Document gbrv3G6q6bVaYypv2KxBV579e
StudyTitle
AnalyticLaalboratorRyeporton PresencaendConcentratioofn Potassium Perfluorooctanesulfonatien Serum and Liver of Sprague-Dawley' Rats
Exposed to PFOS viaGavage
REPORT AMENDMENT
NO. I
Amendment Date: 19 April2000
Performing Laboratory ')M Environmental Technology & SafetyServices
")M EnvironmentalLaboratory 935 Bush Avenue
St.Paul,MN 55106
Laboratory Project ldentirication ET&SS LRN-U2006
ET&SS FACT Tox-012
3M EnvironmentaLlaboratory
FACT Tox-012 ReportAmendment No.1
Thisamendment modifiesthefollowinpgortion(so)fthefinarleport:
1. FINAL REPORT.Iviaktehefollowiandgdititoonthefinarleporttext.
ADD:
Afterthefinarleporwtasissue(dsigneadndarchiveidt)w,asdiscovertehdathepuritvyalue statedfortheanalyticarleferencesubstancewas inaccurateT.his valuewas based only on NIMR analyses.Subsequentchemicalcharacterizatiiosnoccurringto determinetheactual purityof thesessubstances(PFOS Lot 193).Ifnecessaryt,hefinalreportwillbe amended to detaitlheanalyticarlesultussingthepurityvaluefrom thecertificaotfeanalysiswhen itis issuedforPFOS Lot 193.
REASON:
To describethechangesthatwillbe made tothefinalreportwhen thecertificaotfeanalysis isissuedforPFOS Lot 193.
2. FINAL REPORT. Ntakethefollowinca,dditiontotheexceptionslistintheStatementof Compliance section.
ADD:
o The purityand stabiliotfytheanalyticarleferencseubstanceisunknown and was not determinedpriortotheinitiatioofnthisstudy.
REASON:
The Statementof Compliance sectionwas incomplete.
3. FINAL REPORT. The purityof theanalyticarleferencesubstanceis99.28% (Lot 193),as listeidnTable2.
AMEND TO READ:
The purityof theanalyticarleferencesubstance(PFOS Lot 19'))listeidnTable2 isunknown.
REASON:
The purityvaluestatedfortheanalyticarleferencesubstanceisincorrect.
4. FINAL REPORT.i'vfaktehefollowingadditiontothefinalreporttextundercircumstancetshatmay affecthe qualityof thedata.
ADD:
AccordingtotheDilutonSummary Sheet,theextractfsorFO livesramples8243M, 8244M. 8249M, 8250M, and 8255M were dilute0d.001mL intol.OmL(1:1000),butwere calculateads 1:2000.On thesame sheet theextractfsorFO livesrample 8181M and F1 livesrample8937 Pup were diluted 0.001mL intol.OmL (1:1000),butwere calculateads 1:500.
The resulftor8249M livesramplewas quantitatebdasedon peak responsethatexceeds the highestcalibratisotnandardby 20%.
REASON:
To include the circumstances thatwere found afterthe finalreportforthisstudy was issued.
3M EnvironmentalLaboratory
FACT TOX-012 ReportAmendment No.1
5. FINAL REPORT.On page 4 of 16,a printerroorccurretdharteplacesedveralinesoftheExceptiotnosGLP compliancetextwith an "X" characterw,ithoutany change in thespacingbetween the subsequentbulletson therestof thepage.The incorrecptage willbe replacedinthefinal reportand itwillarchivedinthestudyfolder.
AMEND TO READ:
The correctpage 4 of 16 isattachedtothisamendment. The followinglisitsthecomplete textlostintheprinterror:
The identitsyt,rengthp,uritya,nd compositiodnefinintghetestorcontroalrticwlaes notdetermined atthebme ofthestudy;however,theseanalyseswere performedon thelotofPFOS used subsequenttothepresentstudy. Two separatestudydirectorwsere assignedtotheinvivoand theana@ficaplortionosfthisstudy. From 15 September 1998 unti1lOctober1998,theprotocowlas notsponsorapprovedand was duringthatbme periodnotincompliance. Deviationwsere notconsistentalpyprovedby thestudydirectoars requiredby GLP regulabons. Datacorrectonswere notalwaysrecordedas requirebdy GLP regulations. Datawere notalwaysatbibutetdotheindividuraelsponsiblfeorrecordintghedata.
REASON: The finalprintedreportwith the incorrectpage does not reflectthe complete reportdocument as reviewedbN-thestudydirectorand sponsorrepresentative.
Amendment Approval
Marvin T. Case,D. P'-tfP,h.D.,Sponsor Representative
4m -A,)
'Date
4KI, /-@@ KristenJ Hansen. Ph.D., Study Director
0,411,7100 Date
3M EnvironmentaLlaboratory
PotassiuPmerfluorooctaresutbmto CAS Number-2759-30,3
3M EnvironmentLaalboratcxy ReMt No.Tox-012
t-abaatoRreyquestNumber(LRN)-U2006
STATEMENT OF COMPLIANCE
Study Tibe:
AnalyticaLlaboratoryReport on Presenceand ConcentratioonfPotassiumPerfluorooctanesulfonatienSerum and LiverofSprague-DavAeyoRats Exposed toPFOS via Gavage
Study IdentificatNiuomnber:, FACT Tox-012
Thisstudywas conductedincompliancewithFood and Drug AdministratioGnood LaboratoryPractice (GLP) RegulationfsorNonclinicLaalboratorSytudiesPata Requirement(s)2:1 CFR (Part58)]w,ith theexceptionsinthebuileteldisbtelow.Inadditiont,hepresentstudyhas been auditedretrospectively by an independentqualitayssuranceunitA.uditproceduresand findingfsorauditsperformedatthe3M EnvironmentalLaboratoryand atparticipaticnogntractlaboratorieasrehoused withdocumentation pertinenttothisstudyinarchivesatthe3M laboratoraynd willbe retainefdoratleast10 years.The analyticaplortioncompletedatthe3M EnvironmentalLab was performedinaccordancewith3M EnvironmentalTechnologyand SafetyServicesStandardOperatingProcedures.
