Document ExkaYvJmbXXaXxrXevKoY8awV
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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
Testing Certificate #2052.01 This laboratory maintains A2LA accreditation to: ANSI/ISO/IEC 17025:2005for the specific tests/calibrations listed in A2LA Certificate # 2052.01 and meets the principles o fISO 9001:2000. The tests results included in this report are covered by this accreditationfollowing method ETS-8-045.1
Study: ISO12-01-01-01 Analysis for PFOS in Fish Collected from the Tennessee River in April 2012
Ta b l e o f Co n t e n t s
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
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Study: ISO12-01-01-01 Analysis for PFOS in Fish Collected from the Tennessee River in April 2012
Lis t o f Ta b l e s
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 (April2012).....................................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
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Study: ISO12-01-01-01 Analysis for PFOS in Fish Collected from the Tennessee River in April 2012
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
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Study: ISO12-01-01-01 Analysis for PFOS in Fish Collected from the Tennessee River in April 2012
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 C8-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
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Study: ISO12-01-01-01 Analysis for PFOS in Fish Collected from the Tennessee River in April 2012
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 (13C4PFOS) 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
ISO12-01 -01-01 -021 ISO12-01-01-01-022 ISO12-01-01-01-023 ISO12-01-01-01-024 ISO12-01-01-01-025 ISO12-01-01-01-026 ISO12-01-01-01-027 ISO12-01-01-01-028 ISO12-01-01-01-029 ISO12-01-01-01-030 ISO12-01 -01-01-031 ISO12-01-01-01-032 ISO12-01-01-01-033 ISO12-01-01-01-034 ISO12-01-01-01-035 ISO12-01-01-01-036 ISO12-01-01-01-037 ISO12-01-01-01-038 ISO12-01-01-01-039 ISO12-01-01-01-040 ISO12-01 -01-01 -041 ISO12-01-01-01-042
Sample ID
Process Control #1 (Pre-Study) P rocess C ontrol #2 (M id-Study) MSF 01 120403 MSF 02 120403 MSF 03 120403 MSF 04 120403 MSF 05 120403 MSF 06 120403 MSF 07 120403 MSF 08 120403 MSF 09 120403 MSF 10 120403 MSF 21 120404 MSF 22 120404 MSF 23 120404 MSF 24 120405 MSF 25 120405 MSF 26 120405 MSF 27 120403 MSF 28 120404 MSF 29 120405 Process C ontrol #3 (Post Study)
Avg. PFOS Concentration
(ng/g) [b] 0.815 0.909 [a] 4770 4240 70.7 576 471 156 2920 1360 2020 1840 671 4660 52.5 437 306 2090 2120 180 2050 0.912
RPD (%)
2.0% 34% 1.4% 5.3% 1.9% 6.5% 4.9% 5.4% 1.6% 0.13% 5.2% 10% 6.1% 3.5% 0.0056% 12% 3.8% 1.0% 5.4% 9.2% 0.063% 16%
LMS PFOS Recovery
102% 99.1% 96.5% 92.4% 81.0% 108% 100% 76.7% 98.7% 99.8% 104% 105% 86.0% 99.6% 99.7% 90.7% 125% 94.2% 106% 124% 102% 102%
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Study: ISO12-01-01-01 Analysis for PFOS in Fish Collected from the Tennessee River in April 2012
Table 1. PFOS Results for Tennessee River Largemouth Bass Fillet Tissues, April 2012
3M LIMS ID
Sample ID
Avg. PFOS Concentration RPD (%)
LMS PFOS Recovery
(ng/g) [b]
ISO12-01-01-01-043 N IS T S R M 1947
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
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Study: ISO12-01-01-01 Analysis for PFOS in Fish Collected from the Tennessee River in April 2012
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
C h em ical N a m e C h em ical Fo rm u la Use Source E xpiratio n D a te Storage C onditions C h em ical Lot TCR N u m b er
PFO S (linear isom er)
Potassium Perfluoro-n-octane Sulfonate CsF^SOaK
Calibration, LMSs & LCSs
Wellington Laboratories
10/18/2018
Frozen LPFOSKBM06 TCR08-0001-1/1
PFO S (linear + branched isom ers) Potassium Perfluorooctane
Sulfonate C8F17SOaK Linear + Branched Isomers
LCSs Wellington Laboratories
12/1/2014
Frozen br-PFOSK1111
TCR11-0041-1/19, -7/19
M4-PFOS
13c,, - pfos
13C,,12C4F,,SO3Na
Surrogate Wellington Laboratories
12/21/2014 Frozen
MPFOS1211 TCR12-0003-1/14
M 8-P F O S
13C8-PFOS
13C8F,,SOaNa
Internal Standard (IS) Wellington Laboratories
6/17/2012 Frozen
M8PFOS0609 TCR09-0051-9/15
Physical D e scrip tio n
White Powder
White Powder
Methanol Solution [a]
Methanol Solution [a]
P u rity
99.8%
>98%
>98%
>98%
[a] Stable isotope labeled materials are provided as m ethanol 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, Analytics 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.
