Document 7MgK85kY04zJGeMdn3YQG5kwg
DeterminationoftheWater Solubilitoyf PFOS by the Shake FlaskMethod
WATER SOLUBILITY
TEST SUBSTANCE
Identity:Perfluorooctanesulfonatmea;y alsobe referredtoas PFOS or FC-95. (1-Octanesulfonicacid,1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8heptadecafluoro-p,otassium salt,CAS # 2795-39-3)
Remarks: The testsubstance isa whitepowder. Sample was taken from 3M lotnumber 217. Sample was storedunder ambient conditions priortotesting...Puddteytermined to be 90.49% by LC/MS','H-HMR, 19FNMR and elementalanalyses techniques.
METHOD
Method: OECD 105, OPPTS 830.7840, and 40 CFR 796.1840. GLP (Y/N): Yes Year completed: Study completed 1999. Reportcompleted 2000
Remarks: The definitivtestconsistedof equilibratioofnan excess amount oftestsubstance withNANOpure!* water at300C followedby equilibratioant200C and analyzingsubsamples by high performance liquidchromatography withmass spectrometricdetection(LCMS).
NANOpure waterisequivalentto ASTM Type 11DesignationDl 193-91.
RESULTS
Value (mg/L) attemperature OC: 519 mg/L at20 0.50C. Description of solubility:Slightlsyoluble.
Remarks: Triplicatseubsamples were removed from theappropriate boftlesafterone,two and threedays ofshaking ina water bath maintainedat30 1.OOC and followingone day ofa 20 0.50C equilibratipoenriod.Analysisofaqueous subsamples afterone day had a mean analyticarlesultof459 mg/L (SD = 8.96,CV = 1.95%). For subsamples collectedaftertwo and threedays,the mean concentration were 537 mg/L (SD = 27.6,CV = 5.14%) and 501 mg/L (SD = 64.2,CV 12.8%),respectively.
CONCLUSIONS
The Day 2 and Day 3 mean solubilitcyoncentrationwere within15% of each otherand were averaged toobtaintheoverallmean solubility concentration.The overallmean solubilictoyncentratioonfthe test substance inNANOpure waterwas 519 mg/L (SD = 48.3;CV = 9.31%; N = 6).
Submifter: 3M Company, EnvironmentalLaboratory,P.O. Box 33331, St. Paul,Minnesota,55133
DATA QUALITY
ReliabilityK:.i.miscrhanking1.
REFERENCES
Study conducted at the requestof3M Company by WildlifIenternational, Ltd.of Easton,Maryland.
OTHER
Last changed: 5/3/00
DETERMINATION OF THE WATER SOLUBILITY OF PFOS BY THE SHAKE FLASK METHOD
WILDLIFE INTERNATIONAL LTD. PROJECT NUMBEK- 454C-107
OECD GuidelinefortheTestingofChemicals,105 Water Solubility
AUTHORS: Raymond L. VanHoven, Ph.D.
WillardB. Nixon,Ph.D.
STUDY INITIATION DATE: February10,1999 STUDY COMPLETION DATE: May 3,1999 AMENDED REPORT DATE: April26,2000
Submittedto 3M Corporation EnvironmentalLaboratory 1 935 Bush Avenue St.Paul,Minnesota 55144
WildlffeInternationalLtd.
8598 Commerce Drive Easton,Maryland 21601
(410)822-8600
Page Iof 35
AMENDED
WILDLIFE INTERNATIONAL LTD. -2-
PROJECTNO.: 454C-107
GOOD LABORATORY PRACTICE COMPLIANCE STATEMENT
SPONSOR: 3M Corporation
TITLE:
DeterminatioonftheWater SolubilitoyfPFOS by theShake FlaskMethod
WILDLIFE INTERNATIONAL LTD. PROJECT NUMBER: 454C-107
3M LAB REQUEST NO., U2723
STUDY COMPLETION: May 3,1999
AMENDED REPORT: April26,2000
Thisstudywas conductedincompliancewithGood LaboratoryPracticSetandardsaspublishedby the U.S.EnvironmentaPlrotectioAngency in40 CFR Part792,17August 1989 and OECD PrincipleosfGood LaboratoryPracticeO,CDE/GD (92)32,EnvironmentMonograph No. 45,Paris1992,withthefollowing exceptions:
The testsubstancewas not characterizeidnaccordancewithfidlGLP complianceh;owever,the characterizatiwoans performedaccordingto3M StandardOperatingProceduresand Methods,and allraw data arebeingmaintainedinthe3M archives.The testsubstanceisbeingrecharacteriziendaccordancewithGLP.
The stabilitoyf thetestsubstanceunder storageconditionastthetestsitewas not determinedin accordancewithGood LaboratoryPiacticSetandards.
STUDY DIRECTOR.
RYyrdorLd. VanHoven,Ph.D.
Scientist WildlifIenternationLatld. SPONSOR PPROVAL:
Sponsor
DATE
tlla 7
DA'rg
AMENDED
WILDLIFE INTERNATIONAL LTD. -3-
PROJECT NO.: 454C-107
QUALITY ASSURANCE STATEMENT
Thisstudywas examinedforcompliancewithGood LaboratoryPracticSetandardsaspublishedby the U.S.EnvironmentaPlrotectioAngency in40 CFR Part792,17August1989 and OECD PrincipleosfGood LaboratoryPracticeO,CDE/GD (92)32,EnvironmentMonographNo. 45,Paris1992. The datesof all inspectionasnd auditsand thedatesthatany findingwserereportetdothe StudyDirectorand Laboratory
Management were as follows:
ACLTVITY: Test Substance Preparation Sample Preparation Dmft Reportand Data FinalReport Amended Report
DATE CONDUCTED: February15, 1999 February17 and 18,1999 March 16 and 17,1999 May 3,1999 April26,2000
DATE REPORTED TO:
STUDYDIRECTOR.
MANAGENffiNT:
February15, 1999
February15, 1999
February 18,1999
February 18, 1999
March 17,1999
Mamh 23,1999
May 3, 1999
May 3,1999
April26,2000
April26,2000
ia46 H. Coleman QualityAssuranceRepresentative
oo
DATE
AMENDED
WILDLIFE INTERNATIONAL LTD. -4-
PROJECT NO.: 454C-107
REPORT APPROVAL SPONSOR: 3M Corporation TITLE: DeterminatioonftheWater SolubilitoyfPFOS by theShakeFlaskMethod WILDLIFE INTERNATIONAL LTD. PROJECT NUMBER. 454C-107 3M LAB REQUEST NO.: U2723
STUDY DIRECTOR-
Raymon&. 7aai-ove-nP,h.D. Scientist MANAGEMENT:
WillardB. Nixon,Ph.D. Manager,AnalyticaClhemistry
4-io-oo DATE
DATE f
AMENDED
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PROJECT NO.: 454C-107
TABLE OF CONTENTS
Title/CovePrage.........................................................................I.............
Good LaboratoryPracticCeomplianceStatement...............................................2.............
QualityAssuranceStatement...............................................................3.............
ReportApproval.........................................................................4.............
Tableof Contents........................................................................5.............
Summary ...............................................................................7.............
Introducti.o.n..........................................................................8.............
Purpose................................................................................8.............
ExperimentalDesign.....................................................................8.............
Materialsand Methods ....................................................................8............. Test Substance......................................................................9............. InternaSltandar.d...................................................................9............. Reagents...........................................................................9............. PreliminarTyestProcedure............................................................9............. DefmitiveTestProcedure.............................................................9............. CalibratioCnurve ..................................................................1.0............. ReagentBlanks....................................................................1.0............. Example Calculatio.n.s.............................................................1.1.............
Results ..............................................................................1.1............. WaterSolubili.t.y..................................................................1.1.............
