, a low-level continuing calibration verification (CCV) must be run. After
every ten field samples a subsequent CCV must be at Level 4 (same as calibration point 4 above). A
closing CCV must also be run at the end of each analysis. The requirements for the CCVs are 70-130% of
the true value for method analytes.
7.4.3 An instrument blank is required to be run following analysis of the highest-level standard analyzed
(after a calibration in this case). One is also required daily prior to sample analysis. All analytes must be at
>½ the MRL in order to pass.
7.4.4 All isotope dilution analogues (surrogates) must have a recovery between 50-150%
7.4.5 In the case of CCV failure, two consecutive CCVs should be immediately run. If these pass,
analysis can continue. Otherwise, a calibration or tuning and re-analysis of affected samples is required.
7.4.6 A checktune will be run as needed to verify MS operating criteria. This is run through the
MassHunter program using Agilent ESI-L Low Concentration Tuning Mix. If criteria are out of spec, the
parameters set forth in the Agilent 6400 Series Triple Quadrupole LC/MS System Quick Start Guide must
be followed to adjust values. If re-run of checktune does not pass, an autotune must be run and the
instrument must be recalibrated.

7.5 Sample Extraction Procedure

7.5.1 For each required lab QC sample, fill a clean sample bottle with 250 mL DI water. Check verify pH
is 7.0 ± 1.0 pH units with pH paper. If sample pH is not 7.0 ± 1.0 pH units, note on bench sheet and adjust
using Trizma.
7.5.2 For every 20 field samples, a blank and a blank spike must be extracted. (Field blanks are considered
field samples in this consideration as they are treated as such) Ideally, if adequate sample volume is available,
a matrix spike and matrix spike duplicate should be included on every batch.
7.5.3 All polypropylene equipment including graduated cylinders and sample transfer lines/reservoirs
should be washed prior to using with extraction solvent (96:4 Methanol:water), followed by a DI water rinse.
CONTEST, A PACE ANALYTICAL LABORATORY SOP 454 PFAS Water Isotope Dilution Author: BLH
39 Spruce Street Doc 454. Revision No. 7
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Page 12 of 37

7.5.4 Add 5uL of surrogate to each sample and 25ul of 100ppb spike to all BS and MS samples included
on the extraction batch. Cap and invert to mix well.
7.5.5 Take initial weight (in grams) of each bottle and sample with the Sartorius Top Loading Balance.
Observe if any samples have heavy sediment or are very cloudy, and decide if it seems like the cartridge will
clog. These samples can be extracted following a special protocol using a centrifuge outlined in Appendix C.
7.5.6 After SPE system is set up, condition the cartridges first with 5mL methanol, followed by 5mL of
0.1M Sodium Phosphate Buffer.
*Note: The sample cartridges must not be allowed to run dry at any point during conditioning.
If they become dry, the conditioning must be started over.
7.5.7 Next add 2 mL of 0.1M Sodium Phosphate Buffer and attach either sample transfer tube or reservoir
to the cartridge and begin transferring sample. The samples should be passed through the cartridge at
approximately 5mL/min. This equates to a drop wise fashion eluting from the cartridge.
7.5.8 Rinse sample bottle with 7.5mL of reagent water and pass through tubing or reservoir and cartridge.
Repeat once more.
7.5.9 Add 0.5mL of acetonitrile to each cartridge. Remove sample transfer tubes/reservoirs and allow air
to pass through the cartridges for a minimum of 5 minutes at approximately 10-15” Hg.
7.5.10 Turn off vacuum and add tray of labeled collection vials to manifold.
7.5.11 Using a pipette, rinse each respective reservoir into the sample container taking care to rinse the
sides with 6mL of 1% NH4OH methanol. Pour solvent from sample bottles directly into cartridge and allow
to soak in the cartridge for 5 minutes after solvent fully wets the solid phase. Then, allow to elute through
the cartridge at a low vacuum elute with an additional 5-6mL of solvent so the final eluent is ~12mL.
7.5.12 Samples can then be concentrated to ~850uL at room temperature
7.5.13 Add 96:4 methanol:water, taking care to rinse the side of the container until the final volume reaches
1mL.
7.5.14 Determine initial volume by taking the weight (in grams) of the empty container following
extraction. Subtract this from the weight taken in step 7.5.5 to determine the volume by weight of the sample
(it is assumed that 1 gram is equal to 1 mL).
7.6 Data Analysis
7.6.1 The analyst cannot extrapolate beyond the range of the calibration. However, by isotope dilution
analysis, the extract cannot be diluted. If an analyte is outside of the determined range, the sample must be
re-extracted at an appropriate dilution level.
7.6.1.1 There is extrapolation allowed only to determine if there is blank contamination. Since there is no
blank subtraction, any contamination present must be below 1/2 of the MRL for specific analyte.
7.6.1.2 If a sample exceeds the calibration range the sample must be re-extracted. This would involve diluting
the sample with reagent water to be within the calibration range and adding ammonium acetate to be at a
final concentration of 1 g/L.
7.6.1.3 Additionally, if a sample exceeds the calibration range, one or more LRB must be run until the system
meets acceptable criteria. If this occurs during an automated sequence, the samples subsequently must be
evaluated. If the over-range analytes are present in the subsequent samples at or above the RL, the samples
are considered invalid and must be re-run. If the analyte in question does not exceed the RL, the samples can
be reported.
7.6.2 Compounds that have both branched and linear isomers will be reported as total. These
compounds include PFOS, PFHxS, N-Et-FOSAA, N-Me-FOSAA and PFOA. PFOS, PFHxS, N-Et-
FOSAA, and N-Me-FOSAA have the branched and linear compounds available for quantitation. PFOA is a
special case outlined below:
CONTEST, A PACE ANALYTICAL LABORATORY SOP 454 PFAS Water Isotope Dilution Author: BLH
39 Spruce Street Doc 454. Revision No. 7
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Page 13 of 37

7.6.2.1 PFOA will be quantitated by using a qualitative/semi-quantitative approach per EPA guidance.
Since there is no standard available, the calibration will be done using the linear isomer only. A technical
grade standard will be run to identify the retention time of the branched isomer. All samples will be
quantitated using the area of both the linear and branched isomers of PFOA that may be present within the
sample. A branched isomer check for PFOA will be run with every calibration curve to verify the retention
times of the branched isomers for PFOA.

7.6.3 All analytes and surrogates will be calculated based off the initial calibration criteria.

7.6.4 All results for analytes shall be reported as the neutral acid.

7.6.5 Retention time windows are established once per ICAL and at the beginning of each sequence. On
days when an ICAL is not run, the initial CCV is used to set the times. All retention times of analytes and
EIS analytes must fall within 0.4 minutes of the established time. Analytes must also elute within 0.1
minutes of their respective EIS.

7.6.6 In addition to retention time identification criteria, most ions are identified by two ion transitions.
(The following ions are exceptions: PFBA, PFPeA, FBSA, FHxSA, PFMPA, PFMBA, 9Cl-PF3ONS,
11Cl-PF3OUdS, ADONA). The secondary, or qualifier ion, must have a signal to noise of 3:1. The ratio
between the qualifier and the quantifier ion must be averaged from the calibration. For samples to be valid,
the ratio of qualifier to quantifier must be +/-50% from the average ratio from the applicable calibration.

7.7 Instrumentation Procedure

7.7.1 Before any QC or samples can be run, the HPLC must be allowed to purge for at least thirty
minutes. This purge can be done using any combination of the mobile phases, but prior to samples running,
the initial mobile phase conditions used in the method must be allowed to run for 15 minutes or until
pressure has stabilized.
7.7.2 The instrument must be stable in all parameters before a run is started. The following are the
HPLC and ESI-MS Method Conditions. Also, See Appendix A for additional MS/MS Method Conditions.
Time (min) % 5 mM Ammonium Acetate % Methanol Flow Rate
in water (mL/minute)
0.00 95 5 1.0
0.10 65 35 1.0
2.00 50 50 1.0
3.00 25 75 1.0
4.50 1 99 1.0
4.51 1 99 1.0
5.00 1 99 1.0
5.10 95 5 1.0
6.50 95 5 1.0

Injection Volume 6470 10 uL

Injection Volume 6495 5 uL
Column Compartment Temperature 40 °C
Autosampler Compartment Temperature 10 °C
Polarity Negative
Gas Temperature 250 °C
Gas Flow 11 L/min
Nebulizer 50 psi
Sheath Gas Temperature 300 °C
Sheath Gas Flow 12 L/min
CONTEST, A PACE ANALYTICAL LABORATORY SOP 454 PFAS Water Isotope Dilution Author: BLH
39 Spruce Street Doc 454. Revision No. 7
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Page 14 of 37

Capillary Needle Voltage (Negative mode) -3000 V

Cell Accelerator Voltage 5V

7.7.3 An instrument sequence will be made. It will open with a blank and a low level CCV. After the
CCV, the batch can start running. Every 10 field samples (excluding QC and FRBs) a subsequent CCV
must be run, at level 4. The sequence must end with a CCV.
7.7.4 The run can end with a script to put the instrument into standby mode.

8.0 QUALITY CONTROL

8.1 Definitions

For definitions and explanations of quality control measures (blanks, LCS/QC Reference, LFB, Duplicates,
MS/MSD, etc.) refer to the Contest, A Pace Analytical Lab Quality Assurance Manual.

8.2 Quality Control Measures & Acceptance Criteria

8.2.1 Method Blank

The method blank is matrix specific and extracted with every batch or every 20 samples
(whichever is more frequent). The target compounds and ranges must be ≤1/2 the MRL, or <1/10th the
amount measured in any sample, or <1/10th the regulatory limit. If any analytes are present above this
level, the detected analytes are considered invalid for all samples extracted in that batch.

8.2.2 Field Reagent Blank

It is highly recommended to collect a Field Reagent Blank per every sampling event. If
provided, Field reagent blanks only need to be run and reported if there are analytes at or above the
MRL in any associated field samples. Any analyte peaks present in field reagent blanks must be below
1/3 of the MRL of that analyte. If any analytes are present above this level, all samples collected with
said FRB are invalid and must be recollected and reanalyzed. Data will be reported to client as suspect,
noting the field blank contamination.

8.2.3 Laboratory Control Sample/Duplicate (LCS)

A matrix-specific LCS is extracted every 20 samples or per batch. The concentration must be
≥ LOQ and ≤ mid-range of calibration. All analytes recoveries must be within limits specified in
Appendix D. If analyte is not listed in table, acceptance criteria is to remain 50-150% until in-house
limits can be determined. Samples should be re-extracted if criteria are not met, even if outside of hold.
If samples cannot be re-extracted then the failures must be notated in the narrative.

8.2.4 Matrix Spikes

A matrix-specific MS is extracted every 20 samples or per batch. MS spike concentrations

will be at the mid-level of the calibration curve. If historical data is available, the sample will be spike
at a level similar to expected contaminant levels. All analytes recoveries must be within limits
specified in Appendix D. If analyte is not listed in table, acceptance criteria is to remain 50-150% until
in-house limits can be determined. See calculation 3.
CONTEST, A PACE ANALYTICAL LABORATORY SOP 454 PFAS Water Isotope Dilution Author: BLH
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*Note: Matrix spike samples may display matrix bias. If the CCC and LFB samples are passing, but
the MS recoveries are outside the designated range, the recovery is deemed to be matrix biased. A
note on the unfortified sample will indicate the possibility of matrix effects being suspect.

8.2.5 Matrix spike duplicates

Extract a spiked sample duplicate every 20 samples (when enough aliquot is

provided). Matrix Spike Duplicate samples should be calculated to have an RPD ≤30%. See
calculation 4.

8.2.6 Quality Control Samples

A quality control sample must be run from a second source at least quarterly, or after an initial
calibration as an ICV. If a second source is not commercially available, a different lot number from the
same vendor is acceptable. The recoveries must be within 70-130% of the true value.

8.2.7 Isotope Dilution Analogue

Isotope dilution analogues are added to all blanks, standards, samples, and spikes. Analogue
compounds must have an area of 50-150% of the associated compound in level 4 of the calibration on
days when a calibration is run. On days when a calibration is not run, analogue compounds must have
an area of 50-150% of the associated compound in the opening instrument sensitivity check/CCV.

If the surrogate is outside these limits, the extract should be re-analyzed. If the re-analysis
passes, report re-analyzed sample. If this fails, the associated isotope performance standard should be
evaluated. The system may need recalibration or maintenance. If the CCV has surrogate out of range,
the instrument needs to be recalibrated.
If the re-analysis fails, re-extract the sample to confirm failure if an aliquot is available. If the
re-extract fails, report both results with appropriate flagging criteria .If not enough volume is provided
to re-extract the sample then report with appropriate flags.

8.2.8 Calibration Curve

A minimum of a 5-point calibration curve (for linear regression) or a 6-point calibration curve (for
quadratic) is used to calibrate the system.

The curve must be verified with an independent standard (QCS) prior to sample
analysis, (10 ng/L). The curve will be forced through zero and may or may not
be concentration weighted.

If a peak is not properly integrated by the data system, manual integration may be necessary. Manual
integrations must comply with the Pace SOP on Chromatographic Integration Procedures. The integration
of the peaks for the samples and quality control samples must be as consistent as possible with the
integration used with the initial calibration.

8.2.9 Continuing Calibration Checks (CCCs)

The results must be between 70-130% of the true value for all analytes for the initial low level CCV
After every ten field samples, a subsequent CCV must be at level 4. The requirements for the CCVs are 70-
130% of the true value. All analogue compound areas must fall within 50-150% of the appropriate
calibration point or CCV.
CONTEST, A PACE ANALYTICAL LABORATORY SOP 454 PFAS Water Isotope Dilution Author: BLH
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The ending CCV acquisition time must fall within 24 hours of the acquisition starting time of the opening
CCV with the associated analysis batch.
8.2.10 Any failures in QC require reanalysis, even if the samples in question are outside of hold.

9.0 CORRECTIVE ACTIONS/ CONTINGENCIES OF HANDLING OUT-OF-

CONTROL DATA
9.1 Refer to Pace Quality Assurance Manual
9.2 Refer to Pace Corrective Action SOP.

10.0 POLLUTION PREVENTION

10.1 Pollution prevention encompasses any technique that reduces or eliminates the quantity or toxicity
of waste at the point of generation. Many opportunities for pollution prevention exist in laboratory
operation. EPA has established a preferred hierarchy of environmental management techniques that places
pollution prevention as the management option of first choice. Whenever feasible, laboratory personnel
should use pollution prevention techniques to address waste generation. Standards should be prepared in
volumes consistent with laboratory use to minimize the disposal of excess volumes of expired standards.

11.0 WASTE MANAGEMENT

11.1 It is the laboratory’s responsibility to comply with all federal, state, and local regulations
governing the waste management, particularly the hazardous waste identification rules and land disposal
restrictions, and to protect the air, water, and land by minimizing and controlling all releases from fume
hoods and bench operations. Also, compliance is required with any sewage discharge permits and
regulations.

11.2 Acidic samples and waste are dumped into satellite waste containers.

12.0 CALCULATIONS

12.1 Calculation 1: Adding 4 mole equivalents to standards to prevent esterification

12.2 Calculation 2: Mass of the anion

Massacid= Measured Masssalt * (Molecular Weightacid/Molecular Weightsalt)

12.3 Calculation 3: Percent Recovery

CONTEST, A PACE ANALYTICAL LABORATORY SOP 454 PFAS Water Isotope Dilution Author: BLH
39 Spruce Street Doc 454. Revision No. 7
East Longmeadow, MA 01028-0591 Effective Date: 06/30/2021
Page 17 of 37

12.4 Calculation 4: Relative percent deviation calculation

13.0 REFERENCES

13.1 Pace Analytical Chemical Hygiene Plan

13.2 Pace Analytical Quality Assurance Manual.

13.3 Pace Analytical Corrective Action SOP.

13.4 Pace Analytical Controlled Document SOP.

13.5 Agilent 1260 Infinity Binary LC Operators manual

13.6 Agilent MassHunter Study Manager

13.7 Agilent MassHunter Optimizer

13.8 MassHunter Personal Compound Database and Library Manager

13.9 Agilent 6400 Series Triple Quadrupole LC/MS System Quick Start Guide

13.10 MassHunter Data Acquisition Compliance Software Quick Start Guide

13.11 MassHunter Quantitative Analysis Compliance Software Quick Start Guide

13.12 Agilent 6000 Series LC/MS System Maintenance Guide

13.13 EPA Method 537, “Determination of Selected Perfluorinated Alkyl acids in Drinking
Water by Solid Phase Extraction and Liquid Chromatography/Tandem Mass
Spectrometry (LC/MS/MS)”, Version 1.1, September 2009.

13.14 Method ISO 25101:2009, “Determination of perfluorooctanesulfonate (PFOS) and

perfluorooctanoate (PFOA) – Method for unfiltered samples using solid phase extraction
and liquid chromatography/mass spectrometry”, April 30, 2009.

13.15 EPA Technical Advisory-Laboratory Analysis of Drinking Water Samples for

Perfluorooctanoic Acid (PFOA) using EPA Method 537 Rev. 1.1 EPA 815-B-16-021
September 2016
CONTEST, A PACE ANALYTICAL LABORATORY SOP 454 PFAS Water Isotope Dilution Author: BLH
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13.16 Agilent Application note by Peter JW Stone, Linda Cote, Jennifer Gushue, Robert J. Letcher
and Shaogang Chu. A Low Femtogram Target Screen Method for Perfluorinated Compounds in Food
Matrices and Potable Water Using the Agilent 6460 Triple Quadrupole LC/MS System Equipped with
Agilent Jet Stream Technology.

