Document 8VE0RGjv8EypqOxyLJ3Xbx39B
ADIETARY LCS0STUPDroYs:WITH THE MALLARD WILDLIFE INTERNATIONAL LTD. PROJECT NO: 454-102
3MREQUEST NO. U2723
ris 8U3s,0EnEvilroonmientBalooPrsoTeessitoGnuAigeeinceys (df) PPS Number 8502200
FIFRA SubdivisioE,n Section 71-2 BCD Guideline 05
AUTHORS: SCJeooaunn.n,eG5ayBl.elaaCvgaehsreesry RiymonLd Van Hoven
STUDY INITIATION: Aprit21, 1999 STUDY COMPLETION: Apri 26,2000 AMENDED REPORT DATE: March 23, 2004
SUBMITTTEDO
Envir5oMnmeCnotpaolrLsiiboonrstory St.Pa5u5l5,bMiunhneAsovnes 5c5144
Wildlife International, Ltd.
Eas6t5o9n8,CMoamymlearncde D2r1i6v0e1 io) 323600 Page or
AMENDED 600692
----------------e -- ee--
WILDLIFE INTERNATIONAL, LTD
2
" 0740; 483402
GOOD LABORATORY PRACTICE COMPLIANCE STATEMENT
SPONSOR: 3M Corporation
TITLE: PFOS: A Dietary LCS0 Stdy witheMtallhard WILDLIFE INTERNATIONLATDL. PROJECT NO: 454-102 STUDY COMPLETION:April26,2000 AMENDED REPORT DATE: March 23, 2004 the"TUhSs.stEundvyiwraonsmceonntdaulctPerodteinctcioomnplAigaennccey,wi4th0GCoFoRd LPaabrotra1t6o0ryanPrdac7t9i3c,e S1t7anAduagrdussta 1p9u8b3l,isOhEedCDb)y AP1r9iu5n2c)gi,plae1ns9d8o4fJawGpioatonhdtMhLeAaFfbooFlr,laot5woi9rnygNPocrbxaScctaeinpc,ei,oNn(oOtCifDiEc/atGiDo(n9N2o). 3328,5E0n,vAigrroincmueinttraMlonPorogdruacpthioNno.Bu45r,eP.ari1s0, `PrGoLAcPecdhuormeeptsliimaaenntcdeh;Mtehetoshwtoewdvase,sr,catonhnddcahulacrarttcaetwhedridtzaeatstatiosanurbewstabasenicpneegwrfmsoarnimmoeatdiancchecadroarmcdntientrghiezteod39iMnMaaScracchnoidrvdaeasr.ndceOpTweihrteahtiutnlogln substance was recharacteized i accordance ith GLP siandads following stdy completion `was"Tnhoetsdteabtielnitmyionfetdhien atecsctosrudbasntcaencweitahndGoroefderLeanbcoesrtsaonrdyarPrduacntdiecerSctaondnardds.iotfistoornagse atthetest sie accoArnadlaynsceewsiohf GliLvPerstaannddasredrsaansdamspulletssocofnhduscetead bny a3aMelCboeyrnpgosrarteeipoonrscwderseepanroattelpye.rformed in
STUDYDIRECTOR:
DieP Sabladn
SSeneiaBinaClaolgligsh,erAvian Toxicology
oars 333g
SPONSOR'S REPRESENTATIVE
3A 4s S. uAs.Boascnh0 Louch:
ore 34 [oy
AMENDED hos
_--_--
WILDLIFE INTERNATIONAL, LD
3
PROJECTNO: 45-102
QUALITY ASSURANCE STATEMENT
by "tThheisUsSt.udyEnwvaisroenxmaemnitn]edPrfootrecctoimopnliAagnecnecyw,it4h0GoCoFdRLPaabratra1t6o0ry&Pr7a9c2t,ice17StAaungduasrtds1a9s89p;ubOliEsCheDd P1r9i9n2c)i;plaensdofJGaopaondMLAaFboFr,at5o9ryNPorhacStainc,e,N(oOtiCfDiEca/tGiDonY9N2o.) 3328,50E,nvAigrroincmueltnutraMlonPorogdruacpthioNno.Bu4r5e,auP,ari1s0, tAhueguSsttud1y98D4i.recTthoreadnadteLsoabfoarlaltoaruydiMtasnaangdeimnesnpetctwieornes aasndfotlhleowdsa:te tht any findings were reported (0
AcTviTY
DATECONDUCTED~~ STUDYDDAITREECRTEOPRORMTEADNATOG:EMENT
MaTterstixSuFborsttiafniccaetiPorne,p,
Analytical Sampling April 22, 1999
Aprl22,1999 April 23,1999
FeAneadlyCtoincaslumSpamipolninagnd April 27, 1999
APL2I, 1999 Mayd,1999
NLeicvreorpsWye,iBglhtosodnCdolBlielcetion,
Collection
May 14,1999
May 14,1999 May21,1999
AnDarlayftticRaelpoDratta and July7,8,and9, 1999 July9, 1999 July 16, 1999
BiDolroagticRaelpoDratta and Aug. 30,31,Sept 1,199 Sepiember 1, 1999 September, 1999
Final Report
Apri 18, 2000
April 18,2000 April 19,2000
[Am2ended Final March 19, 2004
March 19,2004 March 19,2004
SusaSLnu. Csolaemnan & oleman Senior Quality Assurance Representative
DATE 3-32-04
AMENDED nonay
_------
8 Y-
WILDLIFE INTERNATIONAL, Lo a
fROi OINO Ii
AMENDED REPORT APPROVAL
SPONSOR: 3M Corporation
TITLE: PROS: A Dietary LCSO Sadly with the Mllrd
WILDLIFE INTERNATIONAL LTD, PROJECT NOL: 454.102 3M LAB REQUEST NO: Uz723
STUDY DIRECTOR:
Mi P. Bey,
SSeeannor BGiaolllaoggh,erAvian Toxicology
Due 30
CHEMISTRYPRINCIPAL INVESTIGATOR
Bld
2s
Scbireono,dAnlVyainaHlovCehnm,iPsRnDy
Bite
REPORT APPROVEDBY:
MTiannSgaagaeRr.MofiYcEhWceoletloax tOpeereat!ions
Be23 March 2004.
DirectorlofCNlohoenm,isityA Hin
-- bw P24
AMENDED
C0891
_--
WILDLIFE INTERNATIONAL, Lm
se
PROJENCOT ha454-102
TAOFBCONLTENETS
TITLE PAGE... we -------------- 0
GOOD LABORATORY PRACTICECOMPLIANCE STATEMENT...
Page 2
QUALITY ASSURANCE STATEMENT...
Pag3e
REPORT APPROVAL.........co. wim-- i PE 4
`TOAFCOBNTELNTS.E........
vos
nn PUBS
"TaandbAPlPEeNCsES cr
wee----------
SUMMARY...
