Document ZzYZ0ONJK26EpzyrBYgNXyrY
Jean B. Sweeney Vice President
3M Environmental, Health and Safety Operations
3M Center, Building 0224-05PIVb3 St . Paul, MN 55144-1000 651 7373569
February 11, 2010 CERTIFIED MAIL
ORIGINA-L
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/k J~-a-~ ~ -3 9 ~.~
Document Processing Center EPA East - Room 6428 Attn : Section 8(e) Office of Pollution Prevention and Toxics, U .S . EPA 1200 Pennsylvania Avenue NW Washington, DC 20460-0001
NO CBI c.~
Re : TSCA 8(e) Substantial Risk Notice : Sulfonate-based and Carboxylic-based Fluorochemicals, Docket 8EHQ-0598-373 - Results from 2009 Analyses of Fish from the Mississippi River
Dear Sir or Madam :
3M is submitting this notice to supplement previous submissions on sulfonyl and carboxylie,
based fluorochemicals (FCs), and more specifically our June 20, 2006 and November 9, 200
submittals concerning fish data generated by the Minnesota Pollution Control Agency
(MPCA) . The November 9, 2009 submittal included an Excel file, provided to 3M by the ==~~
MPCA, entitled "PFCs MissPool2 Fish all years.xls" and 3M Environmental Laboratory 0. Report No. E09-0372, Fluorochemical Analysis of Fish Fillet Homogenates Collected from_
the Mississippi River by the Minnesota Pollution Control Agency May 2009 - Phase 1 :
PFOS .
.
The aforementioned Excel file included fish fillet data for 297 fish collected from Pool 2 of"" the Mississippi River in 2009 that were analyzed by the MPCA for C4-C 12 perfluorocarboxylic acids; C4, C6 and C8 perfluorosulfonates ; and
perfluorooctanesulfonamide . The 3M Environmental Laboratory Report presented fish tissue analytical results for PFOS for 30 of the 297 fish samples that were split in order to obtain confirmatory data.
Enclosed with this correspondence is a second 3M report entitled "3M Environmental
Laboratory Report No. E09-0372, Fluorochemical Analysis of Fish Fillet Homogenates
Collected from the Mississippi River by the Minnesota Pollution Control Agency May 2009 Phase 2 : Perfluorocarboxylic Acids (C4-C 12), Perfluorobutane sulfonate (PFBS),
Perfluorohexane sulfonate (PFHS) and Perfluorooctanesulfonamide (FOSA) . This report presents additional corroborative analytical data for the previously referenced 30 fish for the analytes indicated . As noted in the November 9 correspondence, sample identification
numbers in the Excel spreadsheet and the 3M Environmental Laboratory Report can be crossreferenced to compare split sample results.
While 3M does not believe that any of the referenced or submitted data taken alone or cumulatively meet the "substantial risk" reporting threshold, and while 3M was not responsible for the generation of this information (other than the 3M analytical data provided here), we nevertheless recognize the ongoing work by U.S. EPA to assess fluorochemical
CONTAINS NQ (M
U.S . EPA Document Processing Center February 11, 2010 Page 2
exposure pathways. Therefore, we are placing these results in the 8(e) docket as a supplement to previous submissions. If you have any questions, please do not hesitate to contact Deanna Luebker at (651) 737-1374 or djluebker@mmm .com. Sincerely,
,4~ ( 4gpr)
Jean B. Sweeney Vice President Environmental, Health and Safety Operations Enclosure
3A4 ENVIRONMENTAL LABORATORY REPORT NO. EO~;0372 .t
Final Analytical Report
Perfluorochemical Analysis of Fish Fillet Homogenates Collected from the Mississippi River by the Minnesota Pollution Control Agency in May 2009 - Phase II : Perfluorocaryboxylic
Acids (C4-C12), Perfluorobutane sulfonate (PFBS), Perfluorohexane sulfonate (PHFS) and Perfluorooctanesulfonamide (FOSA)
Laboratory Request Number: E09-0372
Method Requirement: ETS 8-45.0
Testing Laboratory 3M EHS Operations 3M Environmental Laboratory
3M Center Building 260-5N-17 Maplewood, MN 55144
Requester Gary Hohenstein 3M EHS Operations
3M Center Building 224-5W-03 Maplewood, MN 55144
111"d WIN`
ACCREOtTED Certificate #2052-01
The testing reported herein meet the requirements of ISOItEC 17025-2005 "General Requirements for the Competence of Testing and Calibration Laboratories", in accordance with the A21A Certificate ait2052-01 . Testing that complies with this International Standard also operate in accordance with ISO 9001 :2000 .
3M ENVIRONMENTAL LABORATORY
PAGE 1 OF 104
3M ENVIRONMENTAL LABORATORY REPORT NO. E09-0372
3M Environmental Laboratory 3M Environmental Laboratory Manager: William K. Reagen, Ph .D . 3M Principal Analytical Investigator and Report Author: Michelle D. Malinsky, Ph.D.
Analytical Report Ea9-t7372 Perfluorochemical Analysis of Fish Fillet Homogenates Collected from the Mississippi
River by the Minnesota Pollution Control Agency May 2009 - Phase II : Perfluorocarboxylic Acids (PFCAs) C4-C12, PFBS, PFHS, and FOSA
Introduction
Report Date: Upon Signing
The Minnesota Pollution Control Agency (MPCA) submitted thirty fish fillet homogenates to the 3M Environmental Laboratory for perfluorochemical analysis under project number E09-0372. The thirty submitted samples represented an approximate ten percent split of the total samples collected during a May 2009 fish sampling campaign from Pool 2 of the Mississippi River. Six samples from five different species comprised the sample set. The fillet homogenates were extracted and analyzed using method ETS 8-45.0 "Determination of Fluorocinemicals via Protein Precipitation of Fish Tissues (Fillet or Whole Body) and Analysis by High Performance Liquid Chromatography with Tandem Mass Spectrometry" . The target analytes for this study included perfiuoroalkane carboxylic acids ranging from C4 to C12, perfluoroalkane sulfonates (C4, C6, and C8), and perfluorooctaresulfonamide. Results for PFOS were provided in the Phase I report. This Phase II report provides the results from the other target analytes. The full chemical names, structures, and acronyms for each analyte are provided in Section 2.1 .
The summarized results have been broken out into four separate tables: " Table 1: perfluorobutanoic acid (PFBA), perfluoropentanoic acid (PFPeA), and perfluorohexanoic acid (PFHxA). " Table 2: perAuoroheptanoic acid (PFHpA), perfiuorooctanoic acid (PFOA), and perfluorononanoic acid (PFNA). " Table 3: perfluorodecanoic acid (PFDA), perHuorouncecanoic acid (PFUnA), and perfluorododecanoic acid (PFDoA) " Table 4: perfluorobutane sulfonate (PFBS), per8uorohexane sulfanate (PFHS), and perfluorooctanesulfonamide (FOSA).
A!I results for quality control samples prepared and analyzed with the samples will be reported and discussed elsewhere in this report.
111"d MFM ACCREDITED
The testing reported herein meet the requirements or tsonEC 1702s-2o06 c,enerai
Requirernents for the Competence of Testing and Calibration Laboratories", in accordance with the A2LA Certificate #2Q52-01 . Testing that complies with this International Standard also operate in accordance with ISO 9001 :2000.
