Document NeDayEk3KgEpeb9eOEpymj8a8
July30,1998
Faxto:Dr.JackMoore Dr.Joe Rodricks
Fax # 202-289-8530 Fax # 703.516.2393
Frorm John L.Butenhoff
Fax # 651.733-1773 Tel # 651.733-1962
I have attachedourrationalfeordose settingon thesixmonth chronicoralstudyof PFOS inCynomolgus Monkeys.
You willalsofindattachedthereportfmm theenvironmentallaboratoryof serum levels forthefourweek rangefmder ammals.
Serum levelsarereportedfor days-7,2,3, 14 and 29. We have done an analysiosf the slopesof the cumulativedose vs.serum concentrationfortheserange finderanimalsatthe low dose of0.02 mg perkg perday. Thisisalsoattached.
By theend of theday,we willhave an analysisof the slopesfbrthetwo highdose animst, I willfaxthatto you when available.
The interestintghingtoknow from thisanalysiisstheextreme linearitoyf thecumulative dose vs.serum concentratiorl
We willneed to produce new chartsbecauseof a slighetrrorinthedataanalysisF.or example: Ifa serum valueisreportedforday two, therehas onlybeen one dose since collectioonf serum isalwayspriorto dosingon the&y of serum collection.
The valuethatI stateintherational(e5.8ppm PFOS in senun per I mg perkg ofPFOS dose),has takenthatfactintoconsideration.
We lookfbrwardto discussintghisdose-settinegxercisewithyou on August 3,at 5:30 pm (Eeastem)duringour conferencecall(Dial-I#n (303)633-6052).
to
Dose-SettiRnagtionalfeorSix-MontChhronicOralStudyinCynomolgusMonkeys
Preparebdy JohnButenhoff July29,1998
RecommendedDoseGroups:0.00,.02,0.a1n5d0.75mg/kg/day
Rationale:
ResultosfanalysefsorsenircnoncentratioofPnFsOS fromthefour-weerkangefmder (attacheds)uggesta highlylinearelationshibpetween totalaccumulated dose and serum concentrationA. significanmtale/femaledifferenciesnot apparent.The consistencyof theratiof serum [PFOS] tototalaccumulateddose ateitherdose leveland alltime pointsgivesconfidenceinusing thispreliminarydatato setdose levelsforthesix-month definitivsetudybased on estimatedserum concentrations.
The dataprovidedby theEnvirom-nentalLab suggestthatserum PFOS concentration increasesincrementallwyitheach dailydose ofPFOS indirectproportiontothe magnitude of thedose. The datagiveus a good degreeof confidenceinderivinga factor relatinagccumulateddose inmg/kg to ppm PFOS serum. This relationshispuggeststhat I mg/kg resultsina serum PFOS concentratioinncreaseof approximately5.8 0.39 ppm. Assuming thatthiszero-orderaccumulationkineticwillhold atthehigherdose overthecourseof a six-monthstudy,thisallowsus topredictserum concentrationtshat would be attainedwith chronicdosing.The apparentprecisionof thisisinvaluablein settindgose levelsthatwillresultinserum PFOS concentrationwshich can be compared directltyo known human serum PFOS concentrationfsorriskcharacterization.
In additionw,e now have thedatatoestimatethesenun concentrationtshatwere associatewdith effectosbserved intheprevious90-dayrhesusstudies.Going back tothe origina9l0-day rhesusstudiest,heLOEL, which we believeto be closeto a NOEL, of 0.5 mg/kg/day would have resultedinan estimatedsenun PFOS concentratioonf 261 ppm. At the1.5mg/kg/day dose,783 ppm would have been attainedafter90 days. Death resultedwithinseven weeks of dosingat4.5mg/kg/day,which isnow estimatedtohave produced senun PFOS concentrationosf approximately1305 ppm.