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Study: ISO12-01-01-01 Analysis for PFOS in Fish Collected from the Tennessee River in April 2012
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 ISO12-0101-01-053.
5.3 Sample Preparation
Each frozen fish tissue homogenate was removed from its polypropylene sample bag and re homogenized 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
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Study: ISO12-01-01-01 Analysis for PFOS in Fish Collected from the Tennessee River in April 2012
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.
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Study: ISO12-01-01-01 Analysis for PFOS in Fish Collected from the Tennessee River in April 2012
Table 3. Instrument Information
Instrument Name(s) Analytical Method ID Liquid Chromatograph Guard column Extraction column Analytical column Column Switching Valve Injection Volume Mass Spectrometer Ion Source Polarity Software
ETS-MaryAnn (m), ETS-Ginger (g), ETS-Jonas () ETS-8-045.1
Agilent 1100 with Binary Pump Pre-Autosampler; Prism RP (2 x 50 mm, 5 mm) Waters Corp. Oasis HLB Online Column (3 x 20 mm, 5 mm); 30C
Betasil C18 (100 x 2.1 mm, 25 mm); 30C 0 to 5 min (waste); 5 minutes to 21 minutes to analytical column
40 mL API 5000 Triple Quadrapole Turbolon Spray (Electrospray)
Negative Ion Analyst 1.4.2
Table 4. Gradient Liquid Chromatography Conditions (ETS-8-045)
Step Number
Total Time [min]
0 0.00 1 3.00 2 3.50 3 13.5 4 15.5 5 16.0 6 18.0 7 18.3 8 21.0
Flow Rate [mL/min]
0.400 0.400 0.400 0.400 0.400 0.400 0.400 0.400 0.400
Percent A [2 mm Aqueous Ammonium Acetate]
97.0 97.0 70.0 40.0 40.0 10.0 10.0 97.0 97.0
Percent B [ Acetonitrile]
3.0 3.0 30.0 60.0 60.0 90.0 90.0 3.0 3.0
Table 5. Mass Transitions
Analyte PFOS
13C 8-P F O S (IS)
Mass Transitions Monitored 499>130, 499>99, 499>80 507>80
13C4-P F O S (Surrogate)
503>80
Note: Multiple mass transitions are summed to give the instrument response for PFOS
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Study: ISO12-01-01-01 Analysis for PFOS in Fish Collected from the Tennessee River in April 2012
Table 6. Analytical Batches for PFOS in Tennessee River Fish (April 2012)
Analytical Batch #
Analytical Run ID
Report Decision
Samples Analyzed/Reported (ISO1201-01-01-xxxp
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 (13C4PFOS), 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 ISO12-01-0101-49 within that range. In analytical run m120423a, the 250 ng/mL standard was not analyzed.
12
Study: ISO12-01-01-01 Analysis for PFOS in Fish Collected from the Tennessee River in April 2012
Table 7. L-PFOS Solvent Calibration Results
Analytical Run ID
STD CONC.
STD 1 0.025
STD 2 0.05
STD 3 0.1
STD 4 0.25
STD 5
STD 6
PFOS (n g /m L ) STD 7 STD 8 STD 9
0.35
0.5
1 2.5 7.5
STD 10
10
STD 11
15
STD 12
25
STD 13
100
STD 14 250
r
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
Average Value
Std Dev.
0.0237 0.0489 0.101 0.253
0.35 0.496 0.997 2.68
0.0231
0.0484 0.0994
0.254
0.357
0.506
1.02
2.61
0.00092 0.0014 0.0015 0.0045 0.0061 0.0064 0.019 0.039
7.42 7.47 0.48
10 9.92 0.15
15.7 15.6 0.55
24 24.6 1.34
101 excluded 0.9996
99.7
253 0.9995
2.21
NA 0.00074
RSD
Average Recovery
n
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%
92.2%
96.7%
99.4%
102%
102%
101%
102%
104% 99.5% 99.2% 104% 98.4% 99.7%
101%
NA
6 6 6 6 6 6 6 677777 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.
13
Study: ISO12-01-01-01 Analysis for PFOS in Fish Collected from the Tennessee River in April 2012
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:
(Determined Concentration o f LCS) LCS Recovery -------------------------------------------------- * 100%
Spike Concentration
14
Study: ISO12-01-01-01 Analysis for PFOS in Fish Collected from the Tennessee River in April 2012
Table 8. L-PFOS LCSs and Matrix Blank Results
A nalytical Run ID
CD
r
T--1
o
(N T--1
_Q
r
tH
<osf
(N T--1
E
_Q
00
tH
<osf
(N
t0H0
LCS S a m p le I.D .