Certificate #2052-01
3M ENVIRONMENTAL LABORATORY
PAGE 2 OF 104
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O- O-'
O- 0_
U Nf Q ~ Q 4 i K Q :
Q Qi
Q Q'
Q Q
g
N N
Nr N~
9 Qi N N-
R~ M Mi
uU W ;
M oMlI
N N~ 9 Q~ N N' R h M Mi
W W;
o~ 't -,r I
N N:
Q 9i N N~ R R' M Mi
o53 ~W' :
o~
M) 1q i N Ni q QI N N R R~ M Mi
ui w ;
<o0I
<O N Ni Q 4 N N h R M Mi
W w;
o~ h I-- :
N N, Q qi N N~ R R; M Mi
W W~
o
OD OD : N fV~ Q Qi fY Ni R R; M Mi
w a
byt~ CO fki ~w
mg0z
Jrx
w~ z
z w m
ar. o a~o g
U
!
k
~~r e
~ Ni N Q
N C4~1i h
Q
2
~
0
U
!
TI U
Z ~
o
V
V= aV
~
o
V
oVl' ~p V
ep y
k Of f ~ N ; to N
f
eV~~
1 y~f
N Ni 1~ p.~ 4
~?~
O C; j K O GOi ? G.'
Q at k r
r
G Y
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~
r
i
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r r
f 1
3
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L
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i
k
m i
{
m ii k
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d 03 I
t~ U~ m ai"
CV (C
fj pi t~ UIf P_,: r;;
?
_I ~~
m a o;
aoi J J' f
R T!' Ek
~k
R O
~'1 k
r,
M
fl Mr
11 M Mk
t`'~' W W ;
W W;
3M ENVIRONMENTAL LABORATORY REPORT NO . E09-0372
dy and Sample Information 2.1 Target Analytes.
Table 5 below provides pertinent information regarding the target analytes, internal standards, and surrogate recovery standards investigated in this study. Surrogate recovery standards were separate isotopically labeled compounds spiked into each sample aliquot prior to extraction . The recovery of the surrogate was used to evaluate the overall extraction process and any potential matrix effects .
Table 5. Target Analyte Summary .
Compound Name
PerfluorobutarKk acid Pa1(uoropentarpic aad Perlfuaohexanoic acid Pertluoroheptanoic acid
~ acid
Perfluorononarxx add PeAluorodecanac acid Perkmroundecarxxc acid
add Pertfuorobutane sutfonate PerAuorohexane suFonate
Synonym r Acronym
PFBA (C4 Acid) PFPeA (C5 Add) PFHxA (C6 Acid) PFHpA (C7 Acid)
PFOA (C8 Add)
PFNA (C9 Add) PFDA (C10 Acid) PFUnA (C11 Acid) PFDoA (C12 Acid) PFBS (C4 Sulfonate) PFHS (C6 Sulfonate)
PerAuaooctane sulfonate
PFOS (C8 S~te)
P~
~ FOSA) (CS
Formula
C,F,COOH C,FoCOOH C5F COOH C,F,COOH G,F COOH CTFCOONI-4' CsFCOOH CoF,qCOOH C,aF2,COOH . C FnCOOH C,FoSO,1C' C,F Sp,-Na'
C!~v+
CeF tT-n~ ~
~1T~~
CeFSOZNHZ
Analytical Purpose Target Target Target Target
T~ Taw Target Target Target Target Target
Target
Target
Reference Standard source
Akhch
Nta Aasar
Oakwood Products
Aldrnch
L-PFOA (~)
W
Labs
ECF PFOA (brandtied) 3M Produclion Lot 332
Oakwood Products
Oakwood Products
Oakwood Products
Oakwood Products
3M Production Lot 2
L-PFHS (knear) Wellington Labs L-PFOS {5ne~) Wellingtaon Labs ECF PFOS (b~ed) 3M Production Lot 217
3M (L-15709)
3M ENVIRONMENTAL LABORATORY
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3M ENVIRONMENTAL LABORATORY REPORT NO . E09-0372
Table 5 Continued .
Compound Name
'3C,-Perfluorobt#8rM acid
'30rPerffuomhexanoic acid '3C4-Perfluoroadanaic add '3Cs-Perfkuoronananoicadd '3Gr-Perfluorodecanoic acid '3CrPerfk,or(xjnde,canoiC ecid '3CrPeftaododecenac acxi 'eOrAmmoriiixn Perfluarobufene sulfonate 'eOrA~mor~m Perfl~nt~aare
s'C,Sodiurn Perfluorooctane sulfonat efonate
'~OrPe&oraoctanesulfonwvde
'3Cz-PertauorooQanaic add
'
Pedimctam
synonym r Acronym [1,2,3,413C4~FBA [1,2 -,3C2]PFHxA [1,2,3,413C4PFOA [1,2,3,4,5~'3C5]PFNA [1,2 -'3C,JPFDA [1,2 -13Cz[PFUnA [1,2 '3C~]PFDoA [,g0APFBS ['~PFHS
[1,2,3,4-'3C,IPFOS ['BOJFOSA
[1,2-'3C,]PFOA ['fl0JPFOS
~u~
13CF~(13CF~13CL10H CF3(CFz},{13CF,),3COOH CF,{CFZ},('3CF2b'3COOH CF3{CFZk,('3CF,),'3COOH CF3(CF2),("CF,)'3COOH CF3(CF2M13CFZ),3COOH CF,(CF,),t13CF2)13COOH
[C4FsS"e0ZOrNH,' [C,F,3S'BOZQjNH,` CF3CFz,'3Nza),'3CF,S03
[CBF S1sOZNHJ CF3(CF2M "CF2'3COOH
[C..FS'e02OrNi-l`
Arratyrpcaf Purpose
IS IS IS is IS IS IS IS
IS
IS IS Sturogate Surrogate
Refererrce Standard source
Welling6on Labs WeAirx,fton Labs Weing6on Labs WellffVton Labs WellirKjton Labs WeAirxjtOn Labs Wel6ngton Labs RTI Inbemational Wellinglon Labs Wellngtm Labs RTllntemationd Perkn Elrner RTIIrrtenzetional
2.2 Sample Collection and Receipt. On August 6, 2009, the 3M Environmental Laboratory received the homogenized fillet samples in Nalgene bottles on dry ice . The samples were logged into the 3M Environmental Laboratory LIMS system under project E09-0372 and placed in frozen storage. No information was provided regarding the sampling date.
It should be noted that these fish samples have been previously handled and analyzed at another laboratory under contract by the MPCA . It is unknown how these samples have been handled and if the integrity ofthese samples has been comprised prior to receipt .
3M Environmental Laboratory personnel removed the homogenized fish tissue from the sample container by cutting the bottle away from the frozen sample mass using a clean utility knife. The frozen sample mass was then rehomogenized before extraction. See Section 3.1 for more information. Based on the sample consistency in the Nalgene bottles, it appeared that the sample homogenates had thawed at some point before they arrived at the 3M Environmental Laboratory. (Homogenized tissue had refrozen into a solid mass that could not be broken up while frozen in the container.)
3 Methods -A
cal and Preparatory
3.9 Tissue Preparation Each submitted sample and bluegill fillet control tissue (TN08-0220-1(1) purchased from Osage Catfisheries, Inc. (Osage Beach, MO) were rehomogenized frozen using dry ice .