13.17 TNI Standard, The NELAC Institute, EL-V1-2009-ISO, 2009.

13.18 Department of Defense (DoD) Department of Energy (DOE) Consolidated Quality Systems Manual
(QSM) for Environmental Laboratories Based on ISO/IEC 17025:2005(E) ISO/IEC 17025:2017(E) and
The NELAC Institute (TNI) Standards, Volume 1, (September 2009).

Appendix A
6470 Transitions and MS Conditions:
Analyte Precursor Product Collision Energy Fragmentor Qualifer/Quantifier
Ion Ion Voltage (V) Voltage (V)
11Cl-PF3OUdS 631 451 24 100
N/A
4-2 FTS 327 307 20 120
Quant
4-2 FTS 327 81 30 120
Qual
6-2 FTS 427 406.9 24 135
Quant
6-2 FTS 427 80 40 125
Qual
8-2 FTS 527 507 28 145
Quant
8-2 FTS 527 80 40 170
Qual
9Cl-PF3ONS 531 351 24 100
N/A
ADONA 377 251 12 100
Quant
ADONA 377 85 12 100
Qual
d3-N-MeFOSAA 573.2 419 20 114
N/A
d5-N-EtFOSAA 589.2 419 20 104
N/A
FBSA 297.99 78 28 115
N/A
FHXSA 398 78.1 30 135
N/A
HFPO-DA 285.1 184.9 5 150
Quant
HFPO-DA 285.1 169 5 150
Qual
M2-4-2-FTS 328.99 309.2 20 135
N/A
M2-6-2-FTS 428.99 409.2 24 160
N/A
M2-8-2-FTS 528.99 509 28 170
N/A
M2PFDA 514.9 469.9 5 102
N/A
M2PFHxA 315 270 4 66
N/A
M2PFOA 415 370 4 69
N/A
M2PFTA 715 670 9 100
N/A
M3HFPO-DA 287 169 2 50
N/A
M3PFBA 216 171.8 4 56
N/A
M3PFBS 301.9 80 45 100
N/A
M3PFHxS 401.9 80 49 100
N/A
M4PFHpA 367 322 4 102
N/A
M5PFHxA 318 273 4 68
N/A
M5PFPeA 268 223 8 120
N/A
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M6PFDA 519 474 4 81

N/A
M7PFUnA 570 525 5 73
N/A
M8FOSA 506 78 36 125
N/A
M8PFOA 421 376 5 65
N/A
M8PFOS 506.9 80 50 100
N/A
M9PFNA 472 427 4 85
N/A
MPFBA 217 172 8 60
N/A
MPFDoA 615 570 5 79
N/A
MPFOS 502.9 80 60 180
N/A
N-EtFOSAA 584 525.9 20 115
Qual
N-EtFOSAA 584 418.9 20 115
Quant
NFDHA 201 85 14 115
N/A
N-MeFOSAA 570 482.9 16 115
Qual
N-MeFOSAA 570 418.9 20 115
Quant
PFBA 213 169 8 60
N/A
PFBS 298.9 98.9 29 100
Qual
PFBS 298.9 80 45 100
Quant
PFDA 513 469 4 81
Quant
PFDA 513 169 16 100
Qual
PFDoA 613 569 5 79
Quant
PFDoA 613 268.7 20 100
Qual
PFDS 598.9 99 60 100
Qual
PFDS 598.9 80 80 100
Quant
PFEESA 315 135 24 110
N/A
PFHpA 362.9 319 8 72
Quant
PFHpA 362.9 169 20 72
Qual
PFHpS 448.9 98.7 44 44
Qual
PFHpS 448.9 79.7 52 52
Quant
PFHxA 313 268.9 8 8
Quant
PFHxA 313 119 18 18
Qual
PFHxS 398.9 99 45 100
Qual
PFHxS 398.9 80 49 100
Quant
PFMBA 279 85.1 8 55
N/A
PFMPA 229 85.1 12 55
N/A
PFNA 463 419 4 66
Quant
PFNA 463 169 17 66
Qual
PFNS 548.9 98.9 40 165
Qual
PFNS 548.9 79.9 40 165
Quant
PFOA 413 369 4 69
Quant
PFOA 413 169 12 69
Qual
PFOS 498.9 99 50 100
Qual
PFOS 498.9 80 50 100
Quant
PFOSA 497.9 77.9 36 125
Quant
PFOSA 497.9 47.9 80 100
Qual
PFPeA 263 218.9 8 60
N/A
PFPeS 348.9 98.9 40 135
Qual
PFPeS 348.9 79.9 40 135
Quant
PFTA 713 669 9 100
Quant
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PFTA 712.9 169 30 100

Qual
PFTrDA 663 619 9 91
Quant
PFTrDA 663 169 30 100
Qual
PFUnA 563 519 5 73
Quant
PFUnA 563 218.7 20 100
Qual

6495 Transitions and MS Conditions:

Analyte Precursor Product Collision Energy Fragmentor Qualifer/Quantifier
Ion Ion Voltage (V) Voltage (V)
11Cl-PF3OUdS 631 451 24 166
N/A
4-2 FTS 327 307 20 166
Quant
4-2 FTS 327 81 30 166
Qual
6-2 FTS 427 406.9 24 166
Quant
6-2 FTS 427 80 40 166
Qual
8-2 FTS 527 507 28 166
Quant
8-2 FTS 527 80 40 166
Qual
9Cl-PF3ONS 531 351 24 166
N/A
ADONA 377 251 12 166
Quant
ADONA 377 85 12 166
Qual
d3-N-MeFOSAA 573.2 419 20 166
N/A
d5-N-EtFOSAA 589.2 419 20 166
N/A
FBSA 297.99 78 28 166
N/A
FHXSA 398 78.1 30 166
N/A
HFPO-DA 285.1 184.9 5 166
Quant
HFPO-DA 285.1 169 5 166
Qual
M2-4-2-FTS 328.99 309.2 20 166
N/A
M2-6-2-FTS 428.99 409.2 24 166
N/A
M2-8-2-FTS 528.99 509 28 166
N/A
M2PFDA 514.9 469.9 5 166
N/A
M2PFHxA 315 270 4 166
N/A
M2PFOA 415 370 4 166
N/A
M2PFTA 715 670 9 166
N/A
M3HFPO-DA 287 169 2 166
N/A
M3PFBA 216 171.8 4 166
N/A
M3PFBS 301.9 80 45 166
N/A
M3PFHxS 401.9 80 49 166
N/A
M4PFHpA 367 322 4 166
N/A
M5PFHxA 318 273 4 166
N/A
M5PFPeA 268 223 8 166
N/A
M6PFDA 519 474 4 166
N/A
M7PFUnA 570 525 5 166
N/A
M8FOSA 506 78 36 166
N/A
M8PFOA 421 376 5 166
N/A
M8PFOS 506.9 80 50 166
N/A
M9PFNA 472 427 4 166
N/A
MPFBA 217 172 8 166
N/A
MPFDoA 615 570 5 166
N/A
MPFOS 502.9 80 60 166
N/A
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N-EtFOSAA 584 525.9 20 166

Qual
N-EtFOSAA 584 418.9 20 166
Quant
NFDHA 201 85 14 166
N/A
N-MeFOSAA 570 482.9 16 166
Qual
N-MeFOSAA 570 418.9 20 166
Quant
PFBA 213 169 8 166
N/A
PFBS 298.9 98.9 29 166
Qual
PFBS 298.9 80 45 166
Quant
PFDA 513 469 4 166
Quant
PFDA 513 169 16 166
Qual
PFDoA 613 569 5 166
Quant
PFDoA 613 268.7 20 166
Qual
PFDS 598.9 99 60 166
Qual
PFDS 598.9 80 80 166
Quant
PFEESA 315 135 24 166
N/A
PFHpA 362.9 319 8 166
Quant
PFHpA 362.9 169 20 166
Qual
PFHpS 448.9 98.7 44 166
Qual
PFHpS 448.9 79.7 52 166
Quant
PFHxA 313 268.9 8 166
Quant
PFHxA 313 119 18 166
Qual
PFHxS 398.9 99 45 166
Qual
PFHxS 398.9 80 49 166
Quant
PFMBA 279 85.1 8 166
N/A
PFMPA 229 85.1 12 166
N/A
PFNA 463 419 4 166
Quant
PFNA 463 169 17 166
Qual
PFNS 548.9 98.9 40 166
Qual
PFNS 548.9 79.9 40 166
Quant
PFOA 413 369 4 166
Quant
PFOA 413 169 12 166
Qual
PFOS 498.9 99 50 166
Qual
PFOS 498.9 80 50 166
Quant
PFOSA 497.9 77.9 36 166
Quant
PFOSA 497.9 47.9 80 166
Qual
PFPeA 263 218.9 8 166
N/A
PFPeS 348.9 98.9 40 166
Qual
PFPeS 348.9 79.9 40 166
Quant
PFTA 713 669 9 166
Quant
PFTA 712.9 169 30 166
Qual
PFTrDA 663 619 9 166
Quant
PFTrDA 663 169 30 166
Qual
PFUnA 563 519 5 166
Quant
PFUnA 563 218.7 20 166
Qual
 CONTEST, A PACE ANALYTICAL LABORATORY SOP 454 PFAS Water Isotope Dilution Author: BLH
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Appendix B

Analyte Acronym Isotope Dilution

Analogue
11- Chloroeicosafluoro-3-oxaundecane-1-sulfonic 11Cl-PF3OUdS M8PFOS
acid
9- Chlorohexadecafluoro-3-oxanonane-1- 9Cl-PF3ONS M8PFOS
sulfonic acid
4,8-Dioxa-3H-perfluorononanoic acid ADONA M4PFHpA
Hexafluoropropylene oxide dimer acid HFPO-DA M3HFPO-DA
Nonafluoro-3,6-dioxaheptanoic acid NFDHA M5PFHxA
Perfluorobutanoic acid PFBA MPFBA
Perfluorobutanesulfonic acid PFBS M3PFBS
1H,1H, 2H, 2H-Perfluorodecane sulfonic acid 8:2FTS M2-8:2FTS
Perfluorodecanoic acid PFDA M6PFDA
Perfluorododecanoic acid PFDoA MPFDoA
Perfluoro(2-ethoxyethane)sulfonic acid PFEESA M3PFBS
Perfluoroheptanesulfonic acid PFHpS M8PFOS
Perfluoroheptanoic acid PFHpA M4PFHpA
1H,1H, 2H, 2H-Perfluorohexane sulfonic acid 4:2FTS M2-4:2FTS
Perfluorohexanesulfonic acid PFHxS M3PFHxS
Perfluorohexanoic acid PFHxA M5PFHxA
Perfluoro-3-methoxypropanoic acid PFMPA MPFBA
Perfluoro-4-methoxybutanoic acid PFMBA M5PFPeA
Perfluorononanoic acid PFNA M9PFNA
1H,1H, 2H, 2H-Perfluorooctane sulfonic acid 6:2FTS M2-6:2FTS
Perfluorooctanesulfonic acid PFOS M8PFOS
Perfluorooctanoic acid PFOA M8PFOA
Perfluoropentanoic acid PFPeA M5PFPeA
Perfluoropentanesulfonic acid PFPeS M3PFHxS
Perfluoroundecanoic acid PFUnA M7PFUnA

N-ethyl perfluorooctanesulfonamidoacetic acid NEtFOSAA d5-N-EtFOSAA

N‐methyl perfluorooctanesulfonamidoacetic acid NMeFOSAA d3-N-MeFOSAA
Perfluoro‐1‐butanesulfonamide FBSA M5PFHxA
Perfluoro‐1‐hexanesulfonamide FHxSA M8PFOA

Perfluorotetradecanoic acid PFTA M2PFTA

Perfluorotridecanoic acid PFTrDA M2PFTA
CONTEST, A PACE ANALYTICAL LABORATORY SOP 454 PFAS Water Isotope Dilution Author: BLH
39 Spruce Street Doc 454. Revision No. 7
East Longmeadow, MA 01028-0591 Effective Date: 06/30/2021
Page 23 of 37

Perfluorooctanesulfonamide FOSA M8FOSA

Perfluorononanesulfonic acid PFNS M8PFOS

Perfluorodecanesulfonic acid PFDS M3PFBS

Appendix C

Requirement Specification Acceptance Criteria

Demonstration of Extract and analyze 7 Percent relative standard

precision replicate laboratory deviation must be
fortified blanks at the </=20%.
mid-range of the
calibration.

Demonstration of Calculate mean recovery Mean recovery within 70-

accuracy for replicated used in 130% of the true value.
demonstration of
precision.

MDL Confirmation Extract and analyze 9 Calculated MDL and

blanks, and 9 laboratory MDL-b is < Proposed
fortified blanks at the reporting limit.
proposed reporting limit Calculation done using
over three days. 40CFR.

Calibration Verification Analyze a mid-level QCS Results must be within

after each initial 70-130% of the true
calibration. value.
CONTEST, A PACE ANALYTICAL LABORATORY SOP 454 PFAS Water Isotope Dilution Author: BLH
39 Spruce Street Doc 454. Revision No. 7
East Longmeadow, MA 01028-0591 Effective Date: 06/30/2021
Page 24 of 37

Appendix D
 CONTEST, A PACE ANALYTICAL LABORATORY SOP 454 PFAS Water Isotope Dilution Author: BLH
39 Spruce Street Doc 454. Revision No. 7
East Longmeadow, MA 01028-0591 Effective Date: 06/30/2021
Page 25 of 37

Appendix E

Table B-15. Per- and Polyfluoroalkyl Substances (PFAS) Using Liquid Chromatography Pace SOP
Tandem Mass Spectrometry (LC/MS/MS) With Isotope Dilution or Internal Standard Section
Quantification in Matrices Other Than Drinking Water
QC Check Minimum Frequency Acceptance Corrective Flagging Comments SOP SECTION
Criteria Action Criteria
Aqueous Sample Each sample and Solid Phase NA. NA. Samples with > 1% solids 7.0 Procedure &
Preparation associated batch QC Extraction (SPE) may require Summary, Scope
samples. must be used unless centrifugation prior to & Application
samples are known SPE extraction.
to contain high Pre-screening of separate
PFAS aliquots of aqueous
concentrations (e.g., samples is
Aqueous Film recommended.
Forming Foam
(AFFF)
formulations). Inline
SPE is acceptable.
Entire sample plus
bottle rinsate must
be extracted using
SPE.
Known high PFAS
concentration
samples require
serial dilution be
performed in
duplicate.
Documented project
approval is needed
for samples
prepared by serial
dilution as opposed
to SPE.
Solid Sample Each sample and Entire sample NA. NA. NA. Soil SOP: 7.5.1
Preparation associated batch QC received by the
samples. laboratory must be
homogenized prior
to subsampling.
Biota Sample Each sample and Sample prepared as NA. NA. NA. N/A
Preparation associated batch QC defined by the
samples. project (e.g., whole
fish versus filleted
fish).
QC Check Minimum Frequency Acceptance Corrective Flagging Comments SOP SECTION
Criteria Action Criteria
 CONTEST, A PACE ANALYTICAL LABORATORY SOP 454 PFAS Water Isotope Dilution Author: BLH
39 Spruce Street Doc 454. Revision No. 7
East Longmeadow, MA 01028-0591 Effective Date: 06/30/2021
Page 26 of 37