ExceptionstoGLP compliance:
Storagecontainersforthereferencesubstance,PFOS, were notlabeledwithname, CAS number, batch number, expiratiodnate,or storageconditions.
Detailsofthepreparationm,aximum storagetime,and stabiliptryopertieosfthereference substance,PFOS, areunknown.
The identitsyt,rengthp,uritya,nd compositiondefinintghetestorcontrolarticlweas notdetermined atthetimeofthestudy;however,theseanalyseswere performedon thelotofPFOS used subsequenttothepresentstudy.
Two separatestudydirectorwsere assignedtotheinvivoand theanalyticaplortionosfthisstudy.
From 15 September 1998 unti1l October1998,theprotocowlas notsponsorapproved and was duringthattimeperiodnotincompliance.
Deviationswere notconsistentlaypprovedby thestudydirectoars requiredby GLP regulations.
Data correctionwsere notalwaysrecordedas requiredby GLP regulations.
Data were notalwaysattributetdotheindividuarlesponsiblfeorrecordingthedata.
Records inelectronifcormdo notmeet thecriterisaetforthunder21 CFR(L 1). A finalizesdample log-i/ntrackinsgystemwas notinplaceatthetimeofthestudy;however,draft
documents didexistand were used tologinand tracksamples.
Prophetary and ConfidenbW
Page 4 of 16
Potassium Peffluorooctanesulfonate CAS Number-2759-39-3
3NI EnvironmentalLaboratory ReportNo. Tox-ol2
LaboratoryRequest Number (LRN)-U2006
ANALYTICAL
LABORATORY
ON THE
REPORT
Determinationof the Presence and Concentration
of Potassium Perfluorooctanesulfonate
(CAS Number: 2759-39-3)
inthe Serum and Liverof Sprague-Dawle Rats Exposed to PFOS viaGavage
Laboratory Report No. <U2006> Requester ProjectNo. <3M TOX 6295.9>
Study Dates
Study Initiatio2n6:May 1998 Study Completion:At Signature
Potmium Perfluoroocianesulfonate CAS Number-2759-39-3
3M EfTvimnnientaLlaboratory
Report No. Tox-012 LaboratoryRequestNunter (LRN)-U2006
TABLE OF CONTENTS
TableofContents....................................................................2................
StudyPersonneland Contributor.s.....................................................3.................
StatementofCompliance..............................................................4.................
QualitAyssuranceStatement..........................................................6.................
Introductio.n.......................................................................7.................
Sample Receipt.....................................................................7.................
Materialasnd Methods.................................................................8................. ChemicalCharacterizabo.n........................................................8................. Method Summaries ..............................................................9................. AnalytcaElquipment............................................................I.I................. ProtocoDleviation.s.............................................................1.2.................
DataSummary, Analyses,and Results..................................................1.2................ Summary ofQualityControAlnalysesResults........................................1.2................. Summary ofSample Results......................................................1.3.................
StatistiMceatlhods ..................................................................1.4................
DataQualityObjectiveasnd DataIntegri.t.y.............................................1.5.................
StatementofConclusion.............................................................1.5.................
References...........................................I..........................................................1..5....................
AttachmentA: ReportSignaturePage
AttachmentB:TableofResultsofAnalysisofL:iveSramples fromFO Male and Female Sprague-DawleyRatsAdryfinistePrFeOdS Dailyby Gavage
AttachmentC:TableofResultsofAnalysisofSeraSamples fromFO Maleand Female Sprague-DawleyRatsAdministeredPFOS Dailyby Gavage
AttachmentD:TableofResultsofAnalysisofUiverSamples fromFl Maleand Female Sprague-DawleyRatsAdministeredPFOS Dailyby Gavage
AttachmentE In-liPfreotocoalnd AnalyticSatludyProtocol
AttachmentF:Preparatoraynd AnalyticMaelthods
AttachmentG: Dose Analyses
Page 2 of 16
Potassium Perfluomoctanesulfonate CAS Number-2759-39-3
STUDY PERSONNEL AND CONTRIBUTORS
AnalyticalChemistry Laboratories
3M Envimnmental Technology and SafetyServices
3M Envimnmental Laboratory FluorineAnalyticaClhemistryTeam (FACT) 2-3E-09 935 Bush Avenue St Paul,MN 55106
KrisHansen, Ph.D.,AnaWal StudyDirector LisaClemen, AnaiyUcalChemist(s)
In-lifTeesting Laboratory
Argus Research LaboratoriesI,nc. 905 Sheehy Drive,BuildingA Horsham, PA 19044
Raymond G. York,Ph.D.,Study Director
Sponsor
3M To)(icologSyervices 3M Center Building220-2E-02 St Paul,MN 55144
MarvinCase, Sponsor Representative
3M EnvironmentalLaboratory
ReportNo. Tox-012 LaboratoryRequest Number (LRN)-U2006
Page3 of16
Potassium Perfluorooctanesuffonate CAS Number-2759-39-3
3M Envimnmental Laboratory Report No. Tox-ol2
LaboratoryRequestNumber (LRN)-U2006
STATEMENT OF COMPLIANCE
StudyTite: -
AnalyticaLlaboratoryReport on Presence and ConcentrabonofPotassiumPernuorooctanesulfbnatienSerum and LiverofSprague-DawleyeRatsExposed toPFOS via Gavage
Study ldentficaboNnumber -FACT Tox-012
Thisstudywas conductedincompliancewithFood and Drug AdministratioGnood LaboratoryP.ractice (GLP) RegulationfsorNonclinicaLlaboratory'Studi[eDsataRequirement(s)2:1 CFR (Part58)],with theexceptionsinthebulleteldisbtelow.Inadditiont,hepresentstudyhas been auditedretrospectively by an independentqualitayssuranceunitAuditproceduresand findingfsorauditsperformedatthe3M EnvironmentalLaboratoryand atparficipaficnogntracltaboratodeasre housed withdocumentation perfinenttothisstudyinarchivesatthe3M laboratoraynd valble retainefdoratleast10 years.The analyticaplortiocnompletedatthe3M EnvironmentalLab was perfomiedinaccordancewith3M EnvironmentalTechnology and SafetyServicesStandardOperatingProcedures.
