Document 91ZnGy8X7arqGJLjnpK8rMgaD
ENVIRONMErNeTiArL LABOERrAtTaOnRY
Analytical Report
Revised August 12, 2005 Initial Quantitative Screening of Commercial Lots of Human Serum Obtained in 2004 for Endogenous Fluorochemicals Using Protein Precipitation and Liquid Chromatography/Tandem Mass Spectrometry
Laboratory Request Number: E05-0120
Testing Laboratory
3M Environmental Health and Safety Operations 3M Environmental Laboratory
3M Environmental Laboratory
Project Lead
Mark E. Ellefson
Building 2-3E-09
PO Box 3331
Saint Paul, MN 55133-3331
Phone: 651-778-5405
Testing Facility Management `William K. Reagen, Ph.D.
Building 2-3E-09 PO Box 3331
Saint Paul, MN 55133-3331
Phone: 651-778-6565
Requestor
Dale Bacon Building 2-3E-09
PO Box 3331 Saint Paul, MN 55133-3331
Phone: 651-778-4736
Page 10018
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MENVIRONMFENRTOALVLEABCEORoTAsTN1O2RG0Y 3M Environmental Laboratory
Analytical Report Project Number: E05-0120
TET tebe
= ERE ERR
The following analytical report presents data from commercially available normal pooled human serum thatwerepurchasedbythe 3M Environmental Laboratory and subsequently screened for endogenous fluorochemicls.
`Thirty-six scparate lotsofpooled normal human serum were extracted using a protein precipitation technique and screened for sixteen different fluorochemicals using liquid chromatography/tandem mass spectrometry (LC/MS/MS). Serum `samples were received from various suppliers between 9/9/04 and 12/2/04. Samples were analyzed between 02/03/05 and 02/09/05.
"Thisrange-finding study was designed to provide lower limitsof quantitation (LLOQs)ofapproximately 100 - 300 ppt. Upon reviewofthe data, it was discovered. that many ofthe target analytes were present at concentrations much lower than the 100 - 300 ppt target level. As an interim solution and unil 2 more definitive study an be completed, signal to noise ratios were evaluated and a limitofdetection (LOD) was defined for each analyte. Asa result, calibration curves were extrapolated between the LLOQ and the newly defined LOD, allowing screening quality data to be determined for some analytes as low as 19.9ppt. Therefore, it mustbe emphasized that much ofthedata presented in his report was derived from extrapolated curves and should be considered as screening quality data only.
SsaammpplleesriensuwlhtiscahretsheurmemwaarsizdeedteicntaTbalbeleana1.lytTeh.eSaavmepralgeevvaalluueesstrheaptowretrederleepsrsetsheannt the limitofdetectionwerenot included in the average value calculations. Please refer to Section 5 for individual sample results.
Page20f18
E--S --------------------------------eeseeSerA---------------- Table1. SuomfPemrfuoarinartedAycid Rosults WENVIROVMP ENTARLTLANGOEORSAC 1T2OR0Y
onLan m1c ]eo) Median
1
[|2 NR[05 954| 450|106 |NR |0660 |<LoD|
High
|708 [7356| [102 [230[ 263 |nee|
Low
[0507|245|
0477| 0337
`NSutmabnedraorfdSaDemvpilaetsiownith|2.17_| 0.627|NR"[0.0933634]| 0571|NR" [02[0Np8"|
DatactabloAnalyto
XDTR NNoott RDeeptoerctteedd.TTihssaanbalyctewwaasdottdedtectnedoneanofy hoeF36ssagmlplebet sppsrs 0be contaminoar
an anomaly th plewillbersnalyzedand therlswillbereportedin hedefinitivesy.
Tabl2e. Summary of Other Perfluorinated and Highly Fluorinated Analyte Results
77 Sa E FnE E R SE RIEE a ECe eSts BFEAerFneEBAaSsiTR]
[ina|osen [osee [aus [ao [sone| ws | sos [Cob Joost[00330 527| 169.
[ wWign
<op|o1e0 [00706]580 |328| <tOD |0.18|5
Fm
io at fosetefm a son| 00480
|FStranadaorcfdSDatemvpiilaetsoiwoinnth | ND" [0.081200122] 152 | 671|ND"| 0.0257
WD NotDetected. This mlywatsenotde3t7ocfhsisaldes
iPseoamkesrwsefroer oPbFsOeSrvaenddinPtFhOeAch.roRmealattoigvreabmrsanccohrerdeasnpdolnidneianorgislomeirraanntdiobesrawanecrehr.ed detormined for both analytes and are presented below (the human serum value representsanaverageof %branched isomers foral ofthe humanserumsamples that contained detectable levelsofeach analyte). The %branched isomer results for individual samarepprelsenetedsin Table 12.Inaddition, well-characterized los of PFOS (Lot 171) and PFOA (Lot 332) were analyzed togeastahmaetrrix spike (QC050203-004)in rabbit serum. Dita from that analysis was evaluated for branchedandlinearisomerdistributionandagreeswellwithpreviously obtained. NMR data.