AFFF and AFFF Each sample and Each field sample NA. NA. Adsorption onto bottle is N/A
Mixture Samples associated batch QC must be prepared in negligible compared to
Preparation samples. duplicate (equivalent sample concentration so
to matrix duplicate). subsampling is allowed.
Serial dilutions must Multiple dilutions will most
be performed to likely have to be reported
achieve the lowest in order to achieve the
LOQ possible for lowest LOQ possible for
each analyte. each analyte.
Sample Cleanup Each sample and ENVI-CarbTM or NA. Flagging is not Cleanup should reduce Soil:7.5.10
Procedure associated batch QC equivalent must be appropriate. bias from matrix
samples. used on each interferences.
Not applicable to sample and batch
AFFF and AFFF QC sample.
Mixture Samples.
Mass Calibration Instrument must have
Calibrate the mass If the mass Flagging is not Problem must be 7.3.2
a valid mass scale of the MS with calibration appropriate. corrected. No samples
calibration prior to any
calibration fails, then may be analyzed under a
sample analysis. compounds and recalibrate. If failing mass calibration.
Mass calibration is
procedures it fails again, The mass calibration is
verified after each
described by the consult updated on an as-needed
mass calibration, prior
manufacturer. manufacture basis (e.g., QC failures,
to initial calibration
Mass calibration r instructions ion masses fall outside of
(ICAL). range must bracket on corrective the ±0.5 amu of the true
the ion masses of maintenance value, major instrument
interest. The most . maintenance is
recent mass performed, or the
calibration must be instrument is moved).
used for every
acquisition in an
analytical run.
Mass calibration
must be verified to
be ±0.5 amu of the
true value, by
acquiring a full scan
continuum mass
spectrum of a PFAS
stock standard.
QC Check Minimum Frequency Acceptance Corrective Flagging Comments SOP SECTION
Criteria Action Criteria
Mass Spectral Each analyte, A minimum of 10 NA. Flagging is not NA. 7.3.1
Acquisition Rate Extracted Internal spectra scans are appropriate.
Standard (EIS) acquired across
Analyte. each
chromatographic
peak.
Calibration, All analytes. Standards NA. Flagging is not Standards containing 7.6.2
Calibration containing both appropriate. both branched and linear
Verification, and branched and linear isomers are to be used
Spiking Standards isomers must be during method validation
used when and when reestablishing
 CONTEST, A PACE ANALYTICAL LABORATORY SOP 454 PFAS Water Isotope Dilution Author: BLH
39 Spruce Street Doc 454. Revision No. 7
East Longmeadow, MA 01028-0591 Effective Date: 06/30/2021
Page 27 of 37

commercially retention times, to ensure

available. the total response is
PFAS method quantitated for that
analytes may analyte.
consist of both Technical grade
branched and linear standards cannot be used
isomers, but for quantitative analysis.
quantitative
standards that
contain the linear
and branched
isomers do not exist
for all method
analytes.
For PFAS that do
not have a
quantitative
branched and linear
standard, identify
the branched
isomers by
analyzing a
qualitative standard
that includes both
linear and branched
isomers and
determine retention
times, transitions
and transition ion
ratios. Quantitate
samples by
integrating the total
response (i.e.,
accounting for
peaks that are
identified as linear
and branched
isomers) and relying
on the initial
calibration that uses
the linear isomer
quantitative
standard.
QC Check Minimum Frequency Acceptance Corrective Flagging Comments SOP SECTION
Criteria Action Criteria
Sample PFAS All analytes detected The chemical NA. PFAS identified For example: Ion Ratio = 7.6.6
Identification in a sample. derivation of the ion with Ion ratios (quant ion abundance/
transitions must be that fail confirm ion abundance)
documented. A acceptance Calculate the average
minimum of two ion criteria must be ratio (A) and standard
transitions flagged. deviation (SD) using the
(Precursor → quant Any ICAL standards. An
 CONTEST, A PACE ANALYTICAL LABORATORY SOP 454 PFAS Water Isotope Dilution Author: BLH
39 Spruce Street Doc 454. Revision No. 7
East Longmeadow, MA 01028-0591 Effective Date: 06/30/2021
Page 28 of 37

ion and precursor quantitation ion acceptance range of ratio

→ confirmation ion) peak that does could be within A ±3SD
and the ion not meet the for confirmation of
transitions ratio per maximization detection.
analyte are required criteria shall be
for confirmation. included in the
Exception is made summed
for analytes where integration and
two transitions do the resulting
not exist (PFBA and data flagged as
PFPeA). “estimated,
Documentation of biased high”.
the primary and
confirmation
transitions and the
ion ratio is required.
In-house
acceptance criteria
for evaluation of ion
ratios must be used
and must not
exceed 50- 150%.
Signal to Noise
Ratio (S/N) must be
≥ 10 for all ions used
for quantification
and must be ≥ 3 for
all ions used for
confirmation.
Quant ion and
confirmation ion
must be present and
must maximize
simultaneously (±2
seconds).
QC Check Minimum Frequency Acceptance Corrective Flagging Comments SOP SECTION
Criteria Action Criteria
Ion Transitions Every field sample, In order to avoid NA. Flagging is not NA. Appendix A
(Precursor-> standard, blank, and biasing results high appropriate
Product) QC sample. due to known
interferences for
some transitions,
the following
transitions must be
used for the
quantification of the
following analytes:
PFOA: 413 → 369
PFOS: 499 → 80
PFHxS: 399 → 80
PFBS: 299 → 80
4:2 FTS: 327 → 307
 CONTEST, A PACE ANALYTICAL LABORATORY SOP 454 PFAS Water Isotope Dilution Author: BLH
39 Spruce Street Doc 454. Revision No. 7
East Longmeadow, MA 01028-0591 Effective Date: 06/30/2021
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6:2 FTS: 427 → 407

8:2 FTS: 527 → 507
NEtFOSAA: 584 →
419
NMeFOSAA: 570 →
419
If these transitions
are not used, the
reason must be
technically justified
and documented
(e.g., alternate
transition was used
due to observed
interferences).
QC Check Minimum Frequency Acceptance Corrective Flagging Comments SOP SECTION
Criteria Action Criteria
Initial Calibration At instrument set-up The isotopically Correct Flagging is not No samples shall be 7.6.4 & 7.3.5
(ICAL) and after ICV or CCV labeled analog of an problem, appropriate. analyzed until ICAL has
failure, prior to analyte (Extracted then repeat passed.
sample analysis. Internal Standard ICAL. External Calibration is not
Analyte) must be allowed for any analyte.
used for quantitation Calibration can be linear
if commercially (minimum of 5 standards)
available (Isotope or quadratic (minimum of
Dilution 6 standards); weighting is
Quantitation). allowed.
Commercial PFAS
standards available
as salts are
acceptable providing
the measured mass
is corrected to the
neutral acid
concentration.
Results shall be
reported as the
neutral acid with
appropriate CAS
number.
If a labeled analog is
not commercially
available, the
Extracted Internal
Standard Analyte
with the closest
retention time or
chemical similarity
to the analyte must
be used for
quantitation.
(Internal Standard
 CONTEST, A PACE ANALYTICAL LABORATORY SOP 454 PFAS Water Isotope Dilution Author: BLH
39 Spruce Street Doc 454. Revision No. 7
East Longmeadow, MA 01028-0591 Effective Date: 06/30/2021
Page 30 of 37

Quantitation)
Analytes must be
within 70-130% of
their true value for
each calibration
standard.
(continued next
page)
QC Check Minimum Frequency Acceptance Corrective Flagging Comments SOP SECTION
Criteria Action Criteria
Initial Calibration ICAL must meet one 7.3.6
(ICAL) of the two options
(Continued) below:
Option 1: The RSD
of the RFs for all
analytes must be ≤
20%.
Option 2: Linear or
non- linear
calibrations must
2
have r ≥ 0.99 for
each
analyte.
Retention Time Once per ICAL and at Position shall be set NA. NA. Calculated for each 7.6.5
window position the beginning of the using the midpoint analyte and EIS.
establishment analytical sequence. standard of the ICAL
curve when ICAL is
performed.
On days when ICAL
is not performed, the
initial CCV is used.
Retention Time Every field sample, RT of each analyte Correct NA. Calculated for each 7.6.5
(RT) window width standard, blank, and and EIS analyte problem and analyte and EIS.
QC sample. must fall within 0.4 reanalyze
minutes of the samples.
predicted retention
times from the daily
calibration
verification or, on
days when ICAL is
performed, from the
midpoint standard of
the ICAL.
Analytes must elute
within 0.1 minutes of
the associated EIS.
This criterion applies
only to analyte and
labeled analog
pairs.
QC Check Minimum Frequency Acceptance Corrective Flagging Comments SOP SECTION
 CONTEST, A PACE ANALYTICAL LABORATORY SOP 454 PFAS Water Isotope Dilution Author: BLH
39 Spruce Street Doc 454. Revision No. 7
East Longmeadow, MA 01028-0591 Effective Date: 06/30/2021
Page 31 of 37

Criteria Action Criteria

Instrument Prior to analysis and Analyte Correct Flagging is not No samples shall be 7.4.1
Sensitivity Check at least once every 12 concentrations must problem, appropriate. analyzed until ISC has
(ISC) hours. be at LOQ; rerun ISC. If met acceptance criteria.
concentrations must problem ISC can serve as the
be within ±30% of persists, initial daily CCV.
their true values. repeat ICAL.
Initial Calibration Once after each ICAL, Analyte Correct Flagging is not No samples shall be 7.3.7
Verification (ICV) analysis of a second concentrations must problem, appropriate. analyzed until calibration
source standard prior be within ±30% of rerun ICV. If has been verified.
to sample analysis. their true value. problem
persists,
repeat ICAL.
Continuing Prior to sample Concentration of Immediately If reanalysis Results may not be 7.4.2, 7.4.5
Calibration analysis, after every analytes must range analyze two cannot be reported without valid
Verification (CCV) 10 field samples, and from the LOQ to the additional performed, CCVs.
at the end of the mid-level calibration consecutive data must be Instrument Sensitivity
analytical sequence. concentration. CCVs. If qualified and Check (ISC) can serve as
Analyte both pass, explained in a bracketing CCV.
concentrations must samples the Case
be within ±30% of may be Narrative.
their true value. reported Apply Q-flag to
without all results for
reanalysis. If the specific
either fails, analyte(s) in all
or if two samples since
consecutive the last
CCVs acceptable
cannot be calibration
run, perform verification.
corrective
action(s) and
repeat CCV
and all
associated
samples
since last
successful
CCV.
Alternately,
recalibrate if
necessary;
then
reanalyze all
associated
samples
since the
last
acceptable
CCV.
QC Check Minimum Frequency Acceptance Corrective Flagging Comments SOP SECTION
Criteria Action Criteria
 CONTEST, A PACE ANALYTICAL LABORATORY SOP 454 PFAS Water Isotope Dilution Author: BLH
39 Spruce Street Doc 454. Revision No. 7
East Longmeadow, MA 01028-0591 Effective Date: 06/30/2021
Page 32 of 37

Instrument Blanks Immediately following Concentration of If Flagging is No samples shall be 7.4.3

the highest standard each analyte must acceptance only analyzed until instrument
analyzed and daily be ≤ ½ the LOQ. criteria are appropriate in blank has met
prior to sample Instrument Blank not met after cases when acceptance criteria.
analysis. must contain EIS to the highest the sample Note: Successful analysis
enable quantitation calibration cannot be following the highest
of contamination. standard, reanalyzed and standard analyzed
calibration when there is determines the highest
must be no more concentration that
performed sample left. carryover does not occur.
using a When the highest
lower standard analyzed is not
concentratio part of the calibration
n for the curve, it cannot be used
highest to extend out the
standard calibration range, it is
until used only to document a
acceptance higher concentration at
criteria is which carryover still does
met. not occur.
If sample
concentratio
ns exceed
the highest
allowed
standard
and the
sample(s)
following
exceed this
acceptance
criteria (>1/2
LOQ), they
must be
reanalyzed.
QC Check Minimum Frequency Acceptance Corrective Flagging Comments SOP SECTION
Criteria Action Criteria
Extracted Internal Every field sample, Added to solid Correct Apply Q-flag Failing analytes shall be 8.2.7, 8.2.10
Standard (EIS) standard, blank, and sample prior to problem. If and discuss in thoroughly documented in
Analytes QC sample. extraction. Added to required, re- the Case the Case Narrative.
aqueous samples, extract and Narrative only EIS should be 96% (or
into the original reanalyze if reanalysis greater) purity. When the
container, prior to associated confirms impurity consists of the
extraction. field and QC failures in unlabeled analyte, the
For aqueous samples. exactly the EIS can result in a
samples prepared If recoveries same manner. background artifact in
by serial dilution are every sample, standard
instead of SPE, acceptable and blank, if the EIS is
added to final for QC fortified at excessive
dilution of samples samples, but concentrations.
prior to analysis. not field
Extracted Internal samples, the
 CONTEST, A PACE ANALYTICAL LABORATORY SOP 454 PFAS Water Isotope Dilution Author: BLH
39 Spruce Street Doc 454. Revision No. 7
East Longmeadow, MA 01028-0591 Effective Date: 06/30/2021
Page 33 of 37

Standard Analyte field

recoveries must be samples
within 50% to 150% must be re-
of ICAL extracted
midpoint standard and
area or area analyzed
measured in the (greater
initial CCV on days dilution may
when an ICAL is not be needed).
performed. Samples
may be re-
extracted
and
analyzed
outside of
hold times,
as
necessary
for corrective
action
associated
with QC
failure.
Method Blank (MB) One per preparatory No analytes Correct If reanalysis Results may not be 8.2.1
batch. detected >½ LOQ or problem. If cannot be reported without a valid
> 1/10th the amount required, re- performed, MB.
measured in any extract and data must be Flagging is only
sample or 1/10th the reanalyze qualified and appropriate in cases
regulatory limit, MB and all explained in where the samples
whichever is QC samples the Case cannot be reanalyzed.
greater. and field Narrative.
samples Apply B-flag to
processed all results for
with the the specific
contaminate analyte(s) in all
d blank. samples in the
Samples associated
may be re- preparatory
extracted batch.
and
analyzed
outside of
hold times,
as
necessary
for corrective
action
associated
with QC
failure.
Examine the
project-
 CONTEST, A PACE ANALYTICAL LABORATORY SOP 454 PFAS Water Isotope Dilution Author: BLH
39 Spruce Street Doc 454. Revision No. 7
East Longmeadow, MA 01028-0591 Effective Date: 06/30/2021
Page 34 of 37

specific
requirement
s. Contact
the client as
to additional
measures to
be taken.
QC Check Minimum Frequency Acceptance Corrective Flagging Comments SOP SECTION
Criteria Action Criteria
Laboratory Control One per preparatory Blank spiked with all Correct If reanalysis Results may not be 8.2.2
Sample (LCS) batch. analytes at a problem, cannot be reported without a valid
concentration ≥ LOQ then re- performed, LCS.
and extract and data must be Flagging is only
≤ the mid-level reanalyze qualified and appropriate in cases
calibration the LCS and explained in where the samples
concentration. all samples the Case cannot be reanalyzed.
A laboratory must in the Narrative.
use the DoD/DOE associated Apply Q-flag to
QSM Appendix C preparatory specific
Limits for batch batch for analyte(s) in all
control if project failed samples in the
limits are not analytes if associated
specified. sufficient preparatory
If the analyte(s) are sample batch.
not listed, use in- material is
house LCS limits if available.
project limits are not Samples
specified. may be re-
extracted
and
analyzed
outside of
hold times,
as
necessary
for corrective
action
associated
with QC
failure.
Examine the
project-
specific
requirement
s. Contact
the client as
to additional
measures to
be taken.
Matrix Spike (MS) One per preparatory Sample spiked with Examine the For the specific For matrix evaluation 8.2.3
batch. Not required all analytes at a project- analyte(s) in only. If MS results are
for aqueous samples concentration ≥ LOQ specific the parent outside the limits, the
 CONTEST, A PACE ANALYTICAL LABORATORY SOP 454 PFAS Water Isotope Dilution Author: BLH
39 Spruce Street Doc 454. Revision No. 7
East Longmeadow, MA 01028-0591 Effective Date: 06/30/2021
Page 35 of 37

prepared by serial and requirement sample, apply data shall be evaluated to

dilution instead of≤ the mid-level s. Contact J-flag if determine the source(s)
SPE. calibration the client as acceptance of difference (i.e., matrix
concentration. to additional criteria are not effect or analytical error).
A laboratory must measures to met and
use the DoD/DOE be taken. explain in the
QSM Appendix C Case
Limits for batch Narrative.
control if project
limits are not
specified.
If the analyte(s) are
not listed, use in-
house LCS limits if
project limits are not
specified.
QC Check Minimum Frequency Acceptance Corrective Flagging Comments SOP SECTION
Criteria Action Criteria
Matrix Spike For MSD: One per For MSD: Sample Examine the For the specific The data shall be 8.2.4
Duplicate (MSD) or preparatory batch. spiked with all project- analyte(s) in evaluated to determine
Matrix Duplicate For MD: Each analytes at a specific the parent the source of difference.
(MD) aqueous sample concentration ≥ LOQ requirement sample, apply For Sample/MD: RPD
prepared by serial and s. Contact J-flag if criteria only apply to
dilution instead of ≤ the mid-level the client as acceptance analytes whose
SPE. calibration to additional criteria are not concentration in the
concentration. measures to met and sample is ≥ LOQ.
A laboratory must be taken. explain in the The MD is a second
use the DoD/DOE Case aliquot of the field sample
QSM Appendix C Narrative. that has been prepared
Limits for batch by serial dilution.
control if project
limits are not
specified.
If the analyte(s) are
not listed, use in-
house LCS limits if
project limits are not
specified.
RPD ≤ 30%
(between MS and
MSD or sample and
MD).
 CONTEST, A PACE ANALYTICAL LABORATORY SOP 454 PFAS Water Isotope Dilution Author: BLH
39 Spruce Street Doc 454. Revision No. 7
East Longmeadow, MA 01028-0591 Effective Date: 06/30/2021
Page 36 of 37

Post Spike Sample Only applies to Spike all analytes When Flagging is not When analyte N/A
aqueous samples reported as < LOQ analyte appropriate. concentrations are
prepared by serial into the dilution that concentratio calculated as < LOQ,
dilution instead of the result for that ns are results may not be
SPE that have analyte is reported calculated reported without
reported value of from. The spike as < LOQ, acceptable post spike
< LOQ for analyte(s). must be at the LOQ and the recoveries.
concentration to be spike
reported for this recovery
sample as < LOQ. does not
When analyte meet the
concentrations are acceptance
calculated as < criteria, the
LOQ, the post spike sample,
for that analyte must sample
recover within 70- duplicate,
130% of its true and post
value. spike
sample must
be
reanalyzed
at
consecutivel
y higher
dilutions until
the criteria is
met.

Appendix F

PFAS Aqueous sample centrifugation protocol (for DoD work in GW/SW/NPW)

Preliminary considerations:
The DoD QSM5.3, Table B-15, states that “[aqueous] samples with >1% solids may require centrifugation
prior to SPE extraction.” Samples should only be centrifuged when the suspended solids content appears
visually high enough, by chemist inspection, that it would cause the SPE cartridge to clog. It is expected
that the solid phase remains in the container when rinsing the container walls with the polar elution solvent.
Rinsing the container walls would therefore also include rinsing of the solids. If removing the solvent
disrupts the solid phase significantly, the container can be centrifuged again before removing the solvent
for use during the elution step of the SPE procedure. When the sample has significant solids, the laboratory
should account for the weight or volume displaced by the solids in the initial sample volume determination.
One or more rinses of polar solvent can be used for quantitative transfers. Rinse the sample bottle and cap
with elution solvent, pour the solvent from each rinse through the SPE cartridge during the elution step, and
collect the filtrate for analysis. Bring to a quantitative final volume with the final injection solvent and
vortex well. Whether or not an individual sample will require centrifugation for proper preparation will be
determined and documented by the preparation analyst.