I
INTRODUCTION...
snrssm--------
OBIECTIVE ccm
Boge
MATERANDIMEATHLODSS.d........coc.
m--------&
TTersetatSmuebnsttGanrcoeups... wm DUFOIf thOeTNS
nm-- wn
rss onDOPEASE9 rrmeanmo--rtgt
TeAsntimBaliDs.i..c.
werweree--tn--e--mptra----B---- ----g
Dia etPreparation... --i.-- Page3 110
AnHaoluystiincgaalnMd eEntvihroonmden.t.al.ConditioA ns...
Observations...
err
SII I vargaeeno
II
een
GArnoismsalNBCoIdOyDSWYei.g.h.isFeed Consumption... in i renPaPaggee1144
Statistical Analyses...
wm--
I page ra
RESULTS...
rss --
P28 14
DIMORAoBRIYraSnSdCaliniicar lOtbscre veatiosns s s ...... BodyWeight and Fecd Consumption...
Jwr em
PPaaggee1145 Page16
GrossNecropsy.
---
B--
A:4
CONCLUSION
NV
wn sm Boge VE
REFERENCES...
s--
s
Pog 19
GOOD
_--
-s o-
NWL ILDLP IFENINETEARNATTIONAL, TLDD "6
PROMCTO. ss4102
TAcOonFBtCiOnuNLeTdENETS TABLES AND APPENDICES
TABLE I: TABLE: TABLES: TABL4E: TABLES:
CSutmuudlyawtiitvhePMoFraOliSty fiom a Mallard Acie Dice ary Tosice ty Pie 20
SMEeUaAnYWBAodRyPWReOiSgh.t.).fioam Moallasrd Acce Dicr tary Teoxictoy e Pag2e1
AMceuatne FDeicadrCyonTsouxmipctioSnr(gyiWridtdhayP)RfOSr.aM.aol.lamrd
Pe 25
DGircoisasryPaTtholoogicSawl OybiseWrivtiahtPioRynOsSfraoMoalelmrdrAcute
Pag2e5
MCumualaiiAlvceutMeloDriaeltaiatryyT(rEosxtiicdmiattyedSCtuumduylwaittihvPeRDOoSs.e.,.mg)+.from Page 26
APPENDIX I APPENDIIXI: APPENDIX II: APPENDIX IV: APPENDIX V:
APPENDIX VE:
APPENDIX VII:
APPENDIX VI: APPENDIX IX: APPENDIXX: APPENDIX XI:
Coriicic ofATBYSs..cccvrco Page 21
Dict Formulaion...........
Bo
Page 28
Analytical MetahndoRdESsURS.c.ccvc
PAGE 20
Diet Preparation oe _--
Page 3
`CAucmuutleDaiteitvaeryMoTroalibiyScPttueydntyfwyihroPMFoaOlSlmard
on Page
`IAncduitveidDuiaeltaBroydTyoxWieciitgyhsSt(u2d)yrwiothmaPRMOaSll.ard
nn Pog 45
MFaeleldarCdonAscutmpitDioinc(aybToisirchydbSyytPude)yn rtohmPa ROS... Pa 0
Changes Protocol...
. Page64
Personnel Involved i heStuy...
ne PORE 65
Revised Study Summary...
nn P4566
Report ATOM...
re Page 71
AMENDED
GOD
_--
WILDLIFE INTERNATIONAL, LTD 2.
PROJECTNOlai456n
SPONSOR: 3M Corporation
SUMMARY
TEST SUBSTANCE: PFOS
WILDLIFE INTERNATIONAL LTD. PROJECT NO: 454-102 STUDY: PFOS: A Dietary LCSO Sy withthe Mallard
RESULTS":
eTh6e2d8ipetamry .L.CSw0itvhaalu5e5f%orcMalolarnds feixnpitoesrevddaltooefP4F4n8OSpcowmaes.d.et1e0r9m5i8npedmt.o
s43q.uarTehveaslluopweaosft2h1e3.conTcehnetrnaotimoonrarleisptoyncsoencceunrtvreatwiaons w3.a6s7 1a4n6dptphemcahii.
Based upon reductions in body weight gain at the 73.2 concentration, the o-obsevedfiet concentration wis 36.6
ppm ppm
ai. a.
test
TEST DATES,
Hatch April 13,1999 Aclimat-iAporinl 16:2, 1999 Experimental Surt Apri 2, 1999 Experimental Terminat-iMoayn 14, 1999
CNOONMCIENNATLRATTEISOTNS: 0,9.1, 18.3, 36.6,73.2, 146,295, 586, and 1171 ppm. TESTANIMALS: Mallrd nas playriynchos) AGE TEST ANIMALS: 10days ofageatfestiiiion SOURCE TEST ANIMALS: PWOrBiosxing1WAings, In.
H11a3nWoavsheIirLng6o1n04St1.3512 STUDY COMPLETION: April 26,2000
AMENDED REPORT DATE: March 23,2004
mSereiAppTendolXlfosthecroemvipetdonno.minal st conenrions ad eusbased upon the reals fs es. AMENDED 600ca7
_--mm
WILDLIFE INTRERNNATTIIOONNAALL,, LTD
eee se
PROJJEECTCTNO.NO: 5-45.102
This
study
was
conducted
by
INTRODUCTION Wildife Intemational Lid.
for
3M
Corporation
at
the
Wildlife
Intemational Lid. toxicology facility in Easton, Maryland. The incite portion of the test was
conducted from April 2,1999to May 14, 1999. Raw data generated at Wildie Inemational Lid.
and a copy of the final report are filed under Project Number 454-102 in archives locaicd on the
Wildlife Intemational Lid. site.
`The objective of this study was to evaluatOeBtJhEeCtToxIiVciEty of a test substance to the mallard (Anas playrhynchos) administered through the dit for five days. An LCSO value will be calculated, if possible.
MATEARNDIMEATHLODSS "The methods used in conducting this study are based upon procedures spesificd in the US. Environmental Protection Agency Series 850 Ecological fects Test Guidelines OPPTS Number 850.2200 (1) Section 71-2 of the Environmental Protection Agency's Registration Guidelines, Pesticide Assessment Guidelines, FIFRA Subdivision E, Hazard Evaluation: Wilde and Aquatic Organisms (2); OECD Guideline 205, Guideline for Testing of Chemicals, Avian Dietary Toricity Test (3; and upon ASTM Standard E8S7-87, "Standard Practice for Conducting Subacute Dietary Toxicity Tests with Avian Species" (4). TestSubstance "The test substance was received from IM Corporation on Octaber 29, 1998 and was assigned `Wilde Ioemmational Li. Identification Number 4675 upon receipt. The tet substance was white powder identified as: FC-95; Lot No:217. The reported purty ofthe est substance was 98.9%, with an expiration dcof2008.Followingtest temmination, the est material wes reanalyzed. The results ofreanalysis indicaatteset substance purity of90.49%. All est concentrations have been adjustetdo reflect the purity reported on the new Certificate of Analysis (Appendix 1). The tet substance was stored under ambient conditions.