12041 7 -2 -0 0 2 M B 12041 7 -2 -0 0 3 M B 12041 7 -2 -0 0 4 M B 120417-2-LowLCS 120417-2-LowLCSD 120417-2-LowLCST 1 2 0417-2-M idLC S 120417-2-MidLCSD 120417-2-M idLC S T 120417-2-HighLCS 120417-2-HighLCSD 120417-2-HighLCST 12041 7 -1 -0 0 1 M B 12041 7 -1 -0 0 2 M B 12041 7 -1 -0 0 3 M B 120417-1-LowLCS 120417-1-LowLCSD 120417-1-LowLCST 1 2 0417-1-M idLC S 120417-1-MidLCSD 120417-1-M idLC S T 120417-1-HighLCS 120417-1-HighLCSD 120417-1-HighLCST 120418-001M B 120418-002M B 120418-003M B 120418-LowLCS 120418-LowLCSD 120418-LowLCST 120418-MidLCS 120418-MidLCSD 120418-MidLCST 120418-HighLCS
120418-HighLCSD
_c
op
g,
322 TMco
S "=
fM
tH00
CO O
fM
<Osf
(N tH
e?
120418-HighLCST
120418-MidLCS 120418-MidLCSD 120418-MidLCST 120418-HighLCS 120418-HighLCSD 120418-HighLCST 12042 0 -1 -0 0 1 M B 12042 0 -1 -0 0 2 M B 12042 0 -1 -0 0 3 M B 120420-1-LowLCS 120420-1-LowLCSD 120420-1-LowLCST 1 2 0420-1-M idLC S
M e a s u re d PFOS C o n cen tratio n (n g /g )
0.545 0.576 0.580 12.8 13.1 11.9
108 123 112 >ULOQ >ULOQ >ULOQ 0.590 0.596 0.598 28.9 27.0 25.6 [a] [a] [a] 953 794 901 0.605 0.621 0.555 12.0 11.8 12.8 115 102 107 933
Endogenous PFOS Conc. (n g /g ) 0.567
0.595
0.594
900
867 113 100 104 929 913 891 0.643 0.537 0.535 12.2 12.1 10.9 100
0.594 0.572
F o rtifie d Conc. (n g /g )
0.00
10.5 10.9 10.1 102 103 101 994 1060 941
0.00
25.7 26.1 23.0 102 99.5 101 1040 933 1050
0.00
10.4 10.3 11.0 106 96.0 103 1040
1050
1050 106 96.0 103 1040 1050 1050
0.00
10.6 10.1 9.88 95
PFOS R eco very
NA
116% 115% 113% 106% 119% 110%
NA NA NA
NA
110% 101% 109%
NA NA NA 91.4% 85.1% 86.0%
NA
110% 110% 111% 109% 106% 104% 90.1%
85.8%
82.5% 106% 103% 100% 89.7% 87.0% 84.8%
NA
109% 114% 105% 105%
A verag e R eco very
NA 114% 111%
NA NA 107% NA 87.5% NA 110% 106%
86.1%
103% 87.2%
NA 109% 108%
Recovery RSD NA 1.5% 6.0% NA NA 4.7% NA 3.9% NA
0.53% 2.4%
4.4%
2.9% 2.8%
NA 4.2% 2.8%
15
Study: ISO12-01-01-01 Analysis for PFOS in Fish Collected from the Tennessee River in April 2012
Table 8. L-PFOS LCSs and Matrix Blank Results
A nalytical Run ID
O<CND <O<sNf
T--1
E
n<CNoD O<N
T--1
E
LCS S a m p le I.D .
M e a s u re d PFOS C o n cen tratio n (n g /g )
Endogenous PFOS Conc. (n g /g )
120420-1-MidLCSD 120420-1-M idLC S T
120420-1-HighLCS 120420-1-HighLCSD 120420-1-HighLCST 120420-1-HighLCS DF:100 120420-1-HighLCSD DF:100 120420-1-HighLCST DF:100 12042 0 -1 -0 0 1 M B 12042 0 -1 -0 0 2 M B 12042 0 -1 -0 0 3 M B 120420-1-LowLCS 120420-1-LowLCSD 120420-1-LowLCST 1 2 0420-1-M idLC S 120420-1-MidLCSD 120420-1-M idLC S T 120420-1-HighLCS 120420-1-HighLCSD 120420-1-HighLCST 120420-1-HighLCS DF:100 120420-1-HighLCSD DF:100 120420-1-HighLCST DF:100 120420-1-HighLCS df:10 120420-1-HighLCSD df:10 120420-1-HighLCST df:10 120417-2-HighLCS df:10 120417-2-HighLCSD df:10 120417-2-HighLCST df:10
120 114
1090 >ULOQ >ULOQ
10.1 10.1 10.0 0.643 0.545 0.508 12.3 11.9 10.7 96.6 117 111 721 816 960 8.88 9.25 10.0 948 953 1030 1050 934 1040
0.565
0.565 [b] 0.571 [b]
Average Recovery (Low LCS)
Standard Deviation (Low LCS)
RSD (Low LCS)
n
Average Recovery (Mid LCS)
Standard Deviation (Mid LCS)
RSD (Mid LCS)
n
Average Recovery (High LCS)
Standard Deviation (High LCS)