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Frozen fish homogenate was added to a pre-chilled stainless steel bawl of a Robot Coupe RSI 2Y-1 vertical batch processor and homogenized with dry ice until a fine powder consistency was reached . After homogenization, the ground fish tissue was transferred to a polyethylene bag which was placed in the freezer at -20C unsealed overnight to allow the residual carbon dioxide to sublime . After sublimation, the bag was sealed and homogenates were kept frozen.
3.2 Sample Extraction
A 0 .25 g sub-sample of the fish tissue homogenate was accurately weighed in a 50 mL polyethylene centrifuge tube. Tissue aliquots were then spiked with internal standards (ISs) and surrogates to produce an approximate concentration of 1 nglg. Samples designated as lab matrix spikes or matrixmatched calibration standards were additionally spiked with the target and surrogate anatytes at appropriate levels to produce the desired tissue concentrations. Samples intended for laboratory QC (laboratory control spikes, LCSs) were spiked with surrogates at 1 nglg and the ottw target analytes at approximately 0.5 nglg, 1 .5 nglg, and 8 rig/g . After spiking, the tissue homogenates were allowed to sit for 30 minutes before 2.5 mL ofacetanitnle was added. The samples were then thoroughly homogenized with the acetonitrile using an Omni Prep multi-place horrtogenizer with disposable plastic probes (15,000 rpm for 2 minutes). The acetonitrile/tissue extracts were then placed in a freezer at 20C for at least one hour. Upon removal from the freezer, sample extracts were centrifuged at a targeted temperature of-5C for 20 minutes at 3000 rpm .
After centrifugation, 1 mL of clarified supernatant was transferred to a 2 mL autovial spiked with 10 uL of 10/fl formic acid. Autovials were then capped and vortex mixed.
Samples were extracted in three separate batches . For each submitted sample, a sample, sample duplicate, and laboratory matrix spike (LMS) were prepared. Each extraction batch included the following quality control samples:
" a thirteen point extracted matrix calibration curve using the purchased bluegill fillet control tissue
" triplicate lab control spikes of the bluegill fillet control tissue at three levels of all target analytes (0.5 nglg, 1 .5 ng/g, and 8 ng/g)
" triplicate lab control spikes of 3M electrochemical fluorination (ECF) PFOA and PFOS at 5 ng/g
" triplicate lab control spikes of PFOS only at 100 nglg " duplicate control matrix blanks with and without internal standards and surrogates " duplicate method (aqueous) blanks with and without internal standards and surrogates
The table below summarizes the preparation dates and samples prepared in each batch .
Table 6. Sample Preparation Summary.
Batch Number
Batch 1 Batch 2 Batch 3
Preparation Dates
8Y1512009 - 8Rfil'I009 812712009 - 8128d2009 8131C1009-9l1I2009
samples
E090372-001 to E09-0372-010 E09-0372-021 to E09-0372-030 E09-0372-011 to E09-03T2-020
3.3 Analysis Analysis for the PFCs listed in Table 5 was performed using high performance liquid chromatographytandem mass spectrometry (HPLCIMSIMS). For this investigation, samples were analyzed using an Agilent 1100 series (Palo Alto, CA) HPLC system interfaced to either a PE SCIEX API 4000 triple
3M ENVIRONMENTAL LABORATORY
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3M ENVIRONMENTAL LABORATORY REPORT NO . E09-0372
quadnapole or SCIEX API 4000 Q-Trap mass spectrometer (Foster City, CA) . Both instruments were equipped with a SClEX Turbo V ion-spray interface operating in the negative ion MS/MS mode using multiple reaction monitoring (MRM) . AnalystTm 1 A.2 software was used for all data collection and reduction. Table 7 lists the MRM transitions monitored for each anatyte . A Thermo Scientific PRISM RP guard column (2 .1mm x 50 mm ; 5u particle size) was placed in-line after the purge valve and before the sample injection port to trap any PFC contaminants coming from the NPLC instrument and/or the mobile phases. This sufficiently separated the elution of the "system" PFC peaks from those present in the sample extracts.
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Table 7. MRM Transition Summary .
Compound
PFBA C4Acid PFPeA (C5 Acid)
A
T T
PFWA C6 Add
T
PFH C7Add
T
PFOA C8 Acid
T
PFNA C9 Add
T
PFDA C10 Acid)
T
PFUnA C11 Acid)
T
PFDoaA C12 Acid ) PFBS C4 Sulfanate PFHS C6 Suffanate
T T t T
PFOS C8 Sulfonate FOSA C8 StdFor~ [1,2,3,4 ?'Cs]PFBA [1,2 "CPFHxA [1 z,3,~3C)PPO,q
[1,2,3,4,5-T3C~)pFNq [1,2 !'Cz]PFDA [1,2 -"CjPFUnA [1,2 !3CJPFDop, ('eO,jPFBS rO,JPFHS [1,2 .3,4 "C~IPFOS ['80JFOSA [1,2 -'3Cz[PFOA ['eOJPFOS
T T IS for PFBA and PFPeA IS for PFHxA IS for PFHpA and PFOA IS for PFNA IS for PFDA IS for PFUnA IS for PFDoA IS for PFBS IS for PFHS IS for PFOS IS for FOSA Suffogate (Acids) Surto9ate(Sulfonabes, FOSA)
MMRM Tra ' s
213>169 263>219 313>209 313>119 363>319 363>169 413>369 413>219 413>169 463>419 463>219 463>169 513>469 513>269 513>219 563>519 563>269 563>219 613>569 613>319 613>169 299>80 299>99 399>80 399>99 499>80 499>99 499>130
498>78 217>172
315>270
417>372
46423
515>470
565>520
615>570
303>&f
403>84
503>80
502>82
415>370
503>84
Dwett 77me ms
1 100 100 100 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 100
100
50
50
50
50
50
50
50
50
50
50
50 ---- I
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Analysis of the short-chain acids (C4 through Cs) was performed using a Thermo Scientific PRISM RP column (2.1mm X 50 mm; 5 u particle size) held at 30C with the mobile phase system consisting of 5 mM ammonium acetate with 0.01 % acetic acid in water (A) and methanol (B) . The gradient used to elute the analytes of interest is presented in Table 8. The flow rate was held at 300 pUmin throughout the run and a 20 pL injection volume was used. The outlet of the analytical column was directed to a column switching valve where the first three minutes of the run were diverted to waste. After three minutes, the valve was switched to direct the effluent to the mass spectrometer for analysis. The large-chain acids (C7 through C12), the sutfonates, and FOSA were analyzed using a Then-no Scientific BetasilT"" C18 analytical column (2.1mm x 100 mm; 5p particle size) held at 30C with the mobile phase system consisting of 2 mM ammonium acetate in water (A) and acetonitrile (B) . The gradient used is also provided in Table 8 . The flow rate was held at 4001zUmin throughout the run and a 25 uL injection volume was used. Samples were injected onto a Waters (Milford, MA) Oasis@ HLB on-line extracaction column (3.0 mm X 20 mm, 25 p particle size) with the outlet directed to a column switching valve where the fust five minutes were diverted to waste. After fire minutes, the valve was switched and the effluent was directed to the analytical column for separation of the target analytes and analysis by the mass spectrometer.
Table 8. LC Gradient Parameters.