Conclusions...........................................................................I.I.............
References............................................................................1.2.............
TABLES
Table 1. TypicalLC/MS OperationalParameters .............................................1.3............
Table2. ReagentBlanksAnalyzedConcurrentlDyuringSampleAnalysis.........................1.4.............
Table3. SolubiliotfyPFOS inNANOpure(D Water...........................................1.5............
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TABLE OF CONTENTS -Continued-
FIGLTRES FigureI -AnalyticamlethodflowchartfortheanalysiosfPFOS inNANOpureS Water ..............1.6............ Figure2 -A typicaclalibraticounrveforPFOS ...............................................1.7.............. Figure3 -A representatiivoenchromatogramofa low-levesltandard..............................1.8............. Figure4 -A representatiivoenchromatogramof ahigh-levesltandar.d............................1.9.............. Figure5 -A representatiivoenchromatogramof a reagentblanksample............................2.0............. Figure6 -A representatiivoenchromatogramofa Day 2 solubilistaymple..........................2.1.............
APPENDICES Appendix I-Protocol....................................................................2.2............ Appendix R -CertificaotfeAnalysi.s.......................................................3.3............. AppendixHI -PersonneIlnvolvedintheStudy.................................................3.4............. Appendix IV -ReportAmendment ..........................................................3.5.............
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PROJECTNO.: 454C-107
SUNEMARY
STUDY TITLE:
WILDLIFE INTERNATIONAL PROJECT NUMBER:
LTD.
SPONSOR: TESTINGFAC=Y:
LOCATION OF STMY, RAW DATA AND A COPY OF THE FINAL REPORT:
DeterminatioonftheWater SolubilitoyfPFOS by theShake Flask Method
454C-107 3M Corporation WildlifIenternationLatlcL Easton,Maryland 21601 WildlifIenternationLatld. Easton,Maryland21601
TEST SUBSTANCE: TESTDATES:
PFOS
ExperimentalStart-February11,1999 ExperimentaTlermination-February20,1999
TEST SYSTEM:
NANOpureg Water
SUMMARY:
The overallmean solubilictoyncmtratioonf PFOS m NANOpureO waterwas 519mg a.i./(LSD 48.3;CV 9.31%; N 6).
AAffiNDED
WILDLIFE INTERNATIONAL LTD. -8-
PROJECT NO.: 454C-107
INTRODUCTION
Tiusstudywas conductedby WildlifIenternationLatld.for3M CorporatioanttheWildlifIenternational Ltd.analyticcahlemistryfacilitiynEaston,Maryland.A testwas conductedtodeterminethewatersolubiliotfy PFOS (perfluorooctasnuelfoniaccid,potassiumsalt)T.he experimentaplortionofthisstudywas conducted betweenFebruary11,1999 and February20,1999. The originarlaw datageneratebdy Wildlif6International Ltd.anda copyofthefinarleportarefileudnderProjecNtumber 454C- 107 inarchivelsocatedon theWildlife InternationLatld.site.
PURPOSE
The purposeofthistudywas todeterminethewatersolubiliotfyPFOS at20 0.5'Cby theshakeflask method. Determinatioonfwatersolubilibtyy theshakeflaskmethod isapplicablteotestsubstanceswithwater solubiliteiqeusaltoorexceeding0.01gram/Lite(rg/L).
EXPERUWENTAL DESIGN
The watersolubiliotfyPFOS was detmm3inedata temperaturoef 20 0.5*C. A preliminartyestwas performedtoestimatethewatersolubilitTyh.e testconsisteodfadditionosfincreasinglyargeamountsof NANOpure(P watertoa known weightoftestsubstanceatroom temperatureT.he definititvestconsisteodf eqxnhbratioonfan excessamount oftestsubstancveathNANOpure@ waterat30"C followedby equilibrataiton 20'C andanalyzinsgubsamplesby highperformanceliquicdhromatographywithmass spectrometridcetectim (LCMS).
MATERL4J,S AND METHODS
Thisstudwyasconducteadccorditnogtheproceduroeustliniendtheprotoco"lDetaminafiomfthWeater SolubiliotfyPFOS by theShakeFlaskMethod."The protocowlas basedon proceduresoutlineidnft OECD GuidelinfeorTestingofChemicals,105: WaterSolubilit(y1).Thisstudymeetsdatarequirementusnderthis guidelinaendunderU.S.EPA ProductPropertieTsestGuidelinesO,PPTS, Series8:30.7840,WaterSolubility:
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ColumnElutioMnethod,S-hakeF7askMetho(d2)forshakeflasmkethodoloagnydTSCA Titl4e0ofthFeederal CodeofRegulatioPnasr,t796,Sectio1n840:WaterSolubjli(t3y).
TestSubstance The testsubstancewas receivedfrom 3M Corporationon October 29, 1998 and was assignedWildlife
InternationaLltd.identificatinounmber 4675 upon receipt.The testsubstance,a whitepowder,was idmitifieads: FC-95, Lot Number: 217, ExpirationDate: 2008 and a reportedpurityof 8.9%. The testsubstancewas reanalyzedby theSponsor and theCertificatoef AnalysisdatedMarch 9,2000 indicateadpurityof90.490/oT.he testsubstancewas storedunder ambient conditions.
InternalStandard The internasltandardwas receivedfrom 3M Corporationon July2, 1998 and was assignedWildlife
InternationaLltcLidentificatinounmber 4526 upon receipt.The internasltandard@a granularmaterial,was identifieads: 1H, 1H, 2K 2H PerfluorooctanSeulfonicAcid,ChemicalAbstractNumber: 27619-97-2. The standardwas storedunder ambient conditions.
Reapnts AU solventsused inthe solubilitaynalyseswere of ACS reagentgrade.Nanopure(D water (equivalento
ASTM Type ]IDesignationD 1193-91)was used (4).
Preliminmy Test Procedure A preliminarytestwas conductedto estimatethewatersolubilitoyf PFOS. The testwas performed at
room temperatureby adding 10..0mg of the testsubstanceto a 50-mL volumetricflask-Increasingvolumes of NANOpures waterwere added (10.0,25.0 and 50.0mL) totheflasl,A-fiffeach additiont,hecontainerwas shaken vigorouslyforapproximately10 minutesand visuallyexamined forundissolvedtestsubstance.Afterthe additionof 50-mL ofNANOpure(D water,no particulatewsere observecl
DefinitivTeest Procedure Approximately 0.500 grams of thetestsubstancewas weighed intoeach of threelabelled16-ounceFrench
squarebottlesN.ANOpure(D water,250 mL equilibrateadt30*C forapproximately30 minutes,was added to AMENDED
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eachbottle.The bottleswere sealedwith screwcapsand wrapped with parafilm.The bottleswere *agitateidna shaker-watr,brath (Lindberg/BlueM,odel No. SWB 1122A) maintainedat30 I.O*C. Afterone day,one bottle was removed and placedin a second constanttemperaturebath (Lindberg/BlueM,odel No. SWB I122A, with agitatiosnwitchedoff)maintainedat 20 0.5'C for 24 hourswith occasionalshaking. Triplicat1e0-mL subsampleswere removed from thebottlec,entrifugedatapproximately3 000 rpm forapproximately15 minutes ina centrifugreegulatedat20'C. Fiftymicroliterfsrom each subsample were volumetdcallryemoved and diluted to 5 00 mL witha solutionof 50% methanol and 50% NANOpure(D waterconmininge0.05% v/v fomiic acidand 0.100 mgtl of theinternalstandard.The dilutionwsere placedintoautosamplervialsforanalysisby reversephase highperformanceliquidchromatography (BPLC) usinga Hewlett-PackardModel 1100 High Performance LiquidChromatograph with a Perldn-ElmerAPI IOOLC Mass Spectrometerequippedwitha Perldn-ElmerTurbo lonsprayionsource.The remainingtwo bottleswere treatedina similamranner followinginitiaelquilibration periodsof 2 and 3 days at 30.0 I.OOC. Eash-umentalparametersfortheanalysisof PFOS'are summarized in Table 1 and a method flowchartisprovided inFigure 1.