ExceptionstoGLP compliance:
Storagecontainersforthereferencesubstance,PFOS, were notlabeledwithname, CAS number, batchnumber,expiratiodnate,orstorageconditions.
Detailsofthepreparationm,aximum storagetime,and stabiliptryopertieosfthereference substance,PFOS, are unknown.
The idenbtys,trengthp,uritya,nd compositiondefinintghetestorcontroalrtclewas notdetermined atthetimeofthestudy;however,analysestodetaitlhecharacterizatiwoenre ongoingatthetimeof thisanalyticasltudy.
Two separatestudydirectowresreassignedtotheinvivoandtheanalyticpaolrtionosfthistudy. From 15 September 1998 unti1lOctober1998,theprotocowlas notsponsorapproved and was
dudng thattimeperiodnotincompliance.
Deviationwsere notconsistentalpyproved by thestudydirectoars requiredby GLP regulations. Data correctionwsere notalwaysrecordedas requiredby GLP regulations.
Data were notalwaysattributetdotheindividuarlesponsiblfeorrecordingthedata. Records inelectronifcormdo notmeet thecdtenasetfortuhnder21 CFR(L 1).
A finalizesdample log-i/ntrackinsgystem was notinplaceatthetimeofthestudy;however,draft documents didexistand were used tologinand tracksamples.
Page 4 of16
Potassium Perfluorooctanesulfonate CAS Nuffiber-2759-39-3
3M EnvironmentalLaboratory RAportNo. Tox-012
LaboratoryRequest Number (LRN)@.U2006
Not allraw data were verifiedby the study director.
No in-phase auditswere conducted dudng the present study. At the time of the present study,personnel filesdid not always containdocumentation of equipment
and method training;however, personnel fileshave been updated, and thisnoncompliance has been resolved.
Signature of Study Director Signature of Study Sponsor
a YI
Page 5 of16
Potassium Pc- -@rooctanesulfonate CAS Number-275-9-39-3
GLP STUDY QUALiTy AssURANCE
STATEMENT
@llr-:nvironmentLaalboratory
Report No. Tox-012 LaboratoryRequestNumber (LRN)-U2006
StudyTitle: AnalyticLalboratoRreyportonPresencaendConcentratoifoPnotassium PerfluorooctanesulfbniantSerum and LiverofSprague-Dawley"R,a,ts Exposed to PFOS viaGavage
Study IDNumber: FACT Tox-O12
Thisstudyhas been inspectebdy the3M EnvironmentaLlaboratorQyualitAyssuranceUnit(QAU) as indicateidnthefollowintgable.The findingwsere reportedtothestudydirectoarnd managemenl
9 August 1999 1 27 September 1999 FinalReport
4 October1999 4 October1999
4U
Page 6 of16
Potassium Perfluorooctanesuffbnate CAS Number-2759-39-3
INTRODUCTION
3M EnvironmentalLaboratory Report No. Tox-ol2
LaboratoryRequestNumber (LRN@U2006
Potassium Perfluorooctanesulfonate CAS Number 2795-39-3
Chemical Formula:C,F17SO3-K+
MolecularWeight 538
Inthepresentstudy,groupsofFO ratswere administered0.1,0.4,1.6,or3.2mg PFOS kg/dayin0.5% TWeen 80.(Thesedoses correspondtcc)oncentrationosf0.02,0.08,0.32,and 0.64mg/mL.)Groups of vehiclecontrolFO ratswere administereodnlyTween 80.Male FO animalswere treated42 days prior tomatng and throughthematng periodf;emaleFO animalswere administeredPFOS daily42 days priortomabng, throughgestationa,nd up to20 days fbIloMnglittdeerliveryF.l male and femalerats were exposed tothechemicalinuteroand duringlactatioFno.llowinwgeaning at21 days ofage, selectedFl animalswere treatedduringdevelopmentand productioonfF2 animals.Variousphysical and biologicaplarameterswere monitoredinFO,Fl,and F2 animals.
Intheanalyticsatludyreportedhere,livearnd serasamples collectefdromtheinitipaolpulatioonf dosed animals(generatioFnO)and theiorffsprin(ggeneratioFnl)were analyzedforthepresenceof PFOS. (Analyseswere perfbrmedtodeterminethepresenceofETFOSE, PFOSA, POAA< PFOSEA, PFOSAA, and themonoester,however,thesedatawere collectefdorinfbrmatonaplurposesonly,and were notreported.)