"The intemal standard was received from 3M Corporation on July 2, 1998 and was assigned `Wildlife Intemational Lid. identification number 4526 upon receipt, The intemal standagararnudla,r
*esFwouelsllo!wOpifunregtaysn,tlutydheyirscoimenpdliecvtasitoteind,puhssreuitmteyemsat8drm6ya.t9ienr%iA.]pwpaeWnhsdirilenechXssrtuamccmeoanrcrzeienzdreaistnitoahncesccohrsadacvncnoefohwiebtehhnGaLanPgtssaienndpaturordrsiealnetthhe AMENDED
GROSDS
_---s mmmn
ee
WILDLIFE INTERNATIONAL, LTD
"9.
PRONOTYD: 454101
material, was identified as: 1H, 1H, 2H, 2H Perfluorooctane Sulfonic Acid, Chemical Abstract Number: 27619.97-2. Thestanwdasasorreddunderambientconditions
TreatmentGroups "The tes consistedof a geometric seriesofeight test concentrations and a control group. Thirty
mallard ducklings were assigned (0 the controlgroupan ten mallard duckwlerieanssiggnsedto each roefptrheesetnrteeadtmweenitgghrtoucplsa.ss fBorirpdlsawceermsenotrinttoebcdyontwreoilgha,nthternescthmoesnetngrionudpiss.criTmhineatbeilrydsfrwoemrewihtohuinseedacihn brooding pens containing five ducklings cach. On Day 0, followin cxpermental sar, the Sponsor vequested the addition fa10 pm est concentration. Birds from the same ot that remained afer test. initiation were usdfo the additonal est concentration. Nominal dietary concentrations used inthis study were 0, 9.1, 18.3, 36.6, 73.2, 146,293, 586 and 171 partspe million active ingredient (ppm 2). The dietary concentrations were established based upon known toxicity data and information supplid by th Sponsor.
Each group was fd the appropriatefetorcontol ditforfive days. During the exposure period
the control group received siven untested basal dit
untreated fed. for three days.
Following On Day 8,
th five-day exposure period all groups half ofth trestment and control birds
were were
cuthanized and liver weight and tissu, blood, and bile samples collected or analysis. The remaining
birds werefe basa raion until Day 22. iver weight and tsue, blood, and bie.
On Day 22, these birds were saiiced and also sampled for
Du"TrheaporftitmhiaeroyTepnshatses of tis est and ther durtions were: 1. Acclimtion-9 days. 2. Exposure 3 days. 3. Postexposure observation 3 or17days Tea Binds
All mallard (nas playriynchos ) were 10 days of age and appeared to be in good health at initiation of the test. The birds were obtained from Whistling Wings Inc., Hanover, IL. and were hatched on April 13, 1999. Birds ranged in weight from 96 0 204 grams at fest inition. The birds used in tis study were immature and could no be differentiated by sex. All birds were from the
CROL99
_--mm---m--mm
WILDLIFE INTERNATIONAL, tT 10
ISRCTN: fon
same hatch, sclmaed
penreared and phenaypially indisingushable from wild 1thcagingandcis fom th day afte hatch un nition
bic. ofthe
All est.
birs
were
AvimalDiet
Throughout acclimation Wide Intemational L's
and testing all test birds specifications (Appendix
were fed a 11). Water
game bird ration from th town of
formisid (0 Eston public
0atefrorsmuopfplayn,tiabinodtiecemdedwiecraetipornovdiudreidngaadcclilbiimtautmiodnurorintgesatcincgl.imation and testing. The birds received
DietPre"Tphaecatiesotnsubstance was mixed irc it the ration. Ming was done witha Hobart mixer
t(hMeodreeplorNtuemdbpeurriAtyS2of0t0h)e.teAtlsudbisettaarnycetesAtlclodnicesnatrryatcioonncsenwterraetiaodn aincd hteo L1C0S0D% vPaFlOueSabraesreedpourptoend
a59pma. inthe diet. 73.2, 146,293,586 and
Nominal dietary test concenteatons 1171 ppm. (Appendix IV)
used
in
tis
studywere
9.1,
18.3,
36.6,
DietSamling
Samples confi the
of the est dics were collected stability and homogeneity of the
0 verity he test test substance in
concentration administered the dies. Homogeneity of
and to the test
ssuabmsptlaenscefoinmtthheed1ie1t7wapspmevaal.u.at.edtebsyt dciactlslcatinpgrepsaixratsiaomnploensDfaryom0. thHeom9.o1gepnoemitay.samip,leasndwesriex
Hcoolmloegcetendeiftryomsatmhpelteos,amlioddsieervaenddabs overtifiocafttihone slafmplaensd. rigOhntesevcetriiofniscaotifonthesammipxliengwavesssceol.llecTtheed
sfrrooumptahterceopnatrraotliodinetonanDdaty0wo. vAeritfhiecaetnidonosfatmhpelxepsowsueree cpoelrlieocdte(dDafyro5m,coancehsraemmpalieniwnagsccaamlecrd
ifhreomtestthesucobnsttroalnanndthtewdoicstam.pTlhees swtearbielictyaslamcpdlesrowemreesccllrcatidmefnrtomgrfouep rtoemdaeitnrimnignien hsiesbfieledyeorsf
aftr being at ambien est pen condions fo fv dsys. Wilde Intemational Li. analytical chemistry.
Samples were wansteed
immediately to
AnslvicalMethod deveTlhopeedmaethWodilseedItfcomatthieoannlalLyis.isanofd ethne eavdia"nMedtihtodsaOmuptlliensewfaorshbeasDeedteeumpionnatmieotnhoofdoPlOogSy
GOO
m _-- mm M
WILDLIFE INTERNATIONAL, to ne
Roms on
in Avian Feed. Avian requisite quantityoffeed
dict samples contained in
were extracted with French square glass
methanol. Methanol was bot. Bots were capped
added to a snd shaken
ionna esdhatkhreebmlee.s Swaitmhplmeesthwaenroelvnatcouutmhelieter.usiTnhgequialtiteatiwveasfiaenspfapeerre.dTthoearevtoailnuedmefesedflwaassk
Eanadchbrsoaumgphltettohvenolwuamse dwiiltihcmdetwhiatnhola. 50A%s pmpetrhoapnroilt:,0sa%mpNleAsNwOrpeurfeurthweartieresodlutwiiotnhcmoetnhtainonl.
a0.l1i0b0rtmigon4HraPngFeOSof GtheerPaRlOSstmaentdhaodrodl.ogya.nd
0.05% formic acid (vs) so they fel within A method flowchart s provided in Appendix
the II,
Figs.