RSD (High LCS)
n
Average Recovery (High LCS-Diluted)
Standard Deviation (High LCS-Diluted)
RSD (High LCS-Diluted)
n
F o rtifie d Conc. (n g /g )
108 106 929 916 950 929 916 950
0.00
10.6 10.1 9.88 95 107.6 106 929 916 950 929 916 950 929 916 950 1040 933 1050
PFOS R eco very
111% 108% 117%
NA NA 108% 110% 105%
NA
110% 112% 103% 101% 108% 105% 77.6% 89.1% 101% 95.5% 101% 105% 102% 104% 108% 101% 100% 99.0%
A verag e R eco very
117%
108%
NA
108%
104%
89.2%
101%
105%
99.9%
110% 4.2% 3.8%
15 107% 4.5% 4.2%
15 89.8%
10% 11% 13 103% 4.4% 4.2% 12
Recovery RSD
NA 2.4%
NA 4.4% 3.4% 13% 4.9% 2.9% 0.82%
16
Study: ISO12-01-01-01 Analysis for PFOS in Fish Collected from the Tennessee River in April 2012
Table 8. L-PFOS LCSs and Matrix Blank Results
A n alytical Run ID
______
. ._
M e asu red PFOS C o n cen tratio n (n g /g )
Endogenous PFOS Conc. (n g /g )
F o rtifie d Conc. (n g /g )
PFOS Recovery
A verag e 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 M ethod Uncertainty fo r L-PFOS)
19%
N ote: The endogenous level is th e average resu lt fro m th re e analyzed m a trix blanks fo r th e bass c o n tro l tissue TN 08-0193 used fo r LCS preparations. NA; not applicable [a] IS a ccidentally was n o t added to th e p re p a ra tio n and LCS value n o t d e te rm in e d [b] Endogenous level was not determ ined in the analytical run, so previous result was used (when m ore than one result was available, preference was given to th e endogenous PFOS resu lt o b taine d on th e same in stru m e n t)
Table 9. brPFOS LCSs and Matrix Blank Results
A nalytical Run ID
CD r T--1
o
<N T--1
_Q r T--1
o
<N T--1
E
_Q 00 T--1
o
<N T--1 00
LCS S a m p le I.D .
12041 7 -2 -0 0 2 M B 12041 7 -2 -0 0 3 M B 12041 7 -2 -0 0 4 M B 120417-2-BLLowLCS 120417-2-BLLowLCSD 120417-2-BLLowLCST 120417-2-BLMidLCS 120417-2-BLMidLCSD 120417-2-BLMidLCST 120417-2-BLHighLCS 120417-2-BLHighLCSD 120417-2-BLHighLCST 12041 7 -1 -0 0 1 M B 12041 7 -1 -0 0 2 M B 12041 7 -1 -0 0 3 M B 120417-1-BLLowLCS 120417-1-BLLowLCSD 120417-1-BLLowLCST 120417-1-BLMidLCS 120417-1-BLMidLCSD 120417-1-BLMidLCST 120417-1-BLHighLCS 120417-1-BLHighLCSD 120417-1-BLHighLCST 120418-001M B 120418-002M B 120418-003M B 120418-BLLowLCS 120418-BLLowLCSD
120418-BLLowLCST
M e a s u re d PFOS C on cen tratio n
(n g /g ) 0.545 0.576 0.580 12.6 10.6 11.4
112 113 122 >ULOQ >ULOQ >ULOQ 0.548 0.549 0.615 12.7 12.5 11.9 119 103 116 >ULOQ >ULOQ >ULOQ 0.605 0.621 0.555 12.8 12.2
11.8
Endogenous PFOS Conc. (n g /g )
F o rtifie d Conc. (n g /g )
PFOS R eco very
A verag e R eco very
Recovery RSD
0.567 0.595 0.594
0.00
10.3 9.29 9.64 95.6 92.0 97.2 1000 934 1020
0.00
10.5 10.8 9.97 104 92.7 105 1020 999 975
0.00
10.7 10.6 10.5
NA
117% 108% 113% 117% 122% 125%
NA NA NA
NA
115% 110% 114% 114% 111% 110%
NA NA NA
NA
114% 110% 107%
NA 113% 121%
NA NA 113% 112% NA NA 110%
NA 4.1% 3.4%
NA NA 2.4% 1.9% NA NA 3.2%
17
Study: ISO12-01-01-01 Analysis for PFOS in Fish Collected from the Tennessee River in April 2012
Table 9. brPFOS LCSs and Matrix Blank Results
A nalytical Run ID
_C op
g>
S "=
(N tH CtO
oCD
<<OsNf
<N
teH?
oCD
<N <sf O (N tH
E
CD no <N <sf O <N tH
E
M e a s u re d PFOS
LCS S a m p le I.D .