Liversamples were homogenized,and livearnd serasamples were extractedby an ion-paidng extractiopnrocedure.The extractwsere quantitativealnyalyzedusinghigh-pressurleiquid chromatography/electrosprtayndem mass spectrometry(HLPC/ESMSMS), and PFOS levelswere evaluatedagainstextractedstandards.Analyticadletailasre includedinthisreportf;urthedretailasre availablienthestudybindermaintainedby the3M FluorAnalyticCahlemistryTeam (FACT).
AnalysesassessingthetoxicologiceaflfectosfPFOS intheliverasnd seraofSprague-Dawleyrats were conductedincompliancewithGood LaboratoryPracbceRegulation(s21CFR 58).Validated methods and standardoperatingprocedureswere followeduringthepreparatioannd analysisofthe samples associatedwiththisstudy.
SAMPLE RECEIPT
Tissuesamples were receivedfromArgus Research Laboratoriessporadicallfyr,omAugust,1998, throughJanuary,1999. Samples receivedwere pa6kaged indryice.Specimens were registerewdith the3M EnvironmentalLaboratoryand transferretdoa freezerfbrstorage.Sample receiptidentficaton, storage,and chainofcustodyprotocolasnd dataarelocatedinthestudyfbldefrbrthisreportt;hefblder islocatedinthe3M archives.
Specimens analyzedatthe3M EnvironmentalLaboratorywillbe maintainedfora periodof10 years and willbe storedatthelaboratorayt-20*C 1O*C.
Page 7 of16
Potassium Perfluorooctanesuffonate CAS Number-2759-39-3
3M EnvironmentalLaboratory Report No. Tox-O 12
LaboratoryRequest Number (LRN)-U2006
MATERIALS AND METHODS
Chemical Characterization
Table 1 presents information regarding characterizationof the test,control,and reference substances used inthe analyticalportionof thisstudy.
Reference Substance Source Preparation Maximum Storage Time Storage Conditions
Chemical Lot Number Physical Descriptionand IdentityAnalyses Purity Stability Control Substance Source
Preparation Maximum Storage Time Storage Conditions Chemical Lot Number
PhysicalDescriptionand Iden* Analyses Purity Stability
Potassium peffluorooctanesulfonate 3M SpecialtyChemical Division Detailsunknown Unknown In a sealed container at room temperature, exposed to light 193 White powder 99.28% Unknown Rat liverand serum matrices Rabbit liver-Argus Research Laboratories,Inc. Rabbit serum-Sigma Chemical Company Unknown Unknown Frozen at -20'C Liver-F.0001 2 Serum: 17H9306 Rabbit liverand serum Unknown Unknown
Table 1.Characterization and Treatment of the Control, Reference, and Test Materials inthe Study of the Presence and Concentration of Potassium Perfluorooctanesuffonate inSerum and Liver of Sprague-Dawley' Rats Exposed to PFOS via Gavage
Page 8 of 16
Potassium Perfluomoctanesuffonate CAS Number-2759-39-3
3M Envimnmental Laboratory
Report No. Tox-01 2 LaboratoryRequest Numt>er (LRN)-U2006
9:1=Fl ill
Test Material Source
Preparation
Maximum Storage Time StorageConditions Chemical Lot Number PhysicalDescriptioannd Ident4 Analyses Purity Stabil@y
Rat serum and liver Sprague-Dawley ratsexposed toPFOS via gavage duringtheinvivoportionof 3M TOX 6295.9 atArgus Research Laboratories
Liverextractiopner FACT M-1.0 (referto AttachmentF) Serum: extractiopner FACT M-3.0 (referto AttachmentF) I week
Frozen at-20'C NA Rat livearnd serum NA NA
Table 1.Characterization and Treatment of the Control,Reference, and Test Materials inthe Study of the Presence and Concentration of Potassium
Perfluorooctanesulfonate inSerum and Liverof Sprague-Dawley' Rats Exposed to PFOS via Gavage (continued)
Followinganalysis,remainingoriginaslpecimens (testand controlmaterialw)ere storedfrozen at -20'C and willbe maintainedfora periodof 10 years.
Method Summaries
Followingisa bdefdescriptioonfthemethods used duringthisanalyticasltudyby the3M EnvironmentalLaboratory.Detailedescriptionosfthesemethods are locatedinAttachment F.
Preparatory Methods:
Method FACT-M-1.0-.ExtractioonfPotassium PerfluorcoctanesulfonaotreOtherAnionic FluorochemicalSurfactantsfrom LiverforanalysisUsing HPLC/ESMS. Inthismethod, an ionpaihngreagentwas added tothe sample and theanalyteionpairwas partitioneidntoMTBE. Four mLs ofthe MTBE extractwas transferretdoa centrifugteube and put onto a nitrogenevaporatoruntidlry.Each extractwas reconstituteidn1.0mL ofmethanol,then filteretdhrougha 3 cc plastiscyringeattachedtoa 0.2pm nylonfilteirntoglassautovials.
Page 9 of 16
Potassium Perfluamoctanesudbnate CAS Number-2759-39-3
3M EnvironmentalLaboratory
Report No. Tox-012 LaboratoryRequest Number OPJ4)-U2006
Method FACT-M-3.0: Extractionof Potassium Peffluorooctanesulfonateor Other Fluorochemical Compounds from Serum or other FluidforAnalysis Using HPLC-Electrospray/Mass Spectrometry. Sera samples were extracted using an ion-paidngextractionprocedure. Insummary, an ion pairing reagent was added to the sample and the analyte ionpairwas partitionedintomethyl-teft-butyl ether (MtBE). Four mL of the MTBE extractwas removed and put onto a nitrogenevaporator until dry. Each extractwas reconstitutedin 1.0mL of methanol and filteretdhrough a 3-cc plasticsyringe attached to a 0.2 pm nylon fifteirntoglass autovials.