perfCoornmcaencnetsliiqounisd cohfrPoFmaOtSogirnaphheysstainndaradsHeanwdatsaPmapclkeasrwdeMroddeelte1r1m0i0neidgbhyrPeevrefrosremdapnhcaeseLiqhuiigdh EChlimreormaTtuorgbraalpohnS(pHrPayL)iownistouharcPe.erkiHnP-ELlCmesreApaPrIat1io0n0sLwCeMreasacShpieetvreodmuestiensgcaaKpepyestdonweitBheatPaesrkCiny. analytical column (100 mm x 2 mm 1D, 3 um particle size). The instrument parameters are summarized in Appendis I, Table 1
Calibration standards of PFOS prepared ina 50% methanol: 50% NANOpure te solution
Containing 0.100mg 4H PROS (memal standsrdyL and 005% formic acid (vb), ranging in
pcroonmcieniernatiopneafkrroems0p.on0s0e22f9or0P0F.O0S457wmsgut.i.l/izLedwetoremoanniatloyrzePdRwOitShitnhealsacmaplliebsra.tiTonh, qsuaalimtyancodntmrooslt,
and study samples. components. Linear
No attempt was made to regression equtons were
quantify PFOS on the generated using peak
basis of individual arc response ratios
isomeric (PFOS
intemal standard) versus the respective concentration ratios (PROS + intemal standard) of the
calibration sandards. A typical calibration curve is presented in Appendix II, Figure 2. The
concentation of PFOS inthe samples ws determined by substituting the pak arc response ratios
into the applicable nar regression equation. Representative ion chromatograms of low and high
Calibration standards sre prscated in Appendi I Figures 3 and 4, respectively,
The method imiof quantitation (LOQ) fo thes analyses was sta 1.15 ppm .. calvlted 55
he the
product of he lowest matrix blank samples
calibration (500).
standard
analyzed
(0.00229
mg
8.1L)
and
the
ition
fcr
of
ovo
_--
WILDLIFE INTERNATIONAL, LD
12
PROJECT NO: 454-102
"Two matix blank samples were analyzed to determine possible interference. No interferences were observed ator above the LOQ during sample analyses (Appendix 1, Table 2). An interference in th feed appeared at approximately the same retention fime as the peak of intrest but it was well below the LOQ. A representative chromatogramof a matrix blaak is presented in Appendix Ill Figur5e
Avian diet was fortified at 4.5, 183 and 1830 ppm ai. and analyzed concurrently with the samples to detenmine the mean procedural recovery (Appendix I, Table 3). Sample concentrations were not comected for the mean procedural recoveryof94.7%. A representative chromatogram ofa matix fortifiscpraetseinotend in Appendix I, Figure 6.
An example calculation is presented for sample number 454-102-2, nominal concentration of 18.3 ppm.i. in avian die,
nial Weight 100g Final Volum: 200 mL Dion actor, 100 nermedintediluionfctor x final dilution fctor) PROS Peak Ave: 1135658 InternalStandardPeak Aves: 413160 Peak Area Ratio: 02749
Calibration cure cqution, Slope: 277397 Inercept: 001894 Curve isweighted (1).
PFOS (mg iat siumeny
= (( PLeEsRaCrV ca0oitooi - (Y-in neSreheoppte)e 1x 1S5r CCoonneceenn) itrrantiioonn BE (02749-001854)%0.100 mp. - 000923mi
Note: 15, nema standard
nan
_---msm
WILDLIFE INTERNATIONAL, 10 "3
PROJECTECTNO:NOS 454salon
ROS Goma nme +IPROLS (mL g 1) st nsrneomeFrintal Voolanme 1(1)) xDDiisonnpFacetor
0009x0200 100
oo
- spas
PercentofNominalConcentration = --FrFoGsUGoPm EaL1)) 0noSmipalle
PLTo E
Housing andEnironmentalCondens
During acclimation and testing al birds were housed indoors in baterics of thermastatically
controlled measured
brooding pens approximately
manufactured 62 X 90 cm
by Safeguard Ceiling heght
Products, Ine. Each pen was approximatly 25.5
hd cm.
floor pace that External wll,
ewelilgnhg,s tahnedn fclhooosrsenweirndeisccornismtirnuactteeldy offogmalwviatnhiiznedcatcchlrewpirerseanntdedshweeeitgihngt. laBsisrdfsorwaesrsigsnomreendt bfyo
pens. Each group of birds was identified by pen number and test concenaton. were identified by wing bunds.
Individual bis
(SD).DurAivnegrhageeeastmbtiheenavoeroamgetteemmppeerraattuurreefoin ththeisbrsotouddiyngwacsom2p5a.r2tmCen+t0o7fthC (pSeDn)s wwiatsh3a8nCaver2ageC
relative Hoursof
humidity light pe
of $3% + 18% (SD). The day during acclimation and
photoperiod throughout
(maintained by a time clock) the test. The igh source was
was sixteen flsrescent
laipgphrtosxiwmhaitcehlycl2o0s7ellyuxapopfrolxliummaitneronno.ondHaouysisnungliagnhtd. huTshbeanbeiyrdpsrawcetriceesexwpeorsedbatosanonagvueirdaegleinoesf
estabishedbythe National Rescarch Council (5).
Observations injurDyurwienrgeacncoltimuasteid.on alFloblilrodwsiwnegreteostbsienrvtetdiodniya.ndBicrodnstienxuhiinbgitiunnglabntoermmianlatboenh,aviaoirobirpdhsyswiecrael
Cong
-_--
WILDLIFE INTERNATIONAL, to "uae
PROJECTTNs O. 4 e
observed at losst once daily. abnormal behavior.
A record was maintained of all morality, signs of toxicity, and
`AnimalBodyWeightsFecdConsumption
bindsI,ndainvdidounalDabyosdy15weaingdhtDsawyer2e2 mfoerasalulrreedmaatintihnegibniirtdisa.tionAvoefratgeeefsete,docnonDsauymp5t,ioonn vDaalyuesfdourisnlg
bhye pexepnosfuorrecpaecrhiodtr(eDsaiymsen0t-5g)raonudp tahnedpotshetxcpoontsruolr gorbosuepr.vatiAodndipteiroinoadll(yD,ayfsee6d)cownesruemdpettieornmiwnaesd
cdeotnesrummipniedonfowraDsadyeste8r-m1i5neadndby15m-e2a2sufrorintghethreemcahiannignegihnatlhfeofwetihgehtorefatthmenfteadndpcroensternotledbtiodst,he bFiercdsd
ovaegivren navadable
period oftim. The accuracy wastage offedby th bids
of
feed
consumptionvaluesmay
have
beenaffectedby
the
GrosAsTNHetcrsobpisnyds ht died during the course ofthe test andl birds remaining a he termination were isulbejewcetreed tco0lalgercotesdsfnreocrmobpisryd.s Aedudtihtainoinzaeldlyo,nivDearyw8eiagnhdt a22n,d nlvdewihsesnuep,soessriublmeanfdrombltohoodseoltdhast,iaendd during th course ofthe study.
StatisMtoircaallAintayldytsseswere analyzed using the computer program ofCE. Stephan (6). The program was
designed to calculate the LCS0 value an the 95% confidence average method or the binomial probability method (7, 5, 9.
interval n this
by probit study the
analysis, moving LCS0 value was
gdeotoednnmeisnsedofbyftthetepstrobairte mreetphoordte.d. TheBosdlyopeweoifghthtecdoantcaenwrearteiocnormepsaproendsebcyurDveuaentd 'resutletsst oufstihneg
TOXSTAT software (10,11). No stsical analyses were applied 0 fecd consumption dts.