Small Acids: C4-C6
Step Total rime (min)
%q
Large Acids: C7-C12, Sulfonatres, FOSA
gbg
Totalf;me stop
YaA
%B
0
0.0
1
3 .0
2
3 .5
3
9 .0
4
15.0
5
15 .1
6
19 .0
90
10
0
0.0
97
3 .0
90
10
1
3.0
97
3_0
30
70
2
3.5
70
30
5 .0
95
3
13 .5
40
60
5 .0
95
4
15 .5
40
60
90
10
5
16 .0
10
90
90
10
6
18.0
10
90
7
18.3
97
3.0
8
21 .0
97
3.0
A
5 mM amrariun acetate with 0 .01 % acetic add (aq)
A
2 mM artrnmm acetate (aq)
B
MeUaid
B
Acetondrde
To manage the large number of transitions required for the large acids, sulfonates, and FOSA, a mulUperiod method was constructed. The "first period (time--0 minutes to 11 minutes) collected the transitions for PFBS, PFHpA, PFOA, {1,2 -'3CJPFOA, PFHS, PFhlA and their respective ISs . The second period (time=11 min to 21 .0 minutes) collected the transitions for PFDA, PFUnA, PFOS, ['$OAPFOS, PFDoA, and FOSA and their respective ISs . For analytes where more than one transition was acquired, a total ion chromatogram (TIC) which summed the respective transitions for that analyte was used to quantitate the results for that analyte .
'Exact location of the period break may vary as individual analy6cal colLnm may perform chifere.ntly or if instrument tubing is dWged .
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4,1 Calibration Extracted *matrix-matched
calibration
standards
and
QCs
were
prepared
by
spiking
known
amounts
of
target analytes, surrogates, and internal standards into individual 0.25 g aliquots of bluegill fillet control
tissue (TN08-0202-111) homogenate . Each spiked fish aliquot was then extracted using the
procedures outlined in Section 3.2. A total of thirteen spiked standards ranging from 025 nglg to 300
nglg (nominal) were prepared. Additionally, an unextracted acetonitrile solvent curve ranging from
0.0025 nglmL to 50 nglmL was also prepared which is equivalent to 0.025 nglg to 500 nglg tissue
concentration .
All sample results reported here were quantitated against the extracted curve except for PFBA and PFPeA. For PFBA, the endogenous concentration present in the bluegill fillet control tissue (historical average = 4.44 nglg t 8.3% for N=6 measurements) was significantly higher than the PFBA levels observed in the study samples . Adjusting the concentration of the extracted calibration standards for the endogenous level would have produced limits of quantitation very dose to the lab matrix spike level rendering the sample results generally urueportable. Therefore, the solvent curve was used for reporting the sample results . As for PFPeA, the solvent curve was used for quantitation because the response (raw area counts) of the [1,2,3,4-13C4]PFBA surrogate internal standard in the study samples more closely approximated the surrogate internal standard response in the solvent curve than it did in the extracted calibration curve. The average [1,2,3,4-'3C4]PFBA area counts in the extracted bluegill control matrix was approximately 80% (Batch 1), 56% (Batch 2), and 77% (Batch 3) of the average area counts in the solvent curve. For the study samples, the average ([1,2,3,4-13C4]PFBA area counts was 97% (Batch 1), 84% (Batch 2), and 91 % (Batch 3) of the average area counts in the solvent curve.
A quadratic, 11x weighted calibration curve was used to ft the data for each analyte . Internal standard quantitation was used. The data was not forced through zero during the fitting process . The accuracy of each curve point was verified by back-calculating the concentration using the area count ratio of the target analyte to the internal standard. Each solvent calibration standard used to generate the final calibration curve met the method calibration accuracy requirement of 100t25% (100t30% for the LOQ). The coefficients of determination (rz) were greater than 0.990 for all analytes. As required by ETS 8-45, a minimum of six calibration points were used to generate the final calibration curve .
For PFHS, PFOS, and PFOA, the reference standard used for calibration and lab control spikes was comprised predominantly of the linear isomer. A separate set of LCSs using an electrochemical fluorination (ECF) source of PFOS and PFOA was also included .
*As fish matrix can vary "Uy from species to species, ft contrd mahix used to generate the extraded calilxation curve and latboratory control QC samples is cormdered to be mahvc4natdied for the bluegl samples onN. The ta"atorY control matrix shaild be considered as a 'surrogate' control matrix for ft dher species investigated in Ctis study .
.4.2 Limit of Quantitation (LOQ) The LOQ as defined in ETS 8-45 is the lowest non-zero calibration standard in the curve in which the area counts of the target analyte are at least twice those of the matrix blank(s) . When unextracted solvent quantitation was used, the area counts of the extracted aqueous method blanks was used to assess the LOQ .
4.3 System Suitability Three replicate injections of a solvent calibration standard were analyzed at the beginning of the analytical sequence to demonstrate overall system suitability. Method criteria states that system suitability injections shall produce a RSD of less than 7% for the ratio of target analyte area counts to
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3M ENVIRONMENTAL LABORATORY REPORT NO. E09-0372
internal standard area counts and an RSD of less than 2% 1w the retention time. All analytes met method acceptance criteria for system suitability with the following exceptions:
" Extraction Batch 1 : Area Count RSDs- PFNA (13%), PFDA (10%), PFUnA (13%), [1,213C~PFOA (8.1Jo), PFBS (9.1%), and PFHS (8.3fo)
" Extraction Batch 2: Area Count RSDs - PFNA (7.5%), PFUnA (19%), PFBS (17%), and PFHS (12%). Additionally, no system suitability injections were analyzed for the short acids (PFBA, PFPeA, and PFHxA).
" Extraction Batch 3: Area Count RSDs - PFOA (10k) A method deviation has been issued and is included in the data package. The overall impact to the sample results is considered minimal . An increase in variability in the area count precision is often observed with an increased number of MRM transitions collected within a given period of the data acquisition method . With a large number of MRMs, fewer scans across the chromatographic peak are collected and overall peak shape can vary more from injection to injection . However, the calibration curve, QCs, and samples all experience the sarne conditions. Any increased variability observed in the system suitability injections would also be observed in the LCS data which is used to determine overall method uncertainty. Thus, the variability is accounted for in the assigned method uncertainty reported with the sample results.
4.4 Continuing Calibration
During the course of the analytical sequence, several continuing calibration verification samples (CCVs) were analyzed to confirm that the instrument response from initial calibration curve was still in control .
For PFCAs (C7-C12), PFBS, PFHS, FOSA, [1,2-13C2]PFOA, and [180JPFOS a8 CCV injections (extracted curve) produced recoveries within 100%t25%, which met method criteria . " For PFBA, and PFPeA, no solvent curve CCVs were analyzed in the second batch (s090831 a) as the solvent curve vials were inadvertently not placed on the autosampler tray until the end ofthe analysis. However, the extracted curve CCVs analyzed throughout the run were compliant except for one injection of PFPeA (74 .1 lo). This indicated that the overall instrument response was stable throughout the run .
" In the third batch, one CCV injection did not meet method acceptance criteria for PFBA (139l0) and PFHxA (198%) . The samples prior to this injection were LCSs . These LCSs were included in the method uncertainty determination and any nonoompliant recoveries essentially increased the overall uncertainty. The sample results after this injection were not reported for PFHxA due to non-compliant LMS recoveries.
A method deviation has been issued and is included in the data package .