Analytical Methods
Method FACT-M-2.0: Analysis of Fluorochemicals in LiverExtractUsing HPLC-Electrospray/Mass Spectrometry The analysiswas perfbrmed by monitoring a singleion characteristiocf a particulafrluorochemical, such as the perfluorooctanesulfonate(PFOS) anion, rri/=z 499. Additionallys,amples were analyzed using a tandem mass spectrometer to furtherverffythe identitoyf a compound by detecting daughter ions of the selected parent ion.
Method FACT M-4.0: Analysis of Potassium Perfluorooctanesuffonateor other Fluorochemical in Serum or other FluidUsing HPLC-Electrospray/Mass Spectrometry The analysiswas perfbrmed by monitoring a singleion charactedsbc of a particularfluorochemical, such as the perfluorooctanesulfonate(PFOS) anion,rrdz=499. Additionallys,amples were analyzed using a tandem mass spectrometer to furtherverifythe idenbtyof a compound by detecting daughter ions of the parent ion.
Confirmatory dose analyses were conducted followingthe end of the study, and the resultsof these analyses are presented inAttachment G.
Page 10 of 16
PotassiumPerfluorooctanesuffonate CAS Number-2759-39-3
3M EnvironmentalLaboratory Report No. Tox-012
L2boratorRyequestNumber (LRN)-U2006
Ana"cal
Equipment
Following are typicalanalyficalequipment settingsforthe procedures used inthe present study. These settingsvary somewhat duringactualdata collecbon.Exact settingsduringallphases ofdata collection are recorded and presented inthe analysissectionofthe study binderforFACT Tox-12.
HPLC System: Hewlett-Packard Series1100 Liquid Chromatograph
Column:
Keystone BetasilC18 Column 2Xl 00 mm, 5pm particlseize
Flow rate:
300 pumin
Solvent A-
2.0 mM ammonium acetate
Solvent B:
Methanol
SolventGradient
InjectonVolume: Run Time:
40% to 90% B in8.5 minutes Hold at 90% B fbr3.0 minutes Return to40% B in1.0 minute Hold at40% B for1.0 minute
10 pL
13.5 minutes
ElectrosprayTandem Mass Spectrometer: Micromass Quattro 11API Mass Spectrometer
Mass Lynx 3.1 Software
Cone Voltage:
30--60V
CollisionGas Energy:
40 eV
Mode:
ElectrosprayNegative
Source Block Temperature: 115'C
Desolvabon Temperature: 2500C
Electrode:-
Z-spray
Primary Ion/DaughterIons: 499/80,99, 130, 180, 230 amu
Page 11 of16
Potassium Perfluorooctanesudbnate CAS Number-2759-39-3
3M EnvironmentalLaboratory
ReportNo. Tox-012 LaboratoryRequest Number (LRN)-U2006
Protocol Deviations
There was one deviabon'totheprotocolT:he protocolcalledfbrtheuse of referencestandardlot number 217; however, lotnumber 193 was used duringthe analyticaplortionofthestudy.
DATA SUMMARY, ANALYSEST AND RESULTS
Summary of QualityControlAnalyses Results
CalibratioCnheck Standards:A mid-levele,xtractedmatrixcalibraticohneck was analyzed at leastevery 10 samples tomonitorinstrumentdriftCalibratiocnheck standardswere compliant, and allcalibratiocnriteriwaere met (within 30%).
Blanks:Four blankswere extractedconcurrentlwyitheach batchof samples.Two extraction blankswere preparedwithwateras a surrogatematrix.Two additionaslamples of blank matrixwere typicallpyreparedwithrabbitliveorr seraand were used forextractedcalibration curves.AJIblankswere nondetectfbrthe oompound(s) ofinterest
Duplicatematrixspikeanalysesincludingalltargetanalyteswere preparedand analyzed for one controlanimal.With a few exceptions,recoverieswere withintheacceptablerange of70130%.
Analysisoftwo livemratrixspikesamples resultedin51% and 61% recoveriesof PFOS. These recoveriesare below the lowerboundary oftheacceptablerange;however, thesecond setof matrixspikesamples were withintheacceptablerange.No actionwas taken tofurther characterizethesematrixspikes.
Analyses ofboth setsofsera matrixspikesresultedinPFOS recoveriesgreatlyabove the high limiotftheacceptablerange.AlthoughI mL ofserawas used forthe analysisof PFOS levels inthesera oftestanimals,onlya smallamount ofserum (500 pL)was availablfeorthe matrix spikepreparationt;hislimiteqduantityofmatrixmay have been a sourceoferror.As the extractioinsscaled-down toaccommodate theavailablematrix,the resultinegxtractis particuladsyusceptiblteolossofsolventdue toevaporation.These evaporativelosses,which would noteffecthe PFOS concentratio(nPFOS has a low vapor pressure)intheextract couldresultina greater-than-expectPeFdOS concentratioinntheextractWhen analysesof sera matrixspikesare performedwitha full1 mL ofsera,highPFOS recoveriesare unusual.
MatrixSpikes: Matrixspikesand matrixspikeduplicatewsere analyzedevery40 samples, witha minimum of two perbatchduringanalyticaalnalyses.The resultosfthese analysesare locatedinthe archivedstudybinder.
Page 12 of 16
Potassium Perfluorooctanesuffonate CAS Number-2759-39-3
3M EnvironmentalLaboratory
ReportNo. Tox-012 LaboratoryRequest Number (LRN)-U2006
Summary of Sample Results
PFOS was detected in the livercsfallcontrol and exposed groups of FO animals and Fl
animals. PFOS was detected inthe sera samples collectedfrom allof the controland dosed FO males and insera samples collectedfrom alldosed FO females. The PFOS levelsfound in the samples from controlanimals were quitelow--generally100-less than levelsdetected in the low-dose group. PFOS concentrationin exposed animals increased with increasingdose group, generallyin a dose-related manner.