Procedure:
1. Inspect the sample and consider the necessity of centrifuging. Consider any visible indications of
particulate matter including settled solids collected on the bottom of the container, cloudiness and/or dark
color of the sample, suspended solids within the sample, increased viscosity, etc. If uncertain, seek a
second opinion from another analyst, supervisor, or operations director.
CONTEST, A PACE ANALYTICAL LABORATORY SOP 454 PFAS Water Isotope Dilution Author: BLH
39 Spruce Street Doc 454. Revision No. 7
East Longmeadow, MA 01028-0591 Effective Date: 06/30/2021
Page 37 of 37

2. If, in the judgement of the preparation analyst, a sample requires centrifugation the analyst will
contemporaneously make a note on the prep batch log indicating this fact.

3. Spike samples requiring centrifugation in the same manner and with the same standard volume as
samples which will not be centrifuged.

4. Label a 500mL polypropylene centrifuge bottle with the sample ID for each sample that will be
centrifuged. Set them in an appropriate rack with the caps removed.

5. Vigorously shake the spiked sample and then quickly pour into the labeled centrifuge bottle. Try
to ensure that the original sample bottle is devoid of any solid material. Be careful to avoid spilling sample
during the transfer process. Tightly cap each centrifuge bottle after transfers are complete.

6. Transfer capped centrifuge bottles to centrifuge, ensuring that the centrifuge carousel is
symmetrically balanced. Close top and centrifuge at 2200 RPM for 20 minutes.

7. Remove centrifuge bottles and decant the centrifuged liquid off of the condensed solids and back
into the original sample bottle. Try to avoid transferring any of the condensed solids from the centrifuge
bottle back to the original sample bottle, while maximizing the amount of liquid decanted off of the solid
portion. Take weight for initial volume of sample without solids in original container.

8. Extract the decanted sample as normal alongside un- centrifuged samples, up to the bottle rinse
and elution steps.

9. When the SPE cartridges have been dried, rinse the original sample bottle as normal.
Additionally, add 4mL of Methanol (MeOH) to each centrifuge bottle to rinse the inside of the centrifuge
bottles as well as the cap. If the condensed solids become re-suspended while rinsing the centrifuge tubes,
re-centrifugation may be required. Using a transfer pipet or mechanical pipet, transfer the MeOH rinse
from the centrifuge bottle into the SPE cartridge and elute with the original sample bottle rinse into a 15mL
conical centrifuge tube.

10. Add an additional volume of MeOH to the elution of all batch QC samples (MB/LCS/LCSD) to
match the volume used for elution for any centrifuged sample in the prep batch. Typically, this will mean
that 4mL of clean MeOH will be added directly to the SPE reservoir and eluted with the normal bottle
rinses.

11. Concentrate samples down to ~0.5 mL and reconstitute as outlined in sample extraction procedure.

12. Add a case narrative onto the work order indicating which samples had to be centrifuged.
CONTEST, A PACE ANALYTICAL LABORATORY SOP 466 PFAS Soils Isotope Dilution Author: BLM.
39 Spruce Street Doc 466. Revision No. 7
East Longmeadow, MA 01028-0591 Effective Date: 06/30/2021
Page 1 of 37

Determination of Selected Per and Polyfluorinated Alkyl

Substances (PFAS) Soil/Solid Samples Isotope Dilution by
Liquid Chromatography/Tandem Mass Spectrometry
(LC/MS/MS)

Approved:

_________________________ _________________________

Tod Kopyscinski Katherine Allen

Laboratory Director QA Officer

Revision Number: 7

DRAFT
 CONTEST, A PACE ANALYTICAL LABORATORY SOP 466 PFAS Soils Isotope Dilution Author: BLM.
39 Spruce Street Doc 466. Revision No. 7
East Longmeadow, MA 01028-0591 Effective Date: 06/30/2021
Page 2 of 37

Change Record

Revision Date Responsible Description of Change

Person
0 2/16/2018 Brianna Original
McLaughlin
Kelly Fellows
1 11/13/2018 Katherine Updates from Sept 2018 NH audit: Sec 3.0 (addition pres. requirements added), Sec 7.2.1
Allen/KAF (deleted ref. to section 7.7.2), Sec 7.2.4 (deleted ref. to sections 7.2.5.5 and 7.7.2), Sec 7.2.5
(surrogate should changed to surrogate must), Sec 7.4.2 (deletion of blank spike duplicate), Sec
7.4.3 (addition of DI water rinse after methanol), Sec 7.4.4 (changed to vortex for 1 min), Sec.
7.5.1.1 (blank subtraction replaces blank extraction), Sec 7.6.3 (deleted to prove instrument is
contamination free), Sec 7.6.6 (deleted ref to App C), and deleted Sec 8.2.12 (deleted response
QC check). Sec 3.1(added temperature note), Sec 3.2(added ‘of collection’), Sec 3.3(Added
Appendix C), added ‘Appendix C’ Sec 7.2.8 added section on isotope dilution technique.
2 03/12/2020 Brianna Update to make entire new procedure: Entire SOP edited.
McLaughlin
James
Constantino
3 05/18/2020 Brianna Updates to procedure: Section 1.0 – added CAS #’s to chart, section 7.5: includes
Henriquez clarification on how to use Envi-carb cleanup cartridges. Edited section 8.2.9 to
reflect correct limits from previous sections.
4 07/15/2020 Brianna Updates: Removal of reagent (Section 5.0) Change to concentration of CCV
Henriquez standards (Section 7.2.6.4). Multiple changes to extraction procedure 7.5.
5 0817/2020 Brianna Updates from July 2020 NH audit: Edit to section 7.3.7 and 7.3.8 to updated
Henriquez procedure, Section 7.3.5 change should to shall. Edit to Section 8.2.1 and 7.4.3 typo
changed all > to <, added Sec 8.2.11 for IDOC and Section 8.2.12 for CDOC.
6 02/26/2021 Katherine Allen Update for MA DEP approval: Sec 3.5, 8.2.13 and Appendix A (field blank is
optional for client), Sec 13.13 (replace EPA 537 with EPA 533), and addition of
13.19 (addition of ASTM D7968-17 ref).
7 06/30/2021 Brianna Updates to procedure: Overall: References to Con-Test changed to Pace. Section 1.1: Updated
Henriquez summary to reflect procedure changes, changed initial WT and FV, added 6495 instrument.
Analyte RLs changed. Section 4: Removed pH paper from reagents (in supplies), removed
cartridge and filter disc. Section 5: 20MM mobile phase changed to 5MM, 70:30 changed to
96:4, mobile phase expiration changed. Section 7: Removed section on separate internal
standard. Section 7.1: Sampling instructions changed. Section 7.2.6: Standards stored at room
temperature. Added 24PAR and certain individual analyte stocks for separate spike sourcing.
Standard prep and calibration tables were changed. CCVs changed to only be run at the mid-
level (level 4) after an opening low-level. Section 7.3: Updated calibration method, technique,
and requirements (removal of IS/surrogate now functioning as both surrogate and IS). Section
7.3.4: Branched compound summation added. Section 7.4.1: Requirement added of IBL and
low-level CCV every 12 hours. Section 7.5: Extraction method changed completely after the
base-addition step. Removed syringe filtering, changed envi-carb instructions, added blow
down step. Removed centrifuge step. Surrogate and spike amounts changed and are now always
the same. Amount of acid changed. Section 7.5.3: replaced Dup with MSD, Section 7.5.10:
changed final to 5mL, Section 7.6.2.1: TPFOA branched check section added. Section 7.7.2:
Instrument method conditions changed. Cell accelerator voltage moved to this section. Section
8.2.3: MS/MSD spike levels held constant. 8.2.6: Updated how isotope dilution technique is
calculated. Appendices: Removed Appendices A and C, changing other appendices letters.
Appendix A: Now has transitions and conditions for both instruments, and qualifier transitions.

Distribution/Training List

See Employee Training Record File for signed training statements for trained users.
 CONTEST, A PACE ANALYTICAL LABORATORY SOP 466 PFAS Soils Isotope Dilution Author: BLM.
39 Spruce Street Doc 466. Revision No. 7
East Longmeadow, MA 01028-0591 Effective Date: 06/30/2021
Page 3 of 37

1.0 SUMMARY, SCOPE, AND APPLICATION

1.1 This method is used to analyze soil and product/solid samples for selected per- and
polyfluorinated alkyl substances (PFAS). A sample of 5.5 grams of soil/solid sample is fortified with spikes
and surrogates. A 2 mL aliquot of the sample is removed and pulled through an Envi-Carb cartridge and
then concentrated to a final volume of 5mL. All samples are analyzed using an Agilent 6470 or 6495 Triple
Quad LC/MS (LC/MS/MS) system. Target analytes are identified by comparing mass spectra and retention
times to reference spectra and retention times of calibration standards. Analytes are quantitated using the
isotope dilution technique explained in the initial calibration section. The following compounds can be
identified by this method.

Analyte Acronym Standard RL Cas Number

(ug/kg)
11- 11‐chloroeicosafluoro‐3‐oxanone‐1‐ 11Cl-PF3OUdS 0.5 763051-92-9
sulfonic acid
9 -Chlorohexadecafluoro-3-oxapentane- 9Cl-PF3ONS 0.5 756426-58-1
1-sulfonic acid
4,8-Dioxa-3H-perfluorononanoic acid ADONA 0.5 919005-14-4
Hexafluoropropylene oxide dimer acid HFPO-DA 0.5 13252-13-6
Nonafluoro‐3,6‐dioxaheptanoic acid NFDHA 0.5 151772-58-6
Perfluorobutanoic acid PFBA 0.5 375-22-4
Perfluorobutanesulfonic acid PFBS 0.5 375-73-5
8:2 Fluorotelomer sulfonic acid 8:2FTS 0.5 39108-34-4
Perfluorodecanoic acid PFDA 0.5 335-76-2
Perfluorododecanoic acid PFDoA 0.5 307-55-1
Perfluoro(2-ethoxyethane)sulfonic acid PFEESA 0.5 113507-82-7
Perfluoroheptanesulfonic acid PFHpS 0.5 375-92-8
Perfluoroheptanoic acid PFHpA 0.5 375-85-9
4:2 Fluorotelomer sulfonic acid 4:2FTS 0.5 757124-72-4
Perfluorohexanesulfonic acid PFHxS 0.5 355-46-4
Perfluorohexanoic acid PFHxA 0.5 307-24-4
Perfluoro-3-methoxypropanoic acid PFMPA 0.5 377-73-1
Perfluoro-4-methoxybutanoic acid PFMBA 0.5 863090-89-5
Perfluorononanoic acid PFNA 0.5 375-95-1
6:2 Fluorotelomer sulfonic acid 6:2FTS 0.5 27619-97-2
Perfluorooctanesulfonic acid PFOS 0.5 1763-23-1
Perfluorooctanoic acid PFOA 0.5 335-67-1
Perfluoropentanoic acid PFPeA 0.5 2706-90-3
Perfluoropentanesulfonic acid PFPeS 0.5 2706-91-4
 CONTEST, A PACE ANALYTICAL LABORATORY SOP 466 PFAS Soils Isotope Dilution Author: BLM.
39 Spruce Street Doc 466. Revision No. 7
East Longmeadow, MA 01028-0591 Effective Date: 06/30/2021
Page 4 of 37

Perfluoroundecanoic acid PFUnA 0.5 2058-94-8

N-ethyl perfluorooctanesulfonamidoacetic NEtFOSAA 0.5 2991-50-6
acid
N‐methyl perfluorooctanesulfonamidoacetic NMeFOSAA 0.5 2355-31-9
acid
Perfluoro‐1‐butanesulfonamide FBSA 0.5 30334-69-1

Perfluoro‐1‐hexanesulfonamide FHxSA 0.5 41997-13-1

Perfluorotetradecanoic acid PFTA 0.5 376-06-7

Perfluorotridecanoic acid PFTrDA 0.5 72629-94-8

Perfluorooctanesulfonamide FOSA 0.5 754-91-6

Perfluorononanesulfonic acid PFNS 0.5 68259-12-1

Perfluorodecanesulfonic acid PFDS 0.5 335-77-3

2.0 INTERFERENCES
2.1 Standards and samples should not come into contact with glass other than standards purchased in
glass ampules. PFAS commonly adsorb to the surface and could result in recovery discrepancies.

2.2 Matrix interferences may be caused by co-extracted contaminants present in the sample.
2.3 Method interferences may be caused by contaminants in solvents, reagents, and other sample
processing hardware.
2.3.1 Other common lab supplies that are associated with PFAAs and should be avoided where possible:
aluminum foil, permanent marker, and PTFE.
2.3.1.1 To eliminate any residual PTFE from the Agilent LC, an inline filter column has been installed to
reduce any background contamination prior to sample introduction into the system. See Equipment and
Supplies Section 4.0.
2.3.2 Organic contaminants can pose a threat of interference due to the high quantities of de-
chlorinating agent added to samples.
2.3.3 Contamination levels should be monitored and all blanks should be free from interferences (less
than 1/2 the MRL) in all Laboratory Reagent Blanks (LRB).
2.3.4 Blank subtraction is not permitted in this method.
2.3.5 There is a possibility of matrix effects due to co-extracted organic material. When high levels of
TOC are present, this can affect the ionization of 4:2 FTS considerably.
CONTEST, A PACE ANALYTICAL LABORATORY SOP 466 PFAS Soils Isotope Dilution Author: BLM.
39 Spruce Street Doc 466. Revision No. 7
East Longmeadow, MA 01028-0591 Effective Date: 06/30/2021
Page 5 of 37

3.0 SAMPLE PRESERVATION/STORAGE/HOLDING TIME

3.1 Samples should be collected in a wide mouth polypropylene bottle fitted with a polypropylene
screw cap capable of holding at least 50 grams of soil.
3.2 Samples cannot exceed 10°C during the first 48 hours following sample collection. Samples must
be received at or below 10°C OR have ice remaining in the cooler.
3.3 Prior to extraction, samples must be stored at or below 6°C and cannot be frozen.
3.4 Samples collected must be extracted within 28 days. Extracted samples must be run within 28
days of extraction and remain stored at room temperature.
3.5 Clients may choose to send a field blank if they want wish to as it is optional. Field reagent blanks
only need to be run and reported if there are analytes at or above the MRL in any associated field samples.

4.0 EQUIPMENT & SUPPLIES

4.1 Triple Quad LC/MS System

4.2 Inline Delay column
4.2.1 Agilent Zorbax Eclipse Plus C18 3.0x50mm 1.8-micron P.N. PFCDELAY or equivalent
4.3 Analytical Column
4.3.1 Agilent Zorbax Eclipse Plus C18 3.0x50mm 1.8-micron P.N. 959757-02 or equivalent
4.4 Auto-pipettors: 0-10uL, 10-100uL, 100-1000uL
4.5 Polypropylene pipet tip: 0-10uL, 10-100uL, 100-1000uL
4.6 Polypropylene transfer pipets
4.7 Polypropylene graduated cylinder: 10mL, 50mL, 100mL, 1000mL
4.8 Vials: 2ml polypropylene vials or 2mL glass vials with polypropylene inserts
4.9 Caps: 11mm polypropylene snap caps
4.10 Sample containers: 250ml wide mouth polypropylene containers
4.11 Polypropylene centrifuge tubes
4.12 Hydrion pH paper strips: 1.0-11.0 B
4.13 Balance: Analytical, capable of accurately weighing 0.0001g
4.14 N-Evap concentrator system
4.15 Polypropylene inserts
4.16 Vortex
CONTEST, A PACE ANALYTICAL LABORATORY SOP 466 PFAS Soils Isotope Dilution Author: BLM.
39 Spruce Street Doc 466. Revision No. 7
East Longmeadow, MA 01028-0591 Effective Date: 06/30/2021
Page 6 of 37

5.0 REAGENTS & STANDARDS

5.1 Reagent Water: interferent free

5.2 Methanol: LC/MS Grade
5.3 Nitrogen: Ultra high purity
5.4 Ammonium Acetate: LC/MS Grade
5.5 Stock Standard Solutions: Purchased as either certified solutions or neat standards.
5.6 Surrogate and ESI-L Low concentration tuning mix purchased as certified solutions
5.7 5 mM Ammonium Acetate reagent water: Prepared by adding 1 mL of 5 M ammonium acetate to
a final volume of 1000 mLs DI water.
5.8 Ammonium Hydroxide
5.9 Acetic acid, glacial
5.10 Agilent ESI-L Low Concentration Tuning Mix
5.11 96:4 Methanol:DI Water- Made fresh every 2 days
5.12 Envi-Carb cartridge or equivalent