Page 301s
-------------------------------------- J-- a --ey
Table. Sumomf PaFOrAayndPEOS lsomor Ratios
lH OS rs a ERE e lrmEve i 1son aai SodPFSSuAan ri| ]
Human Serum, average
LotMT3at5orrix(12SH3pRi(kcesh8(Q0aC03r500a2.0Pc30F-90Ov0)aA4lu)
[ [ssow x||
Lot 171 (WTCRRrpra0S e valu
7RATERESDantes HIERSreeERROE ULES
Tabl4e. Reforance Substances
onee Fase Ttoiekn S 0 aamFmeoTsees0 AetaiTt
[CL omh mLiinAsEE|EEArEePnrirnf +|SOr URS,|
LEE S se ABI
rm |
|FeSoeEsom]
ianccroen|| ammoaere| woomron|| son|| oomm ||
emrornns|| woonm |[rmowo[owmeen||eowmmn]]
eNsiecbturmtraeiesrts|| GOin|s camaro| on | on| om |
[r mon|| m ore s [nom |
t tsTeTty A,se
ro]
TTS
a I ATAS ----T
Troawmemo ||omoenn||oomns]|
| os [ween | om |
Spats
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ro=nan
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Fagssatts
_--_--
WM ENVIRONMPENRTALOLNAEOBOR0CA8T0OT2R0Y
TESultiidrrepividokt FHTEICNEE Het SREP IER
"The samples for this study were prepared as follows: 1. The serum samples were allowed to thaw and warm (0 room temperature. 2. One milliliter ofserum(orwater in the caseofmethod blanks) was added to a 15 mi screw-cappedpolypropylenecentrifuge tube. 3. Matrix spikes and method blanks were preped by spiking he serum (or water) with the analytesof interest. In addition, ""C PFOA was added as a surrogate to all serum samples, calibration standards, matrix blanks, and method blanks ata concentrationof3.01 ng/mL. to evaluate extraction method performance 4. Two milliltesofacetonitrile were added to the serum and the centrifuge tubes were capped and vortex mixed, allowing the serum proteins to precipitate. 5. The tubes were centrifuged at approximately 3000reffor 20 minutes to pellet the precipitate. 6. After removal from the centrifuge, the superatant was transferred to an autovial for analysis.
Quantification was accomplished by high performance liquid chromatography tandem mass spectrometry. The following parameters wer used: Liquid Chromatograph: Hewlett-Packard" Series 1100 Liquid Chromatograph system Analytical column: Keystonc BetasilTM Cis 2x100mm, Sum particle size SCtoolpuTminmet:emp2e0r.a0tumrien:ut3e0s C Flow rate: 300 L/min Injection volume: 5 uL Mobile phase components: Solvent A: 2.0 mM ammonium acetate in ASTM Type I water Solvent B: HPLC Grade Methanol Solvent Gradient:
T0me0 2%0B% 110s s2o0n% 155500 290%% 200 20%
Page Sof 18
-
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Mass Spectrometer: MDX SciexTM API 4000 Q-Trap Mass Spectrometer Detector
SIoonfitzwaatrieo:n AMnoadley:sETMtleVcetrrossipornay1.N4egative
`Temperature: 450 C
GS1:35
GS2: 45
Entrance Potential: 10
Fates. Analysis sessions Type: Multiple Reaction Monitoring (MRM)
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HF RAI
Fs ER EY
4.1 Calibration
4.1.1 Calibration Curve
Alenveelxstarnacdteexdtcraalcitbirnagtitohne ccuurrvveewinasthfeosrammeed mbaynsnpeirkiansgtrhaebbsiatmspleersu.m aSttopcreksscorliubteidons `(weexrceepitndPiFviBdAu,alwlyhipcrehpwaaresdpfrrepoamrneedatinfMliulolrio-cQhewmaitcearlstoapnrdedvielnutteedstienrimfeitcahtainoonlfrom oTchceurrreigngr)e.ssiSoanmapnlaelysswiserweasqupaenrtiffoiremdevdeurssiunsgaan q8u-apdorianttiecxcturravcteefditcawleiibgrhattieodn 1c/uxr.ve.