4.5 Blanks Five types of blanks were prepared and analyzed with the samples: matrix blanks of bluegill fillet control tissue (two with IS and surrogate, two without IS and surrogate), aqueous method blanks (two with 1S and surrogate, two without IS and surrogate), acidified acetonitrile solvent blanks, acetonitrile blanks with internal standards and surrogates, and acetonihile solvent blanks (unmodified) . Each blank result was reviewed and used to evaluate method performance to determine the LOQ for each analyte . Surrogate recoveries of spiked blanks are provided in the Supplemental Information Section 10.3 4.6 Lab Control Spikes (LCSs) Triplicate lab control spikes of the purchased bluegill fillet control matrix at three different levels for all analytes were prepared each extraction day to determine analytical method uncertainty (See Section 4.9) . For PFHS, PFOS, and PFOA, the standard reference material used for the LCS spikes was the linear isomer, which was the same used for the calibration curves. Additional triplicate PFOS LCSs at
3M ENViRONhfENTAL LABORATORY
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3M ENVIRONMENTAL LABORATORY REPORT NO . E09-0372
100 ng/g were prepared as most of the study sample concentrations were anticipated to be around this
concentration level . Separate technical grade (3M electrochemical fluorination) spikes of
branched/linear isomer standards of PFOS and PFOA were prepared to evaluate any potential bias
from quantitation against a linear standard. Lab control spikes were prepared to evaluate method
accuracy and precision and were used to determine analytical method uncertainty (See Section 4.9).
The [1,2-'3Cz]PFOA and [18NPFOS surrogate were spiked at a consistent 1 ng/g (0.1 nglmL) for all control spike recoveries are provided in the Supplemental Information (Section
LCS samples . Lab 10.1) . For ETS 8-45.0,
method
acceptance criteria
for LCS
recoveries
is
104t30%
with
RSD
<20%.
4.7 Sample Concentration Calculation The extracted calibration curve or solvent calibration curve used for quantitation provided extract concentration in units of nglmL . The following equation was used to convert the extract concentration to sample tissue concentration in units of nglg.
Extract Concentration ( ~ )' ExtractionVolume(mL)
Tissue Concentration (2g ) =
E~~~ Mass (g)
Extraction Volume = 2.5 mtExtraded Mass -0 .25 g
4.8 Lab Matrix Spikes (LMSs) For each submitted sample, a separate laboratory matrix spike was prepared by extracting a separate aliquot of the fillet tissue that was spiked with the target anafytes. For each LMS, all target anafytes were spiked at a nominal concentration of 5 ng/g except PFOS, which was spiked at a concentration of approximately 110 nglg. The recovery of the LMS was calculated using the following equation.
(Extract Conc ( mL )' ExtractionVolume(mL)}-(Average Sample C~c.~9 1' LMS Sample Mass (g)} f
1
l 1
f t~
LMS%Recovery= l
Spike Amount (rig)
Although ETS 8-45.0 does not provide specific criteria for the lab matrix spike concentrations, the 3M Environmental Laboratory uses a general guideline that lab matrix spike concentration should be at least half the endogenous concentration before the spike is considered "appropriate" for the given matrix . For PFOS, the LMS was 27 .6 rig (-110 nglg.tissue concentration) except far sample E090372-012 where the LMS was 128 ng (-510 ng/g tissue concentration) . The FOSA LMS forthis sample was also increased to -110 nglg tissue concentration . The [1,2-13CAPFOA and ['BO~]PFOS surrogate was spiked at 0.261 ng (-1 ng/g tissue concentration).
4.9 Analytical Uncertainty ETS 8-45.0 states that the analytical method uncertainty will be determined by statistical evaluation of the individual analyte recoveries in LCSs. Current project QC results, as well as historical values, are used in the evaluation until a maximum of fifty points (minimum of twenty, if possible) are included . For ETS 8-45.0, the historical LCSs are categorized by tissue type: whole-body or fillet unless there is strong evidence suggesting that a species-specific matrix effect is present and that results from these species should be considered separately . The standard deviation of the collective recovery results (in %) is calculated. The expanded uncertainty is then calculated by multiplying the standard deviation by a factor of two, which corresponds to the 95% confidence limit . Table 9 below provides the analytical uncertainty for the target analytes.
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3M ENVIRONMENTAL LABORATORY REPORT NO . E09-0372
Table 9. Analytical Method Uncertainty
Average %Recovery Standard Deviation 2'Standard Deviation
Analytical Uncertain
n)PFBA
99.2 6 .56 13.1 50 13k
"WP-,4
97 .9 24 .0 47 .9 27 d8Yo
PFHxA
103 19 .6 39 .1 50 t39%
PFNPA
91 .9 10 .7 21 .3 50 21%
PFOA
101 9.68 19.4 50 19
PFNA
101 11 .1 22 .2 50 22
PFDA
97 .5 8.65 1Z3 50 17
PFUnA
PFDaA
PFBS
PFNS
FOSA j1,2-"C?jPFOA ('"O~]PFOS
Average %Recovery
101
96 .6
96 .9
99 .1
101
105
Standard Deviation
10 .5
6.34
9.55
13 .3
10 .2
9 .63
2'Standard Deviation
21 .1
12.7
19 .1
26 .6
20 .5
19 .3
50
50
50
50
50
50
~ Analytical Uncertainty I
21
1 t13 1 19i6 1 t27%
20%
+19
(1) A1 LCS reoovenes far PFBA arid PFPeA used to determine uncertainly were calculated
solvent curve. The
PFPeA LCSs from this study ~r were ~d for lhe analytical rnettiod uncertainty deterri~ti~
(2) For N>27, sane of the t>ist0fical LCSs were prepared in walleye fillet.
Data Summary and Discussion
106 8.73 1Z5 50
te%
5.1 All Samples The tables below provide the results for the sample, sample duplicate, and lab matrix spikes for the target anatytes and the two surrogates . Except for PFBA and PFPeA, al! sample results were reported from the extracted calibration curve prepared in bluegill fillet control matrix. In general, most samples produced lab matrix spike and surrogate spike recoveries within method acceptance criteria of 100t30% with RPDs less than 209'o between the sample and sample duplicate. Exceptions have been flagged appropriately in each data table.
The recoveries of the 3M PFOA ECF LCSs prepared with each extraction batch (N=9 for the study) ranged from 82.3% to 113% with a calculated average of 98.0 (11% RSD). See Section 10.1 . Simiiiariy, the recoveries ofthe 3M PFOS ECF LCSs (E09-0372 Phase I Report) ranged from 98.9% to 114lo with a calculated average of 107% (4.7% RSD). The results of the ECF QC spikes demonstrated that there is no discernible bias when PFOA andlor PFOS ECF isomers are quantitated against the predominantly linear reference standard . Therefore, the reported PFOA and PFOS values are considered accurate to within the stated method uncertainty regardless of the degree of branched isomers present in the sample extracts .
5.2 Sample E09-0372-012 (Client ID 090064, Smallmouth Bass River Section 4)
Sample E09-0372-012 exhibited the highest concentration of PFCs collectively: PFOS (570 nglg Phase I report), FOSA (35.3 nglg), PFDA (5.79 nglg), PFDoA (5.40 nglg), PFUnA (4.03 nglg), PFBA (0 .748 ng/g), and PFPeA (0.298 nglg). PFHxA was detected in this sample, but quantitative concentrations could not be reported due to QC failures . For this smallmouth bass sample only (River Section 4), branched isomers of PFUnA and PFDoA were present in the fillet extract. Isomers of PFDA were present, but at a significantly less amount than the PFUnA and PFDoA isomers.