RESULTS
DitAAvmisalnysdiiset samples wer collected from the 9.1, 18.3 and 1171 ppm ai. test concentrations nd analyzed 10 exalate homogeneity of the est substance in the avian diet. The analysis of these samples also served as verification of test substance concenirations. Resulting mean measured
enon
_----
mbro
WILDLIFE INTERNATIONAL, 11D "se
SROMCINO: Hil
WcEonTcen9t.r8a.t5io0n.s9,69stpapnmdaard.de(viat=iCo9ns94aV%nd),coe1f9f.i5cien2t.s13opfpmvaraiia.tio(nC(VCV=)1f0o9r%t)heasndtes1t19c6onc7c0r2tpoonms
i8h3e t(eCstVs=hoSw8ed79n)o irnesepfeecnicveels(aAbpopveentdhiexLIO,Q.TabSlaemp4)l,esCocnotlrleoclteadvidaunridnigttshaemspltes0cvoleleysetdheur36i.n6g,
4730:22,,741.456,,219734,a2n9d15a5n6dp3pm7 pa.pmetai. rseusbpsetsatnicveeclo.nceTnhtersaetivoanlsuehsadrempereasnenmteedas1u1r0e,d10c2o,nc1e1n9t,a9t9i.o3nsanod.f
s9a1m.p6l%esofcotlhleecntoemdifnaolmcofneceednetrrsaftioens,beriensgpehteilvdelsty a(mAbppieennditaempIe,raTtaubrleef5o).fivAenadlayyssisavoefraagveid 1d1i9t,
97:4, SHG
130,104,943, and 1171 ppm
101, ai.
101 test
and 95.0%of he Day alu or the substance concenrsions, respectively
9.1, 183, 36.6, 73.3, 146, (Appendi I, Table),
295, A
representativechromatogram ofa test sample s shown in Appendi I Figure 7.
-
No mortalities occured behavior throughout the st.
in the Ther
control were no
group, and ll bird were samen elated morse
normal or over
in sppearance and signs of toxiy in
he 0.1, 183,366,732, 0 146 Was 20% (2 of 10) mortality in
ppm .. the 293
rcatment groups pm a. restment
(Table group,
and 30%
Appendix 3 of 10)
V). However, thee mrtalty in the 586
PP i. treatment graond u90%p(9 of 10) ithe 1171 ppm a. ncatment group.
Day1. thSeig2n9s3ofptpomxii.cewsetrmeefnittgorbosucprsviengolenmtohretamloimtiinegsowfeDreanoyt4edsond tcohntmionruneidnngsiloftDcamyina7tiaonnd
Dfoaryt2h2o.seAlblinbdsidssacirniftiicsedwaenamDeanyt8evneddiunstpillaytehde sfgenrsmoofonooyf.DeyThesfeoritnhcolsdeebdirrdesdsucaecdiiroeesdtoonn
to simul weakness.
(sound and motion), lossof coordination, ued AI emining birds appeared 1 have recovered
appearance, lethargy, and were normal in
and lower limb appearance and
Sehavior fromthe morningof Day nil test rmination.
At onthe
the 586 moming
pm a. of Day
aeament evel ther 5, Day 6, and Day 7.
wer thre esmentested mortals, Signs oftoxicity wee frst observ on
one cach ned themorningof
Day Day
3 8
and for
continued uni termination for thos bird sacrificed on Day
those 2.
birds sacrificed on Day 8 nd uni the afermoon All birds in ths treatment level displayed signs
of of
toxicity. These included reduced resction to simul (sound and moron), ruffed appearance, lower
no on
_--
WILDLIFE INTERNATIONAL, 10 "16+
(Ronse
tim weakness, appearanceand
an lethargy. behavior fom
All remaining bids the momoifDnayg9
appeared unl test
to have recovered ermination,
and
were
normal
in
In the moraities
1171 ppm ai. were found
otnreatthmeenmtogrrnoiunpgthoefreDawyere4,ifnoellroawetdmebnytrfeoluartemdormtoalritailess,on
The first Day 5,
two two
tmhoertmaloimtiiensgonofDDaayy62anadndtthhefsinianlgmeorstuarlviitvyinognbDiardyc7o.ntOivnueerd s0idginsspolfatyoxsiicgintsyowfterricfirsttyotbhsreoruvgehdtohne
ainfctleumdoeodnroefduDcaeyd 8r.eacStiigonnstoofsttoixmiucliity(osbosunedrveadndammoatnigonbi,rdossisnotfheco1o7rd1inpatpimon,..prwoesattamteentposgtruoruep,
dbierpdreaspspioena,recdon0wshiaovnesr,ercouvfefreeddapapnedsrwaanscen,orlmoawelrilniampbpewaeraaknnceessa,nadndbeehhavairogry.roTmhethoenmeosmuirvnigviongf
LE --
BodyWeightndFeedConsumption
When compared weight among birds
10 in
the the
control group, there were no apparent 0.1, 183, or 36. pom a. treatment
restment related effects on body groups. There was a sutistcally
sigaificant (p<0.05) reduction in weight gain at reflected in significantly lower (p< 0.05) mean
the Day
9.1 5
ppm body
.. level weight.
fo the When
Day 0-5 period that was compared fo the control
srreosuupl,inrgeidnucatisoingsniifnicwaentilgyht(<g0a.i0n5w)erloewealrsoDasytis8timceaallny bsoidgnyifwieciagnhtt.(p<H0o.w0e5v)eor,r dtihfefeDraenyce5s:8frpoemritohde,
c9o:1ntproplmgraio.upleavtelthaen9d.1weprpemnoat.doesveeraespppoenasrivte. beThdeurce0fora,ltohweesremdeiaffnerDeancyes0webroedynowteigchotnsifodrerheed
reatment related.
73.2T1h4e6r,e 2w9a3s, a586m,araknedd,117e1spimpemnt..eleastetdm,encotncgernoturpastidounrriensgphonesipvoeseufrfeepceroinodbo(dDyaywsei0g-5h)t (iTnabtlhee
2 and Appendix VI). Jevel anda p<0.01a
tThhees14e6r,e2d9u3c,ti5o5n6s awnedre11s7t1atipstpimcal.ly.sliegvneilfsi.canDtuarinpg<t0h.e05Dafyor5t-h8ep7o3s.t2-cpxppomsuari.e
preeraitomde,ntwegirgohutpsg.ainThceosnetirnudeudti(oonbsewreerdeucseadtifsoircsaulryvisviignnigfibciarndts aitnpt<h0e.02193,fo$8t6hea2n9d31a1n7d1 5p8p6mpapim
ai. levels. compared.
Differnces at the 1171 ppm a.. Body weight losses and reductions in
level were weight gain
marked bu could at the 293, 586 and
not 1171
be statistically ppm a. levels
wer reflected in significanly (p<0.05 o p<0.01) lowermean bodyweightson Days 5,5, 15, and 22.
-_--
SWILeDLeIFmE INNTEERNATTIoOMNAmLL, wueo a.