J
_--_--
SMENVIRONPENRTAOL LNLAOSEOER0A5CT01OT2R0Y
(CaLlLibOrQat)iownitshtaandcaoredffsiwceiernetowfitdheitne+rm2in0a%ti(o+n2>50.%99f5o.r thLeowL-olweevrelLicmaliitobrfatQiuoanntitation Cstaalnidbarradtsi,onwshtiacnhdawredrsetnaoltwweirtehinno4t2g5re%a,tewretrheannottwiicnecltuhdeedmeitnhtohde cballainbkrsatwieonrecunrovte, included in the calibration curve. 412 ContinuingCalibration Verification (CCV) sAamCpolnetsi,nubirnacgkCeatliinbgrtahteiosnamVeprliefsicwaittihonth(eCcCaVl)ibrwaatsiornucnuratvel.easTthoenCeCeVvreercyov10ery was within 25% for all analytes. 413 SystemSuitability aFinvdeasfyesrttehmesluaisttabCiCliVt.y stTahnedsaerdssamwpelreesahnaadlyazreedabceofuonrtestwhietihniati%al4RcaSlDiboraft<i5on%cuarnved.a. irnestteanntcieosn:time %RSDof 2% when evaluated independently exceptforthe following
sSeyvsetreamlsiunitdaibviildiutaylcrtirtaenrsiiat.ioHnoswmeovneirt,ortehde fdoartathreeCpyoprtAecdiwdasfadielerdivtehde rfertoemnttihoen time
`quantitationofthe TIC, which passed system suitability requirements.
6Al.l2%o,ft7h.e5s%yasntdem8su.i4t%a.bilHiotwyseevtesr,fonroFsBamSpAlaersehaacdoaurnetas cfoaiulnetds. aTbhoev%eRthSeDLsLwOeQrfeor
this analyte therfore the data was accepted.
414 LowerLimitofQuantitation(LLOQ)
The that
mLeLtOaQccfuorraecaycrheqaunialryetmeenwtassofeq7ua5l%t-o1t2h5e%l.owest
standard
in
the
calibration
curve
415 Limitof Detection (LOD)
th`Tahte wLeOreD2f-o3rteiamcehsaanbaolvyteetwhaesmeequtalhtboloathndek.area counts (converted to a concentration)
416 Blanks
Method Blanks
Allof themethodblanksin thisstudyhadareacou<n5t0s%ofthe LLOQ. `Matrix Blanks
O`Tnweommaattrriixxbbllaannkkscwoentraeinperdepeanrdeodgeinnoraubsbiltevseelrsoufm PanFdOaSnatlhaytzewderweitahbtohveestahmepLleLsO.Q. qAunaalvteoragetehendLoLgeOnQoufsorvBalLuKe05fo0r2P03F.O0S03w.as determined by substituting a number
Page 7018
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uencReSASEORATTORY Table 6. Average Endogenous PFOS Value Determination
[ uogmgmy) | oss |
Bosve0ro205004
aoantts
`Theaverage endogPFeOSnvaolueuwassused to correctthematrix spike.
`responsefor endogenous levofePlFOsSinrabbit serum.
4.1.7 Solvent Blanks
Acetonitrile blanks were analyzed throughout the analytical run. Analyte response was
less than Y% the LLOQ in all cases.
418 Surrogate
C PFOA was added as a surrogate to all serum samples, calibration standards,
`matrix blanks, and method blanks at a concentration of 3.01 ng/mL to evaluate the extraction method performance. All surrogate recoveries were within 20%,
4.2 Laboratory Control Spikes (LCS)
Six Laboratory Control Spikes (LCS) were prepared in rabbit serum and analyzed `with the samples. Results were corrected for endogenous levelsofanalyte in the
`matrix (where appropriate), and are presented below.
Table 7. Matrix Spike Recoveries In Rabbit Serum
perry
Trae ER Er St
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Presets
--_--
Table. Matix SpikeRecoveries iRabbitSera cotianed) SuEmROMME[ NTALe Usori ToRY
TE Ee Tre IYCo PDR 2 BE re
J wy [oFEoaoMesmEsoPor[(f 5G0oneSale 0nom|i W icio<rmegtroaanis|y tAo ewAea85Yic|el, |
toTa=AtaCtnsgm.