C on cen tratio n
120418-BLMidLCS
(n g /g ) 125
120418-BLMidLCSD
107
120418-BLMidLCST 120418-BLHighLCS
108 >ULOQ
120418-BLHighLCSD
>ULOQ
120418-BLHighLCST
>ULOQ
120418-BLMidLCS
126
120418-BLMidLCSD
107
120418-BLMidLCST
108
120418-BLHighLCS 120418-BLHighLCSD
1140 1300
120418-BLHighLCST
1250
12042 0 -1 -0 0 1 M B
0.643
12042 0 -1 -0 0 2 M B
0.537
12042 0 -1 -0 0 3 M B
0.535
120420-1-BLLowLCS
11.5
120420-1-BLLowLCSD
10.3
120420-1-BLLowLCST
11.4
120420-1-BLMidLCS
113
120420-1-BLMidLCSD
113
120420-1-BLMidLCST
117
120420-1-BLHighLCS
>ULOQ
120420-1-BLHighLCSD
>ULOQ
120420-1-BLHighLCST
>ULOQ
12042 0 -1 -0 0 1 M B
0.643
12042 0 -1 -0 0 2 M B
0.545
12042 0 -1 -0 0 3 M B
0.508
120420-1-BLLowLCS
11.1
120420-1-BLLowLCSD
9.98
120420-1-BLLowLCST
11.0
120420-1-BLMidLCS
111
120420-1-BLMidLCSD
110
120420-1-BLMidLCST
114
120420-1-BLHighLCS
>ULOQ
120420-1-BLHighLCSD
>ULOQ
120420-1-BLHighLCST
>ULOQ
120420-1-BLHighLCS df:10
1005
120420-1-BLHighLCSD df:10
996
120420-1-BLHighLCST df:10
1030
120417-2-BLHighLCS df:10
1110
120417-2-BLHighLCSD df:10
1050
120417-2-BLHighLCST df:10
979
Average Recovery (Low LCS)
Endogenous PFOS Conc. (n g /g )
0.594
0.572
0.565 0.565 [b] 0.571 [b]
Standard D eviation (Low LCS)
RSD (Low LCS)
n
Average Recovery (M id LCS)
Standard D eviation (M id LCS)
F o rtifie d Conc. (n g /g )
109 95.8 94.9 981 1030 1010 109 95.8 94.9 981 1030 1010
0.00
9.76 9.19 9.95 104 93.5 110 948 958 976
0.00
9.76 9.19 9.95 104 93.5 110 948 958 976 948 958 976 1020 999 975
PFOS R eco very
114% 111% 113%
NA NA NA 98.5% 94.4% 96.5% 115% 125% 123%
NA
112% 107% 109% 108% 120% 106%
NA NA NA
NA
108% 103% 105% 106% 117% 103%
NA NA NA 106% 104% 105% 109% 105% 100%
A verag e R eco very
113%
NA
96.5%
121%
NA
109%
111%
NA
NA
105%
109%
NA
105%
105%
110% 4.0% 3.6%
15 113% 6.3%
Recovery RSD 1.4% NA 2.1% 4.4% NA 2.3% 6.9% NA NA 2.4% 6.9% NA 1.0% 4.1%
18
Study: ISO12-01-01-01 Analysis for PFOS in Fish Collected from the Tennessee River in April 2012
Table 9. brPFOS LCSs and Matrix Blank Results
A nalytical Run ID
LCS S a m p le I.D .
M e a s u re d PFOS C on cen tratio n
(n g /g ) RSD (M id LCS)
Endogenous PFOS Conc. (n g /g )
F o rtifie d Conc. (n g /g )
PFOS R eco very
A verag e R eco very
5.6%
Recovery RSD
n 15
Average Recovery (High LCS)
NA
Standard D eviation (High LCS)
NA
RSD (High LCS)
NA
n0
Average Recovery (High LCS-Diluted)
105%
Standard D eviation (High LCS)
2.8%
RSD (High LCS-Diluted)
2.7%
n6
A v e r a g e R e c o v e ry (A ll Le vels LCSs)
110%
S ta n d a rd D e v ia tio n (A ll Levels LCSs)
5.7%
RSD (A ll Levels LCSs)
5.1%
n 36
2 x S td. D ev. (A n a ly tic a l M e th o d U n c e rta in ty fo r L-PFOS)
11%
N ote: The endogenous level is th e average resu lt fro m th re e analyzed m a trix blanks fo r th e bass c o n tro l tissue TN 08-0193 used fo r LCS preparations. NA; not applicable [a] IS a ccidentally n o t added to th e p re p a ra tio n and LCS value n o t d e te rm in e d [b] Endogenous level was not determ ined in the analytical run, so previous result was used (when m ore than one result was available, preference was given to th e endogenous PFOS resu lt o b taine d on th e same in stru m e n t)
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 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 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
19
Study: ISO12-01-01-01 Analysis for PFOS in Fish Collected from the Tennessee River in April 2012
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:
TL.M,,S ,,Recovery = -(-D--e-t-e--r-m---i-n--e-d---C--o--n-c--e-n--t-r-a--t-i-o--n--o--f--L--M---S------D---e-t-e--r-m---in--e--d---C--o--n-c--e-n--t-r-a--t-i-o-n---o--f--F--i-e-l-d---S--a-m--*pl1e0)0.%. . . . . . . Spike Concentration
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 TN110194-1/1 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 (pre study process control), one near the middle of the process of homogenizing the Tenn. River fish (mid study 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
S am p le ID
Sam ple Description
PFOS (n g /g )
TN11-0194-1/1
Bluegill fille t control tissue TN11-0194-1/1
0.804
ISO12-01-01-01-021
Pre-Study Process Control (TN11-0194-1/1)
0.815
ISO12-01-01-01-022
Mid-Study Process Control (TN11-0194-1/1)
0.909
ISO12-01-01-01-042
Post-Study Process Control (TN11-0194-1/1)
0.912
Relative percent difference (RPD) was calculated fo r the control tissue result relative to the process control result.