TET PROECTNO 450-102
o(f<th0e )est.lowAer hthaen1h46epcopnmtroil. greovuepl. meDaunribngodtyheweDiagyht1s 5on nDdaD8ayaynsd15222pwosetrsaploossiegnipfeirciaondtsl.y ody weigh gain appeared comparable amon ll groups e29s3d,W5ch5o6en,nsaucnmodpmip1o1an7r1epcdpotnmo ntahee.dcron1e0tsrobelengtorbosguerpro,vuepthdserdteuhrrwoisunghtaheDmaaeryxkpeo1ds5urrieendputechrteiioo2dn95(n,Da$sy8es6d05ac)no.dnsuA1m1p7rt1eiduopcnptimionnath11e westment groups.
GrosDsuNreisnrgotphseycouseofthe ts, al is ta died were subjected 03 gros necropsy. Necropsy cmrouenssscuilsdeerfmeoadrssb,irbdreserfaodtumndesnpdteearcdeltcwelde.oe,Dseeitmmaipilatlrsy.ocfrCothpoes,mnemacnardopposbytsefyrivnadgtiainogsnsvsoaieinnpccrlaeuisdneelndtetcdhtinsn,cToandFbiiltn4eidionngslowsseoef fDraoymH2th,e fa1o8tl.3lh,wlsi1un4rg6vie4fvntidngc1i1b7in1rtdspipwnme.r.e.TshubrejeeanctetcmeredonptsygrroeosuspslnweeocrreobwpieodsrnscymcrDukataybbnlieza.endOtonhneerDbeimynadi8inniantnhgdebDciaorymdeo2tsn2 an5escDronposaiynddofaan5beisdblsildo,niwDiahtyhe 53.36lA6ackpsionmfglme.u.bsicrdleaimmatehsen. .g1rIonpuphpmewae7.e3.e2noaptmpedmnwt.i.tghretousip nenedtcryorpl0siudspaoson edDaroydi accrcoouspsnecorontiDcaymatwraaslsomnallt,hwirtihgh iladck oofftshecaebdommiisn,alnwdala.dDau2c13 0t.heonmataubreieosf tcheomlsionngs boibrsdesrvferdominthbierd9s.,1,an3d65t,h30ldat73e.d2 pinpimdeani.ceeoftomcceuregnrcoeu,pshewefiendointgscolinsstieddesrebdoteo ibnecsutehnlized srreoaumpesnwte.reIonnadrdeimtaiornk,abllel. remaining birds in the conto, 9.1, 36, 37d 73:2 ppm a. weston
findiInngthoefa29r3eapionmed.y.olrkasamc.enAgsreocuopn,dobniedbiirndtuhth2a9n3izpedmoni.Dagyroupw,acs rneodpswitohnanDaiynci,denatmasl wneoitgehdtasgaeimnacfioartehdewiltehvela aancdk oafmsusccolnseimdaesrse.d tTchaitsmleonsts orfemluatsedc.leAmansesrwasbicodnsiinsttehnet 2w5it3hpepdmuca treatment group, cutbnized on Day 22, was noted with 2 cm cysts containing yellow fui on the
oo
WILDLIFE INTERNATIONAL, LTD
"se
PROJECT NO: 454-102
anterior portionofth Ie lobe of the fiver. Due 1 the natureofthe sts andth isolated incidence of occurence, this observation was not considered treatment related.
"Three birds in the 586ppm a.. treatment level were cuthanized on Day8. At necropsy, they were noted to be thin and lacking muscle mass. One bird was noted with a slightly pale liver. These findings are considered to be treatment related. Four birds in the 586 ppm ai. treatment level survived until Day 22. Necropsy results for these four birds were no remarkable.
CONCLUSION "The dictary LCS0 value for Mallards exposed to PFOS was determinedfobe 628ppm ai. witha 95% confidence interval of 448 ppma. {0 958 ppm a... The slope of theconcentrationresponse curve was 3.67 and the chi-square value was 2.13. The no morality concentration was 146 ppm .i. Based upon reductions in body weight gain at the 73.2 ppm ai. tet concentration, the no-observedeffect concentration was 36.6 ppm a...
GO 058
WILDLIFE INTERNATIONAL, LTD
"9-
PROJECTNO. 458-102
REFERENCES
| U.S.EnvironmentalProtectionAgency, 1996. Series850- EcologicalEffectsTet Guidlines (draft), OPPTS Number 850.2200: Avian Dietary Toxicity Test.
2 SEUnuSvb.ivroiEnnimvoeinnrtoanlE.mPernHotoeaacliroPdnroAtEgevecalntuicaoytn,ioOnAf:gfeinccoyW.fiPdeis1ft9e8i2c.iadendPPreosAgtgriaucmiasdi.ecAWsOassrehsgisnamgeniniotn,,GuDi.sCdu.eblsienceiso,n.FI7F1R,A
3 OTersg.aniOzEaCtiDonGufiodrelEicneofnoormTiceCosopoeftrCahtieimoinncaagnld. DGeuivdeelloipnmee2nt0.. P1a9i84s. Avian Dietary Tosicty
4
ASmuebraiccuaenDiSeotacriyetTyrifcoirtyTeTsetstisngwiahndAvMiaatnerSipaclis.cs ASTM Standards, Vol. 11.04. Pildelphis, PA.
A19S87T.M
SSttaannddaarrdd BEPSr7a-ct8i7c,e
AfonrnuCaolnBduocotkinogf
5 NWaatsihoinnagltonR,esDe.aC.rchNotCioounnaclllA.cad1e99m6y. PreGsu.ide12f5oprp.the Care and UseofLaboratory Animals
6 SHtaezpahradn,ECvEal.at1o9n77s., MAmereicafntorSCoaclhiceutlyaotifnogrdaTneLstsCiSn0g.anPda.geMs6ate5s.I-Aq8uPautb4.icNToovxicSoTloHgy6a3n4d PhladephisPA,
7 SCtoempmhuanni,caCtEi.on 1975. US. EPA, Environmental Rescarch Labortory, Duh, MN. Persons
8 Finney, Du. 1971. Statistical Methods in Biological Assay,2nded, Grif Press, London.
9 Thompson, WR. 1947. BacteriRevoielwso,Vgoli2c, Naol2: 115145,
10. wDiutnhnaetC,oCnWtr. Jo1u9r5.5.AmeAr:MuStlattisp.leAsCoocm.pa$r0i-s10o9n6'-1P1r3o1cedure for Comparing Several Treaments
31. Gulley, DD. 1990. TOXSTATRelease3.2.The UniveofrWsyoimitngy.
600ng
--,,,------,,------
WILDLIFE INTERNATIONAL, LTD 2.