3M ENVIRONMENTAL LABORATORY
PAGE 32 OF 104
3M ENVIRONMENTAL LABORATORY REPORT NO. E09-0372
Figure 1 (PFUnA) and Figure 2 (PFDoA) display the total ion chromatograms for both E09-0372-012 and a 15 ng/g extracted calibration standard prepared in the bluegill fillet control matrix using predominantly linear reference standards for comparison. Based on raw area counts, the PFUnA branched isomers in E09-0372-012 comprise approximately 15% of the total PFUnA signal and PFDoA branched isomers comprise approximately 29% of the total PFDoA signal . For comparison, Figure 3 displays the same chromatograms for PFDA. The PFDA isomer signature accounts for approximately 1% of the total PFDA signal . Figure 4 shows the individual MRM transitions of PFUnA and PFDoA in E09-0372-012 confirming that smaller peaks preceding the linear isomer are structural equivalents of PFUnA and PFDoA and not unidentified co-extracted matrix components. Representative 3M PFOS production lots have trace levels of PFDA (0.0005% wt), PFUnA (0.0002lo wt), and PFDoA (0.0004r6 wt) with significant degrees of branched isomers: PFDA (51%), PFUnA (46%), and PFDoA (33r6) .1 Figure 5 displays a representative chromatogram of the PFDA, PFUnA, and PFDoA impurities present in a 10 pglmL solvent standard of 3M ECF PFOA and PFOS. For E09-0372-012, the branching profile observed for PFUnA and PFDoA is consistent with the branching profile of PFCA impurities in 3M ECF production PFOS. The small presence of PFDA isomers in this sample is not consistent with 3M ECF production PFOS. In this study, no other fish tissue samples with measurable PFDA, PFUnA, or PFDoA showed the presence of branched isomers . All study samples had levels of PFDA, PFUnA, and PFDoA above the approximate LOO of 0.25 ng/g with the following exceptions:
" E08-0372-005 (Carp Section 4): PFDA = 0.283 ng/g (near the LO0), PFUnA <0.242 nglg, and PFDoA <0.243 nglg
" E09-0372-024 (Freshwater Drum Section 1): PFDA =1 .56 nglg, PFUnA = 0.448 ng/g and PFDoA <0.267 nglg.
Assuming that the branched isomer response factors are equivalent to that of the linear isomer, the estimated branched isomer LOO as a percentage of the linear isomer detected is provided in Table 15 for each sample . The average branching LOQs were 17% (PFDA), 25% (PFUnA), and 34% (PFDoA). Additionally, the PFUnA and PFDoA total ion chromatograms for sample E09-0372-021 (Fresh water drum, Section 4) are displayed in Figure 6 . This sample represents the second highest detected concentrations ofboth PFUnA (3.35 nglg) and PFDoA (2.26 ngtg) for this project; however, the branched isomer signature in E09-0372-021 is not detected at approximately 7.5% and 11% of the linear isomer for PFUnA and PFDoA, respectively . Therefore this sample is not consistent with the branching profile of PFCA impurities in 3M ECF production PFOS . The PFOS concentration of this sample was 23.6 nglg (Phase I report). Additional samples from river sections 1, 2, and 3, with measurable levels of PFDAIPFUru4IPFDoA, could not be quantitatively assessed for isomer profiles due to limitations of branched isomer LODs. 5.3 Specificity The MRMs of 399>80 and 399>99 (PFHS) and 499>80 (PFOS) have been documented as potential matrix interferences in some biological tissues arising from the presence of 5-pregnan-3,20-dio3-3sulfate isomers and taurodeoxycholate isomers (bile saft), respectively? To verify that the reported quantitation was free from significant unknown matrix interferences, the raw area count ratios of 399>80 to 399>99 and 499>80 to 499>99 were evaluated for each sample extract and compared to the average area count ratios observed in the solvent calibration standards and bluegill fillet control matrix extracted calibration curve and LCSs.
' W*mn K. Reagen et al ., 'Characterization of 3M ECF PFCA Byproducts in PFOA and PFOS Production Lots", Milwaukee SETAC North Anenca 28'' Annual Meeting; (2007) . Z Benskin, JP ., Batabeh, M, Martin, J .W., Anal . Chem 2007, 79, 6455t464 .
3M ENVIRONMENTAL LABORATORY
PAGE 33 OF 104
3A4 ENVIRONMENTAL LABORATORY REPORT NO. E09-0372
Table 16 displays the area count ratio of the 499>80 to 499>99 MRM transitions for each sample replicate . The area count ratio for each sample replicate was within 25r6 of the corresponding instrument batch average for the solvent calibration calibration curve . The average ratio varied between the two instruments used (API 4000 Q-trap and API 4000) in this study and is likely due to differences in tuning parameters. Only three samples produced PFHS concentra6ons greater than the stated LOQ: Ea9-0372-001, -004, and -045 . The area count ratio of 399>80 to 399>99 (data not shown, but is provided in the data package) for each replicate of these three samples was also within 25% of the average ratio for the solvent calibration curve .
3M ENVIRONMENTAL LABORATORY
PAGE 34 OF 104
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3M ENVIRONMENTAL LABORATORY REPORT NO . E09-0372
Figure 1 . (A) E09-0372-012 PFUnA Total Ion Chromatogram (sum of 563>519, 563>269, and 563>219). Average PFUnA calculated sample concentration = 4.03 nglg. Inset displays a baseline zoom of the branched isomer region . (S) PFUnA Total Ion Chromatogram of an extracted bluegill fillet calibration standard spiked at 15 nglg with a predominantly-linear reference standard.
3M ENVIRONMENTAL LABORATORY
PAGE 70 OF 104
3M ENVIRONMENTAL LABORATORY REPORT NO . E08-0372
Figure 2. (A) E09-0372-012 PFDoA Total lon Chromatogram (sum of 613>569, 613>169, and 613>319). Average PFDoA calculated sample concentration = 5.40 nglg. Inset displays a baseline zoom of the branched isomer region . (B) PFDoA Total Ion Chromatogram of an extracted bluegill fillet calibration standard spiked at 15 nglg with a predominantly-linear reference standard.
3M ENVIRONMENTALLABORATORY
PAGE 71 OF 104
Branched P_FDA,.,~
3M ENVIRONMENTAL LABORATORY REPORT NO . E09-a372
Linear PFDA
ao Figure 3. (A) E09-0372-012 PFDA Total Ion Chromatogram (sum of 513>469, 513>269,
and 513>269). Average PFDA calculated sample concentration = 5.79 nglg. Inset displays a baseline zoom of the branched isomer region . (B) PFDA Total Ion Chromatogram of an extracted bluegill fillet calibration standard spiked at 15 nglg with a predominantly-linear reference standard .
3M ENVIRONMENTALLABORATORY
PAGE 72 OF 104
3M ENVIRONMENTAL LABORATORY REPORT NO . E09-0372
A, 563>519 "A07 Branched
PFUnA
563>269.
Linear I ( /PFUnA
t
Ab . -N . ".
612>569 Branched PFDoA . .1.7!