RPD NA 1.3% 12% 13%
20
Study: ISO12-01-01-01 Analysis for PFOS in Fish Collected from the Tennessee River in April 2012
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
L IM S S a m p le ID
S a m p le D e s c rip tio n
ISO12-01-01-01-021
ISO12-01-01-01-021 Dup
ISO12-01-01-01-021 LMS
ISO12-01-01-01-022
ISO12-01-01-01-022 Dup
ISO12-01-01-01-022 LMS
ISO12-01-01-01-023 ISO12-01-01-01-023 Dup ISO12-01-01-01-023 LMS ISO12-01-01-01-024 ISO12-01-01-01-024 Dup IS012-01-01-01-024 LMS ISO12-01-01-01-025 IS012-01-01-01-025 Dup IS012-01-01-01-025 LMS IS012-01-01-01-026 IS012-01-01-01-026 Dup IS012-01-01-01-026 LMS IS012-01-01-01-027 IS012-01-01-01-027 Dup IS012-01-01-01-027 LMS IS012-01-01-01-028 IS012-01-01-01-028 Dup IS012-01-01-01-028 LMS IS012-01-01-01-029 IS012-01-01-01-029 Dup IS012-01-01-01-029 LMS IS012-01-01-01-030 IS012-01-01-01-030 Dup IS012-01-01-01-030 LMS IS012-01-01-01-031 IS012-01-01-01-031 Dup IS012-01-01-01-031 LMS IS012-01-01-01-032 IS012-01-01-01-032 Dup IS012-01-01-01-032 LMS IS012-01-01-01-033 IS012-01-01-01-033 Dup
Pre-Study, Process Control #1 (TN11-0194-1/1)
Pre-Study, Process Control #1 Dup (TN11-0194-1/1) Pre-Study, Process Control #1 LMS (TN11-0194-1/1)
Mid-Study, Process Control #2 (TN11-0194-1/1)
Mid-Study, Process Control #2 Dup (TN11-0194-1/1)
Mid-Study, Process Control #2 LMS (TN11-0194-1/1)
MSF01 120403 MSF 01 120403 Dup MSF 01 120403 LMS High MSF02 120403 MSF 02 120403 Dup MSF 02 120403 LMS MSF03 120403 MSF 03 120403 Dup MSF 03 120403 LMS MSF04 120403 MSF 04 120403 Dup MSF 04 120403 LMS MSF05 120403 MSF 05 120403 Dup MSF 05 120403 LMS MSF06 120403 MSF 06 120403 Dup MSF 06 120403 LMS MSF07 120403 MSF 07 120403 Dup MSF 07 120403 LMS MSF08 120403 MSF 08 120403 Dup MSF 08 120403 LMS MSF09 120403 MSF 09 120403 Dup MSF 09 120403 LMS MSF10 120403 MSF 10 120403 Dup MSF 10 120403 LMS MSF21 120404 MSF 21 120404 Dup
PFOS Conc. (n g /g )
0.806
0.823
52.2
1.07
0.752
53.2
4810 4740 9340 4120 4350 8580 71.4 70.0 108 595 558 1150 460 483 955 152 160 581 2900 2950 8140 1350 1360 6270 2070 1970 7480 1740 1930 7170 651 692
Avg. PFOS C o n c e n tra tio n
(n g /g )
RPD (% )
S u rro g ate ( 13C ,, -P F O S ) R e c o v e r y [a]
L M S S p ike C o n c e n tra tio n
(n g /g )
0.815
52.2
0.909 [b]
53.2
4770 9340 4240 8580 70.7 108 576 1150 471 955 156 581 2920 8140 1360 6270 2020 7480 1840 7170 671
2.0%
NA
34%
NA
1.4% NA 5.3% NA 1.9% NA 6.5% NA 4.9% NA 5.4% NA 1.6% NA 0.13% NA 5.2% NA 10% NA 6.1%
100%
100%
102%
105%
105%
107%
98.6% 107% 101% 107% 103% 98.4% 101% 102% 101% 100% 102% 98.9% 102% 100% 100% 103% 105% 107% 103% 101% 101% 101% 104% 100% 104% 106% 100% 94.8% 91.0% 100% 89.7% 96.3%
NA
50.5 [d]
NA
52.8 [d]
NA 4730
NA 4700
NA 46.0 NA 524 NA 482 NA 554 NA 5280 NA 4920 NA 5280 NA 5060 NA
L M S S p ike R e co very (% )
NA
102%
NA
99.1%
NA 96.5%
NA 92.4%
NA 81.0%
NA 108%
NA 100%
NA 76.7%
NA 98.7%
NA 99.8%
NA 104%
NA 105%
NA
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Study: ISO12-01-01-01 Analysis for PFOS in Fish Collected from the Tennessee River in April 2012
Table 11. Individual PFOS Results for Fillet Tissues from Fish Collected in the Tennessee River, April 2012
L IM S S a m p le ID