6.0 SAFETY
See Material Safety Data Sheets (MSDS) and Pace Chemical Hygiene Plan.

7.0 PROCEDURE

7.1 Sampling
7.1.1 Samples are to be collected in lab-provided plastic containers. Guidance for sampling is obtained
through Pace corporate website or preferably local project authorities/municipalities.
7.2 Surrogate/SpikePreparation
7.2.1 All standards must be documented in Element and have Certificate of Analysis forms attached
electronically. All information should be documented and each standard should be given an Element Standard
ID#.
7.2.2 Standards may be received in purchased glass ampoules but any transfer or dilution must be stored
in polypropylene vials with Non-PTFE caps.
7.2.3 All standards purchased from Wellington come pre-treated with sodium hydroxide for compound
stability. If making standards from solid, standards must be stored under basic condition to prevent
esterification of fluorinated carboxylic acids. See calculation 1 in section 12.1.
7.2.4 PFAS Surrogate Preparation
7.2.4.1 All purchased surrogate stock standards are to be stored until expiration date provided by
the manufacturer at 4°C.
CONTEST, A PACE ANALYTICAL LABORATORY SOP 466 PFAS Soils Isotope Dilution Author: BLM.
39 Spruce Street Doc 466. Revision No. 7
East Longmeadow, MA 01028-0591 Effective Date: 06/30/2021
Page 7 of 37

Surrogate Standards
Compound Abbreviation PDS, ng/mL
Perfluoro‐n‐[1,2,3,4‐ MPFBA 1000
13C4]butanoic acid
Perfluoro‐n‐[1,2,3,4,5‐ M5PFPeA 1000
13C5]pentanoic acid
Sodium perfluoro‐1‐[2,3,4‐ M3PFBS 929
13C3]butanesulfonate
Sodium 1H,1H,2H,2H‐ M2‐4:2FTS 935
perfluoro‐1‐[1,2‐13C2]hexane
sulfonate
Perfluoro‐n‐[1,2,3,4,6‐ M5PFHxA 1000
13C5]hexanoic acid
Perfluoro‐n‐[1,2,3,4‐ M4PFHpA 1000
13C4]heptanoic acid
Sodium perfluoro‐1‐[1,2,3‐ M3PFHxS 946
13C3]hexanesulfonate
Sodium 1H,1H,2H,2H‐ M2‐6:2FTS 949
perfluoro‐1‐[1,2‐13C2]‐octane
sulfonate
Perfluoro‐n‐[13C8]octanoic M8PFOA 1000
acid
Perfluoro‐n‐[13C9]nonanoic M9PFNA 1000
acid
Sodium perfluoro‐ M8PFOS 957
[13C8]octanesulfonate
Sodium 1H,1H,2H,2H‐ M2‐8:2FTS 958
perfluoro‐1‐[1,2‐13C2]‐decane
sulfonate
Perfluoro‐n‐[1,2,3,4,5,6‐ M6PFDA 1000
13C6]decanoic acid
Perfluoro‐n‐[1,2,3,4,5,6,7‐ M7PFUnA 1000
13C7]undecanoic acid
2,3,3,3‐Tetrafluoro‐2‐ M3HFPO‐DA 1000
(1,1,2,2,3,3,3‐
heptafluoropropoxy‐13C3‐
propanoic acid
Perfluoro‐n‐[1,2‐ M2PFDoA 1000
13C2]dodecanoic acid
Perfluoro‐n‐[12‐ M2PFTA 1000
13C2]tetradecanoic acid
Perfluoro‐1‐ M8FOSA 1000
[13C8]octanesulfonamidoaceti
c acid
N‐methyl‐d3‐perfluoro‐1‐ d3‐N‐MeFOSAA 1000
octansulfonamidoacetic acid
N‐ethyl‐d5‐perfluoro‐1‐ d5‐N‐EtFOSAA 1000
octansulfonamidoacetic acid
Surrogate standards are ordered from Wellington Labs part PFAC-24ES
CONTEST, A PACE ANALYTICAL LABORATORY SOP 466 PFAS Soils Isotope Dilution Author: BLM.
39 Spruce Street Doc 466. Revision No. 7
East Longmeadow, MA 01028-0591 Effective Date: 06/30/2021
Page 8 of 37

Compound Abbreviation PDS, ng/mL

2,3,3,3‐Tetrafluoro‐2‐ M3HFPO‐DA 1000

(1,1,2,2,3,3,3‐
heptafluoropropoxy‐13C3‐
propanoic acid
Surrogate standard is ordered from Wellington Labs part M3HFPO-DA

7.2.6 Analyte Primary Dilution Standard Preparation

7.2.6.1 All purchased PFAA spike standard stock standards are to be stored until expiration date
provided by manufacturer at 4°C.
7.2.6.2 Prepared and in use PFAA stock standard solutions should be stored at room temperature
and be vortexed prior to usage. These standards will expire 2 months after preparation date or
manufacturer’s expiration date, whichever comes first.
Analyte Standards
Compound Vendor Concentration of Standard
(ng/ mL)
PFAC-30PAR Wellington 1000*
PFAC-24PAR Wellington 2000*
PFEESA Wellington 50000*
NFDHA Wellington 50000
PFMPA Wellington 50000
PFMBA Wellington 50000
HPFO-DA Wellington 50000
NaDONA Wellington 50000*
9ClPF3ONS Wellington 50000*
11ClPF3OUdS Wellington 50000*
FBSA Wellington 50000
FHxSA Wellington 50000
*Individual analyte concentration may vary due to amount of anion present in solution. All
calculations must use the anion concentration, not the salt concentration. See Calculation 2 in section
12.2.
7.2.6.3 PFEESA, NFDHA, PFMPA, PFMBA, HPFO-DA, NaDONA, 9ClPF3ONS, 11ClPF3OUdS,
FBSA, and FHxSA are not included in the 24PAR mixture and are mixed into a separate
“supplemental” 1000 ng/ mL stock. See below table for mixture.
7.2.6.4 The Supplemental Stock is used to make a 500ppb spike dilution with 24PAR so that a
different lot is used for spiking. The calibration stock is made using 30PAR and directly adding
PFEESA, NFDHA, PFMPA, and PFMBA, which are missing from that mixture. See below table
for prep instructions:
CONTEST, A PACE ANALYTICAL LABORATORY SOP 466 PFAS Soils Isotope Dilution Author: BLM.
39 Spruce Street Doc 466. Revision No. 7
East Longmeadow, MA 01028-0591 Effective Date: 06/30/2021
Page 9 of 37

Stock Dilution Prep Table

Volume of Volume of Final Final
Compound/Standard Mixtures Methanol Volume Concentrati
(µL) on
(µL) (µL)
(ng/mL)
Supplemental 100µL – HPFO-DA 4000µL 5000µL 1000
Stock
100µL – NaDONA
100µL – 9ClPF3ONS
100µL – 11ClPF3OUdS
100µL – PFEESA
100µL – PFMBA
100µL – PFMPA
100µL – NFDHA
100µL – FBSA
100µL – FHxSA
500 ng/ mL 1250µL – PFAC24PAR 1250µL 5000µL 500
Spike
2500µL – Supplemental Stock
100 ng/ mL 250µL – PFAC24PAR 4250µL 5000µL 100
Spike
500µL – Supplemental Stock
100 ng/ mL 500µL – PFAC30PAR 4460µL 5000µL 100
Cal Stock 10µL – PFEESA
10µL – PFMBA
10µL – PFMPA
10µL – NFDHA

7.2.6.5 The calibration is prepared as follows using the stock dilutions prepared above:
Calibration Table
Volume Volume Volume Volume of Volume of Volume of Final Final
Surrogate Stock M3HFPODA 96:4 Volume
100ppb 30PAR Supplementa (µL) Surrogate Stock Methanol: Concentration
l Water (µL) (µL)
Stock Stock (ng/mL)
Standard (µL) Standard Stock
(µL) Standard
(µL)

12.5 0 0 25 25 4937.5 5000 0.25*

25 0 0 25 25 4925 5000 0.5*

50 0 0 25 25 4900 5000 1.0*

125 0 0 25 25 4825 5000 2.5*

250 0 0 25 25 4700 5000 5.0*

CONTEST, A PACE ANALYTICAL LABORATORY SOP 466 PFAS Soils Isotope Dilution Author: BLM.
39 Spruce Street Doc 466. Revision No. 7
East Longmeadow, MA 01028-0591 Effective Date: 06/30/2021
Page 10 of 37

0 50 50 25 25 4850 5000 10.0*

0 125 125 25 25 4700 5000 25.0*

0 250 250 25 25 4450 5000 50.0*

*Individual analyte concentration may vary due to the amount of anion present in solution. All
calculations must use the anion concentration, not the salt concentration. See Calculation 2 in section
12.2
7.2.6.6 Continuing calibration verification (CCVs) standards are made at the mid-level, identically
to the 4th calibration level above. The ICV/QCS is made similarly, just like the 5th calibration level
above, except instead of calibration stock 100 ppb Spike is used as shown in the table below:
QCS/ICV Preparation Table
Standard Volume Standards Volume of Final Final
Name 96:4 Volume
Concentration
Methanol:Wa
(µL)
ter (µL) (ng/mL)
QCS/ICV 25µL- PFAC-24ES 4700µL 5000µL 5.0
25µL- M3HPFO-DA
Surrogate Dilution
250µL- 100ppb Spike

7.3 Initial Calibration Criteria

7.3.1 All analytes must first be product ion optimized with the LC/MS/MS system using the
MassHunter Optimizer program to determine optimal fragmentor and collision cell energy for applicable
ions. This optimization should be done using a high-level standard for each analyte and using all of the LC
parameters used in the analytical method. A level 4 standard must then be run to identify all retention time
windows for all compounds of interest (See Section 7.7.2). A minimum of 10 spectra scans are acquired
across each chromatographic peak. See Appendix A for further analyte MS/MS conditions.
7.3.2 Prior to initial calibrations, and when the instrument is having difficulty passing regular
calibrations, a mass calibration will be performed using Agilent ESI-L Low Concentration Tuning Mix.
The MassHunter program has an autotune feature that performs the mass calibration using this mix and
verifies it in a report.
7.3.3 A calibration is to be run when continuing calibration checks or surrogates do not pass QC criteria.
A calibration should also be performed when any hardware is changed or major instrument maintenance is
performed.
7.3.4 The initial calibration must contain a minimum of five points the lowest point being at or below
the MRL. A minimum of six points is required for a quadratic calibration. The total of the branched and
linear isomers must be used for calibration for the following target analytes: PFOS, PFHxS, NEtFOSAA,
NMeFOSAA.
7.3.5 The LC/MS/MS system is to be calibrated using the isotope dilution technique. Therefore, isotope
dilution analogues are added at a constant concentration in all standards prior to injection. Either linear or
quadratic regression can be used, but it must always be forced through zero and can be concentration
weighted. Forcing through zero allows for more sensitivity to detect background contamination within the
system. The calibration shall be done using the same LC conditions as the samples (See Section 7.7.2).
Because the isotope dilution analogues are added in equal concentrations, calibrate for them using an
average response factor. Not all analytes have an exact mass-labeled analogue, in which case the closest
analogue is used (either by chemical properties or retention time). See Appendix B for the list of isotope
analogues and corresponding target analytes.
CONTEST, A PACE ANALYTICAL LABORATORY SOP 466 PFAS Soils Isotope Dilution Author: BLM.
39 Spruce Street Doc 466. Revision No. 7
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Page 11 of 37

7.3.5.1 The isotope dilution technique utilizes extracted compounds to serve as a traditional
internal standard. In this case, the extracted analogues must pass criteria listed in section 8.2.6. The
analogue is then used as the internal standard compound for associated target analytes.
7.3.6 Calibration levels for linear or non-linear analyte targets must have a r2 ≥ 0.99 for each analyte and
the recovery for each analyte must be within 70-130% of the true value. Surrogate and internal standards
must have an RSD of the RFs for all analytes of ≤20%.
7.3.7 A quality control standard (QCS) will serve as an initial calibration verification (ICV) and be run
following initial calibration and all subsequent calibrations. The ICV shall be prepared from a separate lot
of stock standard mix. This sample must be run following a calibration or quarterly, whichever comes first.
The accepted values for the ICV are 70-130% of the true value for each analyte.
7.3.8 After any changes to calibration instrument parameters, an initial demonstration of capability
(IDOC) for the procedure and instrumentation should be performed. See Appendix C.

7.4 Continuing Calibration

7.4.1 An instrument blank (IBL) and a low-level instrument sensitivity check (ISC) must be run before
any other injections and once every 12 hours. The results must be between 70-130% of the true value for all
analytes. The IBL should have no hits greater than ½ the MRL. The ISC can serve as the opening CCV for
the day.
7.4.2 Prior to samples analysis, a low-level continuing calibration verification (CCV) must be run. After
every ten field samples a subsequent CCV must be at the same level as the Level 4 calibration point. A
closing CCV must also be run at the end of each analysis. The requirements for the CCVs are 70-130% of
the true value for method analytes.
7.4.3 An instrument blank is required to be run following analysis of the highest-level standard analyzed
(after a calibration in this case). One is also required daily prior to sample analysis. All analytes must be
<½ the MRL in order to pass.
7.4.4 All isotope dilution analogues (surrogates) must have a recovery between 50-150%
7.4.5 In the case of CCV failure, two consecutive CCVs should be immediately run. If these pass,
analysis can continue. Otherwise, a calibration or tuning and re-analysis of affected samples is required.
7.4.6 A checktune will be run as needed to verify MS operating criteria. This is run through the
MassHunter program using Agilent ESI-L Low Concentration Tuning Mix. If criteria are out of spec, the
parameters set forth in the Agilent 6400 Series Triple Quadrupole LC/MS System Quick Start Guide must
be followed to adjust values. If re-run of checktune does not pass, an autotune must be run and the
instrument must be recalibrated.
7.5 Sample Extraction Procedure
7.5.1 Every sample must be fully homogenized prior to weighing out for extraction.
7.5.2 Weigh out 5.5-6.0 grams of soil/solid sample into a 15mL polypropylene conical tube.
7.5.3 For every 20 field samples, a blank and a blank spike must be extracted using sand. Ideally, if
adequate sample volume is available, a matrix spike and matrix spike duplicate should be included in every
batch.
7.5.4 All polypropylene equipment including graduated cylinders and sample transfer lines/reservoirs
should be washed prior to using methanol.
7.5.5 Add 25µL of M3HFPO-DA Surrogate Stock and 25uL of PFAC24-ES to each sample (and 25uL
500 ppb spike to the BS and MS samples as appropriate), recap and vortex for 1 minute to evenly distribute.
7.5.6 Add 10mL 96:4 reagent methanol:water to the sample.
7.5.7 Add 20µL of Ammonium Hydroxide to raise the samples’ pH to 9-10 and vortex for 2 minutes.
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7.5.8 Prime Envi-Carb cartridge by rinsing twice with straight methanol (approx. 10 mL methanol).
Transfer a 2 mL aliquot of filtered sample to Envi-carb and pull through into clean centrifuge tube. Rinse
cartridge with 4 mLs of methanol.
7.5.9 To the filtered material, add 10uL of acetic acid and verify the pH to be between 5-6 with pH paper.
7.5.10 Blow the extract down at room temperature to a final volume of 5mL, raising the volume with 96:4
methanol:DI water.
7.5.11 Transfer an aliquot to a 2mL vial with polypropylene insert.

7.6 Data Analysis

7.6.1 The analyst cannot extrapolate beyond the range of the calibration. However, by isotope dilution
analysis, the extract cannot be diluted. If an analyte is outside of the determined range, the sample must be
re-extracted at an appropriate dilution level.
7.6.1.1 There is extrapolation allowed only to determine if there is blank contamination. Since there is no
blank subtraction, any contamination present must be below 1/2 of the MRL for specific analyte.
7.6.1.2 If a sample exceeds the calibration range the sample must be re-extracted. This would involve diluting
the sample with reagent water to be within the calibration range and adding ammonium acetate to be at a
final concentration of 1 g/L.
7.6.1.3 Additionally, if a sample exceeds the calibration range, one or more LRB must be run until the system
meets acceptable criteria. If this occurs during an automated sequence, any subsequent samples must be
evaluated. If the over-range analytes are present in the subsequent samples at or above the RL, the samples
are considered invalid and must be re-run. If the analyte in question does not exceed the RL, the samples can
be reported.
7.6.2 Compounds that have both branched and linear isomers will be reported as total. These
compounds include PFOS, PFHxS, N-Et-FOSAA, N-Me-FOSAA and PFOA. PFOS, PFHxS, N-Et-
FOSAA, and N-Me-FOSAA have the branched and linear compounds available for quantitation. PFOA is a
special case outlined below:

7.6.2.1 PFOA will be quantitated by using a qualitative/semi-quantitative approach per EPA guidance.
Until a standard is available, the calibration will be done using the linear isomer only. A technical grade
standard will be run to identify the retention time of the branched isomer. All samples will be
quantitated using the area of both the linear and branched isomers of PFOA that may be present within the
sample. A branched isomer check for PFOA will be run with every calibration curve to verify the retention
times of the branched isomers for PFOA.

7.6.3 All analytes and surrogates will be calculated based on the initial calibration criteria.

7.6.4 All results for analytes shall be reported as the acid form of the compound.

7.6.5 Retention time windows are established once per ICAL and at the beginning of each sequence. On
days when an ICAL is not run, the initial CCV is used to set the times. All retention times of analytes and
EIS analytes must fall within 0.4 minutes of the established time. Analytes must also elute within 0.1
minutes of their respective EIS.