PROJECT NO: 454-1o
TABLE1 Cumsative Morality fom Mallrd Acute ica Toxicity Study with PFOS
- ErGropup] No DeExapdoPseurreNPoeriEovdponed No.PDoestaEdxpPoesruNroe PEexnpoodsed Gwmai) TBw0 DaT DwZ Dy3 Dud DS Due bu ni
Control
0
030 03 030 om 030 03 03 om 030
Treatment
5
ol 00 010 00 00 ol wo wo io
183
o0 00 ol wo ow ow ow wo om
366
00 00 ol 0 om ow ol wo om
n2
00 00 00 00 ow ow om wo oi
16
0 00 on 00 ol ol om wo ono
I
oo oo 00 00 00 mo oo wo 20
sus
mo mo ol ow 00 mo 20 0 0
m
wo om 00 00 20 el wi 90 io
"TRosLCR0 value was auc tbG28pom 1 wih 95%conten mere STRprOa R
NS morales occured in nyof he restment eves om Day 81 Dy 2.
Cnn in
_----------
WILDLIFE INTERNATIONAL, LTO
ae
PROJECT NO: 454-102
TAPaBgLeE| 2
Mean Body Weight (5) from a Mallard AcuteDietaryToxicity Study with PFOS
--E--r ------
pGrmoauip)
Exposure Period Cans? Dio Dus DOS
PostExposCurheaPnegrieod Toul Dus DySS Change Dayo)
Cont0ra Mem Bmw
wom
215
EY
ue 2
5[
3
Treatment
91 sMoem wWo oeae Eoiwe w6 eaon
9ss0
183 EMSen uwe osom 3i]
m2 omon
2215
366 kMoe) n w6 ooson 122s
mws on
E3y0
B2 BMYen w0 owss ar
w0 owa
2E0)00
"6 0Men wwooew 2
W m6
125s
2 M>an wno oxee 5ae 2 ss w
sstane
6 sMoen wEI ee 2ee sw e w
3300
wi Msoen wPE oe aie w- oo
15|
e5Tc, AoBpnpdeTnmitbVei.vladEsaiseicaly sig Dot's is.
SSm=iiXtoendatie fisrTeaaenindfooomn mohoehercss oonpttollggrroacupprat <<p00.0051(D(unDn'usnsicvs)s,
non
_--mm
eee
WILDLIFE INTERNATIONAL, LD
2.
PROJECT NO: 454-102
TPAaBgeL?E2 Mean Body Weight (8) from a Mallard Acute Dietary Toxicity Study with PFOS
TGro-- up Goma)
PostECxhpaonsguere Period Change Dis Days DaySis Dyn Dey:
Con0tra Men 3
a00 w s mrd
wwou m
Trestment 91 sMDen 83 Men 66 IMan
u5s m2s a2m m7e @2s a2a sm3 @" a"w0
3m 8 sis s0 usa mm s mas
52 Men Me MEem
364 s9m m2e1 m5e sSwo o2m
m Ba w as g one
2% kMoe) n
w"e Haige 32ss
oa"e 3s
6 MIen
vas w0ie m3m
aaxe an
nn MbYen
wsd wy;
i"1S=icNcoiodluysoadtbessrevanafdlosemtohsemcaoorcnnt.lgrwoiunpgsDpo<.e.0s5 Donec' ts) "Silly den econ gotpp00) emer i
;
CE:
er
ChaTnogules)
ai2
aEF a@ asni aso 310%s 05 ao wg.
GRO
_--m_ ee --
WILDLIFE INTERNATIONAL, LTD
Ey
PROJECT NO: 454-102
TAFuBgLeE| 3 Mean Feed ConsumptTioonx9ibnirSdtuddayy)wirh FoOMSalml Acute Diciary
Experiment
f--
rma)
I
ew
0 jo)
Treatment
a1
n
1
"
206
ww
na
"
ue
0s
m
"
s16
5
wn
2
CE
PonBron Prod
i7s
w in us 0 15 a 5s x
--
nor
_--m--mmm--
WILDLIFE INTERNATIONAL, LTD 2.
PROJECT NO: 454-102
TAPaBgLeE? Mean Feed Consumpt"iTooxnic(igt/ybSirtdu/ddyayw)itfhroPmFOaSM.allard Acute Dictary
Experimental Grown (ppmai)
BE o Mseoan
"Buo EPsost Expw osure Pern iod m
m3
1280
Treatment 01 183 366 52 146
El ss nn
Tem
m
198
186
204
165
i
14s
m
159
164
109
1m
ns
0
106
154
GRO 2g
EE
WILDLIFE INTERNATIONAL, LD 25.
PROJECNTOo.n 45s4t.er
TABLE4
froam MallaGrrdosAscPuattehoDliocgtircyalToOxbisceirtvyatSitoundsy with PFOS Binds tot died durin the courseofthe study
TTrT e PMiALUre
Finding
NT 2 sNo NIe
Crop empty GELm.acTioaecd, primarily empty Gizzard contents bie sained IoGtizsztaidn,aecmopntyte, blacank tartke P-- Kidneys, poe SLpolseseno.f mgursecylemass Spleen, smal and pale TShplienen,ple
02'
|}
11
41
24
0 0
0 o
| \
002
0
0
|
2 o
2
s .
i
1
2
0| 2i 43
60
I--
Hl:2
gg
1538 23.
ssss8s
33 33 93 33 83 5% 2% sy
22513 z2ss23 538s 53 83 83 ze 2% gy 1
#78 ss5533 23 33 33 33 335 335 8% 3y i
E:l | ssssss 5% 37 5% 3% 35 3% 5 Ha
| Pg0s5 IH9) sssses 22233538 55 I5F75873 i3f8 5kC f4H:
4s
+ 333H5 |1.3
a5 sf cs sc 52 5% 35 243i8t
2s 8558 82 08 EE88 8gy]
IF 384377 sezess 52 5% 5% 5% 85 55 5E adi]
| 4 "|
! sf:
Cod
8 Bs!
S
5|
;
2|
B
ssssss
sessss 55855355
cnmeme
555225 52 53 33 58 sgh Bi
syezeysgszzoeexge2ts 5s%R 5E%O fBEyLiE
32 33 33 3% 33 835 535 554S[1o44gg8y ono on mn mn mn --{35i3h:
z
5
5
2559
E2|
PFEeRiEfe......FreL. =2=2 R3a3 we7 11 88 53 =2H5eldftlid
5Lwi
8
=
2I3:5s
23
COORG
_--mm----m
WILDLIFE INTERNATIONAL, LTD
PRONTO 409
Certificate Of Analysis
FMCa-r95h,9,Lo2t002017 severe pareLA 0, INmsm. ere
Cemex Tow~~)
Fessox 1""iem----
e ge ---- eso --
[30%
it tnt tg SOBtt g de or r L | am ssw
|
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Festi f| en
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_--_--mmm
WILDLIFE INTERNATIONAL, LTD 28.