We have thefollowingprimaryobjectivefsorthesix-monthstudy:1)toestablisah NOAEL forchronicexposure;2)todeterminethresholdeffectsforhealthsurveillanc3e); to providea fi-ameworkforinterspeciecsomparison;and,4) toprovidea referencedose forhuman riskcharacterizatioInt.isnotourobjectivteo studyhigherlevelsof exposure which would resultinseriousillnesisnthetestmonkeys. Dose levelsforthesix-month studyshould be setto allow us to achievethestatedobjectives:
Low Dose: The low dose needs tobe a confu-medNOEL. Ifwe usethelow dose from therangefinderstudy,we expectto achieveapproximately21 ppm PFOS intheserum of thelow dose animalsattheend of sixmonths. Afterthefirsttwo weeks ofdosing,the animalswillbe atand above theaverageserum concentratioonf our workforce.At study term,theseanimalswillhave serum concentrationtshatareapproximatelyan orderof magnitude above averageworker serum concentrationsW.e are suggestingthatwe keep 0.02mglkg/day dose as our low dose.
Mid Dose: The mid-dose isthemost problematicfrom a dose-settinpgerspectiveF.rom therhesusstudy,we know thatminor effect(squestionablseignificanceo)ccurredat0.5 mg/kg/day over90 days. We estimatethatsenim concentrationasttermwere 261 ppm. Ifwe use thisserum concentratioans a referenceforsettingdose on a six-monthstudy, we would assume thatsimilarserum concentrationwsould be reachedwith0.25 mg/kglday. From a riskcharacterizatipoenrspective2,61 ppm senun PFOS istwo orders of magnitude above averageworker serum [PFOS] of 2-3ppm. Achievinga NOAEL at thislevelof exposurewould be desirableh;owever,confidencethat0.25mg&g/day would be a NOAEL isnot as highas confidencethat0.15 mg/kg/day would be a NOAEL. The S.C. Johnson Wax 90-day drinkingwaterstudyinratsyieldedreduced senim triglyceridiensmales at0.3mg/kg/day. Females had increasedliverweightsdown to 0.06mg/kg/day;however, we know thattheratliverespondsmore thanthemonkey to PFOS. Settingthen-iid-dosaet0.15 mg/kg/day would achievean estimatedserum PFOS concentratioonf 156 ppm attheend of sixmonths. This serum levelisstill reasonablyclosetotwo ordersof magnitude above averageemployee serum levelsand closetofourordersof magnitude above currentlyunderstoodgeneralpopulationvalues. Therefore,the0.15 mg/kg/day dose seems to be thebestfitforthemid-dose.
High Dose: The highdose shouldbe designedto producethresholdeffectsA.fterone month of dosingat2.0mg/kg/day (resultinign an averageof 306 ppm PFOS inserum), themale and femalerangefindermonkeys (one persex)demonstratedtheexpected reductionin senun cholesteroalnd triglyceridewsh,ich appearsfrom mt and monkey studiestobe an earlymarker of biologiceffect.There were no remarkable histopathologiecffects.Even though therewere no major disturbanceasfterone month at 2.0mg/kg/day,six-monthsof exposureatthisdose isnow predictedtoresultin lethal body burdens.The rangefinderdatatellus thatthresholdeffectbseginto occuratserum levelsapproximating300 ppm. Using theserum concentrationesstimatedtohave been produced inthe90-day rhesusstudyby 1.5mg/kg/day (78)'ppm) as a referencea,0.75 mg/kg/day highdose would seem appropriatetoachievemarginaleffectosn body weight and generalenergylevelof theanimals.Raisingthedose to 1.0mg/kg/day would result in senun levelsof approximately1044 ppm by term,which would approachfatalserum concentrationsT.herefore,0.75mg/kg/day isthesuggestedhigh dose thatwillclearly takeus beyond thresholdeffectsof serum cholesteroalnd triglygeridreeduction.