PFOS levelsinboth sera and liversamples collectedfrom female FO animals were much less than levelsdetected insamples collectedfrom the corresponding group of FO males.
PFOS concentratons measured inpooled liversamples collectedfrom the exposed groups of Fl animals shortlyafterbirthwere roughly equal to or lower than concentrationsmeasured in the corresponding groups of FO females. No samples collectedfrom FO females thatreceived 3.2 mg PFOS per kg/day were submitted foranalysis. Table 1 summahzes resultsof the quantitativeanalyses performed on the FO liverand sera samples examined.
IF#
0.0 mg/kg/day
0.0
0.1 mg/kg/day
0.1
0.4 mg/kg/day
0.4
1.6 mg/kg/day
1.6
3.2 mg/kg/day
3.2
Samples were notreceived.
0.0 0.02 0.08 0.32 0.64
Female: 0.0307 Male: 0.0244 Female-. 5.28 Male: 10.5 Female-. 18.9 Male-.45.4 Female: 82.0 Male: 152 Female: NR' Male: 273
Female: 0.171 Male: 0.665 Female: 14.8 Male: 84.9 Female: 58.0 Male: 176 Female: 184 Male: 323 Female: NR a Male-.1360
Table 2. PFOS Concentrations inSerum and Liver Samples from FO Male and Female SpragueDawley Rats
Page 13 of 16
Potassium Perfluor(>octanesulfonate CAS Number-2759-39-3
3M EnvironmentalLaboratory Report No. Tox-012
LaboratoryRequest Number (LRN@-U2006
PFOS concentrations measured in pooled liversamples collected from the control and exposed groups of Fl animals shortly after birthwere roughly the equal to or lower than concentrations measured inthe corresponding groups of FO females. N(D samples collectedfrom Fl males or females thatreceived 3.2 mg/kg/day were submitted foranalysis.PFOS was detected inthe liverof allother exposed groups of Fl animals. Table 2 summarizes resultsof the -quanttativeanalyses performed on the Fl sera samples examined.
0.0 mgikg/day
0.0
0.1 mg/kg/day
0.1
0.4 mg/kg/day
0.4
1.6 mg/kg/day
1.6
ly
3.2
a Samples were notrece@ved.
0.0 0.020 0.080 0.320 0.640
0.051 1 619 57.6 70.4 NRa
Table 3. PFOS Concentrations in Liver Samples from Fl Male and Female Sprague-Dawley
Rats
Paper and electronic copies of raw data and study reports generated during the present studies willbe kept forat leasta pehod of time as establishedby regulationand by the 3M Standard Operating Procedures. Paper and electroniccopies of reports,protocols,and methods generated by the 3M Environmental Technology and Safety Services laboratorywillbe retainedwith the study folderforat leasta period of time as establishedby regulationand by the 3M Standard Operating Procedures.
STATISTICAL
METHODS
For data generated by the 3M Environmental Laboratory,means and standard deviationswere calculatedusing MicrosoftExcer, and relativsetandard deviationswere calculatedmanually.
Standard deviationis a measurement of analyticalprecision-t,hus itisused to gauge the precisionof the analyses. Relativestandard deviationpresents a measure of the magnitude of the standard deviation.
2
F n(n - 1)@ Means are calculatedby adding individuaelntitieasnd dividingthe resultantsum by the number of individualentitiesS.tandard deviationswere calculatedusing the equation:
Relativestandard deviationswere calculatedby divisionof the standard deviationby the mean with subsequent multiplir-aboonf the quotientby 100.
Page 14 of16
Potassium Perfluorooctaneustfonate CAS Number-2759-39-3
3M EnvironrmntalLaboratory
Report No. Tox-ol2 LaboratoryRequestNumber (LM)-U2006
DATA QUALITY OBJECTIVES AND DATA INTEGRITY
No circumstancesexisteduringthepresentstudythatwould have affectetdhequalitoyrintegriotfy thedata.The fbilowindgataqualitoybjecfive(sDQOS) were followeduringthestudy:
No circumstancesexistedduringthepresentstudythatwould have afkkctedthequalitoyrintegriotfy thedata.The fbllowindgataqualitoybjecfive(sDQOS) were followeduringthestudy:
Linearity-The correlaticoonefficie(nRt2)ofthestandardcurvewas equaltoorgreaterthan0.98 using1/xweighting
Limitsofdetecbon-PFOS inserum=1.75ppb Limitsofquanbtabor@--Equatlothelowestacceptablestandardinthecalibraticounrve Duplicatferequen4/acceptablperecision-<30% Spikefrequency/acceptabrleecovedes-70% to130% Use ofconfirmatormyethods-no confirmatormyethodswere used Demonstrationofspecificity-specifwiacsitdyemonstratedby chromatographircetentiotnimeand
mass spectradlaughterioncharacterizabon
STATEMENT OF CONCLUSION Statement ofConclusion:
Undertheconditioonfsthepresensttudiepso,tassiupmerfluorooctanesulfwbansaotbeservedinthe liverasnd seraofalglroupsofratsexposedtothechemicadlurintgheinvatoportioonfthestudies.
REFERENCES None
Page 15 of 16
Potassium Peffluomodanesuffonate CAS Number-275c-L-39-3
3M EnvironmentalLaboratory ReportNo. Tox-ol2
LaboratoryRequest Number (U@N)-U2006
ATTACHMENTS AttachmentA Reportsignaturepage
AttachmentB: Table ofresultsofanalysisoflivesramples from FO male and female SpragueDawley ratsadministeredPFOS dailyby gavage.