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Page 100118
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SEMAROMMPENRTAOLLwIABoOEERoAsCTrOTRsYs Table 7. Matrix Spike Recoveries in RabbitSerum (continued)
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-_--
UENROMMEPNTRALOLAJNSOOEERDASCT1OT2RY0 TLoavbells8o. PFOS Matrix Spiko Recovery in Rabbit Serum, Corrected for Endogenous
L~o LPTEL
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|
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4.3 Accuracy T`hmeiacxcusrpiakceyfreocroevaerciheasnfaolryttehrateapnaolyrtet.weaTdshecarlcuelatsaedrbueyptrlaeskleinntgsetdhbeealvowe.rageofthe. Table 9. Method AccuforrInadivciduyal Anaiytes
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4.4 Sample Related Comments `The extraction and analysis ofthe samples was performed utilizing a non-validated `e`wmxehttrehanopedov.laaTlthueadetcmiaanltgirbtirhaxetiasocnpcicukurerarvceeys(uablntesdt,pwreeCecniPstFihoOenAoLsfLutrOhrQeoagsanatmdepLlreeOcDorvo)essruhiletos.u,ladnbde tchoen.sidered
EaR 1 R eat ea
ee
`Tables10and 11 summarize individualsampledata.
Page 120618
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Table 10. Perfuorinated Acid Resuts (continued) suenRROemLEatoIssaTTrOARY
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Sw rowENeTALSLABOERAoTrOReY
Table 1. Sample Results or Additional Perfluornated and Highly Fuoinated Analtes
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Page 150118
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uewmOwEr NTALo AsORe ATORY
Table 1. Sample ResultsforAdttonal Porlsorinatet and Highly Fluorinated
Analytes (continued)
rm om ono Si EA
|LBHsE S08
[D0
ET
moAL)t slob |t GmtL e bohnomee
[EOCuo Tmoin [oe Te
[boitos aoni]e papTeeoraebe Foie 0Ta
ad bo se0To ee] oo
[tS I on TnTT oreTT oomeT r|CsEer ses0| iTaS Tor1i3|
Le e soe rs10ioes U|e100mtTrwe10|0. wnTw ow] Band oN wraomano a eoe1c0e0i 1.3
Table 12. soreRatos fo ndvidual Samples
Serine15 | CSoemAUNTE SeDEn Do ECt Y A)
[wI ea-- meer
tens [am [ --w--e||
weess
ovaetres|e om |o| w |
[wea
[aeos wen
Tw ircggeaisnp [e | a ae w |
wme ewT n eo d em [eaxm]
Honea, usbes
[ax
rmoeee | vareesss To aco am ucmaw| T|sae wewww ]|
Tinegoss Ter
TGs |T aonTN aw] [eoamn
mom
ees[me em |
[ mr e a em eseev ]Ii Se r
Ce | evr |soa]
Page toot
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Table 12,lamer Ratiosfor individual Samples sntnuse)
[a LE bh Biol LEPEITEeE pay oFEE
Bah EE IHR
[mass THAGos)
T[ T wew n [ o ox m on | | am |
THA6oS5.
[eh | ao ox |
[wee TT [wes TTT
ee [aw| am | on [se [aw
[ [om we en s T TTe eye [ [l e on m | |s asm]|
[o[ wew s e TeTs Tr sea s--ec--se en [||woreex
Tax | aw
|
[mee Tem | ex[ew |
TNAG04
[Fe aw [a m w |
[ NAGSw [ e emsF | s a e owm | | m wx |
TT
[Aw | m Ten o | se ew |
SNaENgG nE AG37 ets sete of prc rum. --
FSAI TTY SIL EE rs
PE Lt All original raw data andtheanalytical report have been archived at the 3M
Environmental Laboratory. The test materials and analytical reference standard reserve samples, aswellas the samples pertainitnog this projectarearchived at the
Reserve samples, digital copiesoforiginal data and all `original paper data will be retained in the archives of 3M Environmental Laboratory for a periodofat least 10 years following report signing.
Eh E eii EeFL
`Uprnedseerntithne aclolnsdaimtpiloensso.fPthFeHSs,tuPdyF,OSen,daongdenToHuPsFfDluSorwoecrheemdiectaelcstwederienefvoeurnydltootobfe
serum while PFOA was present in detectable levels in all but one sample. Conversely, FBSA, THPFOS, and the C\; acid were not detected in anyofthe.
samples using the present methodology. "C PFOA surrogate recoveries were all
`within 20%.
Page 170018
-- er -- --------------------------------------------m ---------------------------------------- ENVIRONMPENRTOALVLAEBOCERGTASTN1O2RG0Y
SimrTEaT d EAA oTRe
ison, Projec Load,
Dai
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Wika Reegen, Loboratory Manager
28/2/05 Dole
`The 3M EnvironmentalLaboratory'sQuality AssasanceUnithasauditedthedataand report fortisproject
CaitAssurance Ropresoniaive
--
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Page 180118
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