Comment: Itisinterestintgo notethattheconsistenltinearitoyf increasinsgerum [PFOS] and apparentlackof significanetliminatiomnay make itpossibletogivemuch
lessfrequentdosesor even to givea singledose atthebeginningof the studyand adjust thatdose periodicalltyomaintaina relativelcyonstantserum level.We willnotreacha steadystatein thestudyas itiscurrentlydesignedand willbe studyingtheeffectof increasinbgody burdenover time. Istherealsovaluein attemptingto hold animalsata relativelcyonstantserum levelovertime? Also,itwould be interestintgo look atserum concentratioantextremelylow levelexposureover time;e.g.,0.1to 1.0ug/kg/day.
AMDI'# 041599.1 CovanceN 6329-222
Study-.
ProductNumber(Tea Subua=): Matrix: mcow"cvisioa: AnalyticaElquipmentSystemNumber.
famument Softwarc/vaam Filenam: R-Squamd Value:
s4c: Y-Imercept.
Dun ofExb=IWWAiWyst. Datesof Analysis/Awlyst:
4-Wcck Capsok Toxkky StudyvdahPffflucroociam'SulfoAncicd PotassiuSmak inCynmoigus Monkeys
T-6295(PFOS)
Monkey Scrwn
FACT-M-3.0A FACT-M4.0
Amelia062498-
MAULYNX 3.1 SeeAtWAummu
SeeAitwismou
SteAnwhmeab SeeAnwhaKim
7116AS RWW 7117/99 HOJ
7f2219SRWW 7/24/92[IOJ
Group I corwoi 0.0IFS&&MAY
Sample 0 105349M 1053SOM 105366F 105367F
Ilaqw[m
Day -7 Day 2 Day 3 Day 7 Day 14 Day 29 Day .7 Day 2 Day 3 Day 7 Day 14 Day 29 Day -7 Day 2 Day 3 Day 7 Day 14 Day 29 Day .7 Day 2
ri4y3 Day 7 Day 14 Day 29
PFOS
CWAW.
ug!nl 0.0173 0.0223 0.172 0.181
O.IS4 0.160
0.0121 0.0207 0.0202 0.0239 0.017$ 0.0220 O.Otis 0.0214 D.On2 0.0194 0.0183 0.0269 0.0135
0.0213 0.0175 0.0153 0,0333 0.118
Group 2 Low Desc 0.02mg&glday
Sample# Timepoint PFOS
Conc.
ug/=L
105344M Day -7 0.0120
Day 2 0.157
Day 3 0.174
Day 7 0.449
Day 14 1.37
Day 29 2.79
105347M Day -7 0.0140
Day 2 0.147
Day 3 0.297
Day 7 0.776
Day 14 1.36
Day 29 2.56
10534&M Day -7 0.0100
Day 2 Day 3
0.161 0.299
Day 7 0,674
Day 14 1.46 Day 29 3.57
105364F Day -7 0.0161
Day 2 0.131
Day 3 0.292
Day 7 0.736 Day 14 1.59
Day 29 3.76
105363F Day -7 0.0144
rimy2 0.117 Day 3 0.267
Day 7 0.629
Day 14 1.22
thy 29 2.65
105370F Day -7 0.0111
Day 2 0.100
Day 3 0.213
Day 7 0.664 Day 14 1.54
Day 29 3.20
PracticQa"lntitadoiUsmit (PQL): PPOS 2S a&/oL.PFOSA 0.3RWM4 PFOSAA 10 ag/mL
PFOS - Peflumooclawsulfonate
Sample 0 nmtp
Group3 ifigohm
105345M Day Day
2.0mg4olday
Day
Day Day
Day
105369F Day
DAY Day
Day
DAY
y
AMD'RN 041598.1 CovanceN 6329-222
Method Ddection[Amit(MDL): PFOS - 12.5ng/mL.PFOSA - 0.25ng/mL.PFOSAA - 3 agImL
Chartl
6.00-
105344M
5.00--
4.00--
0 3.00-LL 9L E 2 2.00-c,oa
