3 

Testing Certificate #2052.01 

This laboratory maintains A2LA accreditation to: ANSI/ISO/IEC 17025:2005 for the specific 

tests/calibrations listed in A2LA Certificate # 2052.01 and meets the principles of ISO 9001:2000. The tests 

results included in this report are covered by this accreditation following method ETS-8-045.1

Final Report

Concentrations of PFOS in Fish Fillet Tissues from 

Largemouth Bass Collected in the Tennessee River in April 2012

Study Title

Analytical Study ISO12-01-01-01: Analysis of PFOS from Tennessee River Fish – Bakers 

Creek Area: April 2012 

Data Requirement 

ISO17025 

Project Director 

Charles Young 

Weston Solutions, Inc. 

Principal Investigator

Cleston C. Lange, Ph.D. 

3M Environmental Laboratory 

Report Completion Date 

May 02, 2012 

Analytical Laboratory 

3M Environmental Health and Safety Operations 

Environmental Laboratory 

3M Center, Bldg 260-05-N-17 

St. Paul, MN 55144 

3M LIMS Project Identification

ISO12-01-01-01 

Total Number of Pages 

25 
Study: ISO12-01-01-01 

Analysis for PFOS in Fish Collected from the Tennessee River in April 2012 

2 

TABLE OF CONTENTS

Table of Contents ....................................................................................................................................... 2

List of Tables .............................................................................................................................................. 3

1 Study Information ................................................................................................................................ 4

2 Summary ............................................................................................................................................. 5

3 Introduction .......................................................................................................................................... 7

4 Reference Substances ....................................................................................................................... 8

5 Method Summary ............................................................................................................................... 8

5.1 Methods .................................................................................................................................. 8

5.2 Sample Receipt ...................................................................................................................... 9

5.3 Sample Preparation ................................................................................................................ 9

5.4 LC/MS/MS Analysis .............................................................................................................. 10

6 Analytical Results .............................................................................................................................. 12

6.1 Calibration ............................................................................................................................. 12

6.2 Limits of Quantitation (LOQs) .............................................................................................. 13

6.3 Continuing Calibration .......................................................................................................... 14

6.4 Blanks .................................................................................................................................... 14

6.5 Laboratory Control Spikes (LCSs) ....................................................................................... 14

6.6 Laboratory Matrix Spikes (LMSs) ........................................................................................ 19

6.7 NIST SRM 1947 Results ...................................................................................................... 20

6.8 Individual Sample Results .................................................................................................... 21

7 Conclusion ......................................................................................................................................... 23

8 Data/Sample Retention .................................................................................................................... 24

9 List of Attachments ........................................................................................................................... 24

10 Signatures ......................................................................................................................................... 24
Study: ISO12-01-01-01 

Analysis for PFOS in Fish Collected from the Tennessee River in April 2012 

3 

LIST OF TABLES 

Table 1. PFOS Results for Tennessee River Largemouth Bass Fillet Tissues, April 2012 .................. 6

Table 2. Analytical Reference Substances .............................................................................................. 8

Table 3. Instrument Information .............................................................................................................. 11

Table 4. Gradient Liquid Chromatography Conditions (ETS-8-045) .................................................... 11

Table 5. Mass Transitions ....................................................................................................................... 11

Table 6. Analytical Batches for PFOS in Tennessee River Fish (April 2012) ...................................... 12

Table 7. L-PFOS Solvent Calibration Results ........................................................................................ 13

Table 8. L-PFOS LCSs and Matrix Blank Results ................................................................................. 15

Table 9. brPFOS LCSs and Matrix Blank Results ................................................................................. 17

Table 10. Homogenization Process Control Results ............................................................................. 20

Table 11. Individual PFOS Results for Fillet Tissues from Fish Collected in the Tennessee 

River, April 2012 ................................................................................................................................ 21
Study: ISO12-01-01-01 

Analysis for PFOS in Fish Collected from the Tennessee River in April 2012 

4 

1 Study Information 

Sponsor 

3M Company 

Project Requestor 

Gary Hohenstein 

Environmental Manager 

Special Projects, EHS Operations 

Bldg 224-5W-03 

St. Paul, MN 55144 

Phone: (651) 737-3570 

gahohenstein@mmm.com

Sampling Coordinator

Charles Young 

Weston Solutions, Inc. 

1400 Weston Way 

West Chester, PA 19380 

610-701-3787 

charles.young@westonsolutions.com

Principal Analytical Investigator 

Cleston Lange, Ph.D. 

3M Environmental Laboratory 

3M Center, Bldg 260-5N-17 

St. Paul, MN 55144 

Phone: (651)-733-9860 

clange@mmm.com 

Analytical Testing Facility 

3M Environmental Laboratory 

Building 260-5N-17 

St. Paul, MN 55144 

Study Personnel (3M Environmental Laboratory) 

Marlene Heying, Analyst (Pace Analytical, Professional Services) 

Kelly Ukes, Analyst (Pace Analytical, Professional Services) 

Jon Steege, Analyst (Pace Analytical, Professional Services) 

William Reagen, Ph.D., 3M Environmental Laboratory Management 

Study Dates 

Study Initiation: April 10, 2012 

Interim Analytical Initiation (PFOS): April 16, 2012 

Interim Analytical Completion (PFOS): April 24, 2012 

Interim Report Completion: April 30, 2012 
Study: ISO12-01-01-01 

Analysis for PFOS in Fish Collected from the Tennessee River in April 2012 

5 

2 Summary 

This analytical report provides the perfluorooctane sulfonate (PFOS) concentrations measured for 

twenty-nine (29) homogenized fillet tissues (skin removed) that were obtained from twenty-six (26) 

largemouth bass collected from the Tennessee River in April 2012 near Decatur, Alabama. This 

analysis was conducted in support of the Weston Solutions, Inc. project plan, titled “Work Plan for Fish 

Sampling and Analysis for PFOS in Largemouth Bass from the Tennessee River in the Vicinity of the 3M 

Decatur Facility, Decatur, Alabama”. The analytical aspects reported herein were conducted in the 3M 

Environmental Laboratory as 3M study ISO12-01-01-01. 

The 26 largemouth bass were collected from the Tennessee River in early April 2012 and were filleted in 

the field (skin removed) by Weston Solutions, Inc. personnel. The 26 right side fillets from each fish, and 

6 left side fillets from six random fish as blind field duplicates, were sent to the Michigan State University 

Wildlife Toxicology Laboratory (MSU-WTL) for homogenization. The processed fish homogenates from 

26 right side fillets and 3 of the left side fillets were then packaged in baggies and shipped frozen with 3 

process controls and 1 standard reference material (NIST SRM 1947) to the 3M Environmental 

Laboratory for quantitative analyses of PFOS. The other 3 left side fillet homogenates (skin removed), 

along with one aliquot of NIST SRM 1947, were sent to Axys Labs for analysis of PFOS as independent 

verification of the 3M Environmental Laboratory results. The remaining 20 whole fillets, skin on and not 

homogenized, were sent directly to the 3M Environmental Laboratory by Weston Solutions, Inc. field 

personnel to be retained frozen as “archive” samples for potential future use. 

The twenty-nine (29) homogenized fish fillet tissues were received on April 14, 2012 and April 17, 2012 

as two separate shipments. The analytical laboratory was blind to the true identities of the Tennessee 

River fish with regard to sex, age and specific sampling location, which was only known to Weston 

Solutions, Inc. and were only labeled with unique, generic IDs. In addition to the largemouth bass 

tissues, three (3) process control fish tissues (pre-study, mid-study and post-study controls) and one (1) 

standard reference material (NIST SRM 1947; Lake Michigan fish tissue) were received with the fish 

samples from MSU-WTL. Those control materials were previously provided by the 3M Environmental 

Laboratory to MSU-WTL prior to fish collection and processing to serve as controls for evaluating the 

homogenization process and evaluating the PFOS results relative to a NIST SRM 1947 fish tissue 

PFOS reference value. In contrast to Tennessee River fish samples, the four control tissues were 

identifiable by the laboratory in order to evaluate them for quality assurance purposes, as discussed 

herein. 

Once received at the 3M Environmental Laboratory, samples were logged into the electronic laboratory 

information management system (LIMS) where each sample was assigned a unique 3M LIMS sample 

ID under project ISO12-01-01-01. Fish homogenates were stored frozen until preparation for analysis. 

Fish homogenates were re-homogenized, extracted by protein precipitation method and analyzed by 

liquid chromatography with triple quadrapole mass spectrometric detection (LC/MS/MS) following the 

validated and published 3M Environmental Laboratory analytical method ETS-8-045.1. 

Quantitation was performed by internal standard (IS) method using stable isotope labeled 13C8-PFOS as 

the IS and utilizing linear isomer of PFOS (L-PFOS) solvent calibration. Each sample was extracted in

duplicate along with corresponding laboratory matrix spikes (LMSs) to evaluate sample-specific PFOS 

recovery. As additional quality control during the analysis, each sample received a fixed quantity of a 

stable isotope labeled surrogate reference standard (SRS) 13C4-PFOS prior to extraction; results of 

which were used as a secondary measure of PFOS recovery from each sample. 