CalibratioCnurve Calibrationstandardsof PFOS, rangingin concentratiofnrom 9.15 to 45.7 gg a.i./Lor 9.15 to
68.6gga.i./]Lw,ere analyzedwith each sample set. Each calibratiosntandardcontained0.100 mgAL of the internasltandard.A linearregressionequationwas generatedusingthepeak arearesponses(ratiosof PFOS to theinternasltandard)versustherespectiveconcentrationosf thecalibratiosntandardsA. typicaclalibratiocnurve ispresentedinFigure2.The concentrationofPFOS inthesamples was detmmined by substitutintghepeak area responseratiosintotheapplicablelinearegressionequation.Representativeioncbromatogmm oflow and lugh calibratiosntandardsarepresentedinFigures3 and 4,respectively.
Reagent Blanks Reagent blanks were analyzedto assessthe presenceof potentialinterferencesN.o chromatographic
interferencewsere observedin the blanks (preparedfrom a solutionof 50*/omethanol and 50% NANOpureO watercontaining0.05% v/v formicacidand 0.100 mg/L of theinternasltandard)at or above the limitof quantitatioonf 9.15gg a.i.A(LTable2).A representativieonchromatogram of a reagentblank ispresentedin Figure5.
AMENDED
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PROJECT NO.: 454C-107
Ex=le Calculations Samplenumber 454C-107-2A,nominalconcentrationf2000 mgtl inNANOpure(& water.
InitiVaollume:0.0500mL FinalVolume:500 mL DilutioFnactor:10,000 PFOS Peak Area:687581 InternaSltandardPeak Area:603048 Peak Area Ratio:1.140
Calibratiocnurveequation. Slope:0.020 hdacept:0.068 Curveisweighted(:I/x)
Calculatecdoncentratio5n3:9 mg a.i./L Note:manualcalculatiomnasy differ.
RESULTS
Water Solubil@y Triplicastuebsampleswereremoved from theappropriatbeotuesafteorne,two and threedaysofshaldng
inawaterbathmaintainedat30 1.OOCandfollowinognedayofa 20 0.5*Cequilibratipoenriod(Table3)., Analysisofaqueoussubsamplesafterwe dayhad amean analyticraelsulotf459 mg a.i./(LSD = 8.96,CV = 1.95%).Forsubsamplescollecteadftertwo andthreedays,themean concentratiownesre 537 mg a.i./(LSD = 27.6,CV.= 5.14-/oa)nd 501 mg a.i./(LSD = 64.2,CV = 12.8*/or)e,spectivelAy.rcpresentative.ion chromatogramof a Day 2 solubilistaympleispresentedinFigure6.
CONCLUSIONS
The Day 2 andDay 3 mean solubilictoyncentratiownesrewithin15% ofeachotherandwere averagedto
obtaint.heoveralmlean solubilictoyncentrationT.he overalmlean solubilictoyncentrationf PFOS in
NANOpure(D waterwas 519 mg a.i./(LSD = 48.3;CV = 9.31%;N = 6).
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REFERENCES
1. Organisatiofnor Economic Cooperationand Development. 1995. GuidelinfeorTestingof Chemicals1,05:WaterSolubility.
2. ProductPropertieTsestGuidelines1.996.OPPTS 830.7840.WaterSolubilitCyo:lumnElution Method,ShakeFlaskMethod
3. TSCA Titl4e0 oftheFederalCode ofRegulations1.994.Part796,Section1840:WaterSolubility.
4. American SocietyforTestingand Materials.1991. StandardSpecificatifoonrReagentWater. DI 193-91,ASTM SectionH Water andEnvironmentaTlechnology,Vol. 11.01:45-47.
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INSTRUMENT:
TableI
TypicalLC/MS OperationaPlarameters
Hewlett-PackarMdodel 1100High PerformanceLiquidChromatograph witha Perldn-EhnerAPI 10OLC Mass Spectrometeerquippedwitha Perkin-ElmeTrurbo IonSprayionsource.Operatedinselectivieon monitorinmgode (SM.
ANALYTICAL COLUMN:
KeystoneBetasiCllscolumn(100mm x 2 nun,3 @= particlseize)
OVEN TEMPERATURE:
300C
STOP TM:
10 minutes
FLOW RATE:
0.220mL/minute
MOBILE PHASE:
72% Methanol:28% NANOpureOD Water containing0.1% Formic Acid
INJECTION VOLUME:
50.0 tLL
PFOS RETENTION TIME:
Approximately7.07minutes
INTERNAL STANDARD RETENTION TM:
Approximately4.85minutes
PFOS MONITORED MASS:
INTERNAL STANDARD MONITORED MASS:
498.6amu 426.7amu
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PROJECT NO.: 454C-107
Table2
ReagentBlanksAnalyzedConcurrentlDyuring Sample Analysis
Number (454C-107-)
Blankt24
Sample
Type ReagentBlank
MeasuredConcentratioonf PFOS
(gga.i./L)'
< LOQ
Blankt4s
ReagentBlank
< LOQ
Blankt72
ReagentBlank
<LOQ
The limitofquanfitafi(oLnOQ) of9.15gg a.i.AwLas basedupon theproductofthelowestcalibration
standaradnalyzed(9.15gg a.i./La)ndthedilutiofnactoroftherespectivbelanksample(1.00).
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PROJFCTNO.: 454C-107
Table3
SolubilitoyfPFOS inNANOpure(& Water
Sample Number (454C-107-)
Day of Test
1A
1
1B
1
ic
1
ConcentratioonfPFOS
Fortified
Measured
(mg/L)
(mg a.i./L)
2000
464
2000
465
2000
449
Mean SD cv
Mean = 459 SD = 8.96
CV = 1.95%
2A
2
2000
2B
2
2000
2C
2
2000
539
Mean = 537
SD = 27.6
508
CV = 5.14%
563
3A
3
2000
3B
3
2000
562
Mean = 501
SD = 64.2
434
CV 12.8%
3C'
3
2000
507
Totals: Mean (Days2 and3)=
519
SD=
48.3
CV
9.31%
N
6
Note: Resultsand correctionwsere generatedusingMacQuan version8.3.3softwareand nmmua
calculationVsa.lueshavebeenroundedforEp2!@29purposes.
AMENDED
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PROJFCTNO.: 454C-107
MIETHOD OUTLINE FOR THE PROCESSING OF PFOS IN NANOPURE(D WATER
Preparecalibratisotnandardisna solutioonf50% methanoland 50% NANOpure(D watercontaining 0.05% v/vformicacidand0.10 mg/L oftheinternasltandard usingvolumetrifclasksand gas-tighstyringes. I
Preparesolubilistaymplesby weighing0.500grams intothree16-ounceFrenchsquarebottles andadding250 mL ofNanopure(&watertoeach. I
Aftertheappropnateequilibratipoenriodsc,mtn&ge triplica1t0e-mL subsamples atapproximatel3y,000rpm forapproximatel1y5 minutesina centrifugregulatetdo20'C. Dilutesubsamplesto500 mL witha solutioonf 50% methanoland50% NANOpure(& watercontaining
0.05% v/vfonnicacidand 0.10 mg/L oftheinternasltandard.
Transfersamplesand standardtso autosamplevrialsforanalysibsy LC/MS.
Figure1. Analyticamlethod flowchartfortheanalysiosfPFOS inNanopureS water.