Attachment C: Table ofresultsofanalysisofsera samples from FO male and female SpragueDawley ratsadministeredPFOS dailyby gavage.
Attachment D: Table ofresultosfana@sis oflivesramples from Fl male and female SpragueDawley ratsadministeredPFOS dailyby gavage.
Attachment E: In-lifperotocoland studyprotocol
Attachment F: Preparatoryand analyticamlethods
Attachment G: Dose Analyses
Page 16 of 16
Potassium Peduomoctonesugonate CAS Number-2759-39-3
ATTACHMENT A REPORT SIGNATURE PAGE
3M EnvirorumntalLaboratory ReportNo. Tox-012
LaboratoryRequest Number OPJ4@WOOS
KriitenJ. Hansen, Ph.D., Study Director
MarvinT. Case, b.vm., Ph.D.,Sbidy Sponsor
Dafe L. Bac4n, Laborarory Director
Date
Date
Date
Page 17 of16
Potassium Perfluorooctanesuffonate CAS Number-2759-39-3
ATTACHMENT B
3M Environmental Laboratory Report No. Tox-012
LaboratoryRequest Number (LRN)-U2006
Method Blank Method Blank Matdx Blank Matdx Blank QC - 100 ppb
0.0 mgfkg/day
0.1 mgfkg/day (0.02mg/mL)
H20 Blank 1 H20 Blank 2 Rabbit Liver (Blank1) Rabbit Liver (Blank2) 8801 F-MS 8801 F-MSD 880IM-MS 8801 M-MSD 8801 F 8807F 8808F 8821 F 8833F 8101M 8103M 8104M 8107M 8108M 8838F 884OF 8842F 8864F 8870F 8138M 8140M 8142M 8145M 8146M
N/A N/A
N/A
N/A 1.0169 1,0169 1.0525 1.0525 1.0169 1.0384 1,0177 1.0439 1.0048 1.0525 1.0407 1.0568 1.0104 1.0321 1.0374 1.0168 l.C)489 1.0608 1.0225 1.0617 1.0509 0.9961 1.036 0.9788
<MDL <MDL
<MDL
<MDL 60 72 100 93 674 158 165 197 166 910 600 577 593 643 17251 14433 15557 13807 12784 80270 76629 92106 87938 87451
<MDL <MDL
<MDL
<MDL 50% 60% 87% 81% 0.674 0.158 0.165 0.197 0.166 0.910 0.600 0.577 0.593 0.643 17.3 144 15.6 13.8 12.8 80.3 76.6 92.1 87.9 87.5
<MDL 'ONO
<MDL
<MDL 55% 84%
0.171' 0.272
<MDL
18%
7%
10.11 0.0172' 82.9 0.225
0.665
21.0 0.139
11.6
14.8
1.71
84.9
7.39 6.28
Table
4. Analyses of Liver Samples from
and Concentration
of Potassium
Sprague-Dawleyo
Rats Exposed
FO Sprague-Dawley
Rats
Perfluorooctanesulfonate
to PFOS via Gavage
in the Study of the Presence in Serum and Liver of
Proprietaryand Confidential
Potassium Perfluorooctanesuffonate CAS Number-2759-39-3
ATTACHMENT B (CONTINUED)
3M EnvironmentalLaboratory
Report No. Tox-O 12 LaboratoryRequest Number (LRN)-U2006
0.4 mglkg/day (0.08mg/mL)
1.6mg/kg/day (0.32mg/mL)
3.2 mglkg/day (0.64mg/mL)
889OF 8893F 8895F 8902F 8905F 8172M 8174M 8175M 8176M 8181 M 8919F 8921 F 8926F 8934F 8937F 8209M 8213M 8219M 8221M 8225M NR NR NR NR NR 8243M 8244M 8249M 8250M 8255M
1.0346 1.0247 1.0154 1.0039 1.0426 1.0234 1.0067 1.0471 1.0335 1.0197 1.0756 1.0544 1,0219 1.022 1.654 1.0016 1.0211 l@0421 1,0153 1.0316 NR NR NR NR NR 1,OD65 1.0397 1.0107 1.0522 0.9908
61853 62547 49543 64081 51922 137715 189256 183050 173288 198173 200313 212684 282001 183576 40958 374773 325087 279320 335349 301145 NR NR NR NR NR 1271156 1116251 2077414 1168784 1149853
61.9 62.5 49.5 64.1 51.9 138 189 183 173 198 200 213 282 184 41.0 375 325 279 335 301 NR NR NR NR NR 1271 1116 2077 1169 1150
58.0 176 184 323 NR 1360
11.6 6.73
13.3 23.4
48.0 88.3
11.2 36.1
NR NR
30.0 40.7
a Sample 8801 F valuewas excludedfrom thecalculations.