Laboratory control spikes (LCSs) were employed to evaluate the method performance. Two sets of 

laboratory control spikes (LCSs) were prepared with each analytical batch in a control bass fillet 

homogenate; each set with three levels of PFOS fortification at nominal 10.0, 100 and 1000 ng/g, each 

level prepared in triplicate. One set of LCSs was prepared with L-PFOS, and the second was prepared 

with a mixed linear plus branched isomer PFOS (brPFOS) to evaluate potential isomer effects on 

quantitation. Additionally, for analytical batches which contained diluted samples, high-level LCSs were 

similarly diluted to incorporate some measure of dilution accuracy and precision into the method 

performance evaluations. The LCS results were used for determining the method performance during 
Study: ISO12-01-01-01 

Analysis for PFOS in Fish Collected from the Tennessee River in April 2012 

6 

the study. The LCS (L- PFOS) results are reported in Table 8 and LCS (br PFOS) results are shown in 

Table 9 and demonstrated excellent analytical method performance with accuracies of within 100 + 25% 

for all LCS results and RSDs < 20%. Statistical evaluation of the LCS results provided method 

uncertainties (95% CI) of + 19% for L-PFOS and + 11% for brPFOS. 

The concentrations of PFOS in pre-study, mid-study and post-study homogenization process controls 

were determined at 0.815 ng/g, 0.909 ng/g and 0.912 ng/g respectively. Those values compared well to 

the determined 0.804 ng/g average PFOS concentration for the same control tissue which did not travel 

to MSU-WTL and was not homogenized by MSU-WTL. The RPDs between results for the unprocessed 

versus MSU-processed control tissues were less than 15% for each and showed that PFOS was not 

significantly introduced to samples as a result of any sample contamination during processing or 

shipping of the fish tissue samples from MSU-WTL to the laboratory. 

The average determined PFOS concentration for the duplicate prepared analytical samples are shown 

in Table 1, along with the RPD of the duplicate results for each. The measured concentrations of 

PFOS ranged from 52.5 ng/g to 4770 ng/g. The recoveries of fortified PFOS from the LMS prepared for 

each at a level comparable to the endogenous PFOS level in the corresponding Tennessee River fish 

sample ranged from 76.7% to 125%, with an overall mean recovery and standard deviation of 98.9% +

10%, as shown in Table 1. The overall recovery of PFOS demonstrated excellent sample specific 

PFOS measurement accuracy. Additionally, the precision of each result was excellent with RPDs less 

than 20%, and typically less than 10%. The recoveries of the added stable isotope labeled SRS (13C4-

PFOS) were within 100 + 20% for all samples, and with excellent precision (RPDs < 20%) for all 

samples, as shown in Table 11. 

The measured PFOS concentration for a NIST SRM 1947 was 6.71 ng/g (4.0% RPD, 95.3% LMS 

recovery), and was accurate to 13% RPD with respect to the published SRM 1947 mean reference 

value and standard deviation of 5.90 ng/g. 

Table 1. PFOS Results for Tennessee River Largemouth Bass Fillet Tissues, April 2012 

3M LIMS ID Sample ID 

Avg. PFOS 

Concentration 

(ng/g) [b]

RPD (%) 

LMS PFOS 

Recovery 

ISO12-01-01-01-021 Process Control #1 (Pre-Study) 0.815 2.0% 102% 

ISO12-01-01-01-022 Process Control #2 (Mid-Study) 0.909 [a] 34% 99.1% 

ISO12-01-01-01-023 MSF 01 120403 4770 1.4% 96.5% 

ISO12-01-01-01-024 MSF 02 120403 4240 5.3% 92.4% 

ISO12-01-01-01-025 MSF 03 120403 70.7 1.9% 81.0% 

ISO12-01-01-01-026 MSF 04 120403 576 6.5% 108% 

ISO12-01-01-01-027 MSF 05 120403 471 4.9% 100% 

ISO12-01-01-01-028 MSF 06 120403 156 5.4% 76.7% 

ISO12-01-01-01-029 MSF 07 120403 2920 1.6% 98.7% 

ISO12-01-01-01-030 MSF 08 120403 1360 0.13% 99.8% 

ISO12-01-01-01-031 MSF 09 120403 2020 5.2% 104% 

ISO12-01-01-01-032 MSF 10 120403 1840 10% 105% 

ISO12-01-01-01-033 MSF 21 120404 671 6.1% 86.0% 

ISO12-01-01-01-034 MSF 22 120404 4660 3.5% 99.6% 

ISO12-01-01-01-035 MSF 23 120404 52.5 0.0056% 99.7% 

ISO12-01-01-01-036 MSF 24 120405 437 12% 90.7% 

ISO12-01-01-01-037 MSF 25 120405 306 3.8% 125% 

ISO12-01-01-01-038 MSF 26 120405 2090 1.0% 94.2% 

ISO12-01-01-01-039 MSF 27 120403 2120 5.4% 106% 

ISO12-01-01-01-040 MSF 28 120404 180 9.2% 124% 

ISO12-01-01-01-041 MSF 29 120405 2050 0.063% 102% 

ISO12-01-01-01-042 Process Control #3 (Post Study) 0.912 16% 102% 
Study: ISO12-01-01-01 

Analysis for PFOS in Fish Collected from the Tennessee River in April 2012 

7 

Table 1. PFOS Results for Tennessee River Largemouth Bass Fillet Tissues, April 2012 

3M LIMS ID Sample ID 

Avg. PFOS 

Concentration 

(ng/g) [b]

RPD (%) 

LMS PFOS 

Recovery 

ISO12-01-01-01-043 NIST SRM1947 6.71 4.0% 95.3% 

ISO12-01-01-01-044 MSF 11 12-04-03 2940 3.9% 93.5% 

ISO12-01-01-01-045 MSF 12 12-04-03 526 4.2% 101% 

ISO12-01-01-01-046 MSF 13 12-04-03 950 1.7% 99.0% 

ISO12-01-01-01-047 MSF 14 12-04-03 3370 0.50% 102% 

ISO12-01-01-01-048 MSF 15 12-04-03 754 9.6% 99.4% 

ISO12-01-01-01-049 MSF 16 12-04-04 1520 3.2% 86.9% 

ISO12-01-01-01-050 MSF 17 12-04-04 165 0.75% 84.2% 

ISO12-01-01-01-051 MSF 18 12-04-04 701 2.7% 103% 

ISO12-01-01-01-052 MSF 19 12-04-04 675 0.86% 104% 

ISO12-01-01-01-053 MSF 20 12-04-04 1180 0.82% 104% 

Notes: The average result and relative percent difference (RPD) was determined from duplicate analytical samples for 

each fish homogenate. All reported concentration results are rounded to three significant figures while statistical values 

are rounded to two significant figures. 

The analytical method data uncertainty was + 11% for brPFOS (30.6% branched isomer) and + 19% for L-PFOS (>99% 

linear isomer) based on LCS results. 

[a] RPD > 20% 

[b] The PFOS result is the cumulative concentration of branched and linear isomers of PFOS. 

 

3 Introduction 

A scientific investigation of PFOS levels in fish and surface water collected from the Tennessee River

near the 3M Decatur facility was conducted in the Spring of 2012 per the study work plan titled: “Work

Plan for Fish Sampling and Analysis for PFOS in Largemouth Bass from the Tennessee River in the 

Vicinity of the 3M Decatur Facility, Decatur, Alabama”. The analytical aspects reported herein are for the 

analyses of the fish tissues as conducted by the 3M Environmental Laboratory under the 3M study 

ISO12-01-01-01. The overall purpose of this study was to provide additional site-specific information on 

PFOS level in fish and surface waters of the Tennessee River near the confluence of Baker’s Creek, 

located near the 3M Decatur facility. 

This report describes the sample receipt, extraction and quantitative LC/MS/MS analysis procedures 

performed for the measurement of PFOS in the fish tissues harvested from largemouth bass collected in 

early April of 2012 from the Tennessee River, and provides the PFOS concentration results for those 

samples. Additionally included in this report are the assessment of the precision and accuracy of the 

results based on several parameters: (1) The precision of sample results was determined from 

preparation and analysis of duplicate analytical samples for each homogenate; (2) the recovery of PFOS

from laboratory matrix spikes (LMS) samples prepared for each sample was a measure of samplespecific accuracy; (3) two LCS sets were prepared with each analytical batch, one fortified with L-PFOS 

and one with br-PFOS in a control largemouth bass fillet homogenate (Osage Catfisheries, Inc.) to 

evaluate analytical method precision and accuracy; and (4), the recovery of a stable isotope labeled 

SRS (13C4-PFOS) from each sample was determined and used as a secondary measure of samplespecific analyte recovery, and accuracy and precision. 

Personnel from Weston Solutions, Inc were responsible for coordinating all field aspects of the study 

such as site selection, site parameters, fish and water collection procedures, handling and storage of

collected specimens, documentation of individual fish information, filleting, and shipment of intact fish 

fillets to the MSU-WTL for homogenization. The MSU-WTL personnel were responsible for proper 

homogenization of fillets, relabeling and distribution of homogenates to the appropriate analytical 

laboratories. The 3M Environmental laboratory was responsible for ensuring samples were received 
Study: ISO12-01-01-01 

Analysis for PFOS in Fish Collected from the Tennessee River in April 2012 

8 

appropriately, stored frozen and prepared and analyzed for PFOS. In addition, Axys Laboratories was 

to receive a sub-set of duplicate fish tissue homogenates for 3 of the Tennessee River fish (10% split) 

from MSU-WTL and one standard reference material (NIST SRM 1947) for independent verification of 

the PFOS results attained by 3M, and will be reported separately. 

This was a laboratory-blinded study and the fish ages, sizes and specific river collection locations were 

not provided to the analytical laboratories. Tennessee River fish samples were only identifiable at the 

laboratory by coded IDs provided to them by Weston Solutions, Inc. personnel. Weston Solutions, Inc. 

is responsible for all final analysis of the data for associating PFOS concentrations with collection 

location, fish species, age, weight, etc. This report focuses solely on the extraction and quantitative 

LC/MS/MS analysis performed at the 3M Environmental Laboratory and reports the determined 

concentration results for PFOS in the fish tissues, along with a measure of analytical accuracy and 

precision of the results based on several quality control aspects. 