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PROJECT NO.: 454C-107
1.40 1.201.000.80 Cd CD 0.60 0.40 0.20 0.00
0.0
0 0 0
20.0
40.0
60.0
80.0
Concentratim(ug&iAL)
Figure2.
A typicaclalibraticounrveforPFOS. Slope 0.020;Intercept0.068;R@ 0.990, Weighted(1/x).
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PROJECT NO.: 454C-107
1 90. so 70@ 60" 50. 40, 30 2& I or 0'
intensity8:955 cps 169
148
-70
1 2125
si . , ii . 91-,
1.30
2.65
3.81
121
5.06
151
6.32
-92
1 l'
7.57
214-
2il 8.83
SM Tkne
Figure3. A represmtativieonchromatog= ofa low-leve(l9.15jiga.i.ALs)tandarcl ANFENDED
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PROJECTNO.: 454C-107
100.
Intensity3:2211 cps 169
go. so.
70"
60. 50
40-
30-
147
20
lo. a
0.1 .
26 ...
31 1.30
AS; 7A 03
. .....
61
ol
2.65
3.81
124
... 121 5.06
.. . 151 6.32
. iii 7.57
2.11 8.83
F'igur4e. A representativoenchromawg= ofa high-low(l45.7Itga.L/L)standarcl AMENDED
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PROJECT.NO.: 454C-107
I0()@ 13
9(@ 8070, ao-
71
-86; @ 85
112 189
intensft5r8.q)s
227-
132 ISO
II 2101
40302&
ot
31
61
91
iii
lal 211
1.30
2.55
3.81
6.06
6.32
7.67
8.83
sm Ttm.
Figure5. A representatiivoen
ofa reagentblanksample(454C-107-Blank-ta)T.he
arrowindicatetsheretentiotnirwof PFOS.
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PROJECT NO.: 454C-107
1 oc@ 90. so. 7& 6050. 40302&
Z 24
47 RZ
31 1.30-
61 2.65
q'i 3.81
intensity4:2163 cps 168
145,
124
iii 6.06
161 6.32
201
181 7.57
211 8.83
Tiine
Figure 6.
A representativieoncbromatogram ofa Day 2 solubilitsyample,454C-107-2A (nominal concentratioonf 2000 mg/L).
I
WILDLIFE
INTERNATIONAL
LTD.
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APPENDIX I Protocol
PROJECTNO.: 454C-107
WILDLIFE INTERNATIONAL LTD.
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PROJECTNO.: 454C-107
I
PROTOCOL
DETERMINATION OF THB WATER SOIJJBUM BY THE SHAKE FLASK WMOD
OF PFOS
OEM Gdddint fordieTestiPgofCb=icole 105 Wdw SolubWdy
3ML@bR*P%tNo6U=
Submittedto
3MC=pmdim
EwAramncntidLaboratuy
P.O.B= 33331
SLPoLMmamU
$5133
WILDLIFE
INTF-RMUONAL
Eadw Mwylnd 21601 (410)SZ"M
LTD.
Dwm2ber2,1998 PP,OTOCOL NO.: 454/12629LWO.794WSLn3454
3M IAB REQUEST NO. U2723
WILDLIFE INTERNATIONAL LTD.
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PROJECTNO.: 454C-107
WILDUFE INTERNA-NONAL LTD.
-2-
vEL MhMWi LON OF THE WATEIL SOLLTBU= BY THE SHAKE PIASK MMOD
OF PFOS
SPONSM
SPONSOR.-S REM TESTING FArRrrY, S71lnYTITR
3M Capmldm BukameaWL&bmiitmy P.O.Box 33331 SL PauL hfmnesoft 55133
ML SusanA. Bea&
VrddUc bftmadmd IAd. 8599 C4mmem Dtive ELqton,Mwjlmd 21601
T-ARORATORYMALWA
VrdladB.Nm^ Ph.D. Monqw ofChemistry
FOR LABORATOITY USE ONLY
Pw poseddatw.
BRM&wow stutD4ft
-3ji6iqq
Ptojodww
Tea Substme No.:
Exp=nmW TamhmfimDdm
RacciptDdm
gloolig oil
PROTONOLAPPP-OVAL
&TEZYbUtWMR
DATE
OSLRATORY ORS
MAN4VEhONT
rA
IMTE
la 171qg
DAIM
PROTOCOL NO.-.454/12029LWO.7940MUB454
3M LAB REQUEST NO. U2723
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PROJFCTNO.: 454C-107
WILDLIFE INTERNATIONAL LrD.
-3-
TNMODUCNON
WUDM hd=aboad lAd.wM evaunowly detumm die wata sobibditoyf PFOS
Couffimmoamsdfamemdmtmsmsdt@
llza*wMbecovAx;tWattheVrddbhlnt=aumd
Ltd.malybw &wmtty &=Uy m Eaton.Maykm& The sb* willbe puftmad Mowmg &c
guidm= presenteidn*a OEM GuiddineforTestiigofChamicileIOS: WaterSohtbibt(y1)for
&balmBa* mWwdoloV. INs a* isbknded tomod datanWhommU uda dm rweline and
mder EPA Prodad hqmlm TestGmddonk OWM 830 7840 WaterS"IAW CDh=Muam
Method, MkLb FZ=kMe" (2)ad TSC& Tfflo40 ofdo FWCW Code ofRcpbdoos6 Put 796,
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wMboveffWatWgdffobkmatioo-IIA Pim dda forag=&Mfirmcd atwddffo bftnadonal
Ltd.mdaaWofftftdrcportwMbofiWbyproedumdwinacWv
bcatedondwWUdH&
latmudwd Lt&.siWoratonshanatinkcs6ontobe specifiiend&a ftd rcparl
PUP@PM
Ilopmposeofddssk* istodemmint ihcwdm whbft ofPPOS at20 * 0.5*C by do shdw Bask mcft& Ddm=xbm of atu muft by diedWm Bak methodn Whcable to &ad
abbn= with atixwl"ities eqmd toOr
0.01gma" (gL).Tinmwwdolou MAY
notbe applkabletourfim outhvorYoh*& ted u&stmm
RYPFRTMMWALDMGN Mc da whmity ofPFOS wM bed*miwd ata ampwbm
of20* O.S*C A wehmins@y
of adM= offi=cnio* bup amouna of wda toa
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sobm=timis affecteadrdw sok&'W iscsdmdodto balm d=0.01 VIL Forsubmancwui&an
esdmdedwdusokibMWIp,--- ik-O.OIVUadefiaMatolvAbopufmmo&
The defidtivteest
ofoquftcadm ofam can amout ofug whe@vdffi wda atan dmted kmpamom
30-C,Mowedbyeqml&sdmat2O'C
MATMTAlr-q ANn MRMODS
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PRDR=L NO.: 454/12029030.7840/SUB454
3M IAB RBQUEST NO. U2723
WILDLIFE INTERNATIONAL LTD.
-26-
PROJECT NO.: 454C-107
WILDUFF- INTERNA-noNAL LTD.
-4-
ChemicalNam
Fomada odor MOIGcwar Wei& C4esuw inWmw
Ddo = ,,oAanfdysis
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VWNRUM St=VC=dW=
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OF TEST SUBSTANCE BY SPOMOR (Anmdm 1),
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andUW=g ofdketea oubsm= Mm Spmw spu toacceptaW =and tm substm= =&or wa
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atdw end af*c audy.
Emma Wdwdnd me* ASTM Type B amdar& VMM
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sobun wM beACS mgmt pu& orbcuw. and vM be dct=dwd tobe fiveofcontmbmmb dut
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t=pcmmm AppmWmoiy 10mg ofdo tea radam wM be pbcadina pladc bgm bureasiq vahmm ofwaur wM be addedstqnm asdmn bdow.