S a m p le D e s c rip tio n
ISO12-01-01-01-033 LMS ISO12-01-01-01-034 ISO12-01-01-01-034 Dup ISO12-01-01-01-034 LMS ISO12-01-01-01-035 ISO12-01-01-01-035 Dup ISO12-01-01-01-035 LMS ISO12-01-01-01-036 ISO12-01-01-01-036 Dup IS012-01-01-01-036 LMS IS012-01-01-01-037 IS012-01-01-01-037 Dup IS012-01-01-01-037 LMS IS012-01-01-01-038 IS012-01-01-01-038 Dup IS012-01-01-01-038 LMS IS012-01-01-01-039 IS012-01-01-01-039 Dup IS012-01-01-01-039 LMS IS012-01-01-01-040 Dup IS012-01-01-01-040 IS012-01-01-01-040 LMS IS012-01-01-01-041 IS012-01-01-01-041 Dup IS012-01-01-01-041 LMS
IS012-01-01-01-042
IS012-01-01-01-042 Dup
IS012-01-01-01-042 LMS
IS012-01-01-01-043 IS012-01-01-01-043 Dup IS012-01-01-01-043 LMS IS012-01-01-01-044 IS012-01-01-01-044 Dup IS012-01-01-01-044 LMS IS012-01-01-01-045 IS012-01-01-01-045 Dup IS012-01-01-01-045 LMS IS012-01-01-01-046 IS012-01-01-01-046 Dup IS012-01-01-01-046 LMS IS012-01-01-01-047 IS012-01-01-01-047 Dup IS012-01-01-01-047 LMS IS012-01-01-01-048 IS012-01-01-01-048 Dup IS012-01-01-01-048 LMS IS012-01-01-01-049 IS012-01-01-01-049 Dup
IS012-01-01-01-049 LMS IS012-01-01-01-050
MSF 21 120404 LMS MSF 22 120404 MSF 22 120404 Dup
MSF 22 120404 LMS MSF 23 120404 MSF 23 120404 Dup MSF 23 120404 LMS MSF 24 120405 MSF 24 120405 Dup MSF 24 120405 LMS MSF 25 120405 MSF 25 120405 Dup MSF 25 120405 LMS MSF 26 120405 MSF 26 120405 Dup MSF 26 120405 LMS MSF 27 120403 MSF 27 120403 Dup MSF 27 120403 LMS MSF 28 120404 Dup MSF 28 120404 MSF 28 120404 LMS MSF 29 120405 MSF 29 120405 Dup MSF 29 120405 LMS Post Study, Process Control #3 Post Study, Process Control #3 Dup (TN11-0194-1/1)
Post Study, Process Control #3 LMS (TN11-0194-1/1) Reference Tissue (NIST SRM Reference Tissue Dup (NIST Reference Tissue 1/2 LMS MSF 11 12-04-03 MSF 11 12-04-03 Dup MSF 11 12-04-03 LMS MSF 12 12-04-03 MSF 12 12-04-03 Dup MSF 12 12-04-03 LMS MSF 13 12-04-03 MSF 13 12-04-03 Dup MSF 13 12-04-03 LMS MSF 14 12-04-03 MSF 14 12-04-03 Dup MSF 14 12-04-03 LMS MSF 15 12-04-03 MSF 15 12-04-03 Dup MSF 15 12-04-03 LMS MSF 16 12-04-04
MSF 16 12-04-04 Dup
MSF 16 12-04-04 LMS MSF 17 12-04-04
PFOS Conc. (n g /g )
1080 4570 4740 9650 52.4 52.5 552 464 409 884 312 300 999 2100 2080 6830 2060 2180 7380 172 188 783 2050 2050 7100
0.984
0.839
Avg. PFOS C o n c e n tra tio n
(n g /g )
1080 4660 9650 52.5 552 437 884 306 999 2090 6830 2120 7380 180 783 2050 7100
0.912
RPD (% )
NA 3.5% NA 0.0056% NA 12% NA 3.8% NA 1.0% NA 5.4% NA 9.2% NA 0.063% NA
16%
S u rro g ate ( 13C 4 -P F O S ) R e c o v e r y [a]
90.1% 90.7% 94.1% 98.3% 97.8% 102% 91.9% 102% 87.1% 98.8% 95.8% 101% 91.0% 94.0% 100% 100% 95.1% 96.1% 102% 98.2% 107.9% 97.1% 92.0% 94.4% 102%
101%
99.0%
L M S S p ike C o n c e n tra tio n
(n g /g )
478 NA 5010 NA 502 NA 494 NA 554 NA 5030 NA 4950 NA 487 NA 4953
NA
56.3
6.85 6.58 54.9 2990 2880 7580 537 515 5450 942 958 5810 3360 3370 8270 718 790 5670 1550 |e|
1500 [e|
1950 [e| 165
56.3
6.71 54.9 2940 7580 526 5450 950 5810 3370 8270 754 5670 1520 1950 165
NA
4.0% NA 3.9% NA 4.2% NA 1.7% NA
0.505 NA 9.6% NA 3.2% NA
0.75%
103%
98.8% 102% 100% 100% 101% 101% 103% 99.0% 104% 96.5% 102% 100% 100% 97.6% 100% 98.7% 102% 102% 100%
99.0%
100% 101%
54.1 [d|