4 Reference Substances 

The reference substances used during the analysis of the fish tissues and the consumable supplies 

are listed in Table 3. 

Table 2. Analytical Reference Substances 

Reference Substance PFOS (linear isomer) PFOS (linear +

branched isomers) M4-PFOS M8-PFOS 

Chemical Name Potassium Perfluoro-n-octane 

Sulfonate 

Potassium Perfluorooctane 

Sulfonate 

13C4-PFOS 13C8-PFOS 

Chemical Formula C8F17SO3K C8F17SO3K 13C4

12C4F17SO3Na 13C8F17SO3Na 

Use Calibration, LMSs &

LCSs 

Linear + Branched Isomers

LCSs Surrogate Internal Standard (IS) 

Source Wellington Laboratories Wellington Laboratories Wellington Laboratories Wellington Laboratories 

Expiration Date 10/18/2018 12/1/2014 12/21/2014 6/17/2012 

Storage Conditions Frozen Frozen Frozen Frozen 

Chemical Lot LPFOSKBM06 br-PFOSK1111 MPFOS1211 M8PFOS0609 

TCR Number TCR08-0001-1/1 TCR11-0041-1/19, -7/19 TCR12-0003-1/14 TCR09-0051-9/15 

Physical Description White Powder White Powder Methanol Solution [a] Methanol Solution [a] 

Purity 99.8% >98% >98% >98% 

[a] Stable isotope labeled materials are provided as methanol solutions by the vendor (source); the vendor’s reported concentration is used 

for calculating concentrations in samples. 

5 Method Summary 

5.1 Methods 

The re-homogenization of fish tissue homogenates from MSU-WTLs, and the sample extractions and 

quantitative LC/MS/MS analyses were performed following validated and previously published 3M 

Environmental Laboratory method ETS-8-045.1 [Malinsky et. al. 2011, Analytica Chemica Acta

683:pp248-257]. Sample analyses for fish samples occurred from April 17th through April 24, 2012. 

The analytical runs are identified by the initial of the instrument name used and the date and a letter 

describing the order of the runs on that day (i.e. j120417a describes instrument Jonas on April 17, 2012 

and was the first analytical run of the day on that instrument; j120417b would describe the second run of 

the day on that same instrument). The identities of the instruments used are shown in Table 3. A 

summary of the analytical runs for reporting is shown in Table 6. 
Study: ISO12-01-01-01 

Analysis for PFOS in Fish Collected from the Tennessee River in April 2012 

9 

5.2 Sample Receipt 

On April 12, 2012 the 3M Environmental Laboratory received twenty (20) frozen whole fillets (skin on, 

not homogenized) directly from the field as “archive” samples which were stored frozen upon receipt. 

Those samples were received with 3M chains of custody (COCs) # 19436 and #19437. The archive 

samples were not analyzed as part of this study and were retained frozen for potential future use, but

associated with this study. The whole fillet samples were logged into the laboratory LIMS system and

consecutively numbered based on their order on the COCs as ISO12-01-01-01-001 through -020. 

On April 14, 2012 nineteen (19) homogenized fillets and two process controls (pre-study and mid-study 

controls) were received at the 3M Environmental Laboratory from MSU-WTL with COCs #14933 and 

#14940 and one MSU-WTL COC for the process controls. On April 17, 2012 an additional ten (10) 

Tennessee River fish tissue homogenates were received with 3M COC #14934, and the post-study 

process control and NIST SRM 1947 material were received with a MSU-WTL COC. 

The three process control tissue homogenates (3 ~10-25 gram aliquots of a control bluegill fillet 

homogenate TN11-0184-1/1) had been provided to MSU-WTL earlier by the 3M Environmental 

Laboratory for quality assurance purposes to monitor potential sample contamination during the 

homogenization process. Additionally, two ~5-10 gram aliquots of a National Institutes of Standards and 

Technology (NIST) standard reference material SRM 1947 (Lake Michigan fish tissue homogenate) was 

also provided to MSU-WTL, to be re-packaged and one aliquot sent to each laboratory for analysis. The

twenty-nine Tennessee River fish tissue homogenates, three process controls and one NIST SRM 1947 

reference were logged into the Laboratory’s electronic LIMS system in the consecutive order they were 

received, and in the order listed on the chains of custodies as ISO12-01-01-01-021 through ISO12-01-

01-01-053. 

5.3 Sample Preparation 

Each frozen fish tissue homogenate was removed from its polypropylene sample bag and rehomogenized with dry ice, then transferred back into the same sample bag and left to stand overnight in 

a refrigerator to allow the dry ice to sublime (dissipate). Then for each sample, duplicate nominal 0.5 

gram aliquots of tissue homogenate were each accurately weighed into a centrifuge tube for extraction. 

Additionally, a third and fourth aliquot of each was aliquoted and one fortified with L-PFOS at nominal 50 

ng/g and the other at 500 ng/g as laboratory matrix spikes (LMSs). Later, some samples were 

determined to be above the calibration range and were re-prepared with a single higher LMS at 5000 

ng/g and re-analyzed after being appropriately diluted. A stable isotope labeled PFC internal standard 

(

13C8-PFOS) and stable isotope labeled surrogate (13C4-PFOS) were added to each sample aliquot prior 

to solvent extraction, final concentration of each was nominal 1.0 ng/mL in final extract. Samples were 

extracted by addition of 5.0 mL acetonitrile, briefly mixed and then placed into a freezer for 

approximately 1 hr to facilitate precipitation of fat and protein solids. Then, samples were centrifuged to 

remove the solids. A 1.0 mL aliquot of each extract was transferred to an autosampler vial with 0.010

mL formic acid, capped and then analyzed by LC/MS/MS. In some instances, samples were diluted 10-

fold or 100-fold appropriately with acetonitrile containing IS and SRS at the same levels as previously 

added to samples during extraction. 

The first sample preparations began on April 16, 2012 and the last preparations occurred on April 23, 

2012. Samples were always prepared as analytical batches, with one analytical batch typically 

comprised of a set of14 solvent calibration standards ranging from 0.025 ng/mL to 250 ng/mL; two sets 

of LCSs as described below, several method and solvent blanks, and sample extracts representing 

approximately10 study samples (an analytical sample, analytical sample duplicate and LMSs were 

included per sample). All the analytical batches for this study are listed in Table 6. 

The typical calibration range for each analytical batch data set was from 0.025 to 100 ng/mL (equivalent 
Study: ISO12-01-01-01 

Analysis for PFOS in Fish Collected from the Tennessee River in April 2012 

10 

to nominal 0.25 to 1000 ng/g of fish tissue concentration), with dropping of the highest prepared 

calibration standard of 250 ng/mL. In several instances the sample results exceeded the response of 

the 100 ng/mL calibrant and in those instances the 50 ng/g and 500 ng/g LMS fortification levels were 

determined to be insufficient relative to the endogenous analyte level because typically the ideal LMS

fortifications are between 0.5-times and 10-times the endogenous analyte level per 3M Environmental 

SOP requirements. In those instances, the samples were re-prepared with a higher LMS fortification at

nominal 5000 ng/g concentration with dilutions, as appropriate, and the original sample results ignored 

and only the re-prepared sample results reported. 

Exception(s): In one instance in run g120418b, the sample result for ISO12-01-01-01-049 sample and 

its LMSs were above the revised upper limit of quantitation (ULOQ) of 100 ng/mL (i.e. > 1000 ng/g), and 

a dilution with a 5000 ng/g LMS was accidentally not prepared for that sample. In that instance, the 

original analysis result along with the high-LMS result (500 ng/g fortification) was reported, despite the 

fortification level being slightly less than 0.5-times the endogenous level and determined from a high

range calibration by inclusion of the 250 ng/mL standard and exclusion of the calibration standards less 

than 7.5 ng/mL (eq. 75 to 2500 ng/g fish tissue); see analytical run/batch data for g120418b-high range 

results for that sample’s results. This fact was footnoted in the appropriate results tables. 

Laboratory control spike (LCS) samples were prepared in sets with three levels of nominal 10.0, 100 and 

1000 ng/g of PFOS and each level prepared in triplicate. Each batch prepared had two sets of LCSs, 

normal LCSs fortified with L-PFOS the same as calibration standards, and the second set prepared with 

brPFOS to verify the quantitative accuracy for PFOS typically found in environmental samples which are

comprised of a mixture of linear and branched isomers. The LCSs were prepared by fortifying nominal 

0.5 gram aliquots of control largemouth bass fillet homogenate (TN08-0193-1/1), purchased from Osage 

Catfisheries, Inc. (Osage Beach, MO). Equivalent prepared non-fortified method blanks were also 

prepared with the control bass tissue to establish the endogenous control matrix PFOS level for 

subtraction from LCS results during LCS recovery calculations. Results of LCSs were used to evaluate 

the overall method performance during the analyses.

Exception(s): In analytical batch m120423a, the typical set of LCSs and method blanks was not 

prepared and analyzed with the diluted samples analyzed. That run was conducted as a cleanup set for 

analysis of diluted samples which earlier results were greater than the ULOQ. This analytical batch/run 

included only equivalently diluted high-level LCSs from the batches those samples were originally 

prepared and analyzed. Those dilution LCS results are included in the LCS tables and the statistics for 

the method performance evaluations. 