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0.1 0.5
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ry -'toalweautdow and -do addedft a ftd vd@amof 100=IL Foamh* cappingand
kilem a wda sotubMW -cOlVL PtdkWnKy testinogfte mob*ft atbwer
(.0.1
sOL)may be* 9 1by unsfirdogdw omkm ofdm 100.mLvmd toa I-Lvmd oricia@
dwWvMakawwwadofWKm6dumre461mg
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PITOTOCOL NO.: 45Ml2MM30.7840/SUB4$4
3M IAB REQUEST NO. U2723
WILDLIFE INTERNATIONAL LTo.
-27-
WILDLIFF- INTF-RNA-NONAL LTD.
PROJECT NO.: 454C-107
Definiem TestPfoced=
Mo quan&y aftetm
necusay tosatanftdiedeshvdvdmc of wdu isesfimuedfiam
ft prdbnionytosl Ma volumeof wda usedwilldependan themab" umthodand wgubft
rmp. Cl.temdun (orequalto)fivetimestheqmmtky ofted
mq*W toacbl-m vvmu
oobbTdywMboemsfuredtom-fith--offtwica
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reAWdod at200C. 7be omombudw of
*etmtsdxtamima-w-oftwoafiqu*ofdwaqunnphmwfflbodftmimi
luraniubg
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7be mdyded method forquudtdim ofthetW nbstnw wM be basedan pmcodurespwvidad
by die Spoubw. 7be mdW mM be wpai&d to gib patocolinAppOndl3LH and nftukwd indo
mpat IU methodnaybovaifieda put efdopmHmimyusKs) =dmaybomomadto mommm
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wmbo wpovedbydo Spmw and
StedyDhoolar,waddocvn=W ifdfw raw datl
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15% of eachotbw
The SponsorwM kkadfy oW speciahl-mi- or ufctypmcvaians to be usedwithdieabove rda=codtwrdwtmm Allnormalpnodions w& Pippy-tou-ntusuddompwmbotdm
PROTOCOLNO.: 45VI2029LWO.7940/SUB454
3M LAB REQLMST NO. U2723
WILDLIFE INTERNATIONAL LTD.
-28-
PROJECT NO.: 454C- 107
WILDUFE: INTERNANONAL LrD.
-6-
SnopkandTestSWUU=Rctmfion Up= completionofbsbn& porboasofdittea rabsta= used aspartofdus sWy willbe
&posed ofinmmimm withftkml stateand kmd mpktiom Any =used portionofthetest m*ftm wM be rcuumodtodo Sponsor.
RFZORM TO BE MAWRAINM
P=ar& tobe maktoinedfordatagma*ed
'Wikdtcr
1. AaWofhcOpodprotoc&
by WildHo latmadow
L4& v&lWcWdc6butnabe
Z Madfics6on nd chwadmizadm aftm testsubstoo, ifFavided by&a Sponsor.
3. Dan ofinitiatiaonndcompkdm ofdn stody.
4 Data ofawmiedd sM andtmswabw
S. Stmp om&dm af*ctesotbsto=
6. Tea obsta= uselog.
7. ICbbccntrafioticakW-goanngd --,,IlofsobstimpvpwadoiL
"a. Insbumcat@opc@@"&dma@ddwomdogmsif
Vplicabit
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11. A copy ofditfmd rcpoit
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by dw Spmw.
6. Douiptim o(dicw nw&W orrefamm todke method und akmg wi& =W modificationsnuml-.
PROTOCOLNO.: 454/1202981930.7840/SUB454
3M IAB REQUEST NO. U2723
WILDLIFE INTERNATIONAL LTD.
-29-
PROJECT NO.: 454C-107
WILDLIFF- INTE:RNAIIONAL LTD.
-7-
7. The indivb&W cmcmtmdoas of eachsample. S. llm means and standarddeviaboasof solubildyddermwabms atca& mkrval forewh
t=Fre= ustal 9. Do=iptim ofanypmblms cqwkncod md how dwy wm resolve& 10. A ddwxft preparedby dkeQualiLyAnumwe Unitlbft dw datesdut sb* bLVectiomand
audits"a ran& md fi"&;n reportetdo&e SV* Di=W andkinnagemml
CffANGMOFPROTOCOT.
Plannedch-go to diepwtocd wM be itdLkefam ofvrbm amendments dped by dw Study
Dnctor anddieSpmw.
vE be coaddcreda partof&c protocoland wM be aftachad
totefinalpawool Azwodicr&aqp wmbo in&e form ofwithm &viatim god withtheraw datil
AILchangestDdo protocolwM be bwkawd indiefiW tVarL
GOON T@ORATORY PITACI=
Thisgt*vm be cm&mw wmfmgto *0 GoodL&bm*my hudm d=rpocdin OECD
(ISBN 92-84-12367-9)=d EPA (40 CFR Put 792). Each t-to-odoyn@&xdad by WMM hdmwdamal
LA& ismutia*
- IbyftW&r&bWmWimdlA&
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compsomirm
Ldwaitmy Pracfimwillbel p- p4formapordm afdw sb*cw&cw by
Wad[& MW=bmd LAL 7be Spomw wM be mWwsUo foromphnw wilhGood L&m*my
P -4forpmoo&m podarmad by cebw bboratmial Paw dds foran wok perfoamw atWOdUt
kdmmimd U& md a awafdo fiwnpixt wM beEW bypr*ct nmbcr inadiva beatedan&a
VIM% htcmadmd LA& siteoratan shemativelocatimtobe spedw in&a findrcpmt
PILOTOCOLNO., 4-WI202M30.7940/SUB454
3M LAB P.EQUEST NO. U2723
WILDLIFE INTERNATIONAL LTD.
-30-
PROJECTNO.: 454C-107
WILDUFF- INTF-RNA-NONAL LTD.
-8-
I OrganisationforEconomic Cooperafionand DeMopment. 1995.GuiddincforTestmg of
Chemicab6 105: WaterSoh&btftty.
2 Product PropertiesTest GuidermaL IM OPM Ehaton Medwd, Shake R=kMeOwd
930.7840.Water SolubiZity.Cohmm
3 TSCA ride 40 of the Fedwd Code of Regulafions.1994.Part796,Swdm 1&4o.water Solubilify-
4 AofticanSoddy forteaft and MdwWL 1991.SWulad Sp=Mofm forRaqpdWdw.
D1193-91,-ASTUSecdmUWatw=dEwkmncnWTecbnoloU,VoL
11.01:45-47.
t
PROTOCOLNO.: 454112029LI930.7940/SUB454
3M LAB REQUEST NO. U2723
WILDLIFF- INTF-RNATIONAL LTD.
-31-
PROJECT NO.: 454C-107
WILDLIFE
INTF-RNATIONAL
LTD.
-9-
APPENDIXI IDENTIFIC.KTION OF TEST SUBSTANCE BY SPONSOP.
To be Cmvbftd by Spmw
L TestSubstanceMenfiLy(nmm tobe used indiereport):PFQS (Pcr&xwoomm SulfotiiAecid PoUL-,simSalt
RefcreweStwmiffd(ifapplicableA)n:akuW Stmxbx& N/A
T"h-WStandw&- 1-1 RHER Pefauamoctam SulfacieAcid
Tat SubstanceSampk Code or BatchN=iber Lot 217 TestSubstancPeudLy(*AAcd" bgufiat)- 98.9
Bq*vfim Datm 2008
IL TestSubstow Chwwtedzdka
Usrnd
dwadmistka
bOsMtVimm@,=ttmuludbom
*W tDib usein9& studyinammbnw wi& W SW&ff&?
yes
No X
M TestSaWtow Stmp Camfibm
Pie= ubdiufftt rowmnuum swW coaften-sttvrddl&btcmdmd M
Ambientroom toogr=
Hasthedabft ofac scstsubstancuenderdww xmn4p ewditm been detmvmima in womdmw vi& GLP Stmadards?