Table4. Analyses ofLiverSamples from FO Sprague-DawleyRats intheStudy ofthePresence
and Concentration of Potassium Perfluorooctanesulfonate
in Serum and Liver of
Sprague-Dawley'
Rats Exposed to PFOS via Gavage (continued)
Prophetaryand Confidential
Potassium Perfluomoctanesuffonate CAS Number-2759-39-3
ATTACHMENT C
3M EnvironmentalLaboratory Report No. Tox-012
LaboratoryRequest Number (LRN)-U2006
Method Blank H20 Blank 1
<MDL
Method Blank H20 Blank2
<MDL
<MDL
MatrixBlank
Rabbit Liver
(Blank1)
<MDL
<MDL
MatrixBlank
RabbitLiver
(Blank2)
<MDL
<MDL
QC - 100 ppb 8801 F-MS
60
50%
8801 F-MSD
72
60%
8801 M-MS
100
84%
8801 M-MSD
93
78%
0.0 mgfkg/day 8801 F/M
369
8807F/M
51.5
8808F/M
53.2
8821 F/M
52.7
8833F/M
46.9
0.1mg/kg/day 8838F/M
5872
(0.02mg/mL)
8840FIM
5149
8942F/M
7390
8864F/M
6745
8870FIM
5788
0.369 0.0515 0.0532 0.0527 0,0469 5.87 5.15 7.39 6.74 5.79
0.4mglkg/day 889OF/M
62050
62.1
(0.08mg/mL)
8893F/M
61745
61.7
8895F/M
47738
47.7
8902F/M
63057
63.1
8905F/M
53492
53.5
1.6mglkg/day 8919F/M
78376
78.4
(0.32mg/mL)
8921 F/M
58893
58.9
8926F/M
91956
92.0
8934F/M
65023
65.0
8937F/M
57897
57.9
a Sample 8801F valuenot includedinthiscalculation
<MDL <MDL 55% -81% 0.0511, 0.115
6.19
57.6
70.4
<MDL
<MDL 18% 7% 5.583 0.002851 124 0.142
14.2 0.879
11.7 6.72
20.6 14.5
Table 5. Analyses of Liver Samples from
and Concentration of Potassium
Sprague-Dawley'
Rats Exposed
Fl Sprague-Dawley
Rats
Perfluorooctanesulfonate
to PFOS via Gavage
in the Study of the Presence in Serum and Liver of
Propbetaryand Confidential
Potassium Perfluorooctanesulfonate CAS Number-2759-39-3
ATTACHMENT D
3M EnvironmentalLaboratory
Report No. Tox-O 12 LaboratoryRequest Number (LRN)-U2006
Method Blank Method Blank MatrixBlank MatrixBlank QC - 100 ppb
0.0 mglkg/day
0.1 mg/kg/day (0.02mg/mL)
0.4mg/kg/day (0.08mg/mL)
H20 Blank 1 H20 Blank2 RabbitLiver(Blank1) RabbitLiver(Blank2)
8801 F-MS 8801 F-MSD 8801M-MS 8801M-MSD 8801 F 8807F 8808F 8821 F 8833F 8101M 8103M 81 C)4M 8107M 8108M 8838F 884OF 8842F 8864F 887OF 8138M 8140M 8142M 8145M 8146M 8890F 8893F 8895F 8902F 8905F 8172M 8174M 8175M 8176M 8181 M
<MDL <MDL <MDL <MDL 136% 133% 119% 121% 0.127 0,0229 0.0291 0.0445 0.0265 0.0306 0.0311 0.0177 0.0213 0.0214 5.14 4.93 5.24 5.23 5.89 11.8 9.54 9.65 11.0 10.4 20.9 18.6 18.4 17.4 19.1 40.7 54.1 41.6 47.4 43.3
<MDL <MDL 135% 120% 0.037' 0.0500
0.0244
5.28
10.5
18.9
45.4
<MDL <MDL 1% 1% 30.9 0.00950 a 87.6 0.044
2.48 0.00605
6.78 0.358
9.02 0.946
6.89 1.30
12.1 5.49
Table6. Analyses ofSeraSamples from FO Sprague-Dawley Rats in the Study of the Presence
and Concentration of Potassium Perfluorooctanesulfonate
in Serum and Liver of
Sprague-Dawley'
Rats Exposed to PFOS via Gavage
Proprietaraynd Confidential
Potassium Perfluorooctanesuffonate CAS Number-2759-39-3
3M EnvironmentalLaboratory
ReportNo, Tox-012 LaboratoryRequest Number (LRN)-U20D6
lq:li"
1.6mglkg/day (0.32mg/mL)
3.2mgikg/day (0.64mg/mL)
8919F 8921 F 8926F 8934F 8937F 8209M 8213M 8219M 8221 M 8225M NR NR NR NR NR 8243M 8244M 8249M 8250M 8255M
75.3 79.2 113 74.4 68.4 144 151 148 165 153 NR NR NR NR NR 249 242 257 361 255
Sample 8801F valuenot includedinthiscalculation
82.0 152 NR 273
21.4 17.5
5.20 7.91
NR NR
18@3 49.8
Table 6.
Analyses of Sera Samples from FO Sprague-Dawley
Rats in the Study of the
Presence and Concentration of Potassium Perfluorooctanesulfonate
in Serum
and Liver of Sprague-Dawleyo
Rats Exposed to PFOS via Gavage (continued)
Proprietaraynd Confidential
Potassium Peffluorooctanesulfonate CAS Number-2759-39-3
3M EnvironmentalLaboratory ReportNo. Tox-012
LaboratoryRequest Number (LRN@U2006
ATTACHMENT E IN-LIFE PROTOCOL
AND ANALYTICAL STUDY PROTOCOL
Proprietaraynd Confidential
Potassium Perfluorooctanesulfonate CAS Number-2750,39-3
ATTACHMENT F: PREPARATORY AND ANALYTICAL METHODS
3M Envimnmental Laboratory ReportNo. Tox-012
LaboratoryRequest Number (LRN@U2006
Proprietaraynd Confidental
Potassium Perfluorooctariesuffonate GAS Number-2759-39-3
3M Envimnmental Laboratory
ReportNo. Tox-M2 LaboratoryRequest Number (LRN)-U2006
ATTACHMENT G: RESULTS OF CONFIRMATORY
DOSE ANALYSES
Proprietaraynd Confidential