5.4 LC/MS/MS Analysis 

The analysis of the fish extracts for PFOS was performed by LC/MS/MS as described in method 

ETS-8-045.1. The relative percent difference (RPD) of the duplicate sample results was a 

measure of the sample-specific analytical precision. The determined recovery of fortified PFOS 

from LMS samples was used as a measure of sample-specific analytical accuracy. Recovery of 

the surrogate (13C4-PFOS) from duplicate analytical samples was additionally used as an indirect 

measure of PFOS recovery and data precision from each sample result. 

Details of the specific instrument parameters, the liquid chromatography gradient program, and the 

specific mass transitions analyzed are detailed in the raw data, and are briefly described below in 

Table 3, Table 4 and Table 5. Additionally, a summary of all analytical runs conducted for collection of 

the PFOS data reported herein are shown in Table 6. 
Study: ISO12-01-01-01 

Analysis for PFOS in Fish Collected from the Tennessee River in April 2012 

11 

Table 3. Instrument Information 

Instrument Name(s) ETS-MaryAnn (m), ETS-Ginger (g), ETS-Jonas (j) 

Analytical Method ID ETS-8-045.1 

Liquid Chromatograph Agilent 1100 with Binary Pump 

Guard column Pre-Autosampler; Prism RP (2 x 50 mm, 5 μm) 

Extraction column Waters Corp. Oasis HLB Online Column (3 x 20 mm, 5 μm); 30°C 

Analytical column Betasil C18 (100 x 2.1 mm, 25 μm); 30°C 

Column Switching Valve 0 to 5 min (waste); 5 minutes to 21 minutes to analytical column 

Injection Volume 40 μL 

Mass Spectrometer API 5000 Triple Quadrapole 

Ion Source TurboIon Spray (Electrospray) 

Polarity Negative Ion 

Software Analyst 1.4.2 

Table 4. Gradient Liquid Chromatography Conditions (ETS-8-045) 

Step 

Number 

Total Time 

[min] 

Flow Rate 

[mL/min] 

Percent A 

[2 mm Aqueous 

Ammonium Acetate] 

Percent B 

[ Acetonitrile] 

0 0.00 0.400 97.0 3.0 

1 3.00 0.400 97.0 3.0 

2 3.50 0.400 70.0 30.0 

3 13.5 0.400 40.0 60.0 

4 15.5 0.400 40.0 60.0 

5 16.0 0.400 10.0 90.0 

6 18.0 0.400 10.0 90.0 

7 18.3 0.400 97.0 3.0 

8 21.0 0.400 97.0 3.0 

Table 5. Mass Transitions 

Analyte Mass Transitions Monitored 

PFOS 499>130, 499>99, 499>80 

13C8-PFOS (IS) 507>80 

13C4-PFOS (Surrogate) 503>80 

Note: Multiple mass transitions are summed to give the instrument response for PFOS 

 
Study: ISO12-01-01-01 

Analysis for PFOS in Fish Collected from the Tennessee River in April 2012 

12 

Table 6. Analytical Batches for PFOS in Tennessee River Fish (April 2012) 

Analytical 

Batch # Analytical Run ID Report Decision Samples Analyzed/Reported (ISO12-01-01-01-xxx)[a]

1 j120417a Data Reported -033, -035, -036, -037, -040 

2 m120417b Data Reported -022, -025 

3 g120418b Data Reported -021, -042, -043, -045, -046, -048, -050 

4 g120418b-high range Partial Data Set 

Reported -049 

5 g120420a Data Reported -038, -039, -041, -044, -047, -051, -052, 

-053 

6 m120420a Data Reported -023, -024, -026, -027, -028, -029, -030, 

-031, -032, -034 

7 m120423a Data Reported Select dilutions 

[a] Samples listed only indicate which analytical run the reported result for the analytical sample and 

sample duplicate were determined and may not indicate where the corresponding LMS results were 

attained since several required dilution and re-analysis in different analytical batches. See the raw data 

for the identity of the exact analytical batch used for the reported LMS results. 

6 Analytical Results 

All reported concentration results are rounded to three significant figures. Statistical values are

rounded to two significant figures. Because of rounding to obtain the limited significant figure values, 

reported results may vary slightly from those results found in the raw data.

6.1 Calibration 

Calibration standards were prepared by spiking varying known quantities of L-PFOS and SRS (13C4-

PFOS), and a fixed quantity of IS (13C8-PFOS) into acetonitrile. A set of 14 calibration standards ranging 

from nominal 0.0250 ng/mL to 250 ng/mL were prepared; equivalent to nominal 0.250 ng/g to 2500 ng/g 

for fish tissue quantitation. Following analysis of the calibration standards, the nominal concentration of 

each standard was plotted versus the measured analyte peak area/IS peak area response ratio. A 

quadratic equation with 1/x weighting was used to fit the calibration response data. Calibration curves 

were not forced through zero. Calculating the determined standard concentration using the resultant 

calibration curve and comparing to the actual known concentration confirmed accuracy of each curve 

point within 100 ± 25% (+ 30% at the LLOQ). The correlation coefficient (r) was greater than 0.999, and 

coefficient of determination (r2) was greater than 0.990 for all calibration curves. The summary for the 

calibration standards for each of the analytical data sets used for reporting is shown in Table 7. 

Exceptions: Typically all calibration standards were analyzed, but the 250 ng/mL standard dropped 

during construction of the calibration curve. In analytical run g120418b-high range, only the 7.5 ng/mL 

to 250 ng/mL standards were used to construct the calibration curve to quantitate sample ISO12-01-01-

01-49 within that range. In analytical run m120423a, the 250 ng/mL standard was not analyzed. 

 
Study: ISO12-01-01-01 

Analysis for PFOS in Fish Collected from the Tennessee River in April 2012 

13 

Table 7. L-PFOS Solvent Calibration Results

Analytical 

Run ID 

PFOS (ng/mL) 

STD 1 STD 2 STD 3 STD 4 STD 5 STD 6 STD 7 STD 8 STD 9 r 

STD 

10 

STD 

11 

STD 

12 

STD 

13 STD 14 

STD CONC. 0.025 0.05 0.1 0.25 0.35 0.5 1 2.5 7.5 10 15 25 100 250 

j120417a 0.0222 0.0459 0.0989 0.261 0.367 0.509 1.04 2.58 7.94 9.92 15 24.3 100 excluded 0.9998 

m120417b 0.0234 0.049 0.0977 0.254 0.354 0.502 1.01 2.6 7.71 9.85 15.8 23.7 101 excluded 0.9995 

jg120418b 0.0219 0.0475 0.0999 0.255 0.361 0.515 1.04 2.61 7.6 9.99 15.4 24.1 100 excluded 0.9998 

g120418bhigh range excluded excluded excluded excluded excluded excluded excluded exclude

d 6.43 9.63 16.7 27.6 94.8 253 0.9978 

g120420a 0.0228 0.0488 0.101 0.254 0.354 0.505 1.01 2.61 7.62 9.93 15.4 24.2 100 excluded 0.9998 

m120420a 0.0243 0.05 0.0977 0.247 0.355 0.506 0.999 2.57 7.54 10.1 15.3 24.3 101 excluded 0.9999 

m120423a 0.0237 0.0489 0.101 0.253 0.35 0.496 0.997 2.68 7.42 10 15.7 24 101 excluded 0.9996 

Average 

Value 0.0231 0.0484 0.0994 0.254 0.357 0.506 1.02 2.61 7.47 9.92 15.6 24.6 99.7 253 0.9995 

Std Dev. 0.00092 0.0014 0.0015 0.0045 0.0061 0.0064 0.019 0.039 0.48 0.15 0.55 1.34 2.21 NA 0.00074 

RSD 4.0% 3.0% 1.5% 1.8% 1.7% 1.3% 1.9% 1.5% 6.5% 1.5% 3.5% 5.4% 2.2% NA 0.074% 

Average 

Recovery 92.2% 96.7% 99.4% 102% 102% 101% 102% 104% 99.5% 99.2% 104% 98.4% 99.7% 101% NA 

n 6 6 6 6 6 6 6 6 7 7 7 7 7 1 7 

6.2 Limits of Quantitation (LOQs) 

The lower limit of quantitation (LLOQ) was the lowest non-zero calibration standard in the curve that

met linearity and accuracy requirements (100 + 30%) and for which the area counts were at least 2X 

that of the average response determined from solvent blanks. The LLOQ for PFOS in this study was 

typically the nominal concentration 0.0250 ng/mL acetonitrile standards, and correlating to a 0.25 ng/g 

LLOQ in fish tissues when accounting for the ~10-fold sample dilution which occurs during extraction 

(nominal 0.5 g tissue diluted with 5 mL acetonitrile). 

Typically, the 250 ng/mL standard was dropped and the upper limit of quantitation (ULOQ) was revised 

to the 100 ng/mL calibration standard (i.e. upper limit of nominal 1000 ng/g for PFOS in fish tissues). 

However, for one run (g120418b) a 6-point high range calibration from 7.5 ng/mL to 250 ng/mL was 

employed to quantify the results for one sample (ISO-01-01-01-049) which was accidentally not diluted 

to be captured with a 100 ng/mL ULOQ. That curve met the same calibration curve requirements as 

defined in section 6.1 and showed good LMS recovery for the high-LMS for that result and was therefore

reported. 
Study: ISO12-01-01-01 

Analysis for PFOS in Fish Collected from the Tennessee River in April 2012 

14 

6.3 Continuing Calibration 

During the course of each analysis, continuing calibration verifications (CCVs) were performed by 

regular injection of low- to mid-level calibration standards during the analytical run. The back calculated 

concentration results were used to verify that the instrument response and the initial calibration curve 

were still in control during the analysis of the entire batch. All CCVs bracketing reported data for PFOS 

met the method criteria of 100% ± 25% accuracy and all CCVs for 13C4-PFOS surrogate were within 100 

+ 25%. The results of CCVs are summarized for each analytical batch in the raw data. 