Yes-No
X
Odiewrhom dm)ft mftnabm
IV. TestCantcobadmr.
A4* textconecatmdm to100% &L X bded Wm do pu* (%)Om aboft
V. T06cityinformofm-
DoaotadiWW9
101006
-a.L Tedowmatm:W0A*LMft,@
MamanuaL
RatLDSO 251 mgft
Mom LDSO N/A
AqWk
kmtft TowftooLcso)
irghTad&Y (LC")
macm 27MML
RainbowTmut IImgl
lbgbwamumSOM&L
FMhudKmnow, 3$moJL
Odaw Todcitybfomufm Cmdaft
ofcbmdc ad m*dkodo to*.
PROTOCOLNO.: 45VI20298/930.7840/SUB454
3M LAB REQUEST NO. U2723
WILDLIFE INTERNATIONAL
-32-
PROJFCT NO.: 454C-107
WILDLIFE INTERNATIONAL
LTt). -10APPENDDCII
ANALYTICAL hffMOD
AnalytiedMethod fordw Dammkofica ofPPOS inWata
PFOS may be deumnizodinwdw usinghighpaftmm hquw &uuorvby withmassgommct& detamon 0,CMS) Sampleprcpmzbm fordw fa%ficahm -awpl vallmvolvediivtm(usintgheSam mwm waw) udo thecowenwm rangeofmshummw rwp=L Matax bbtaimvaU be dffo* zyectc& Mit fonowwg pmsmu qvromaw duamdopvhic coaftm fi)LrCLE qwmuwm ofPFOS. Chwmdogmphic condiuonmsay be changedtoprovidceondeam mm optwzmd Ew womb= and/odruomawgrapbw sepusum Of PFOS.
hwbvmcw Hcwlctt-PukarMdodel 1090A BPLC
C4h=n.-Krist= Bma C-19,100mm x 2.0uma,3;zm MobBe Phasc McdonoL Ammomm AcetacBWExed Wdw FlowRat= 2501AAnimAt CohmmTaMwatum 40C iqwdmvobnw. iooUL Im Soum: Turbo ImSproy Mm Spadmmater Paddn.ElmwAPI)OO Rm Timm 6.0mimfts
65:35(v-v)u,*cmtw,
PROTOCOLNO.: 434/12029UB30.7840/SUB454
3M LAB REQUEST NO. U2723
WILDLIFE INTERNATIONAL LTD.
-33-
PROJECT NO.: 454C-107
APPENDIX 11 CertificaotfeAnalysis
FC-959Iot 217
Mmh 9,2000
RW=d bL Poyft
Thm =14p]aym mdrmd =tog LC&S, IH-Me..
nd ek=mw &twym twbwg=
offtmtmWkowftjuvlotocmtgnftgbuowb*vrd& tC=P*dd=
9&12LT, CCFISC)I"R
CFtisc)llc
c4pmq-2lr
ctrisc),Ic
0.04%
am % 138 ve
130%
3.71%
9OA9% 0.49%
ciffnsollr gk2?L.W-
0.04% 0.05%
0.35%
lim mub
AddkbnaUy, ou i
dWnV@*uoftbow=pbwuddmdwdusing
thefoboiftnx& pecom cook.,oskl=
"F4*Muobdqmmdtmdto
705%
coma MMIUWUNRWL
------o-d6
Ml%
ub= wilIsumbb S.andxo 0.v p01
2-a- -Mil swr
OWNOO]D
n@er -IBM
ciawcp(cps)-Bwlu
(AlAnbconob- Kkoft6)
(MWI"CFIS)wL-KT
02%
t
Cipi<CFSYA4(l
g-1
vkmm+y,
02%
bnmidv44L.' ZOO)
AMENDED
WILDLIFE INTERNATIONAL LTD.
-34-
PROJECTNO.: 454C-107
APPENDIX M
PersonnelInvolvedintheStudy
The followmgkeyWildlifIenternationLatld.persomelwereinvolvedintheconductormanagement of thisstudy:
1 . WillardB.Nixon,Ph.D.,Manager,AnalyticaClhemistry 2. Raymond VanHoven, Pli.D.S,cientist 3. Norman J.GlassbrookM,.S.,SeniorChemist 4. KeriLeach,B.S.,Technician
WILDLIFE INTERNATIONAL LTD.
-35-
PROJECTNO.: 454C-107
APPENDIX IV ReportAmendment
I. OriginaRleport: Pages 1-4and 6
Amendment:
The p@geswere changedtoincludetheamended reportdate,revisedpagenumbers,and new signatureasnddatesdue totheadditioonfthereportamendment asAppendixVI.
Reason:
To reflecttheissuinogf an amended report.
2. OriginaRleport: Page 9
Amendment:
Informatiopnrovidedby theSponsorreflectitnhgereanalysiosf thetestsubstance, includintghereanalysidsateand thepurityw,as addedtotheTestSubstancesection.
Reason:
To reflectthecurrentestsubstanceinformatiopnrovidedby theSponsor.
3. OriginaRleport: Entirereport
Amendment:
Alltestsubstanceconcentrationwsere changedtoreflecttheptiriovfthetestsubstance asdeterminedby theSponsorina reanalysiosfthetestsubstance(FC-95,Lot217). Testconcentratioonnsginau were basedon the portedpurityof 98.9%. The cerfificaotfeanalysi(sia@tVeadrg 9.2000 indicatedp'uarityof90.490/oT.b=fore, all datasuchastestsubstanceconcentrationisn,cludinngoniindconcentratiomnesa,sured concentrationasn,d statisticvaluesw,ere recalculateadnd reporteadsmg &iJL based
onthe90.49% purity.
Reason:
To reporttheresultosf thetestbasedon thetestsubstancepurityof 90.49% atthe requestof theSponsor.
NT SIGNATURES:
AMT-
RayKwhd,'V.-VanH6Vd4Ph.D. StudyDirEctor
-oa DATE
WillardB.Nixon,Ph.D.f Manager,AnalyticaClhenustry
REVIEWED BY:
f4. go.,Vmo
v QualityAssurance
DATE
DATE AMENDED
Ul
PROTOCOL DETERMINATION OF THE WATER SOLUBILITY OF PFOS
BY THE SHAKE FLASK METHOD
OECD GuidelinefortheTestingof Chemicals,105 Water Solubility
3M Lab RequestNo. U2723
Submittedto 3M Corporation EnvironmentalLaboratory P.O.Box 33331 St Paul,Minnesota 55133
WILDLIFE
INTERNATIONAL
8598 Commerce Drive Easton,Maryland 21601
(410)822-8600
LTD.
December 2, 1998
PROTOCOL NO.: 454/120298/830.7840/SUB454
3M LAB REQUEST NO. U2723
WILDLIFE INTERNATIONAL LTD.
-2-
DETERMINATION OF THE WATER SOLUBILITY OF PFOS BY THE SHAKE FLASK METHOD
SPONSO
3M Corporation EnvironmentalLaboratory P.O. Box 33331 St.Paul,Minnesota 55133
SPONSOR!S REPRESENTATIVE:
Ms. Susan A. Beach
TESTING FACILITY:
WildlffeInternationaLltd. 8598 Commerce Drive Easton,Maryland 21601
STUDY DIRECTO
R=rdajacobz, get2ioerheniistS-e-i.,ri4c-@
LABORATORY MANAGEMENT:
WillardB. Nixon, Ph.D. Manager of Chemistry
FOR LABORATORY
USE ONLY
Proposed Dates:
Experimental StartDate:
ProjectNo..
lio/qq
45q-6
Test Substance'No.:
r,ri5
Experimental TerminationDate: IntfDate: ReceiptDate:
31 iol 0 ]to197 n f.)q gr,
PROTOCOL APPROVAL
Sm@fVCTU4TL(CTOR
DATE
SpZLOSOORRTAOTROYR'SYPF MAN*GEMENT
MAN
SP SOR!S REPPRRESENTATIVE
DATE DATE
PROTOCOL NO.: 454/120298/830.7840/SUB454
3M LAB REQUEST NO. U2723
WILDLIFE INTERNATIONAL
LTD.