1.00--
0.00-
-1.00-
0
0.1
105347M
AMDT# 041598.1
105348M
- 105364F
0.2
0.3
0.4
Accumlative Dose
105365F 0.6
Page I
105344M Acc Dose mg/kg 0.00 0.04 0.06 0.14 0.28
0.58
[serum] ug/mL
o.uizu 0.1570 0.1740
0.4490 1.3700 2.7800
105347M Acc Dose ma/ka 0.00 0.04 0.06 0.14 0.28
0.58
[serum] ua/mL
0.0140 0.1470 0.2870 0.7760 1.3600
2.5600
105348M Acc Dose
mg/kg 0.00 0.04 0.06 0.14 0.28 0.58
(serum] ug/mL
0.0100 0.1610 0.2890 0.8740 1.4600 3.5700
105364F
Acc Dose [serum)
mg/kg
ug/mL
0.00- 0.0161
0.04 0.1580
0.06
0.2920
0.14
0.7360
0.28
1.5800
0.58
5.7600
Sheetl
Slope se (slope) r-square F-value ss (reg)
4.92 0.21 0.99
540.118 5.749
-0.08 0.06 o.lo .@ 4
0.043
y-Int se (Y-int) se (y-est)
df ss (resid)
Slope se (slope) r-square F-value ss (reg)
4.43 0.17
0.99 667.749
4.671
0.04 0.05 0.08
4 0.028
y-int se (y-int)
se (Y-est) df
ss (resid)
slope se (slope) r-square F-value ss (reg)
6.13 0.24 0.99
642.745 8.944
-0.06 0.07 0.12
4 0.056
y-int se (y-int) se (y-est)
df ss (resid)
slope se (slope) r-square F-value ss (reg)
9.85 1.06 0.96 86.129 23.092
-0.38 0.29 0.52
4 1.072
y-int se (y-int) se (Y-eSt)
df ss (resid)
Page 1
105365F Acc Dose mg/kg
0.00
0.04 0.06 0.14 0.28 0.58
[serum] ug/mL
O.Ul 44
0.1170 0.2670 0.6280 1.2200 2.6500
105370F Acc Dose
mglkg _
0.00 0.04 0.06 0.14
0.28 0.58
[serum] ug/mL
o.oill 0.1000 0.2130
0.6640 1.5400
3.2000
Shoetl
slope se (slope) r-square F-value ss (reg)
4.58 0.08 1.00 3683.682 4.989
;0.02 0.02 0.04
4 0.005
y-int se (y-int) se Cy-est)
df ss (resid)
slope se (slope) r-square F-value ss (rog)
5.66 0.13 1.00 1886.988 7.623
-0.08 0.04 0.06
4 0.016
y-int se (y-int) so (y-est)
df ss (resid)
Page 2
Dose-SettinRgationaleforSix-MonthChronicOral Study inCynomo%w Record ofAugust 3,1998 ConferenceCall
Monkeys
Pati@ants:
Dr.John Butenhoix3m Dr.Marv Case,3M
Dr.J@ffMandeL 3M M JohnMoore,EM Dr.Geary Olsen,3M Dr.Joe RodricimM,NMONDr.Andrew Seacat3M
Outcome:
Datafrom therangefindewras discussedincludintghestrongrelationshibpetween
=mdative dose and serumPFOS concentratio(nav* of5.27+/-0.61ppm serum PFOS
co
on permg/kg PFOS don). DiscussiovnerifieJduly29,1998 writtenrationale
fbrsettindgoseswithone chang& Dr.Moore &Wasted thatthelow dosebe raisedfrom
0.02mg&g/day to 0.03mglkg(daytokeep dosesatmultipleosfS. Ile suggestiownas
accepted.The studywillproceedon August 12@,1998 withft fouowingdose levels:
Control Low-Dose Md-Dose legh-dose
0.00mgwday 0.03mgwday 0.ISmg/Wday 0.75mg>day
Copies:
Par6cipants Dr.Lany Zobel Dr.BillWeppner StudyFile(T-6295.7)