Exception(s): In analytical batch/run g120418b, one CCV result for PFOS was 163% recovery due to 

an irregularly low IS response which was apparently not reflected in the PFOS response for that 

injection. That CCV immediately followed the analysis of the calibration standards and LCSs in the 

beginning of the run and only flanked one side of a sample set. The remaining CCVs in that run were all 

acceptable and therefore the data was reported with a documented method deviation. The reason for 

the unusual low IS response was not determined. 

6.4 Blanks 

Three types of blanks were analyzed during this analysis: method blanks were prepared with control 

largemouth bass fillet homogenate (TN08-0193-1/1) and were spiked with IS and surrogate to determine 

the endogenous PFOS concentration in the control matrix for the LCSs in each analytical batch; (2) a 

bluegill “sunfish” control matrix (TN11-0194-1/1) was analyzed in analytical runs g120420a and 

m120420a to ascertain endogenous PFOS levels in that control matrix, to compare to process control 

from MSU-WTL which were the same matrix that passed through the homogenization process; and (3) 

acetonitrile solvent blanks (with and without IS & surrogate) were also included for evaluations. 

All blank results were carefully evaluated to determine any effects from instrument contamination and 

analytical carry over and served to aid in defining the LLOQs for quantitation. All of the blank sample 

results were considered acceptable upon evaluation of the raw data based on the intended use of the 

different types of blanks. 

The PFOS results for the bluegill (TN11-0194-1/1) control matrix blank tissue analyses was 0.804 +

0.089 ng/g (n = 6) and that result was used for comparing the homogenization process control sample 

results. 

6.5 Laboratory Control Spikes (LCSs) 

Laboratory control spikes (LCSs) were prepared by fortifying known quantities of PFOS into 0.5 gram 

aliquots of a control bluegill fillet tissue. The LCS sets were prepared at three levels (nominal 10, 100 

and 1000 ng/g fish tissue), and each level was prepared in triplicate. Two sets of LCSs were prepared

for these analyses, one with L-PFOS (same as calibration standards) and the second set fortified with 

brPFOS (30.6% branched isomer content). The LCS results are used to evaluate the method 

performance during the study and overall used for calculation of the analytical method uncertainty for the 

study, as discussed in the conclusion section. The LCS results and the analytical method uncertainties 

determined from the statistical evaluation of LCS with L-PFOS and with br-PFOS are shown in Table 8 

and Table 9, respectively. The method acceptance criteria of 100 ± 30% was met for all LCSs, with a 

few exceptions as noted in the LCS results tables. 

The LCS recoveries were calculated using the following equation: 

*100%

Spike Concentration

(Determine d Concentration of LCS) LCS Recovery =
Study: ISO12-01-01-01 

Analysis for PFOS in Fish Collected from the Tennessee River in April 2012 

15 

Table 8. L-PFOS LCSs and Matrix Blank Results

Analytical 

Run ID LCS Sample I.D. Measured PFOS Concentration (ng/g) 

Endogenous PFOS 

Conc. (ng/g) 

Fortified 

Conc. (ng/g) 

PFOS 

Recovery 

Average 

Recovery 

Recovery 

RSD 

j120417a 

120417-2-002MB 0.545 

0.567 

120417-2-003MB 0.576 0.00 NA NA NA 

120417-2-004MB 0.580 

120417-2-LowLCS 12.8 10.5 116% 

120417-2-LowLCSD 13.1 10.9 115% 114% 1.5% 

120417-2-LowLCST 11.9 10.1 113% 

120417-2-MidLCS 108 102 106% 

120417-2-MidLCSD 123 103 119% 111% 6.0% 

120417-2-MidLCST 112 101 110% 

120417-2-HighLCS >ULOQ 994 NA 

120417-2-HighLCSD >ULOQ 1060 NA NA NA 

120417-2-HighLCST >ULOQ 941 NA 

m120417b 

120417-1-001MB 0.590 

0.595 

120417-1-002MB 0.596 0.00 NA NA NA 

120417-1-003MB 0.598 

120417-1-LowLCS 28.9 25.7 110% 

120417-1-LowLCSD 27.0 26.1 101% 107% 4.7% 

120417-1-LowLCST 25.6 23.0 109% 

120417-1-MidLCS [a] 102 NA 

120417-1-MidLCSD [a] 99.5 NA NA NA 

120417-1-MidLCST [a] 101 NA 

120417-1-HighLCS 953 1040 91.4% 

120417-1-HighLCSD 794 933 85.1% 87.5% 3.9% 

120417-1-HighLCST 901 1050 86.0% 

g120418b 

120418-001MB 0.605 

0.594 

120418-002MB 0.621 0.00 NA NA NA 

120418-003MB 0.555 

120418-LowLCS 12.0 10.4 110% 

120418-LowLCSD 11.8 10.3 110% 110% 0.53% 

120418-LowLCST 12.8 11.0 111% 

120418-MidLCS 115 106 109% 

120418-MidLCSD 102 96.0 106% 106% 2.4% 

120418-MidLCST 107 103 104% 

120418-HighLCS 933 1040 90.1% 

86.1% 4.4% 120418-HighLCSD 900 1050 85.8% 

120418-HighLCST 867 1050 82.5% 

g120418b-High 

Range 

120418-MidLCS 113 

0.594 

106 106% 

120418-MidLCSD 100 96.0 103% 103% 2.9% 

120418-MidLCST 104 103 100% 

120418-HighLCS 929 1040 89.7% 

120418-HighLCSD 913 1050 87.0% 87.2% 2.8% 

120418-HighLCST 891 1050 84.8% 

G120420a 

120420-1-001MB 0.643 

0.572 

120420-1-002MB 0.537 0.00 NA NA NA 

120420-1-003MB 0.535 

120420-1-LowLCS 12.2 10.6 109% 

120420-1-LowLCSD 12.1 10.1 114% 109% 4.2% 

120420-1-LowLCST 10.9 9.88 105% 

120420-1-MidLCS 100 95 105% 108% 2.8% 
Study: ISO12-01-01-01 

Analysis for PFOS in Fish Collected from the Tennessee River in April 2012 

16 

Table 8. L-PFOS LCSs and Matrix Blank Results

Analytical 

Run ID LCS Sample I.D. Measured PFOS Concentration (ng/g) 

Endogenous PFOS 

Conc. (ng/g) 

Fortified 

Conc. (ng/g) 

PFOS 

Recovery 

Average 

Recovery 

Recovery 

RSD 

120420-1-MidLCSD 120 108 111% 

120420-1-MidLCST 114 106 108% 

120420-1-HighLCS 1090 929 117% 

120420-1-HighLCSD >ULOQ 916 NA 117% NA 

120420-1-HighLCST >ULOQ 950 NA 

120420-1-HighLCS DF:100 10.1 929 108% 

120420-1-HighLCSD DF:100 10.1 916 110% 108% 2.4% 

120420-1-HighLCST DF:100 10.0 950 105% 

m120420a 

120420-1-001MB 0.643 

0.565 

120420-1-002MB 0.545 0.00 NA NA NA 

120420-1-003MB 0.508 

120420-1-LowLCS 12.3 10.6 110% 

120420-1-LowLCSD 11.9 10.1 112% 108% 4.4% 

120420-1-LowLCST 10.7 9.88 103% 

120420-1-MidLCS 96.6 95 101% 

120420-1-MidLCSD 117 107.6 108% 104% 3.4% 

120420-1-MidLCST 111 106 105% 

120420-1-HighLCS 721 929 77.6% 

120420-1-HighLCSD 816 916 89.1% 89.2% 13% 

120420-1-HighLCST 960 950 101% 

120420-1-HighLCS DF:100 8.88 929 95.5% 

120420-1-HighLCSD DF:100 9.25 916 101% 101% 4.9% 

120420-1-HighLCST DF:100 10.0 950 105% 

m120423a 

120420-1-HighLCS df:10 948 

0.565 [b]

929 102% 

120420-1-HighLCSD df:10 953 916 104% 105% 2.9% 

120420-1-HighLCST df:10 1030 950 108% 

120417-2-HighLCS df:10 1050 

0.571 [b]

1040 101% 

120417-2-HighLCSD df:10 934 933 100% 99.9% 0.82% 

120417-2-HighLCST df:10 1040 1050 99.0% 

Average Recovery (Low LCS) 110% 

Standard Deviation (Low LCS) 4.2% 

RSD (Low LCS) 3.8% 

n 15 

Average Recovery (Mid LCS) 107% 

Standard Deviation (Mid LCS) 4.5% 

RSD (Mid LCS) 4.2% 

n 15 

Average Recovery (High LCS) 89.8% 

Standard Deviation (High LCS) 10% 

RSD (High LCS) 11% 

n 13 

Average Recovery (High LCS-Diluted) 103% 

Standard Deviation (High LCS-Diluted) 4.4% 

RSD (High LCS-Diluted) 4.2% 

n 12 
Study: ISO12-01-01-01 

Analysis for PFOS in Fish Collected from the Tennessee River in April 2012 

17 

Table 8. L-PFOS LCSs and Matrix Blank Results

Analytical 

Run ID LCS Sample I.D. Measured PFOS Concentration (ng/g) 

Endogenous PFOS 

Conc. (ng/g) 

Fortified 

Conc. (ng/g) 

PFOS 

Recovery 

Average 

Recovery 

Recovery 

RSD 

Average Recovery (All Levels LCSs) 103% 

Standard Deviation (All Levels LCSs) 9.7% 

RSD (All Levels LCSs) 9.4% 

n 55 

2 x Std. Dev. (Analytical Method Uncertainty for L-PFOS) 19% 

Note: The endogenous level is the average result from three analyzed matrix blanks for the bass control tissue TN08-0193 used for LCS 

preparations. 