-3-
INTRODUCRION WildlifeInternationLatld.willexperimentalldyeterminethe water solubilitoyf PFOS (perfluorooctasnuelfoniaccidp,otassimnsalt)T.he studywillbe conductedattheWildlifeInternational Ltd. analyticaclhemistryfacilitiynE aston,Maryland. The studywillbe performedfollowingthe guidancepresentedintheOECD GuidelineforTestingof Chemicals,105: Water Solubilit(y1)for shake flaskmethodology. This studyisintendedtomeet datarequirementsunderthisguidelineand tmderEPA ProductPropertieTsestGuidelinesO,PPTS 830.7840,Water SolubilityC.olumn Elution Method,- Shake FlaskMethod (2)and TSCA Title40 oftheFederalCode of RegulationsP,art796, Section1840: Water Solubilit(y3).The method ofanalysifsorquantitatipornovidedby theSponsor willbe verifieadtWildlifIenternationLatld.Raw dataforallwork performedatWildlifeInternational Ltd. and a copy of thefmal reportwillbe filedby projecntut@berinarchiveslocatedon theWildlife InternationLatld.siteoratan alternatilvoecatiotnobe specifieidnthefinalreport.
PURPOSE The purposeofthissb* istodeterminethewatersolubilitoyfPFOS at20 0.5'C by theshake flaskmethod. Determinatioonf watersolubilibty theshakeflaskmethod isapplicablteo test substancewsith watersolubilitieeqsualtoorexceeding0.01grmn/Liter(g/L).The methodologymay notbe applicablteo surfaceactiveorvolatilteestsubstances.
EXPERIMENTAL DESIGN The water solubilitoyfPFOS willbe determinedata tmnperatureof20 --0h.5C. A preliminary testwillbe performedto estimattehewatersolubiliattyroom temperature.The preliminartyestconsists of additionsof increasingllyargeamounts of waterto a known weightof testsubstanceuntil solubflizatimseffecteodrthesolubiliitsye,stimatetdobe lessthan0.01 g/L. For substanceswithan estimatewdatersolubilitgyreatetrhan0.01 g/L,adefinitivtestwillbe performeclThe definitivteest consistsof equilibrationf an excessamount oftestsubstancewithwateratan elevatedtemperature, 30'C, followedby equilibratiaotn20'C.
MATERLKLS AND METHODS TestSubstance
The testsubstancewillbe thehighestpurityavailableT.he testsubstanceshouldbe atleast99% pure by weight. The Sponsorwillbe askedtoprovidethefollowinignformatiocnoncerningthetest substance,ifavailable:
PROTOCOL NO.: 454/120298/830.7840/SUB454
3M LAB REQUEST NO. U2723
WILDLIFE INTERNATIONAL LTD.
-410
Chemical Name
Lot or Batch Number
EmpiricalFormula and/orMolecularWeight
Chemical Purity:
In
DissociatioCnonstantinWater
Vapor Pressure
ExpirationDate
StorageConditions
Method ofAnalysis
The attachedformIDENTIRCATION OF TEST SUBSTANCE BY SPONSOR (Appendix 1), alongwitha MaterialSafetyData Sheet,willbe used toprovideinformationnecessaryforthe handling and testingof the testsubstance.The Sponsor agreesto acceptany unused testsubstanceand/ortest substancecontainersatthe end of the study.
Reag=s Water thatmeets ASTM Type ]Istandards(ASTM D 1193-91)willbe used (4).Othersolvents
may be needed fortheanalyticamlethod or preparationof stocksolutionsand analyticasltandards.All solventswillbe ACS reagentgradeor better,and willbe determinedto be freeofcontaminantsthat interferweith thequantitatioonf thetestsubstance.
PreliminM TestProcedure A prelin-dnartyestwillbe conductedto estimatethe soliibiliotfythetestsubstanceat room
temperature.Approximately 10 mg ofthetestsubstancewillbe placedina plastitcube. Increasing volumes of waterwillbe added stepwiseas shown below:
Totalvolume ofwateradded (mL) Approximatesolubili(tgy/L)
0.1 0.5
1
>100
20
10
2
10
5
1
Aftereach additionofwatertoyieldthetotalvolume indicatedt,hecontainerwillbe capped and shaken vigorouslyforapproximately10 minutes and visuallyexamined forundissolvedtestsubstance.Ifthe testsubstanceisnotdissolvedafteradditionof 10 mL ofwater,thecontentsofthecontainerwillbe tramfmTW toa largecrontainerand water added fora finalvolwne of 100 mL. Followingcapping and shaldng thecontainewrillbe visuallcyheckedforundissolvedmaterial,i.e.u,ndissolvedmaterialwould indicataewatersolubilit<y0.I g/L. Preliminarytestinogfthesolubilitaytlowerconcentration(s<O.I g/L)may be performedby transferrintghecontentsof the 100-mL vesselto a I-L vesselor reinitiating thetestwith a lesseramount of testsubstance,e.g.,I mg. Amount s and volumes noted above areonly guidesforcorductofthepreliminarystudy,largeramounts of testsubstancewith proportionalllyarger volumes of water may be used.
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3M LAB REQUEST NO. U2723
WILDLIFE INTERNATIONAL L-rD.
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DefinitivTeest Procedure The quantityof testsubstancenecessaryto saturatethedesiredvolume of water isestimatedfrom
the preliminarytest.The volume of water used willdepend on theanalyticamlethod and solubility range. Greaterthan (orequalto)fivetimesthequantityof testsubstancerequiredto achievewater solubiliwtiyllbe transferretdoa minirninnofthreetestvessels(plastiacnd/orothersuitablecontainer material)T.he desiredvolume of water willbe added to each vesseland the vesselssealed.The closed vesselswillbe agitate(dshakenorsfirreda)t30 =1:I.O'C in a waterbath. After I day,one of thevessels willbe removed and re-equilibratfeodr24 hours at20:L 0.5*C with occasionalshaldng or sfirringT.he contentsof thevesselwillthenbe centriffigienda centrifugreegulatedat20*C. The concentratioonf thetestsubstanceina minimum of two ahquotsof theaqueousphase willbe determined. The rernaimmg two vesselsare treatedsimilarlfyollowinginitiaelquilibratiopneriodsof 2 and 3 days at30 -1.OOC. The testwill-beterminatediftheconcentrationosf thetestsubstanceinthe aqueous phase of two consecutivevesselsagreewithin 15%. Ifgreatervariationisobserved and/orattherequestof the Sponsor,the testmay be reinitiatewdith longerequilibratiopneriods.
Method The analyticamlethod forquantitatioonf thetestsubstancewillbe based on proceduresprovided
by the Sponsor. The method willbe appended to thisprotocolinAppendix R and referencedinthe reporl The method may be verifieadspartofthepreliminarytest(sa)nd may be modifiedtomeasure theobservedtestsubstanceconcentrationsM.ajor modificationswillbe approved by theSponsor and Study Director,and documented inthe raw data.
Calculations The concentrationof thetestsubstanceineach vesselwillbe expresseding/L,mg/L or ggtlin
'wateras appropriate.The averagesolubilitfyortestvesselswith solubilitiweisthin 15% of each other willbe averaged and reportecl
Sa=16 Handling and S The Sponsor willidentifyany specialhandlingorsafetyprecautionstobe used with the above
referencedtestsubstance.All normal precautionswith respecttohandlingand storagewillbe taken.