.a1 612>169
Linear ~ IPFDoA
Ilt U.Y
7tA G.J~ ~ -J
563>219
t
Ya . OFi7 p.p.
fOJJf 612>319,; ~
MAlI ,
II
W7J~
I I
Rm{
i9DJ ~
I I' 1
7
Figure 4. Individual MRM transitions of E09-0372-012 . (A) PFUnA . (B) . PFDoA .
3M ENVIRONMENTAL LABORATORY
PACE 73 OF 104
3M ENVIRONMENTAL LABORATORY REPORT NO. E09-Q372
Figure 5. Representative Total Ion Chromatograms of (A) PFDA. (B) PFUnA, and (C) PFDaA Impurities in a 'f0 pglmL solvent standard of technical grade (3M ECF) PFOA and PFOS.
3M ENVIRONMENTALLABORATORY
PAGE 74 OF 104
A
Linear
~/ PFUnA
,A,x.. 3
,
3M ENVIRONMENTAL LABORATORY REPORT NO. E09-0372
B ., ;
., . . . ., ._ :. . .... . .,. ..: : ._ ._ _,
Figure 6. Total Ion Chromatograms of E09-0372-021 . (A) PFUnA - average calculated sample concentration 3.35 nglg. (B) PFDoA - average calculated sample concentration 2.26 nglg.
3M ENVIRONMENTAL LABORATORY
PAGE 75 OF 104
3M ENVIRONMENTAL LABORATORY REPORT NO. E08-0372
Table 15. Branched Isomer Summary for PFDA, PFUnA, and PFDoA.
3M LlMS ID
E09-0372-001 E09-0372-002 E09-0372-003 E09-0372-)04 E09-0372-005 E09-0372-006 E09-0372-OU7 E09-0372-008 E09-0372-009 E09-0372-010 E09-0372-011 E09-0372-012 E09-0372-013 E08-0372-014 E09-0372-015 E09-0372-016 E09-0372-017 E09-0372-018 E09-0372-019 E09-0372-020 E09-0372-021 E09-0372-022 E09-0372-023 EOi9-0372-024 E09-0372-025 E09-0372-026 E09-0372-027 E09-0372-028 E09-0372-029 E09-0372-030
Species
Carp Carp Carp Carp Carp Carp Smallmouth Bass Smatlmouth Bass Smailmauth Bass Smallmouth Bass Smallmouth Bass SmaAmouth Bass White Bass White Bass White Bass White Bass White Bass White Bass Fresh Water Drum Fresh Water Drum Fresh Water Drum Fresh Water Drum Fresh Water Drum Fresh Water Drum Bluegill Sunfish Bluegill Sunfish Bluegill Sunfish Bluegill Sunfish Bluegill Sunfish Blue ill Sunfish
River Location
Section 3 Section 1 Section 1 Section 4 Section 4 Section 2 Section 3 Section 1 Section 2 Section 2 Section 1 Section 4 Section 2 Section 2 Section 2 Section 1 Section 1 Section 1 Section 3 Section 3 Section 4 Section 2 Section 2 Section 1 Section 3 Section 3 Section 2 Section 4 Section 4 Section 1
"'Estimated Branching as % of Linear Detected
PFDA
<15 <16 <36 <10 <88 <18 <12 <22 <27 <30 <20 ("1% of total <7 .4 <7 .0 <7-2 <8 .9 <7 .7 <8.7 <5.7 <7 .4 <6.9 <16 <18 <16 <13 <8 .3 <14 <12 <21 <15
PFl1nA
<38 <27 <77 <21 NA <23 <16 <25 <33 <45 <20 12)15% of total <15 <14 <13 <76 <13 <19 <19 <17 <7 .5 <33 <31 <56 <25 <14 <17 <16 <24 <26
PFDoA
<67 <54 <48 <44 NA <37 <25 <23 <22 <46 <12 n)29% of total <19 <22 <19 <25 <26 <36 <38 <35 <11 <61 <54 NA <48 <27 <28 <22 <34 <48
(1) Branching LOQs estimated using ft approx~ttate linear LOO for PFDA, PFUnA, and PFDoA (0 .25 rig/g) .
(2) Branched iscirners detected in this sample. Reported value represents the percentage of the total PFUM and PFDoA quantifaGed .
3M ENVIRONMENTAL LABORATORY
PAGE 76 OF 104
3M ENVIRONMENTAL LABORATORY REPORT NO. E09-0372
Table 16. PFOS sample specificity: Comparison of Area Count Ratio of 499>80 to 499>99 .
Instrument Batch
o09O826a (API 4M Q-
Solvent Standards (0.025 nplmL - 50 ng/ml)
Blu ill Fillet Control Tissue Extracted Curve and LCSs
Sample ID
E09-0372-001 E09-0372-001 Dup_ E09-0372-002 _E09=0372-002 Dup E09-0372-003 -E09-0372-003 Qup.. ...._.E09-0372-004 -E09-0372-004 Dup--... .-. .. E09-0372-005 ED9-0372-005 Dup. ._ ... .E09-0372-006 E09-0372-006 Dup E09-0372-007 E09-0372-007 Dup E09-0372-(Hl8 E09-0372-008 Dup .... . E09-0372-009 E09-0372-009 Dup_-. E09-037722--00110 E09-0372-010 Dup
Sam ple Description
Axys ID- L12916-14, Client ID- 090191 Axys ID- L12916-14, Client ID- 090191 Dup Axys ID- L12916-24, Client ID- 090054 Axys ID- L12916-24, Client ID- 090054 Dup Axys ID- L12916-26, Client ID- 090056 Axys ID- L12916-26, Client ID- 090056 Dup Axys ID- L12917-3, Client ID- 090275 Axys ID- L12917-3, Client ID- 090275 Dup Axys ID- L12917-10, Client ID- 090282 Axys ID- L12917-10, Client ID- 090282 Dup Axys ID- L12917-17, Client ID- 090123 Axys ID- L12917-17, Client ID- 090123 Dup
Axys ID- L12931-6, Client ID- 090173 Axys ID- L12931-6, Client ID- 090173 Dup Axys ID- L12931-14, Client ID- 090032 Axys ID- L12931-14, Client ID- 090032 Dup Axys ID- L12932-2, Client ID- 090094 Axys ID- L12932-2, Client ID- 090094 Dup Axys ID- L12932-4, Client ID- 090096
Axys ID- L129324, Client ID- 090096 Dup
Average Area Count Ratio 499>80:499>99
3. 13
3.32
species Carp
Carp
Carp
Location Section 3
_.-.__.---------- _--_-_ .. .. . .... ._-._. Section 1
-- --. ._ . ._. .. . . .- . . ._. . ..___ ..-Section 1
Carp
Section 4 .
Carp
Section 4
-- .... ..................... _.--- ---- --
Carp
Section 2
Smallmouth Bass
Section 3
SmaAmouth Bass Smallmouth Bass
Section 1
_ . ..--- ... .. . .. . ..... .. ._ .._ .__ .. ._ .-. Section 2
SmaHmaith Bass
Se--c--t-i-o-n---2
%RSD 6.5 8.8
499>80:499>99 Area Count Ratio
3 .36 3.46 3 .43 3.45 3.58 3.20 3.08 3 .37 3 .31 3-50
3 .37 3-48
3 .42 3 .23 2 .92 3-18 3 .08 3-10 3 .22 2.93
3M ENVIRONMENTAL LABORATORY
PAGE 77 OF 104
3M ENVIRONMENTAL LABORATORY REPORT NO. E09-0372
Table 16 Continued .