NA; not applicable 

[a] IS accidentally was not added to the preparation and LCS value not determined 

[b] Endogenous level was not determined in the analytical run, so previous result was used (when more than one result was available, preference 

was given to the endogenous PFOS result obtained on the same instrument) 

Table 9. brPFOS LCSs and Matrix Blank Results

Analytical 

Run ID LCS Sample I.D. 

Measured PFOS 

Concentration 

(ng/g) 

Endogenous PFOS 

Conc. (ng/g) 

Fortified 

Conc. (ng/g) 

PFOS 

Recovery 

Average 

Recovery 

Recovery 

RSD 

j120417a 

120417-2-002MB 0.545 

0.567 

120417-2-003MB 0.576 0.00 NA NA NA 

120417-2-004MB 0.580 

120417-2-BLLowLCS 12.6 10.3 117% 

120417-2-BLLowLCSD 10.6 9.29 108% 113% 4.1% 

120417-2-BLLowLCST 11.4 9.64 113% 

120417-2-BLMidLCS 112 95.6 117% 

120417-2-BLMidLCSD 113 92.0 122% 121% 3.4% 

120417-2-BLMidLCST 122 97.2 125% 

120417-2-BLHighLCS >ULOQ 1000 NA 

120417-2-BLHighLCSD >ULOQ 934 NA NA NA 

120417-2-BLHighLCST >ULOQ 1020 NA 

m120417b 

120417-1-001MB 0.548 

0.595 

120417-1-002MB 0.549 0.00 NA NA NA 

120417-1-003MB 0.615 

120417-1-BLLowLCS 12.7 10.5 115% 

120417-1-BLLowLCSD 12.5 10.8 110% 113% 2.4% 

120417-1-BLLowLCST 11.9 9.97 114% 

120417-1-BLMidLCS 119 104 114% 

120417-1-BLMidLCSD 103 92.7 111% 112% 1.9% 

120417-1-BLMidLCST 116 105 110% 

120417-1-BLHighLCS >ULOQ 1020 NA 

120417-1-BLHighLCSD >ULOQ 999 NA NA NA 

120417-1-BLHighLCST >ULOQ 975 NA 

g120418b 

120418-001MB 0.605 

0.594 

120418-002MB 0.621 0.00 NA NA NA 

120418-003MB 0.555 

120418-BLLowLCS 12.8 10.7 114% 

120418-BLLowLCSD 12.2 10.6 110% 110% 3.2% 

120418-BLLowLCST 11.8 10.5 107% 
Study: ISO12-01-01-01 

Analysis for PFOS in Fish Collected from the Tennessee River in April 2012 

18 

Table 9. brPFOS LCSs and Matrix Blank Results

Analytical 

Run ID LCS Sample I.D. 

Measured PFOS 

Concentration 

(ng/g) 

Endogenous PFOS 

Conc. (ng/g) 

Fortified 

Conc. (ng/g) 

PFOS 

Recovery 

Average 

Recovery 

Recovery 

RSD 

120418-BLMidLCS 125 109 114% 

120418-BLMidLCSD 107 95.8 111% 113% 1.4% 

120418-BLMidLCST 108 94.9 113% 

120418-BLHighLCS >ULOQ 981 NA 

120418-BLHighLCSD >ULOQ 1030 NA NA NA 

120418-BLHighLCST >ULOQ 1010 NA 

g120418b-High 

Range 

120418-BLMidLCS 126 

0.594 

109 98.5% 

120418-BLMidLCSD 107 95.8 94.4% 96.5% 2.1% 

120418-BLMidLCST 108 94.9 96.5% 

120418-BLHighLCS 1140 981 115% 

120418-BLHighLCSD 1300 1030 125% 121% 4.4% 

120418-BLHighLCST 1250 1010 123% 

G120420a 

120420-1-001MB 0.643 

0.572 

120420-1-002MB 0.537 0.00 NA NA NA 

120420-1-003MB 0.535 

120420-1-BLLowLCS 11.5 9.76 112% 

120420-1-BLLowLCSD 10.3 9.19 107% 109% 2.3% 

120420-1-BLLowLCST 11.4 9.95 109% 

120420-1-BLMidLCS 113 104 108% 

120420-1-BLMidLCSD 113 93.5 120% 111% 6.9% 

120420-1-BLMidLCST 117 110 106% 

120420-1-BLHighLCS >ULOQ 948 NA 

120420-1-BLHighLCSD >ULOQ 958 NA NA NA 

120420-1-BLHighLCST >ULOQ 976 NA 

m120420a 

120420-1-001MB 0.643 

0.565 

120420-1-002MB 0.545 0.00 NA NA NA 

120420-1-003MB 0.508 

120420-1-BLLowLCS 11.1 9.76 108% 

120420-1-BLLowLCSD 9.98 9.19 103% 105% 2.4% 

120420-1-BLLowLCST 11.0 9.95 105% 

120420-1-BLMidLCS 111 104 106% 

120420-1-BLMidLCSD 110 93.5 117% 109% 6.9% 

120420-1-BLMidLCST 114 110 103% 

120420-1-BLHighLCS >ULOQ 948 NA 

120420-1-BLHighLCSD >ULOQ 958 NA NA NA 

120420-1-BLHighLCST >ULOQ 976 NA 

m120423a 

120420-1-BLHighLCS df:10 1005 

0.565 [b]

948 106% 

120420-1-BLHighLCSD df:10 996 958 104% 105% 1.0% 

120420-1-BLHighLCST df:10 1030 976 105% 

120417-2-BLHighLCS df:10 1110 

0.571 [b]

1020 109% 

120417-2-BLHighLCSD df:10 1050 999 105% 105% 4.1% 

120417-2-BLHighLCST df:10 979 975 100% 

Average Recovery (Low LCS) 110% 

Standard Deviation (Low LCS) 4.0% 

RSD (Low LCS) 3.6% 

n 15 

Average Recovery (Mid LCS) 113% 

Standard Deviation (Mid LCS) 6.3% 
Study: ISO12-01-01-01 

Analysis for PFOS in Fish Collected from the Tennessee River in April 2012 

19 

Table 9. brPFOS LCSs and Matrix Blank Results

Analytical 

Run ID LCS Sample I.D. 

Measured PFOS 

Concentration 

(ng/g) 

Endogenous PFOS 

Conc. (ng/g) 

Fortified 

Conc. (ng/g) 

PFOS 

Recovery 

Average 

Recovery 

Recovery 

RSD 

RSD (Mid LCS) 5.6% 

n 15 

Average Recovery (High LCS) NA 

Standard Deviation (High LCS) NA 

RSD (High LCS) NA 

n 0 

Average Recovery (High LCS-Diluted) 105% 

Standard Deviation (High LCS) 2.8% 

RSD (High LCS-Diluted) 2.7% 

n 6 

Average Recovery (All Levels LCSs) 110% 

Standard Deviation (All Levels LCSs) 5.7% 

RSD (All Levels LCSs) 5.1% 

n 36 

2 x Std. Dev. (Analytical Method Uncertainty for L-PFOS) 11% 

Note: The endogenous level is the average result from three analyzed matrix blanks for the bass control tissue TN08-0193 used for LCS 

preparations. 

NA; not applicable 

[a] IS accidentally not added to the preparation and LCS value not determined 

[b] Endogenous level was not determined in the analytical run, so previous result was used (when more than one result was available, preference 

was given to the endogenous PFOS result obtained on the same instrument) 

6.6 Laboratory Matrix Spikes (LMSs) 

Laboratory matrix spikes (LMSs) were generated by fortifying known quantity of L-PFOS to an aliquot of

fish fillet tissue and then preparing the sample the same as regular non-fortified samples. For this study, 

each sample had at least two LMSs prepared, and several had a third prepared at a higher level 

following initial analyses. The original two LMSs were fortified at nominal 50 and 500 ng/g, and for 

approximately 1⁄2 of the samples either the 50 ng/g or 500 ng/g LMS was sufficient relative to the 

endogenous level to evaluate the sample-specific PFOS recovery. However, for the other 1⁄2 of samples 

the levels were significantly higher and required a LMS fortified at 5000 ng/g to sufficiently evaluate 

recovery. The most appropriate LMS fortification for evaluating recovery is typically between 0.5-times 

and 10-times the endogenous level determined for the non-fortified sample. 

Exception(s): For the three homogenization process control fish tissues, the nominal 50 ng/g LMS was 

used to evaluate PFOS recovery, but was ~ 50-times the endogenous PFOS level in the corresponding 

non-fortified samples. Additionally, for sample ISO-12-01-01-049, the 500 ng/g LMS result was used to

evaluate PFOS recovery, but was slightly less than 0.5-times the endogenous level of the corresponding

non-fortified sample result. A SOP deviation was included in the raw data to accommodate these 

exceptions. 

Overall, the LMS recoveries for L-PFOS fortified into the fish samples ranged from 76.7% to 125%, with

an overall mean recovery + standard deviation of 98.9% + 10% (n=29 Tennessee river fish fillet 

homogenates). The LMS recoveries for the pre-, mid- and post-study process controls were 102%, 

99.1% and 102%, respectively. The LMS recovery from the analysis of the NIST SRM-1947 Lake 
Study: ISO12-01-01-01 

Analysis for PFOS in Fish Collected from the Tennessee River in April 2012 

20 

Michigan fish tissue homogenate was 95.3%. The method acceptance criteria of 100 ± 30% was met 

for all LMS results. The LMS recovery results are summarized in Table 1 and the individual sample 

results used for calculating LMS recoveries are shown in Table10. The calculation of PFOS recoveries 

used the following equation: 

*100%

Spike Concentration

(Determine d Concentration of LMS Determined Concentration of Field Sample) LMS Recovery −

=

6.7 NIST SRM 1947 Results 

The MSU-WTL was provided two aliquots of a Lake Michigan trout standard reference material (SRM 

1947) purchased from the National Institutes of Standards and Technology (NIST). The reference value 

of 5.9 + 0.39 ng/g PFOS was recently published for NIST SRM 1947 [Reiner et. al. 2012. Anal Bioanal 

Chem; DOI 10.1007/s00216-012-5943-5]. 