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3M LAB REQUEST NO. U2723
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ol WILDLIFEINTERNATIONALLTD. -6-
El
S=Dle and Test SubstanceRetenti Upon completion of testingp,ortionsof thetestsubstanceused as partof thisstudywillbe
disposedof in accordancewith federals,tateand localregulationsA.ny unused portionof the test substancewillbe returnedto the Sponsor.
RECORDS TO BE MAINTAINED Records tobe niamtainedfordatageneratedby WildlifeInternationaLltd.willinclude,but not be
Ilimitedto: . A copy ofthesignedprotocol. 2. Identificatioand characterizatiofn thetestsubstance,ifprovidedby the Sponsor. 3. Dates ofinitiatioannd completionof thestudy. 4. Dates of experimentalstartand termination. 5. Storageconditionsof thetestsubstance. 6. Test substanceuse log. 7. Concentrationcalculationasnd recordsof solutionpreparation. 8. Instnment operatingconditionsand chromatograms,ifapplicable. 9. Statisticcaallculations. 10. Testconditions. 11. A copy of thefinalreport.
FINAL REPORT A finalreportoftheresultosfthestudywillbe preparedby Wildlifehdm-nationalLtd. The reportwillinclude,butnotbe limitedtothefollowingw,hen applicable: 1. Name and addressof the facilitpyerformingthe study, 2. Dates upon which the studywas initiateadnd completed. 3. A statementof compliance signedby the Study Directoraddressingany exceptionsto Good Laboratory PracticeStandards. 4. Purpose and procedure,as statedintheapprovedprotocolm,cludmg allamendments and deviations to the protocol. 5. The testsubstanceidentificatioin,cludingname or code number, purity,empiricalformula, molecularformula,lotor batch number, method of analysisa,nd any otherinformationprovided
by the Sponsor. 6- Descriptionofthetestmethod orreferenceto themethod used alongwith any modificationsmade.
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WILDLIFE INTERNATIONAL
LTD.
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7. The individuacloncentrationosf eachsample. 8. The means and standarddeviationosf solubilitdyeterminationast each intervaflor each
temperaturetested. 9. Descriptionof any problemsexperiencedand how theywere resolved. 10. A statementpreparedby theQualityAssuranceUnit listintghedatesthatstudyinspectionasnd
auditswere made and findingsreportedtotheStudyDirectorand Management.
CHANGING OF PROTOCOL Planned changestotheprotocolwillbe inthefor-xonf writtenamendments signedby theStudy DirectoarndtheSponsor.Amendments willbe consideredaspartoftheprotocoland willbe attached tothefinaplrotocolAny odur changeswillbe intheform ofwrittendeviationfsiledwiththeraw data. Allchangestotheprotocolwillbe indicated'tihnefinalreport.
GOOD LABORATORY PRACRICES This studywillbe conductedaccordingtotheGood LaboratoryPracticedsescribedinOECD (ISBN 92-84-12367-9)andEPA (40CFR Part792). Each studyconductedby WildlifIenternational LtcLisroutnelycommed by theWildlifIentmmhonalLtd.QualityAssuranceUnit forcompliancewith Good LaboratoryPracticesS,tandardOperatingProceduresand thespecifiepdrotocol.A statementof comphar= withGood LaboratoryPracticewsillbe preparedforallportionsofthestudyconductedby WildlifeInternationLatld. The Sponsorwillbe responsiblfeorcompliancewithGood Laboratory Practicefsorproceduresperformedby otherlaboratoriesR.aw dataforallwork performedatWildlife InternationLatlcLand acopy ofthefinarleportwillbe filedby proj=t number inarcluveslocatedon the WildlifeInternationLatld.siteor atan alternatilvoecatiotnobe specifieidnthefinalreport
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REFERENCES
I Organisation for Economic Cooperation and Development. 1995.GuidelineforTestingof Cheniicals1,05: Water Solubility.
2 Product PropertiesTest Guidelines.1996. OPPTS 830.7840. Water Solubility.C-olumn ElutionMethod, Shake FlaskMethod
3 TSCA Title40 of the Federal Code of Regulations.1994.Part796, Section1840. Water Solubility.
4 American SocietyforTestingand Materials.1991. StandardSpecificatiofnorReagent Water. Dl 193-91,ASTM Section]IWater and EnviromnentalTechnology,Vol. 11.01:4547.
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3M LAB REQUEST NO. U2723
WILDLIFE INTERNATIONAL
LTD.
APPENDIX I IDENTIFICATION OF TEST SUBSTANCE BY SPONSOR
To be Completedby Sponsor
I. TestSubstancIedentit(ynametobe usedinthereport)P:FOS (PerfluorooctaSnuelfonicAcid PotassiumSalt
ReferenceStandard(ifapplicableA)n:alyticaSltandard: N/A
InternaSltandard: 1.1.2.2Ha.H.HPerfluorooctaSnuelfoniAccid
TestSubstanceSample Code or BatchNumber: Lot217
TestSubstancePurity(1/AoctiveIngredient): 98.9
ExpiratioDnate: 2008
ii. TestSubstanceCharacterization Have theidentitys, gemty andcompositionorothercharacteristics which appropriatcl,rMde-geet4estsubstanceand referencsetandardbeen detenniiiiOdriort6itsuseinthisstudyinaccordancewithGLP Standards? Yes
No X
III. TestSubstanceStorageConditions
Pleaseindicatteherecommended storageconditionastWildlifIenternationLatld.
Ambientroom =erature
Has thestabilitoyf thetestsubstanceunderthesestorageconditions been determinedinaccordancewithGLP Standards?
Yes
No X
Otherpertinensttabiliitnyformation:
IV. TestConcentrations:
Adjusttestconcentratiotno I00*/oa.i. x basedupon thepurity(*/og)ivenabove.
Do notad' sttestconcentratiotno 100% a.i.Testge materiaAlS IS.
V. ToxicityInformation:
Mammalian: Aquatic:
RatLD50 251mg&g - MouseLD50 N/A
luvertebraTtoexicit(yEC/LC50)
FishToxicit(yLC50)
DgpbWa magga: 27 mpJL
Rainbow Trout:IImgJL
DgpWa M=: 50 mg/L
FatheadMinnow: 38 mg!L
OtherToxicityInformatio(nincludinfgindingosfchronicand subchronitcests):
PleaseseeMSDS
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3M LAB REQUEST NO. U2723
WILDLIFE INTERNATIONAL LTD.
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APPENDIX II
ANALYTICAL METHOD
AnalyticalMethod fortheDeterminationof PFOS inWater
PFOS may be determinedinwater usinghigh performance liquidchromatography with mass spectrometric detection(LCMS). Sample preparatiofnorthe fortificatisoanmpleswillinvolvedilution(usingthe same matrix water) intothe concentratiornange of instnunentarlesponse.Matrix blankswillbe directliy;ijectedT.he followingpresentsapproximatechromatographicconditionsforLCMS quantitatioonf PFOS. Chromatographic conditionsmay be changed toprovideconditionsmore opfimizedforretentionand/orchromatographicseparation of PFOS.
Instrument:Hewlett-PackardModel 1090A BPLC. Colurrm: Keystone BetacilC-18, 100 mm x 2.0mm, 3mm Mobile Phase: Methanol:Ammonium AcetateBufferedWater 65:35 (v:v)i,socratic. Flow Rate: 250 AUniinute Coluirm Temperature:40C InjectioVnolume: 100 AL Ion Source: Turbo IonSpray Mass Spectrometer:Perldn-ElmerAPI 100 Run Time: 6.0minutes
PROTOCOL NO.: 454/120298/830.7840/SUB454
3M LAB REQUEST NO. U2723