7.6.6 In addition to retention time identification criteria, most ions are identified by two ion transitions.
(The following ions are exceptions: PFBA, PFPeA, FBSA, FHxSA, PFMPA, PFMBA, 9Cl-PF3ONS,
11Cl-PF3OUdS, ADONA). The secondary, or qualifier ion, must have a signal to noise of 3:1. The ratio
between the qualifier and the quantifier ion must be averaged from the calibration. For samples to be valid,
the ratio of qualifier to quantifier must be +/-50% from the average ratio from the applicable calibration.
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7.7 Instrumentation Procedure

7.7.1 Before any QC or samples can be run, the HPLC must be allowed to purge for at least thirty
minutes. This purge can be done using any combination of the mobile phases, but prior to samples running,
the initial mobile phase conditions used in the method must be allowed to run for 15 minutes or until
pressure has stabilized.
7.7.2 The instrument must be stable in all parameters before a run is started. The following are the
HPLC and ESI-MS Method Conditions. Also, See Appendix A for additional MS/MS Method Conditions.

Time (min) % 5 mM Ammonium Acetate % Methanol Flow Rate

in water (mL/minute)
0.00 95 5 1.0
0.10 65 35 1.0
2.00 50 50 1.0
3.00 25 75 1.0
4.50 1 99 1.0
4.51 1 99 1.0
5.00 1 99 1.0
5.10 95 5 1.0
6.50 95 5 1.0

Injection Volume 6470 10 uL

Injection Volume 6495 5 uL
Column Compartment Temperature 40 °C
Autosampler Compartment Temperature 10 °C
Polarity Negative
Gas Temperature 250 °C
Gas Flow 11 L/min
Nebulizer 50 psi
Sheath Gas Temperature 300 °C
Sheath Gas Flow 12 L/min
Capillary Needle Voltage (Negative mode) -3000 V
Cell Accelerator Voltage 5V

7.7.3 An instrument sequence will be made. It will open with an instrument blank and a low level CCV.
After the CCV, the batch can start running. Every 10 field samples (excluding QC and FRBs) a subsequent
level 4 CCV must be run.
7.7.4 The run can end with a script to put the instrument into standby mode.

8.0 QUALITY CONTROL

8.1 Definitions

For definitions and explanations of quality control measures (blanks, LCS/QC Reference, LFB, Duplicates,
MS/MSD, etc.) refer to the Contest, A Pace Analytical Quality Assurance Manual.
CONTEST, A PACE ANALYTICAL LABORATORY SOP 466 PFAS Soils Isotope Dilution Author: BLM.
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8.2 Quality Control Measures & Acceptance Criteria

8.2.1 Method Blank

The method blank is matrix specific, and extracted with every batch or every 20 samples
(whichever is more frequent). The target compounds must be ≤1/2 the MRL, or <1/10th the amount
measured in any sample, or <1/10th the regulatory limit. If any analytes are present above this level,
the detected analytes are considered invalid for all samples extracted in that batch.

8.2.2 Laboratory Control Sample/Duplicate (LCS)

A matrix-specific LCS is extracted every 20 samples. The concentration must be ≥ LOQ and
≤ mid-range of calibration . All analytes recoveries must be within limits specified in Appendix D. If
analyte is not listed in table, acceptance criteria is to remain 50-150% until in-house limits can be
determined. Samples should be re-extracted if criteria are not met, even if outside of hold. If samples
cannot be re-extracted then the failures must be notated in the narrative.

8.2.3 Matrix Spikes

A matrix-specific MS is extracted every 20 samples. All analytes recoveries must be within limits
specified in Appendix D. If analyte is not listed in table, acceptance criteria is to remain 50-150% until
in-house limits can be determined. See calculation 3.
*Note: Matrix spike samples may display matrix bias. If the CCC and LFB samples are passing,
but the MS recoveries are outside the designated range, the recovery is deemed to be matrix biased.
A note on the unfortified sample will indicate the possibility of matrix effects being suspect.

8.2.4 Matrix spike duplicates

Extract a spiked sample duplicate every 20 samples. Matrix Spike Duplicate samples should be
calculated to have an RPD ≤30%. See calculation 4.

8.2.5 Quality Control Samples

A quality control sample must be run from a second source at least quarterly, or after an initial
calibration as an ICV. If a second source is not commercially available, a different lot number from the
same vendor is acceptable. The recoveries must be within 70-130% of the true value.

8.2.6 Isotope Dilution Analogue

Isotope dilution analogues are added to all blanks, standards, samples, and spikes. Analogue
compounds must have an area of 50-150% of the associated compound in level 4 of the calibration on
days when a calibration is run. On days when a calibration is not run, analogue compounds must have
an area of 50-150% of the associated compound in the opening instrument sensitivity check/CCV.

If the surrogate is outside these limits, the extract should be re-analyzed. If the re-analysis
passes, report re-analyzed sample. If this fails, the associated isotope performance standard should be
evaluated. The system may need recalibration or maintenance. If the CCV has surrogate out of range,
the instrument needs to be recalibrated.
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If the re-analysis fails, re-extract the sample to confirm failure if an aliquot is available. If the
re-extract fails, report both results with appropriate flagging criteria. If not enough volume is provided
to re-extract the sample then report with appropriate flags.

8.2.7 Calibration Curve

A minimum of a 5-point calibration curve (for linear regression) or a 6-point

calibration curve (for quadratic) is used to calibrate the system.

The curve must be verified with an independent standard (QCS) prior to sample
analysis, (10 ng/L). The curve will be forced through zero and may or may not
be concentration weighted.

If a peak is not properly integrated by the data system, manual integration may
be necessary. Manual integrations must comply with the Pace SOP on
Chromatographic Integration Procedures. The integration of the peaks for the
samples and quality control samples must be as consistent as possible with the
integration used with the initial calibration.

8.2.8 Continuing Calibration Verification Checks (CCVs)

The results must be between 70-130% of the true value for all analytes for the initial low level CCV
After every ten field samples, a subsequent CCV must at level 4. The requirements for the CCVs are 70-
130% of the true value. All analogue compound areas must fall within 50-150% of the appropriate
calibration point or CCV.
The ending CCV acquisition time must fall within 24 hours of the acquisition starting time of the opening
CCV with the associated analysis batch.
8.2.10 Any failures in QC require reanalysis, even if the samples in question are outside of hold.

8.2.11 Initial Demonstration of Capability (IDOC): An Initial demonstration of capability (IDOC)

must be made prior to performing any test method, and at any time there is a significant change in
instrument type, personnel, or test method.

In general, this demonstration does not test the performance of the method in real world samples, but in
applicable and available clean matrix (a sample of a matrix in which no target analytes or interferences are
present at concentrations that impact the results of a specific test method). Before any results are reported
by a new analyst they need to perform and IDOC. See Appendix C.

All demonstrations shall be documented through the use of the IDOC form, which also lists SOP, method
associated with the test, certification statement, and authorized signatures.

8.2.12 Continuing Demonstration of Capability (CDOC): An on-going demonstration of capability

must be performed on an annual basis to document the quality of the data produced. On-going data quality
checks are compared with established performance criteria to determine if the results of analyses meet the
performance standards for the method.

In general, this demonstration does not test the performance of the method in real world samples, but in
applicable and available clean matrix (a sample of a matrix in which no target analytes or interferences are
present at concentrations that impact the results of a specific test method). Extract and analyze 4 replicate
laboratory fortified blanks at level 4 of the calibration with acceptable recoveries between 70-130%.
CONTEST, A PACE ANALYTICAL LABORATORY SOP 466 PFAS Soils Isotope Dilution Author: BLM.
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All demonstrations shall be documented through the use of the CDOC form, which also lists SOP, method
associated with the test, certification statement, and authorized signatures.

8.2.13 Field Reagent Blank:

Clients may choose to send a field blank if they want wish to as it is optional. Field reagent blanks only
need to be run and reported if there are analytes at or above the MRL in any associated field samples. Any
analyte peaks present in field reagent blanks must be below the MRL of that analyte.

9.0 CORRECTIVE ACTIONS/ CONTINGENCIES OF HANDLING OUT-OF-

CONTROL DATA

9.1 Refer to Pace Quality Assurance Manual

9.2 Refer to Pace Corrective Action SOP.

10.0 POLLUTION PREVENTION

10.1 Pollution prevention encompasses any technique that reduces or eliminates the quantity or toxicity
of waste at the point of generation. Many opportunities for pollution prevention exist in laboratory
operation. EPA has established a preferred hierarchy of environmental management techniques that places
pollution prevention as the management option of first choice. Whenever feasible, laboratory personnel
should use pollution prevention techniques to address waste generation. Standards should be prepared in
volumes consistent with laboratory use to minimize the disposal of excess volumes of expired standards.

11.0 WASTE MANAGEMENT

11.1 It is the laboratory’s responsibility to comply with all federal, state, and local regulations
governing the waste management, particularly the hazardous waste identification rules and land disposal
restrictions, and to protect the air, water, and land by minimizing and controlling all releases from fume
hoods and bench operations. Also, compliance is required with any sewage discharge permits and
regulations.

11.2 Acidic samples and waste are dumped into satellite waste containers.

12.0 CALCULATIONS

12.1 Calculation 1: Adding 4 mole equivalents to standards to prevent esterification

12.2 Calculation 2: Mass of the anion

Massacid= Measured Masssalt * (Molecular Weightacid/Molecular Weightsalt)

CONTEST, A PACE ANALYTICAL LABORATORY SOP 466 PFAS Soils Isotope Dilution Author: BLM.
39 Spruce Street Doc 466. Revision No. 7
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12.3 Calculation 3: Percent Recovery

12.4 Calculation 4: Relative percent deviation calculation

12.5 Calculation 5: MRL Statistics

CONTEST, A PACE ANALYTICAL LABORATORY SOP 466 PFAS Soils Isotope Dilution Author: BLM.
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13.0 REFERENCES

13.1 Pace Analytical Chemical Hygiene Plan

13.2 Pace Analytical Quality Assurance Manual.

13.3 Pace Analytical Corrective Action SOP.

13.4 Pace Analytical Controlled Document SOP.

13.5 Agilent 1260 Infinity Binary LC Operators manual

13.6 Agilent MassHunter Study Manager

13.7 Agilent MassHunter Optimizer

13.8 MassHunter Personal Compound Database and Library Manager

13.9 Agilent 6400 Series Triple Quadrupole LC/MS System Quick Start Guide

13.10 MassHunter Data Acquisition Compliance Software Quick Start Guide

13.11 MassHunter Quantitative Analysis Compliance Software Quick Start Guide

13.12 Agilent 6000 Series LC/MS System Maintenance Guide

13.13 EPA Method 533, “Determination of Selected Per- and Polyfluoralkyl Substances in Drinking
Water by Isotope Dilution Anion Exchange Solid Phase Extraction and Liquid
Chromatography/Tandem Mass Spectrometry (LC/MS/MS)”, November 2019, EPA Document
#815-B-19-020.

13.14 Method ISO 25101:2009, “Determination of perfluorooctanesulfonate (PFOS) and

perfluorooctanoate (PFOA) – Method for unfiltered samples using solid phase extraction
and liquid chromatography/mass spectrometry”, April 30, 2009.

13.15 EPA Technical Advisory-Laboratory Analysis of Drinking Water Samples for

Perfluorooctanoic Acid (PFOA) using EPA Method 537 Rev. 1.1 EPA 815-B-16-021
September 2016

13.16 Agilent Application note by Peter JW Stone, Linda Cote, Jennifer Gushue, Robert J. Letcher
and Shaogang Chu. A Low Femtogram Target Screen Method for Perfluorinated Compounds in Food
Matrices and Potable Water Using the Agilent 6460 Triple Quadrupole LC/MS System Equipped with
Agilent Jet Stream Technology.

13.17 TNI Standard, The NELAC Institute, EL-V1-2009-ISO, 2009.

13.18 Department of Defense (DoD) Department of Energy (DOE) Consolidated Quality Systems Manual
(QSM) for Environmental Laboratories Based on ISO/IEC 17025:2005(E) ISO/IEC 17025:2017(E) and
The NELAC Institute (TNI) Standards, Volume 1, (September 2009).

13.19 ASTM Method D7968-17a, “Standard Test Method for Determination of Polyfluorinated
Compounds in Soil by Liquid Chromatography Tandem Mass Spectrometry (LC/MS/MS)”, ASTM
International, West Conshohocken, PA, 2017.
CONTEST, A PACE ANALYTICAL LABORATORY SOP 466 PFAS Soils Isotope Dilution Author: BLM.
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Appendix A

6470 Transitions and MS Conditions:

Analyte Precursor Product Collision Energy Fragmentor Qualifer/Quantifier
Ion Ion Voltage (V) Voltage (V)
11Cl-PF3OUdS 631 451 24 100
N/A
4-2 FTS 327 307 20 120
Quant
4-2 FTS 327 81 30 120
Qual
6-2 FTS 427 406.9 24 135
Quant
6-2 FTS 427 80 40 125
Qual
8-2 FTS 527 507 28 145
Quant
8-2 FTS 527 80 40 170
Qual
9Cl-PF3ONS 531 351 24 100
N/A
ADONA 377 251 12 100
Quant
ADONA 377 85 12 100
Qual
d3-N-MeFOSAA 573.2 419 20 114
N/A
d5-N-EtFOSAA 589.2 419 20 104
N/A
FBSA 297.99 78 28 115
N/A
FHXSA 398 78.1 30 135
N/A
HFPO-DA 285.1 184.9 5 150
Quant
HFPO-DA 285.1 169 5 150
Qual
M2-4-2-FTS 328.99 309.2 20 135
N/A
M2-6-2-FTS 428.99 409.2 24 160
N/A
M2-8-2-FTS 528.99 509 28 170
N/A
M2PFDA 514.9 469.9 5 102
N/A
M2PFHxA 315 270 4 66
N/A
M2PFOA 415 370 4 69
N/A
M2PFTA 715 670 9 100
N/A
M3HFPO-DA 287 169 2 50
N/A
M3PFBA 216 171.8 4 56
N/A
M3PFBS 301.9 80 45 100
N/A
M3PFHxS 401.9 80 49 100
N/A
M4PFHpA 367 322 4 102
N/A
M5PFHxA 318 273 4 68
N/A
M5PFPeA 268 223 8 120
N/A
M6PFDA 519 474 4 81
N/A
M7PFUnA 570 525 5 73
N/A
M8FOSA 506 78 36 125
N/A
M8PFOA 421 376 5 65
N/A
M8PFOS 506.9 80 50 100
N/A
M9PFNA 472 427 4 85
N/A
MPFBA 217 172 8 60
N/A
MPFDoA 615 570 5 79
N/A
MPFOS 502.9 80 60 180
N/A
N-EtFOSAA 584 525.9 20 115
Qual
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N-EtFOSAA 584 418.9 20 115

Quant
NFDHA 201 85 14 115
N/A
N-MeFOSAA 570 482.9 16 115
Qual
N-MeFOSAA 570 418.9 20 115
Quant
PFBA 213 169 8 60
N/A
PFBS 298.9 98.9 29 100
Qual
PFBS 298.9 80 45 100
Quant
PFDA 513 469 4 81
Quant
PFDA 513 169 16 100
Qual
PFDoA 613 569 5 79
Quant
PFDoA 613 268.7 20 100
Qual
PFDS 598.9 99 60 100
Qual
PFDS 598.9 80 80 100
Quant
PFEESA 315 135 24 110
N/A
PFHpA 362.9 319 8 72
Quant
PFHpA 362.9 169 20 72
Qual
PFHpS 448.9 98.7 44 44
Qual
PFHpS 448.9 79.7 52 52
Quant
PFHxA 313 268.9 8 8
Quant
PFHxA 313 119 18 18
Qual
PFHxS 398.9 99 45 100
Qual
PFHxS 398.9 80 49 100
Quant
PFMBA 279 85.1 8 55
N/A
PFMPA 229 85.1 12 55
N/A
PFNA 463 419 4 66
Quant
PFNA 463 169 17 66
Qual
PFNS 548.9 98.9 40 165
Qual
PFNS 548.9 79.9 40 165
Quant
PFOA 413 369 4 69
Quant
PFOA 413 169 12 69
Qual
PFOS 498.9 99 50 100
Qual
PFOS 498.9 80 50 100
Quant
PFOSA 497.9 77.9 36 125
Quant
PFOSA 497.9 47.9 80 100
Qual
PFPeA 263 218.9 8 60
N/A
PFPeS 348.9 98.9 40 135
Qual
PFPeS 348.9 79.9 40 135
Quant
PFTA 713 669 9 100
Quant
PFTA 712.9 169 30 100
Qual
PFTrDA 663 619 9 91
Quant
PFTrDA 663 169 30 100
Qual
PFUnA 563 519 5 73
Quant
PFUnA 563 218.7 20 100
Qual
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6495 Transitions and MS Conditions:

Analyte Precursor Product Collision Energy Fragmentor Qualifer/Quantifier
Ion Ion Voltage (V) Voltage (V)
11Cl-PF3OUdS 631 451 24 166
N/A
4-2 FTS 327 307 20 166
Quant
4-2 FTS 327 81 30 166
Qual
6-2 FTS 427 406.9 24 166
Quant
6-2 FTS 427 80 40 166
Qual
8-2 FTS 527 507 28 166
Quant
8-2 FTS 527 80 40 166
Qual
9Cl-PF3ONS 531 351 24 166
N/A
ADONA 377 251 12 166
Quant
ADONA 377 85 12 166
Qual
d3-N-MeFOSAA 573.2 419 20 166
N/A
d5-N-EtFOSAA 589.2 419 20 166
N/A
FBSA 297.99 78 28 166
N/A
FHXSA 398 78.1 30 166
N/A
HFPO-DA 285.1 184.9 5 166
Quant
HFPO-DA 285.1 169 5 166
Qual
M2-4-2-FTS 328.99 309.2 20 166
N/A
M2-6-2-FTS 428.99 409.2 24 166
N/A
M2-8-2-FTS 528.99 509 28 166
N/A
M2PFDA 514.9 469.9 5 166
N/A
M2PFHxA 315 270 4 166
N/A
M2PFOA 415 370 4 166
N/A
M2PFTA 715 670 9 166
N/A
M3HFPO-DA 287 169 2 166
N/A
M3PFBA 216 171.8 4 166
N/A
M3PFBS 301.9 80 45 166
N/A
M3PFHxS 401.9 80 49 166
N/A
M4PFHpA 367 322 4 166
N/A
M5PFHxA 318 273 4 166
N/A
M5PFPeA 268 223 8 166
N/A
M6PFDA 519 474 4 166
N/A
M7PFUnA 570 525 5 166
N/A
M8FOSA 506 78 36 166
N/A
M8PFOA 421 376 5 166
N/A
M8PFOS 506.9 80 50 166
N/A
M9PFNA 472 427 4 166
N/A
MPFBA 217 172 8 166
N/A
MPFDoA 615 570 5 166
N/A
MPFOS 502.9 80 60 166
N/A
N-EtFOSAA 584 525.9 20 166
Qual
N-EtFOSAA 584 418.9 20 166
Quant
NFDHA 201 85 14 166
N/A
N-MeFOSAA 570 482.9 16 166
Qual
N-MeFOSAA 570 418.9 20 166
Quant
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PFBA 213 169 8 166

N/A
PFBS 298.9 98.9 29 166
Qual
PFBS 298.9 80 45 166
Quant
PFDA 513 469 4 166
Quant
PFDA 513 169 16 166
Qual
PFDoA 613 569 5 166
Quant
PFDoA 613 268.7 20 166
Qual
PFDS 598.9 99 60 166
Qual
PFDS 598.9 80 80 166
Quant
PFEESA 315 135 24 166
N/A
PFHpA 362.9 319 8 166
Quant
PFHpA 362.9 169 20 166
Qual
PFHpS 448.9 98.7 44 166
Qual
PFHpS 448.9 79.7 52 166
Quant
PFHxA 313 268.9 8 166
Quant
PFHxA 313 119 18 166
Qual
PFHxS 398.9 99 45 166
Qual
PFHxS 398.9 80 49 166
Quant
PFMBA 279 85.1 8 166
N/A
PFMPA 229 85.1 12 166
N/A
PFNA 463 419 4 166
Quant
PFNA 463 169 17 166
Qual
PFNS 548.9 98.9 40 166
Qual
PFNS 548.9 79.9 40 166
Quant
PFOA 413 369 4 166
Quant
PFOA 413 169 12 166
Qual
PFOS 498.9 99 50 166
Qual
PFOS 498.9 80 50 166
Quant
PFOSA 497.9 77.9 36 166
Quant
PFOSA 497.9 47.9 80 166
Qual
PFPeA 263 218.9 8 166
N/A
PFPeS 348.9 98.9 40 166
Qual
PFPeS 348.9 79.9 40 166
Quant
PFTA 713 669 9 166
Quant
PFTA 712.9 169 30 166
Qual
PFTrDA 663 619 9 166
Quant
PFTrDA 663 169 30 166
Qual
PFUnA 563 519 5 166
Quant
PFUnA 563 218.7 20 166
Qual
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Appendix B
Analyte Acronym Isotope Dilution
Analogue
11- Chloroeicosafluoro-3-oxaundecane-1-sulfonic 11Cl-PF3OUdS M8PFOS
acid
9- Chlorohexadecafluoro-3-oxanonane-1- 9Cl-PF3ONS M8PFOS
sulfonic acid
4,8-Dioxa-3H-perfluorononanoic acid ADONA M4PFHpA
Hexafluoropropylene oxide dimer acid HFPO-DA M3HFPO-DA
Nonafluoro-3,6-dioxaheptanoic acid NFDHA M5PFHxA
Perfluorobutanoic acid PFBA MPFBA
Perfluorobutanesulfonic acid PFBS M3PFBS
1H,1H, 2H, 2H-Perfluorodecane sulfonic acid 8:2FTS M2-8:2FTS
Perfluorodecanoic acid PFDA M6PFDA
Perfluorododecanoic acid PFDoA MPFDoA
Perfluoro(2-ethoxyethane)sulfonic acid PFEESA M3PFBS
Perfluoroheptanesulfonic acid PFHpS M8PFOS
Perfluoroheptanoic acid PFHpA M4PFHpA
1H,1H, 2H, 2H-Perfluorohexane sulfonic acid 4:2FTS M2-4:2FTS
Perfluorohexanesulfonic acid PFHxS M3PFHxS
Perfluorohexanoic acid PFHxA M5PFHxA
Perfluoro-3-methoxypropanoic acid PFMPA MPFBA
Perfluoro-4-methoxybutanoic acid PFMBA M5PFPeA
Perfluorononanoic acid PFNA M9PFNA
1H,1H, 2H, 2H-Perfluorooctane sulfonic acid 6:2FTS M2-6:2FTS
Perfluorooctanesulfonic acid PFOS M8PFOS
Perfluorooctanoic acid PFOA M8PFOA
Perfluoropentanoic acid PFPeA M5PFPeA
Perfluoropentanesulfonic acid PFPeS M3PFHxS
Perfluoroundecanoic acid PFUnA M7PFUnA

N-ethyl perfluorooctanesulfonamidoacetic acid NEtFOSAA d5-N-EtFOSAA

N‐methyl perfluorooctanesulfonamidoacetic acid NMeFOSAA d3-N-MeFOSAA
Perfluoro‐1‐butanesulfonamide FBSA M5PFHxA
Perfluoro‐1‐hexanesulfonamide FHxSA M8PFOA

Perfluorotetradecanoic acid PFTA M2PFTA

Perfluorotridecanoic acid PFTrDA M2PFTA
CONTEST, A PACE ANALYTICAL LABORATORY SOP 466 PFAS Soils Isotope Dilution Author: BLM.
39 Spruce Street Doc 466. Revision No. 7
East Longmeadow, MA 01028-0591 Effective Date: 06/30/2021
Page 24 of 37

Perfluorooctanesulfonamide FOSA M8FOSA

Perfluorononanesulfonic acid PFNS M8PFOS

Perfluorodecanesulfonic acid PFDS M3PFBS

Appendix C

Requirement Specification Acceptance Criteria

Demonstration of Extract and analyze 4 Percent relative standard

precision (Requirement of replicate laboratory deviation must be
Analyst IDOC and fortified blanks at the </=20%.
CDOC) level 4 of the calibration.

Demonstration of Calculate mean recovery Mean recovery within 70-

accuracy (Requirement of for replicated used in 130% of the true value.
Analyst IDOC and demonstration of
CDOC) precision.

MDL Confirmation Extract and analyze 9 Calculated MDL and

blanks, and 9 laboratory MDL-b is < Proposed
fortified blanks at the reporting limit.
proposed reporting limit
over three days.

Calibration Verification Analyze a Level 5 QCS Results must be within

after each initial 70-130% of the true
calibration. value.
CONTEST, A PACE ANALYTICAL LABORATORY SOP 466 PFAS Soils Isotope Dilution Author: BLM.
39 Spruce Street Doc 466. Revision No. 7
East Longmeadow, MA 01028-0591 Effective Date: 06/30/2021
Page 25 of 37

Appendix D
 CONTEST, A PACE ANALYTICAL LABORATORY SOP 466 PFAS Soils Isotope Dilution Author: BLM.
39 Spruce Street Doc 466. Revision No. 7
East Longmeadow, MA 01028-0591 Effective Date: 06/30/2021
Page 26 of 37

Appendix E

Table B-15. Per- and Polyfluoroalkyl Substances (PFAS) Using Liquid Chromatography Pace SOP
Tandem Mass Spectrometry (LC/MS/MS) With Isotope Dilution or Internal Standard Section
Quantification in Matrices Other Than Drinking Water
QC Check Minimum Frequency Acceptance Corrective Flagging Comments SOP SECTION
Criteria Action Criteria
Aqueous Sample Each sample and Solid Phase NA. NA. Samples with > 1% solids 7.0 Procedure &
Preparation associated batch QC Extraction (SPE) may require Summary, Scope
samples. must be used unless centrifugation prior to & Application
samples are known SPE extraction.
to contain high Pre-screening of separate
PFAS aliquots of aqueous
concentrations (e.g., samples is
Aqueous Film recommended.
Forming Foam
(AFFF)
formulations). Inline
SPE is acceptable.
Entire sample plus
bottle rinsate must
be extracted using
SPE.
Known high PFAS
concentration
samples require
serial dilution be
performed in
duplicate.
Documented project
approval is needed
for samples
prepared by serial
dilution as opposed
to SPE.
Solid Sample Each sample and Entire sample NA. NA. NA. Soil SOP: 7.5.1
Preparation associated batch QC received by the
samples. laboratory must be
homogenized prior
to subsampling.
Biota Sample Each sample and Sample prepared as NA. NA. NA. N/A
Preparation associated batch QC defined by the
samples. project (e.g., whole
fish versus filleted
fish).
QC Check Minimum Frequency Acceptance Corrective Flagging Comments SOP SECTION
Criteria Action Criteria
AFFF and AFFF Each sample and Each field sample NA. NA. Adsorption onto bottle is N/A
Mixture Samples associated batch QC must be prepared in negligible compared to
Preparation samples. duplicate (equivalent sample concentration so
to matrix duplicate). subsampling is allowed.
 CONTEST, A PACE ANALYTICAL LABORATORY SOP 466 PFAS Soils Isotope Dilution Author: BLM.
39 Spruce Street Doc 466. Revision No. 7
East Longmeadow, MA 01028-0591 Effective Date: 06/30/2021
Page 27 of 37

Serial dilutions must Multiple dilutions will most

be performed to likely have to be reported
achieve the lowest in order to achieve the
LOQ possible for lowest LOQ possible for
each analyte. each analyte.
Sample Cleanup Each sample and ENVI-CarbTM or NA. Flagging is not Cleanup should reduce Soil:7.5.10
Procedure associated batch QC equivalent must be appropriate. bias from matrix
samples. used on each interferences.
Not applicable to sample and batch
AFFF and AFFF QC sample.
Mixture Samples.
Mass Calibration Instrument must have
Calibrate the mass If the mass Flagging is not Problem must be 7.3.2
a valid mass scale of the MS with calibration appropriate. corrected. No samples
calibration prior to any
calibration fails, then may be analyzed under a
sample analysis. compounds and recalibrate. If failing mass calibration.
Mass calibration is
procedures it fails again, The mass calibration is
verified after each
described by the consult updated on an as-needed
mass calibration, prior
manufacturer. manufacture basis (e.g., QC failures,
to initial calibration
Mass calibration r instructions ion masses fall outside of
(ICAL). range must bracket on corrective the ±0.5 amu of the true
the ion masses of maintenance value, major instrument
interest. The most . maintenance is
recent mass performed, or the
calibration must be instrument is moved).
used for every
acquisition in an
analytical run.
Mass calibration
must be verified to
be ±0.5 amu of the
true value, by
acquiring a full scan
continuum mass
spectrum of a PFAS
stock standard.
QC Check Minimum Frequency Acceptance Corrective Flagging Comments SOP SECTION
Criteria Action Criteria
Mass Spectral Each analyte, A minimum of 10 NA. Flagging is not NA. 7.3.1
Acquisition Rate Extracted Internal spectra scans are appropriate.
Standard (EIS) acquired across
Analyte. each
chromatographic
peak.
Calibration, All analytes. Standards NA. Flagging is not Standards containing 7.6.2
Calibration containing both appropriate. both branched and linear
Verification, and branched and linear isomers are to be used
Spiking Standards isomers must be during method validation
used when and when reestablishing
commercially retention times, to ensure
available. the total response is
PFAS method quantitated for that
analytes may analyte.
 CONTEST, A PACE ANALYTICAL LABORATORY SOP 466 PFAS Soils Isotope Dilution Author: BLM.
39 Spruce Street Doc 466. Revision No. 7
East Longmeadow, MA 01028-0591 Effective Date: 06/30/2021
Page 28 of 37

consist of both Technical grade

branched and linear standards cannot be used
isomers, but for quantitative analysis.
quantitative
standards that
contain the linear
and branched
isomers do not exist
for all method
analytes.
For PFAS that do
not have a
quantitative
branched and linear
standard, identify
the branched
isomers by
analyzing a
qualitative standard
that includes both
linear and branched
isomers and
determine retention
times, transitions
and transition ion
ratios. Quantitate
samples by
integrating the total
response (i.e.,
accounting for
peaks that are
identified as linear
and branched
isomers) and relying
on the initial
calibration that uses
the linear isomer
quantitative
standard.
QC Check Minimum Frequency Acceptance Corrective Flagging Comments SOP SECTION
Criteria Action Criteria
Sample PFAS All analytes detected The chemical NA. PFAS identified For example: Ion Ratio = 7.6.6
Identification in a sample. derivation of the ion with Ion ratios (quant ion abundance/
transitions must be that fail confirm ion abundance)
documented. A acceptance Calculate the average
minimum of two ion criteria must be ratio (A) and standard
transitions flagged. deviation (SD) using the
(Precursor → quant Any ICAL standards. An
ion and precursor quantitation ion acceptance range of ratio
→ confirmation ion) peak that does could be within A ±3SD
and the ion not meet the for confirmation of
transitions ratio per maximization detection.
 CONTEST, A PACE ANALYTICAL LABORATORY SOP 466 PFAS Soils Isotope Dilution Author: BLM.
39 Spruce Street Doc 466. Revision No. 7
East Longmeadow, MA 01028-0591 Effective Date: 06/30/2021
Page 29 of 37

analyte are required criteria shall be

for confirmation. included in the
Exception is made summed
for analytes where integration and
two transitions do the resulting
not exist (PFBA and data flagged as
PFPeA). “estimated,
Documentation of biased high”.
the primary and
confirmation
transitions and the
ion ratio is required.
In-house
acceptance criteria
for evaluation of ion
ratios must be used
and must not
exceed 50- 150%.
Signal to Noise
Ratio (S/N) must be
≥ 10 for all ions used
for quantification
and must be ≥ 3 for
all ions used for
confirmation.
Quant ion and
confirmation ion
must be present and
must maximize
simultaneously (±2
seconds).
QC Check Minimum Frequency Acceptance Corrective Flagging Comments SOP SECTION
Criteria Action Criteria
Ion Transitions Every field sample, In order to avoid NA. Flagging is not NA. Appendix A
(Precursor-> standard, blank, and biasing results high appropriate
Product) QC sample. due to known
interferences for
some transitions,
the following
transitions must be
used for the
quantification of the
following analytes:
PFOA: 413 → 369
PFOS: 499 → 80
PFHxS: 399 → 80
PFBS: 299 → 80
4:2 FTS: 327 → 307
6:2 FTS: 427 → 407
8:2 FTS: 527 → 507
NEtFOSAA: 584 →
419
 CONTEST, A PACE ANALYTICAL LABORATORY SOP 466 PFAS Soils Isotope Dilution Author: BLM.
39 Spruce Street Doc 466. Revision No. 7
East Longmeadow, MA 01028-0591 Effective Date: 06/30/2021
Page 30 of 37