Two aliquots of NIST SRM 1947 were provided to the MSU-WTL prior to the commencement of the 

study. One non-processed aliquot was subsequently returned to the 3M Environmental Laboratory with 

the processed Tennessee River fish samples and process controls for analysis. The second aliquot of 

NIST SRM 1947 (not-processed) was sent to Axys Laboratories by MSU-WTL to be analyzed with the 

three field duplicates. 

The 3M Environmental Laboratory’s determined average + RPD value for the NIST SRM 1947 sample 

was 6.71 ng/g + 0.27 ng/g PFOS for duplicate analytical sample results. The nominal 50 ng/g LMS 

recovery for the SRM 1947 was 95.3%. The results show that the average PFOS concentration 

determined for SRM 1947 was accurate to 114% with respect to the NIST SRM 1947 reference value. 

6.8 Process Control Results 

The MSU-WTL was provided three ~25 g aliquots of 3M-homogenized control bluegill fillet tissue TN11-

0194-1/1 that was purchased from Osage Catfisheries, Inc. (Osage Beach, Mo) prior to the study. One 

aliquot was planned to be homogenized at MSU prior to the homogenization of Tenn. River Fish (prestudy process control), one near the middle of the process of homogenizing the Tenn. River fish (midstudy process control), and one after the homogenization of the Tenn. R. fish was completed (post-study 

process control). Those homogenization process controls were returned with the sample shipments for 

analysis of PFOS and compared to the PFOS results of for the same control bluegill tissue, but which 

did not travel to MSU-WTL for processing. The results of those comparisons are shown in Table 10. 

The results showed that the RPD was less than 15% for each and that no significant contamination 

occurred during the homogenization process and shipment process that would have affected the PFOS 

results of this study. 

Table 10. Homogenization Process Control Results

Sample ID Sample Description PFOS (ng/g) RPD 

TN11-0194-1/1 Bluegill fillet control tissue TN11-0194-1/1 0.804 NA 

ISO12-01-01-01-021 Pre-Study Process Control (TN11-0194-1/1) 0.815 1.3% 

ISO12-01-01-01-022 Mid-Study Process Control (TN11-0194-1/1) 0.909 12% 

ISO12-01-01-01-042 Post-Study Process Control (TN11-0194-1/1) 0.912 13% 

Relative percent difference (RPD) was calculated for the control tissue result relative to the process control result. 
Study: ISO12-01-01-01 

Analysis for PFOS in Fish Collected from the Tennessee River in April 2012 

21 

6.9 Individual Sample Results 

The individual analytical sample results are shown in Table 11. A summary of the average 

concentrations with their RPDs and LMS recoveries determined from these results are provided in 

Table 1. 

Table 11. Individual PFOS Results for Fillet Tissues from Fish Collected in the Tennessee River, April 2012

LIMS Sample ID Sample Description 

PFOS 

Conc. 

(ng/g) 

Avg. PFOS 

Concentration 

(ng/g) 

RPD (%) 

Surrogate 

(

13C4-PFOS) 

Recovery [a]

LMS Spike 

Concentration 

(ng/g) 

LMS Spike 

Recovery (%) 

ISO12-01-01-01-021 Pre-Study, Process Control #1 

(TN11-0194-1/1) 0.806 

0.815 2.0% 

100% 

NA NA 

ISO12-01-01-01-021 Dup Pre-Study, Process Control #1 

Dup (TN11-0194-1/1) 0.823 100% 

ISO12-01-01-01-021 LMS Pre-Study, Process Control #1 

LMS (TN11-0194-1/1) 52.2 52.2 NA 102% 50.5 [d] 102% 

ISO12-01-01-01-022 Mid-Study, Process Control 

#2 (TN11-0194-1/1) 1.07 

0.909 [b] 34% 

105% 

NA NA 

ISO12-01-01-01-022 Dup Mid-Study, Process Control 

#2 Dup (TN11-0194-1/1) 0.752 105% 

ISO12-01-01-01-022 LMS Mid-Study, Process Control 

#2 LMS (TN11-0194-1/1) 53.2 53.2 NA 107% 52.8 [d] 99.1% 

ISO12-01-01-01-023 MSF 01 120403 4810 

4770 1.4% 

98.6% 

NA NA 

ISO12-01-01-01-023 Dup MSF 01 120403 Dup 4740 107% 

ISO12-01-01-01-023 LMS 

High

MSF 01 120403 LMS High 9340 9340 NA 101% 4730 96.5% 

ISO12-01-01-01-024 MSF 02 120403 4120 

4240 5.3% 

107% 

NA NA 

ISO12-01-01-01-024 Dup MSF 02 120403 Dup 4350 103% 

ISO12-01-01-01-024 LMS 

High

MSF 02 120403 LMS 8580 8580 NA 98.4% 4700 92.4% 

ISO12-01-01-01-025 MSF 03 120403 71.4 

70.7 1.9% 

101% 

NA NA 

ISO12-01-01-01-025 Dup MSF 03 120403 Dup 70.0 102% 

ISO12-01-01-01-025 LMS MSF 03 120403 LMS 108 108 NA 101% 46.0 81.0% 

ISO12-01-01-01-026 MSF 04 120403 595 

576 6.5% 

100% 

NA NA 

ISO12-01-01-01-026 Dup MSF 04 120403 Dup 558 102% 

ISO12-01-01-01-026 LMS MSF 04 120403 LMS 1150 1150 NA 98.9% 524 108% 

ISO12-01-01-01-027 MSF 05 120403 460 

471 4.9% 

102% 

NA NA 

ISO12-01-01-01-027 Dup MSF 05 120403 Dup 483 100% 

ISO12-01-01-01-027 LMS MSF 05 120403 LMS 955 955 NA 100% 482 100% 

ISO12-01-01-01-028 MSF 06 120403 152 

156 5.4% 

103% 

NA NA 

ISO12-01-01-01-028 Dup MSF 06 120403 Dup 160 105% 

ISO12-01-01-01-028 LMS MSF 06 120403 LMS 581 581 NA 107% 554 76.7% 

ISO12-01-01-01-029 MSF 07 120403 2900 

2920 1.6% 

103% 

NA NA 

ISO12-01-01-01-029 Dup MSF 07 120403 Dup 2950 101% 

ISO12-01-01-01-029 LMS MSF 07 120403 LMS 8140 8140 NA 101% 5280 98.7% 

ISO12-01-01-01-030 MSF 08 120403 1350 

1360 0.13% 

101% 

NA NA 

ISO12-01-01-01-030 Dup MSF 08 120403 Dup 1360 104% 

ISO12-01-01-01-030 LMS MSF 08 120403 LMS 6270 6270 NA 100% 4920 99.8% 

ISO12-01-01-01-031 MSF 09 120403 2070 

2020 5.2% 

104% 

NA NA 

ISO12-01-01-01-031 Dup MSF 09 120403 Dup 1970 106% 

ISO12-01-01-01-031 LMS MSF 09 120403 LMS 7480 7480 NA 100% 5280 104% 

ISO12-01-01-01-032 MSF 10 120403 1740 

1840 10% 

94.8% 

NA NA 

ISO12-01-01-01-032 Dup MSF 10 120403 Dup 1930 91.0% 

ISO12-01-01-01-032 LMS MSF 10 120403 LMS 7170 7170 NA 100% 5060 105% 

ISO12-01-01-01-033 MSF 21 120404 651 

671 6.1% 

89.7% 

NA NA 

ISO12-01-01-01-033 Dup MSF 21 120404 Dup 692 96.3% 
Study: ISO12-01-01-01 

Analysis for PFOS in Fish Collected from the Tennessee River in April 2012 

22 

Table 11. Individual PFOS Results for Fillet Tissues from Fish Collected in the Tennessee River, April 2012

LIMS Sample ID Sample Description 

PFOS 

Conc. 