NA 50.6 NA 4970 NA 4880 NA 4910 NA 4810 NA 4940 NA 493 [c| NA
L M S S p ike R e co very (% )
86.0% NA
99.6% NA
99.7% NA
90.7% NA
125% NA
94.2% NA
106% NA
124% NA
102%
NA
102%
NA 95.3%
NA 93.5%
NA 101%
NA 99.0%
NA 102%
NA 99.4%
NA 86.9%
NA
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Study: ISO12-01-01-01 Analysis for PFOS in Fish Collected from the Tennessee River in April 2012
Table 11. Individual PFOS Results for Fillet Tissues from Fish Collected in the Tennessee River, April 2012
L IM S S a m p le ID
S a m p le D e s c rip tio n
PFOS Conc. (n g /g )
Avg. PFOS C o n c e n tra tio n
(n g /g )
RPD (% )
S u rro g ate ( 13C ,, -P F O S ) R e c o v e r y [a]
L M S S p ike C o n c e n tra tio n
(n g /g )
L M S S p ike R e co very (% )
IS012-01-01-01-050 Dup IS012-01-01-01-050 LMS IS012-01-01-01-051 IS012-01-01-01-051 Dup IS012-01-01-01-051 LMS IS012-01-01-01-052 IS012-01-01-01-052 Dup IS012-01-01-01-052 LMS IS012-01-01-01-053 IS012-01-01-01-053 Dup IS012-01-01-01-053 LMS
MSF 17 12-04-04 Dup MSF 17 12-04-04 LMS MSF 18 12-04-04 MSF 18 12-04-04 Dup MSF 18 12-04-04 LMS MSF 19 12-04-04 MSF 19 12-04-04 Dup MSF 19 12-04-04 LMS MSF 20 12-04-04 MSF 20 12-04-04 Dup MSF 20 12-04-04 LMS
166 581 710 691 6080 672 678 5620 1170 1180 6150
581 701 6080 675 5620 1180 6150
NA 2.7% NA 0.86% NA 0.82% NA
100% 100% 100% 101% 99.0% 101% 102% 100% 101% 100% 101%
493 NA 5220 NA 4780 NA 4780
84.2% NA
103% NA
104% NA
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 % 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.
23
Study: IS012-01-01-01 Analysis for PFOS in Fish Collected from the Tennessee River in April 2012
The results for the three process controls showed that contamination during homogenization and shipping from MSU-WTL to the 3M Environmental Laboratory did not occur at any significant level that would affect the results.
The analytical method uncertainty for the data set was determined based on LCS results. Sets of LCSs were prepared with each extraction batch by spiking L-PFOS or br-PFOS into control fish tissue homogenates at nominal 10, 100 and 1000 ng/g, each level in triplicate. The method uncertainty is the range about the mean with which it is expected to statistically find the reported results with 95 out of 100 measurements, referred to as the 95% confidence interval (95% Cl), and is + 2 Std. Deviations of the LCS data. For L-PFOS the method uncertainty (95% Cl) was + 19% and for brPFOS (30.6% branched isomers) the method uncertainty (95% Cl) was + 11%.
8 Data/Sample Retention
All fish samples and associated project data (hardcopy and electronic) will be archived according to 3M Environmental Laboratory standard operating procedures as project IS012-01-01-01.
9 List of Attachments
Attachment A: Preparation Forms, Raw Data and Chromatograms
10 Signatures
Report Approval:
Cleston C. Lange, Ph.D., Principal Analytical Investigator
^ /z bl2
Date
William K. Reagen, Ph.D., 3M Environmental Laboratory Management
3 / n / q y cXo/ Date
5 - 2 - IX . Date
24
Study: ISO12-01-01-01 Analysis for PFOS in Fish Collected from the Tennessee River in April 2012
Attachment A Preparation Forms, Raw Data and Chromatograms
(Available Upon Request)
25