NMeFOSAA: 570 →
419
If these transitions
are not used, the
reason must be
technically justified
and documented
(e.g., alternate
transition was used
due to observed
interferences).
QC Check Minimum Frequency Acceptance Corrective Flagging Comments SOP SECTION
Criteria Action Criteria
Initial Calibration At instrument set-up The isotopically Correct Flagging is not No samples shall be 7.6.4 & 7.4.1
(ICAL) and after ICV or CCV labeled analog of an problem, appropriate. analyzed until ICAL has
failure, prior to analyte (Extracted then repeat passed.
sample analysis. Internal Standard ICAL. External Calibration is not
Analyte) must be allowed for any analyte.
used for quantitation Calibration can be linear
if commercially (minimum of 5 standards)
available (Isotope or quadratic (minimum of
Dilution 6 standards); weighting is
Quantitation). allowed.
Commercial PFAS
standards available
as salts are
acceptable providing
the measured mass
is corrected to the
neutral acid
concentration.
Results shall be
reported as the
neutral acid with
appropriate CAS
number.
If a labeled analog is
not commercially
available, the
Extracted Internal
Standard Analyte
with the closest
retention time or
chemical similarity
to the analyte must
be used for
quantitation.
(Internal Standard
Quantitation)
Analytes must be
within 70-130% of
their true value for
 CONTEST, A PACE ANALYTICAL LABORATORY SOP 466 PFAS Soils Isotope Dilution Author: BLM.
39 Spruce Street Doc 466. Revision No. 7
East Longmeadow, MA 01028-0591 Effective Date: 06/30/2021
Page 31 of 37

each calibration
standard.
(continued next
page)
QC Check Minimum Frequency Acceptance Corrective Flagging Comments SOP SECTION
Criteria Action Criteria
Initial Calibration ICAL must meet one 7.3.6
(ICAL) of the two options
(Continued) below:
Option 1: The RSD
of the RFs for all
analytes must be ≤
20%.
Option 2: Linear or
non- linear
calibrations must
2
have r ≥ 0.99 for
each
analyte.
Retention Time Once per ICAL and at Position shall be set NA. NA. Calculated for each 7.6.5
window position the beginning of the using the midpoint analyte and EIS.
establishment analytical sequence. standard of the ICAL
curve when ICAL is
performed.
On days when ICAL
is not performed, the
initial CCV is used.
Retention Time Every field sample, RT of each analyte Correct NA. Calculated for each 7.6.5
(RT) window width standard, blank, and and EIS analyte problem and analyte and EIS.
QC sample. must fall within 0.4 reanalyze
minutes of the samples.
predicted retention
times from the daily
calibration
verification or, on
days when ICAL is
performed, from the
midpoint standard of
the ICAL.
Analytes must elute
within 0.1 minutes of
the associated EIS.
This criterion applies
only to analyte and
labeled analog
pairs.
QC Check Minimum Frequency Acceptance Corrective Flagging Comments SOP SECTION
Criteria Action Criteria
Instrument Prior to analysis and Analyte Correct Flagging is not No samples shall be 7.4.1
Sensitivity Check at least once every 12 concentrations must problem, appropriate. analyzed until ISC has
(ISC) hours. be at LOQ; rerun ISC. If met acceptance criteria.
 CONTEST, A PACE ANALYTICAL LABORATORY SOP 466 PFAS Soils Isotope Dilution Author: BLM.
39 Spruce Street Doc 466. Revision No. 7
East Longmeadow, MA 01028-0591 Effective Date: 06/30/2021
Page 32 of 37

concentrations must problem ISC can serve as the

be within ±30% of persists, initial daily CCV.
their true values. repeat ICAL.
Initial Calibration Once after each ICAL, Analyte Correct Flagging is not No samples shall be 7.3.7
Verification (ICV) analysis of a second concentrations must problem, appropriate. analyzed until calibration
source standard prior be within ±30% of rerun ICV. If has been verified.
to sample analysis. their true value. problem
persists,
repeat ICAL.
Continuing Prior to sample Concentration of Immediately If reanalysis Results may not be 7.4.2, 7.4.5
Calibration analysis, after every analytes must range analyze two cannot be reported without valid
Verification (CCV) 10 field samples, and from the LOQ to the additional performed, CCVs.
at the end of the mid-level calibration consecutive data must be Instrument Sensitivity
analytical sequence. concentration. CCVs. If qualified and Check (ISC) can serve as
Analyte both pass, explained in a bracketing CCV.
concentrations must samples the Case
be within ±30% of may be Narrative.
their true value. reported Apply Q-flag to
without all results for
reanalysis. If the specific
either fails, analyte(s) in all
or if two samples since
consecutive the last
CCVs acceptable
cannot be calibration
run, perform verification.
corrective
action(s) and
repeat CCV
and all
associated
samples
since last
successful
CCV.
Alternately,
recalibrate if
necessary;
then
reanalyze all
associated
samples
since the
last
acceptable
CCV.
QC Check Minimum Frequency Acceptance Corrective Flagging Comments SOP SECTION
Criteria Action Criteria
Instrument Blanks Immediately following Concentration of If Flagging is No samples shall be 7.4.3
the highest standard each analyte must acceptance only analyzed until instrument
analyzed and daily be ≤ ½ the LOQ. criteria are appropriate in blank has met
prior to sample Instrument Blank not met after cases when acceptance criteria.
 CONTEST, A PACE ANALYTICAL LABORATORY SOP 466 PFAS Soils Isotope Dilution Author: BLM.
39 Spruce Street Doc 466. Revision No. 7
East Longmeadow, MA 01028-0591 Effective Date: 06/30/2021
Page 33 of 37

analysis. must contain EIS to the highest the sample Note: Successful analysis
enable quantitation calibration cannot be following the highest
of contamination. standard, reanalyzed and standard analyzed
calibration when there is determines the highest
must be no more concentration that
performed sample left. carryover does not occur.
using a When the highest
lower standard analyzed is not
concentratio part of the calibration
n for the curve, it cannot be used
highest to extend out the
standard calibration range, it is
until used only to document a
acceptance higher concentration at
criteria is which carryover still does
met. not occur.
If sample
concentratio
ns exceed
the highest
allowed
standard
and the
sample(s)
following
exceed this
acceptance
criteria (>1/2
LOQ), they
must be
reanalyzed.
QC Check Minimum Frequency Acceptance Corrective Flagging Comments SOP SECTION
Criteria Action Criteria
Extracted Internal Every field sample, Added to solid Correct Apply Q-flag Failing analytes shall be 8.2.7, 8.2.10
Standard (EIS) standard, blank, and sample prior to problem. If and discuss in thoroughly documented in
Analytes QC sample. extraction. Added to required, re- the Case the Case Narrative.
aqueous samples, extract and Narrative only EIS should be 96% (or
into the original reanalyze if reanalysis greater) purity. When the
container, prior to associated confirms impurity consists of the
extraction. field and QC failures in unlabeled analyte, the
For aqueous samples. exactly the EIS can result in a
samples prepared If recoveries same manner. background artifact in
by serial dilution are every sample, standard
instead of SPE, acceptable and blank, if the EIS is
added to final for QC fortified at excessive
dilution of samples samples, but concentrations.
prior to analysis. not field
Extracted Internal samples, the
Standard Analyte field
recoveries must be samples
within 50% to 150% must be re-
of ICAL extracted
 CONTEST, A PACE ANALYTICAL LABORATORY SOP 466 PFAS Soils Isotope Dilution Author: BLM.
39 Spruce Street Doc 466. Revision No. 7
East Longmeadow, MA 01028-0591 Effective Date: 06/30/2021
Page 34 of 37

midpoint standard and

area or area analyzed
measured in the (greater
initial CCV on days dilution may
when an ICAL is not be needed).
performed. Samples
may be re-
extracted
and
analyzed
outside of
hold times,
as
necessary
for corrective
action
associated
with QC
failure.
Method Blank (MB) One per preparatory No analytes Correct If reanalysis Results may not be 8.2.1
batch. detected >½ LOQ or problem. If cannot be reported without a valid
> 1/10th the amount required, re- performed, MB.
measured in any extract and data must be Flagging is only
sample or 1/10th the reanalyze qualified and appropriate in cases
regulatory limit, MB and all explained in where the samples
whichever is QC samples the Case cannot be reanalyzed.
greater. and field Narrative.
samples Apply B-flag to
processed all results for
with the the specific
contaminate analyte(s) in all
d blank. samples in the
Samples associated
may be re- preparatory
extracted batch.
and
analyzed
outside of
hold times,
as
necessary
for corrective
action
associated
with QC
failure.
Examine the
project-
specific
requirement
s. Contact
the client as
 CONTEST, A PACE ANALYTICAL LABORATORY SOP 466 PFAS Soils Isotope Dilution Author: BLM.
39 Spruce Street Doc 466. Revision No. 7
East Longmeadow, MA 01028-0591 Effective Date: 06/30/2021
Page 35 of 37

to additional
measures to
be taken.
QC Check Minimum Frequency Acceptance Corrective Flagging Comments SOP SECTION
Criteria Action Criteria
Laboratory Control One per preparatory Blank spiked with all Correct If reanalysis Results may not be 8.2.2
Sample (LCS) batch. analytes at a problem, cannot be reported without a valid
concentration ≥ LOQ then re- performed, LCS.
and extract and data must be Flagging is only
≤ the mid-level reanalyze qualified and appropriate in cases
calibration the LCS and explained in where the samples
concentration. all samples the Case cannot be reanalyzed.
A laboratory must in the Narrative.
use the DoD/DOE associated Apply Q-flag to
QSM Appendix B preparatory specific
Limits for batch batch for analyte(s) in all
control if project failed samples in the
limits are not analytes if associated
specified. sufficient preparatory
If the analyte(s) are sample batch.
not listed, use in- material is
house LCS limits if available.
project limits are not Samples
specified. may be re-
extracted
and
analyzed
outside of
hold times,
as
necessary
for corrective
action
associated
with QC
failure.
Examine the
project-
specific
requirement
s. Contact
the client as
to additional
measures to
be taken.
Matrix Spike (MS) One per preparatory Sample spiked with Examine the For the specific For matrix evaluation 8.2.3
batch. Not required all analytes at a project- analyte(s) in only. If MS results are
for aqueous samples concentration ≥ LOQ specific the parent outside the limits, the
prepared by serial and requirement sample, apply data shall be evaluated to
dilution instead of ≤ the mid-level s. Contact J-flag if determine the source(s)
SPE. calibration the client as acceptance of difference (i.e., matrix
concentration. to additional criteria are not effect or analytical error).
 CONTEST, A PACE ANALYTICAL LABORATORY SOP 466 PFAS Soils Isotope Dilution Author: BLM.
39 Spruce Street Doc 466. Revision No. 7
East Longmeadow, MA 01028-0591 Effective Date: 06/30/2021
Page 36 of 37

A laboratory must measures to met and

use the DoD/DOE be taken. explain in the
QSM Appendix B Case
Limits for batch Narrative.
control if project
limits are not
specified.
If the analyte(s) are
not listed, use in-
house LCS limits if
project limits are not
specified.
QC Check Minimum Frequency Acceptance Corrective Flagging Comments SOP SECTION
Criteria Action Criteria
Matrix Spike For MSD: One per For MSD: Sample Examine the For the specific The data shall be 8.2.4
Duplicate (MSD) or preparatory batch. spiked with all project- analyte(s) in evaluated to determine
Matrix Duplicate For MD: Each analytes at a specific the parent the source of difference.
(MD) aqueous sample concentration ≥ LOQ requirement sample, apply For Sample/MD: RPD
prepared by serial and s. Contact J-flag if criteria only apply to
dilution instead of ≤ the mid-level the client as acceptance analytes whose
SPE. calibration to additional criteria are not concentration in the
concentration. measures to met and sample is ≥ LOQ.
A laboratory must be taken. explain in the The MD is a second
use the DoD/DOE Case aliquot of the field sample
QSM Appendix B Narrative. that has been prepared
Limits for batch by serial dilution.
control if project
limits are not
specified.
If the analyte(s) are
not listed, use in-
house LCS limits if
project limits are not
specified.
RPD ≤ 30%
(between MS and
MSD or sample and
MD).
Post Spike Sample Only applies to Spike all analytes When Flagging is not When analyte N/A
aqueous samples reported as < LOQ analyte appropriate. concentrations are
prepared by serial into the dilution that concentratio calculated as < LOQ,
dilution instead of the result for that ns are results may not be
SPE that have analyte is reported calculated reported without
reported value of from. The spike as < LOQ, acceptable post spike
< LOQ for analyte(s). must be at the LOQ and the recoveries.
concentration to be spike
reported for this recovery
sample as < LOQ. does not
When analyte meet the
concentrations are acceptance
calculated as < criteria, the
LOQ, the post spike sample,
CONTEST, A PACE ANALYTICAL LABORATORY SOP 466 PFAS Soils Isotope Dilution Author: BLM.
39 Spruce Street Doc 466. Revision No. 7
East Longmeadow, MA 01028-0591 Effective Date: 06/30/2021
Page 37 of 37

for that analyte must sample

recover within 70- duplicate,
130% of its true and post
value. spike
sample must
be
reanalyzed
at
consecutivel
y higher
dilutions until
the criteria is
met.
 APPENDIX E

westonandsampson.com
 CTDEEP STATEWIDE PFAS INITIATIVE
SAMPLING OF PUBLICLY OWNED TREATMENT WORKS

SENSITIVITY TABLE - PFAS

Aqueous Solid Tissue

SOP 455 SOP 467 MIN4-0178 (2 g)
Parameter Acronym CAS No.
LOQ DL LOQ DL LOQ DL
ng/L ng/L ug/kg ug/kg ug/kg ug/kg
Perfluoroalkyl carboxylic acid (PFCA)
Perfluorobutanoic acid PFBA 375-22-4 2 0.57 1 0.34 0.250 0.0458
Perfluoropentanoic acid PFPeA 2706-90-3 2 0.66 1 0.08 0.250 0.0490
Perfluorohexanoic acid PFHxA 307-24-4 2 0.75 1 0.19 0.250 0.0695
Perfluoroheptanoic acid PFHpA 375-85-9 2 0.62 1 0.23 0.250 0.0906
Perfluorooctanoic acid PFOA 335-67-1 2 0.38 1 0.15 0.250 0.0581
Perfluorononoic acid PFNA 375-95-1 2 0.48 1 0.17 0.250 0.0294
Perfluorodecanoic acid PFDA 335-76-2 2 0.38 1 0.16 0.250 0.106
Perfluoroundecanoic acid PFUnA 2058-94-8 2 0.49 1 0.22 0.250 0.0644
Perfluorododecanoic acid PFDoA 307-55-1 2 0.29 1 0.10 0.250 0.0738
Perfluorotridecanoic acid PFTrDA 72629-94-8 2 1.20 1 0.23 0.250 0.0502
Perfluorotetradecanoic acid PFTA 376-06-7 2 0.82 1 0.27 0.250 0.0706
Perfluorohexadecanoic acid PFHxDA 67905-19-5 0.250 0.0317
Perfluorooctandecanoic acid PFODA 16517-11-6 0.250 0.0542
Perfluoroalkane sulfonic acid (PFSA)
Perfluorobutanesulfonic acid PFBS 375-73-5 2 0.32 1 0.15 0.221 0.0429
Perfluoropentanesulfonic acid PFPeS 2706-91-4 2 0.51 1 0.26 0.235 0.0575
Perfluorohexanesulfonic acid PFHxS 355-46-4 2 0.58 1 0.22 0.228 0.0437
Perfluoroheptanesulfonic acid PFHpS 375-92-8 2 1.30 1 0.54 0.238 0.0338
Perfluorooctanesulfonic acid PFOS 1763-23-1 2 0.38 1 0.18 0.231 0.0338
Perfluorononesulfonic acid PFNS 68259-12-1 2 0.89 1 0.40 0.240 0.0475
Perfluorodecanesulfonic acid PFDS 335-77-3 2 0.60 1 0.41 0.241 0.0395
Perfluorododecanesulfonic acid PFDoS 79780-39-5 0.243 0.0294
Perfluoroalkane sulfomides (FASA) and derivatives
Perfluorooctanesulfomide PFOSA 754-91-6 2 0.44 1 0.22 0.250 0.0338
N-ethyl perfluorooctane sulfomidoethanol NEtFOSE 1691-99-2 0.250 0.0483
N-methyl perfluorooctane sulfomidoethanol NMeFOSE 24448-09-7 0.250 0.0497
N-ethyl perfluorooctane sulfomide NEtFOSA 4151-50-2 0.250 0.0338
N-methyl perfluorooctane sulfomide NMeFOSA 31506-32-8 0.250 0.0377
N-ethyl perfluorooctanesulfomidoacetic acid NEtFOSAA 2991-50-6 2 0.93 1 0.34 0.250 0.0829
N-methyl perfluorooctanesulfomidoacetic acid NMeFOSAA 2355-31-9 2 0.96 1 0.25 0.250 0.0251
Fluorotelomer sulfonic acid (FTSA)
4:2 Fluorotelomer sulfonic acid 4:2 FTS 757124-72-4 2 1.00 1 0.22 0.234 0.0421
6:2 Fluorotelomer sulfonic acid 6:2 FTS 27619-97-2 2 1.10 1 0.24 0.238 0.113
8:2 Fluorotelomer sulfonic acid 8:2 FTS 39108-34-4 2 1.40 1 0.49 0.241 0.0913
10:2 Fluorotelomer sulfonic acid 10:2 FTS 120226-60-0 0.241 0.0410
Perfluoroalkyl ether carboxylic acid (PFECA)
Perfluoro-3-methoxypropanoic acid PFMPA 377-73-1 2 0.37 1 0.23
Perfluoro-4-methoxybutanoic acid PFMBA 863090-89-5 2 0.65 1 0.09
1/
Hexafluoropropylene oxide dimer acid HFPO-DA 13252-13-6 2 1.60 2 0.95 0.250 0.250
Nofluoro-3,6-dioxaheptanoic acid NFDHA 151772-58-6 2 0.53 1 0.26
4,8-dioxa-3H-perfluorononoic acid ADONA 919005-14-4 2 0.60 1 0.13 0.236 0.0655
Polyfluoroalkyl ether sulfonic acid (PFESA)
Perfluoro(2-ethoxyethane)sulfonic acid PFEESA 113507-82-7 2 0.36 1 0.07
9-Chlorohexadecafluoro-3-oxanone-1-sulfonic acid 9Cl-PF3ONS 756426-58-1 2 0.36 1 0.15 0.233 0.0305
11-chloroeicosafluoro-3-oxaundecane-1-sulfonic acid 11Cl-PF3OUdS 763051-92-9 2 0.54 1 0.20 0.235 0.0358
Perfluoro-1-butanesulfonamide FBSA 30334-69-1 2 0.55 1 0.12
Perfluoro-1-hexanesulfonamide FHxSA 41997-13-1 2 0.70 1 0.16

Compound specified in EPA Method 537.

Notes:
LOQ Limit of Quantitation
DL Detection Limit
ng/L nanograms per liter
ug/kg micrograms per kilogram
1/ The lab will report this compound in biota to the LOQ. Because of the unique biota reference matrix used for batch QC, the labeled EIS used (13C3-HFPO-DA) may
exhibit poor recovery in the blank and LCS results. The lab uses canola oil as the reference matrix for batch QC, in which HFPO-DA is known to exhibit poor recovery.
We anticipate acceptable recovery of 13C3-HFPO-DA in the actual tissue samples.