(ng/g) 

Avg. PFOS 

Concentration 

(ng/g) 

RPD (%) 

Surrogate 

(

13C4-PFOS) 

Recovery [a]

LMS Spike 

Concentration 

(ng/g) 

LMS Spike 

Recovery (%) 

ISO12-01-01-01-033 LMS MSF 21 120404 LMS 1080 1080 NA 90.1% 478 86.0%

ISO12-01-01-01-034 MSF 22 120404 4570

4660 3.5% 

90.7%

NA NA 

ISO12-01-01-01-034 Dup MSF 22 120404 Dup 4740 94.1% 

ISO12-01-01-01-034 LMS MSF 22 120404 LMS 9650 9650 NA 98.3% 5010 99.6% 

ISO12-01-01-01-035 MSF 23 120404 52.4

52.5 0.0056% 

97.8%

NA NA 

ISO12-01-01-01-035 Dup MSF 23 120404 Dup 52.5 102%

ISO12-01-01-01-035 LMS 

Low

MSF 23 120404 LMS 552 552 NA 91.9% 502 99.7%

ISO12-01-01-01-036 MSF 24 120405 464

437 12% 

102%

NA NA 

ISO12-01-01-01-036 Dup MSF 24 120405 Dup 409 87.1%

ISO12-01-01-01-036 LMS MSF 24 120405 LMS 884 884 NA 98.8% 494 90.7%

ISO12-01-01-01-037 MSF 25 120405 312

306 3.8% 

95.8%

NA NA 

ISO12-01-01-01-037 Dup MSF 25 120405 Dup 300 101%

ISO12-01-01-01-037 LMS MSF 25 120405 LMS 999 999 NA 91.0% 554 125%

ISO12-01-01-01-038 MSF 26 120405 2100

2090 1.0% 

94.0%

NA NA 

ISO12-01-01-01-038 Dup MSF 26 120405 Dup 2080 100%

ISO12-01-01-01-038 LMS MSF 26 120405 LMS 6830 6830 NA 100% 5030 94.2%

ISO12-01-01-01-039 MSF 27 120403 2060

2120 5.4% 

95.1%

NA NA 

ISO12-01-01-01-039 Dup MSF 27 120403 Dup 2180 96.1%

ISO12-01-01-01-039 LMS MSF 27 120403 LMS 7380 7380 NA 102% 4950 106%

ISO12-01-01-01-040 Dup MSF 28 120404 Dup 172 180 9.2% 98.2% NA NA 

ISO12-01-01-01-040 MSF 28 120404 188 107.9%

ISO12-01-01-01-040 LMS MSF 28 120404 LMS 783 783 NA 97.1% 487 124%

ISO12-01-01-01-041 MSF 29 120405 2050

2050 0.063% 

92.0%

NA NA 

ISO12-01-01-01-041 Dup MSF 29 120405 Dup 2050 94.4%

ISO12-01-01-01-041 LMS MSF 29 120405 LMS 7100 7100 NA 102% 4953 102%

ISO12-01-01-01-042 Post Study, Process Control 

#3

0.984 

0.912 16% 

101% 

NA NA 

ISO12-01-01-01-042 Dup Post Study, Process Control 

#3 Dup (TN11-0194-1/1) 0.839 99.0% 

ISO12-01-01-01-042 LMS Post Study, Process Control 

#3 LMS (TN11-0194-1/1) 56.3 56.3 NA 103% 54.1 [d] 102% 

ISO12-01-01-01-043 Reference Tissue (NIST SRM 

1947)

6.85

6.71 4.0% 

98.8%

NA NA 

ISO12-01-01-01-043 Dup Reference Tissue Dup (NIST 

SRM 1947)

6.58 102%

ISO12-01-01-01-043 LMS Reference Tissue 1/2 LMS 

(NIST SRM 1947)

54.9 54.9 NA 100% 50.6 95.3%

ISO12-01-01-01-044 MSF 11 12-04-03 2990

2940 3.9% 

100%

NA NA 

ISO12-01-01-01-044 Dup MSF 11 12-04-03 Dup 2880 101%

ISO12-01-01-01-044 LMS MSF 11 12-04-03 LMS 7580 7580 NA 101% 4970 93.5%

ISO12-01-01-01-045 MSF 12 12-04-03 537

526 4.2% 

103%

NA NA 

ISO12-01-01-01-045 Dup MSF 12 12-04-03 Dup 515 99.0%

ISO12-01-01-01-045 LMS MSF 12 12-04-03 LMS 5450 5450 NA 104% 4880 101%

ISO12-01-01-01-046 MSF 13 12-04-03 942

950 1.7% 

96.5%

NA NA 

ISO12-01-01-01-046 Dup MSF 13 12-04-03 Dup 958 102%

ISO12-01-01-01-046 LMS MSF 13 12-04-03 LMS 5810 5810 NA 100% 4910 99.0%

ISO12-01-01-01-047 MSF 14 12-04-03 3360

3370 0.505 

100%

NA NA 

ISO12-01-01-01-047 Dup MSF 14 12-04-03 Dup 3370 97.6%

ISO12-01-01-01-047 LMS MSF 14 12-04-03 LMS 8270 8270 NA 100% 4810 102%

ISO12-01-01-01-048 MSF 15 12-04-03 718

754 9.6% 

98.7%

NA NA 

ISO12-01-01-01-048 Dup MSF 15 12-04-03 Dup 790 102%

ISO12-01-01-01-048 LMS MSF 15 12-04-03 LMS 5670 5670 NA 102% 4940 99.4%

ISO12-01-01-01-049 MSF 16 12-04-04 1550 [e]

1520 3.2% 

100%

NA NA ISO12-01-01-01-049 Dup MSF 16 12-04-04 Dup 1500 [e] 99.0% 

ISO12-01-01-01-049 LMS MSF 16 12-04-04 LMS 1950 [e] 1950 NA 100% 493 [c] 86.9% 

ISO12-01-01-01-050 MSF 17 12-04-04 165 165 0.75% 101% NA NA 
Study: ISO12-01-01-01 

Analysis for PFOS in Fish Collected from the Tennessee River in April 2012 

23 

Table 11. Individual PFOS Results for Fillet Tissues from Fish Collected in the Tennessee River, April 2012

LIMS Sample ID Sample Description 

PFOS 

Conc. 

(ng/g) 

Avg. PFOS 

Concentration 

(ng/g) 

RPD (%) 

Surrogate 

(

13C4-PFOS) 

Recovery [a]

LMS Spike 

Concentration 

(ng/g) 

LMS Spike 

Recovery (%) 

ISO12-01-01-01-050 Dup MSF 17 12-04-04 Dup 166 100%

ISO12-01-01-01-050 LMS MSF 17 12-04-04 LMS 581 581 NA 100% 493 84.2%

ISO12-01-01-01-051 MSF 18 12-04-04 710

701 2.7% 

100%

NA NA 

ISO12-01-01-01-051 Dup MSF 18 12-04-04 Dup 691 101%

ISO12-01-01-01-051 LMS MSF 18 12-04-04 LMS 6080 6080 NA 99.0% 5220 103%

ISO12-01-01-01-052 MSF 19 12-04-04 672

675 0.86% 

101%

NA NA 

ISO12-01-01-01-052 Dup MSF 19 12-04-04 Dup 678 102%

ISO12-01-01-01-052 LMS MSF 19 12-04-04 LMS 5620 5620 NA 100% 4780 104%

ISO12-01-01-01-053 MSF 20 12-04-04 1170

1180 0.82% 

101%

NA NA 

ISO12-01-01-01-053 Dup MSF 20 12-04-04 Dup 1180 100%

ISO12-01-01-01-053 LMS MSF 20 12-04-04 LMS 6150 6150 NA 101% 4780 104%

All concentration values are rounded to 3 significant figures for reporting. Statistical results are rounded to 2 significant figures. As a result, raw data values 

may vary slightly from those reported 

NA; not applicable 

[a]Surrogate was spiked into sample aliquots at nominal 1.00 ng/g, result is shown for the least diluted sample result 

[b] RPD > 20% 

[c] The LMS spike concentration was less than 1⁄2 of the endogenous PFOS concentration for the non-fortified sample results 

[d] The LMS spike concentration was greater than 10-times of the endogenous PFOS concentration for the non-fortified sample results 

[e] Value was determined from a six point high-range calibration curve from 7.5 ng/mL to 250 ng/mL for quantifying PFOS in tissues from nominal 75.0 to 2500 

ng/g. All other reported results were obtained with calibration from 0.025 ng/mL to 100 ng/mL and the 250 ng/mL standard excluded, and samples were 

typically diluted to within that range when they exceeded 1000 ng/g. 

7 Conclusion 

The determination of perfluorooctane sulfonate (PFOS) concentrations for 29 largemouth bass fillet 

tissues (skin removed) collected from the Tennessee River in April 2012 near Decatur, Alabama was 

successfully completed. In addition, 3 homogenization process controls and one NIST SRM 1947 

standard reference material were analyzed. This analysis was conducted in support of the Weston 

Solutions, Inc. project plan, titled “Work Plan for Fish Sampling and Analysis for PFOS in Largemouth 

Bass from the Tennessee River in the Vicinity of the 3M Decatur Facility, Decatur, Alabama”. The 

analytical aspects reported herein were conducted in the 3M Environmental Laboratory as 3M study 

ISO12-01-01-01. 

Fish fillets (skin removed) were prepared in the field by Weston Solutions, Inc. personnel after being

collected from the Tennessee River and the shipped to the Michigan State University Wildlife Toxicology 

Laboratory where fish fillet homogenates were prepared. The frozen homogenates were then shipped 

to the 3M Environmental Laboratory in two shipments that were received on April 14, 2012 and April 17,

2012. Quantitative analysis for PFOS was performed from April 16, 2012 through April 24, 2012 

following analytical method ETS-8-045.1. 

The determined average concentrations of PFOS in the Tennessee River fish tissue samples ranged 

from 52.5 ng/g to 4770 ng/g. The recoveries of fortified PFOS from the LMS samples prepared for each

Tennessee River fish tissue samples were within 100 +25% recovery, ranging from 76.7% to 125%, 

with an overall average + standard deviation of 98.9% + 10%. Additionally, the average recovery and 

standard deviation for a surrogate recovery standard (13C4-PFOS) for all of the Tennessee River fish 

samples was 99.5% + 4.3%. 

The determined average PFOS value and relative difference for the NIST SRM 1947 fish tissue that 

was also received was 6.71 + 0.27 ng/g (4.0% RPD); a 114% recovery relative to the NIST reference 

value of 5.90 + 0.39 ng/g. 

Study: ISO12-01-01-01 

Analysis for PFOS in Fish Collected from the Tennessee River in April 2012 

25 

Attachment A 

Preparation Forms, Raw Data and Chromatograms 

(Available Upon Request)