Instrument Batch
s09D9fl9a API 4000
Solvent Stds (0.025 ng/mL - 50 ng/m!)
Blu ill Fillet Control Ttssue Extracted Curve and LCSs
-- -
Average Area Count Ratio 499>80:499>99
1 .48
1 .48
Sampile !a E09-0372-011 E09-0372-011 Dup
E09-0372-012
_E09-D372-012 Dup E09-0372-013 E09-0372--01.3...Qup .,_ E09-0372-014 E09-0372-014 Dyk E09-0372-015 EO_9-03_72_-0_15 D~ E09-0372-016 E09-03 72-016 Dup __
E09-0372-017 E09-0372-017 Dup E09-0372-018 E09-0372-018 Dup E09-0372-019 _E09-0372-019 Du ~-E09-0372-020 E09-0372-020 Du
Sample DescrP 'on Axys ID- L12932-9, Client ID- 090042 Axys ID- L12932-9, Client ID- 09042 Pup .... .. . .
Axys ID- L12932-26, Client ID- 090248
Axys ID- L12932-26, Client ID- 090248 Dup Axys ID- L12933-18, Client ID- 090064 Axys ID- L12933-18, Client ID- 090(~4 Dup Axys ID- L12933-20, Client ID- 090066 Axys ID- L12933-20, Client ID- 090066 Dup Axys ID- L12933-25, Client ID- 090071 Axys ID- L12933-25, Client ID- 090071 Dup Axys ID- t-12934-10, Client ID- 090024 Axys ID- L12934-10, Client ID- 090024 Dup Axys ID- L12934-12, Client ID- 090026 Axys ID- L12934-12, Client ID- 0. 9....0. .... 0.. .2. 6. . . D.... ...u. .. ...p.... ..... . Axys ID- L12934-18, Client ID- 090017 Axys ID-,L12934-18, Client ID- 090017 Pup Axys ID- L12945-1, Client ID- 090153 Axys ID- L12945-1, Client ID- 090153 Dup Axys ID- L12945-4, Client ID- 090167
s ID- L12945-4Client ID- 090167 D
S pecies Smallmouth Bass SmaAmouth Bass White Bass White Bass White Bass White Bass White Bass White Bass Fresh Water Drum Fresh Water Drum
Location Section 1 Section 4 Section 2 Section 2 Section 2 Section 1 Section 1 Section 1 Section 3 Section 3
hRSD 3 .7 8 .9
499>80 :499>9 9
Area Count Ratio 1 .44 1 .49
1 .50
_1 . .:47._ .1 .46 _ _ 1 .44 1 .50 _ ....... ... ,_ .. .,. 1 .51 1 .44- __ 1 .49 _ 1 .50 1 .51 1 .43 1 .49 1 .45 1 .44 ,1 .45 1 .46 1 .43
3M ENVIRONMENTAL LABORATORY
PAGE 78 OF 104
3M ENVIRONMENTAL LABORATORY REPORT NO. E09-0372
Table 16 continued .
Instrument Batch
o090828a (API 4000 Q-trap)
Solvent Stds (0.025 nglmL - 50 ng1mL)
Blu ill Fillet Control Tissue Extracted Curve and LCSs
Average Area Count Ratio 499>80 :499>99
3. 19
2.97
Sample ID E09-0372-021 E09-0372-021 Dyp . -E09-0372-022 _.-E09-0372-022-Dyp... .... _ E09-0372-023 -E-0-9---0-3-7-2---0-2-3-D-u---E09-0372-024 E09-0372-024 Dup E09-0372-025 E09-0372-025 Dup E09-0372-026 E09-0372-026._Dup_ - . E09-0372-027 E09-0372-027 .Dup_ E09-0372-028 E09-0372-028 Dup E09-0372-029 E09-0372-029 Dup E09-0372-030 E09-0372-030 Du
Sample Description Axys ID- L12945-14, Client ID- 090241 Axys ID- L12945-14, Client ID- 090241 Dup Axys ID- L12945-16, Client ID- 090078 Axys ID- L12945-16, Client ID- 090078 Dup Axys ID- L12945-19, Client ID- 090081 Axys ID- L12945-19, Client ID- 090081 Dup Axys ID- L12945-30, Client ID- 090009 Axys ID- L12945-30, Client ID- 090009 Dup Axys ID- L12947-7, Client ID- 090266 Axys ID- L12947-7, Client ID- 090266. Dup Axys ID- L12947-0, Client ID- 090267 Axys ID- L12947~, Client ID- 090267 Dup Axys ID- L12947-13, Client ID- 090111 Axys ID- L12947-13, Client ID- 090111 Dup Axys ID- L12947-25, Client ID- 090205 Axys ID- L12947-25, Client ID- 090205 Dup Axys ID- L12947-26, Client ID- 090206 Axys ID- L12947-26, Client ID- 090206 Dup Axys ID- L12948-17, Client ID- 090061
s ID- L12948-17 Client ID- 090061 Du
specks Fresh Water Drum Fresh Water Drum Fresh Water Drum Fresh Water Drum BluegiA Sunfish Bluegill Sunfish Bluegill Sunfish BluegiH Sunfish Bluegill Sunfish
. Bluegill Sunfish
Location Section 4 Sec6on 2 Section 2 Section 1 Section 3 Section 3 Section 2 Section 4 Section 4 Section 1
%RSD 8.0 11
499>80:499>99 Area Count Ratio 2 .95 2 .83 3 .19 3 .36 3 .01 2 .64 3.22 2 .82 2.97 3 .07 3 .02 3 .12 2 .87 3 .11 3 .16 2 .99 2 .93 2 .98 3 .03 3 .12
3M ENVIRONMENTAL LABORATORY
PAGE 79 OF 104
3M ENVIRONMENTAL LABORATORY REPORT NO. E09-0372
Conclusions
Sample results presented in Tables 1-4 and Tables 10 -14 are considered accurate within the stated overall method uncertainty for each analyte . The accuracy (% recovery) and precision (%RSD) of 3M ECF (branched) PFOA and PFOS laboratory control spikes quantitated against the predominantly linear reference standard demonstrated that the isomers produce no discemable quantitative bias. Area count ratios for 399>80 :399>99 and 499>80:499>99 for each sample extractwere within 25% of the average area count ratio observed for the solvent calibration curve analyzed within the given instrument batch thus verifying that the reported quantitation was free from significant matrix interferences.
atalSample Retention
All remaining sample and associated project data (hardcopy and electronic) will be archived according to 3M Environmental Laboratory standard operating procedures .
lytical Personnel
8.1 3M Environmental Laboratory Michelle D. Malinsky, Ph.D Research Specialist
8.2 Pace Lab Ops Jonathan Steege
3M ENViRONMENTAL LABORATORY
PACE 80 OF 104
3M ENVIRONMENTAL LABORATORY REPORT NO. E09-0372
eg 7/2
-_--
Yi f,~
.
Mict+e D. Malinsky, Ph .D, ilu Researjdpecialist, Principal Analytical Invesdgator
.--
William K. Reagen, Ph.D., Environmental Laboratory Management
Date
The 3M Environmental Laboratory's Quality Assurance Unit has audited the data and report for this project.
QAI Represeoive
_ -
W)5-/,q
Date
3M ENVRONMEKTAL LABORATORY
PAGE 81 OF 104
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