ROIECTNO: dsia102
APPENDIX 1
WILDLIFE INTERDNIAETTIFOONRAMLULLTDA.TIGOANME BIRD RATION'
INGREDIENTS SFionyeBCeoanmMMeeaall, 48% Protein WPrhoetaeitnMBiadsdes `AAlgtuaaiy MSpeeacilal2,06%0P%roPtreoitncin DGrrioeudnWdhLeimyestone EMahsitomnainnCealPPrhoesm+ Liquid VGLi.taFmeirnma(nFdeMrmiantecro)aProm scebelow) TSoalullodized
PERCENT (6) w30u6s5 665000 130000 203500 00630s 00325 7000.1000
VITAMIN AND MINERAL PREMIX
AMOUNT ADDED PER TON
VViiaommiinnDA; RNiPibaaoncfoiltnahvniinc Acid VFoiltiacmiAnciBd; PyBriiodtoinxine ThViiaammiinne VMainagmainnesKe(Menadione Dimethylpyrimidino Bisulfc) ZCionpeper liorodn e Selenium
27.0000000000001LCU..U. 41060 ggrrraaammmsss 0Smmeess 12megsrams
200001120g,rams 10255grgarmasms 4768grgarmasms 5115grgarmasms 182 mgs
"ThSeongucarruadnetcfiebdera.nalysis is a minimum of 27% procin, a minimum of 2.5% crude ft and a maximum of "Fermentation By-Products (SourceofUnidenified Growth actos)
Cos
_--_-- --mm---
WILDLIFE INTERNATIONAL, Lo
oo
29.
PROJECT NO: 454-102
APPENDIX 0 ANALYTICAL METHODS AND RESULTS
choo
_--_--mnmee m------m------
WILDLIFE INTERNATIONAL, LD 30.
PROJECT NO: 454-102
TRSTRUMENT.
ANALYTICAL COLUMN: OVEN TEMPERATURE: STOPTIME: FLOW RATE MOBILE PHASE: INJECTION VOLUME: PFOS RETENTION TIME: IRNETTEERNNTAILONSTTIAMNED:ARD PROS MONITORED MASS: --IM_NO--TNEIRTNOARLESDTMAANSDSA:RD.
ApTpaebnldeix| lit `Typical LC/MS Operational Parameters F2TeuwPrelbroeklnionPn-aSEpclrkmaeayrrdAfoPMnIos1doe0url0cLe1.C0MO0apeFsrgahtSepPdeecrtfrsooermlmeeatcentircveeeqLu1iionvpmpioedndiCtwhoirtrohimnPagermrookgidrnea-pElomrer (5M). Keystone Beas Cs column (100 mmx 2 mm LD, 3 pm price size) nc 10.0 minutes 0220 minute 72.0% Methanol: 280% NANOpure Water contsining 0.1% Formic Acid 200 Approximately 7.0 minutes
Approximately 4.8 minutes 498.6 amu 4267 amu
Cn.
WILDLIFE INTERNATIONAL, LTO 31-
PROJECT NO: 454-102
ApTpaebnld2eIilxl
`Matrix Blanks Analyzed Concurrently During Sample Analysis
--_--
ViewC ed Po FOSno ot n
Nomber (854-102
MAB-L
Type `Matrix Blank
(opmai) <0
"qTiuTn mtMAotBf-2i00mig22o1(0L)OniGnd)whaesdi1l1Mu5iaotpnipfxSm&BaTlp aocnfTkehteer doaapero o bThaend ksampu les Bt (0e0)Tao w<e1s 0cQartonstandard
analyzed 0.
CRO
WILDLIFE INTERNATIONAL, LTO 2.
PROJECTNO: 454-102
AppTeanbldeix3 Il Matrix Fortifications Analyzed Concurrently During Sample Analysis
-- Sa(maplse41N0u2m)ber Fortes pms Femara Repceorcveenrtsd
--_--MA--S--A--
as
aasm
105
MMAASsS2
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MMAASSSS
11583300
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553671.
-- Nm WT
Standard Deviatoiovn=e
69398
STd maT nual ce alculaTtions TVoalues hS ave beS en rouni ded for reporpinegepudrpowseisgN-Maen ern5 13 shat
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PROECTNO: 454102
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--_--m
WILDLIFE INTERNATIONAL, LTO
se Appendix Il
PROJECT NO: 454-102
10 187 intonaty: 1995cps. 9 8 7 s
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--_
WILDLIFE INTERNATIONAL, LTD
Appendix
PROJECTNOs 454102
10 10s [pr
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WILDLIFE INTERNATIONAL, LTD
"0. AppendiIxl
PROJECT NO: 454-102
"0
sof 24 80 70 50 50
' 180 ntana23t0cyp:s 188
30 |
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--_--
WILDLIFE INTERNATIONAL, LTD
are AppendiIlxl
PROJECT NO: 454-102
" 188 Itonaity:6238cps
s
8
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5
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3
144
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301 82155 9s1m 1s2o1s 168x02 17817s 2s1s1s | TSiamne
Figur6e. A represenative chromatogramofamarx forificaton sample (454-102-MAS-1A) [RUERS
_
WILDLIFE INTERNATIONAL, LTD ae
Appendix Il
PROJECTNOL: 454-102
"
167 intensity: 7248cps
9 8 7 6 5 4
145
2
1
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197 220 i761s 2s1s1s | TSiamne
Figare?. Arepresentative chromatogram oatfest sample (454-102-2).
conn
_
WILDLIFE INTERNATIONAL, LTO
ae
PROJECT NO: 454-102
APPENDIX IV
DIET PREPARATION
`Weightand volumeofconstituents used o prepare est dicts:
ConNceonmtirnaatlions | SubTsotatnce RBaatsiaoln
0 9.1
00910 5809090909
183 366
0013861388 88999999..86
n2 146
o1.4m659
8999.3 89985
293 586
2598122339 88999974.21
171
11.6483 8988.4
Diets were prepared as follows: 5000.0gofbasal ration was weighed into a ared Hobart mixing bowl.
The test substance was weighed inatared weigh boat Approximately 100 g ofbaal ration was taken from the mixing bowl and placed in a Waring
blender. Theittehsttshuebrsitnasneceaswoasbeaidndgedpl0acetdheinbltehnedbelrenadnedr.the weigh boat was rinsed with additonal ration, The"Tbhleebnldeenrdceorntweanstsriwnesreed wbiltehndaedddifotriaonpraptiron,owxithi6t0mhseaercitnosneedasllasnoydbetirnagnspflearrceeddtio htheemmiixxiinnggbboowwll The"Tbheorwemwaaisnipnlgacreatdioonn aasHsodbdaerdt 1m0ixtheerbaonwdlthaendcotnhteenctosntweenrtse wmeirxeedmifxoredspfporrosxiixmmaolcrleymsiixnumisn.utes.
<The diet was transferred 0a labelled paper feed bag.
CeO 1a
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APPENDIX VI
In"dAicvuitdeuDailtBaordyyTWoxeiicgithytsS(t)udryowmitah MPaFlOlaSrd Pag|e
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Change
Change
Gps) Bid Do Dus Dw0S Dys Dw3S
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454102
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--------------------------------------------------------
WILDLIFE INTERNATIONAL, LTD
4
APPENDIX VI
--_--Experimental eGrmoaupi.
In`dAicvuitdeuaDlieBtoardyyTWoeciigthys Sgt)udfyrowimtah MPaFlOlSa.rd Page?
Bid Dsy0 Days DChaayn0gSe Dus
o
21 121067 227 n1as 2360
hl3 10i7m 00 956 22684
Me5 n iTmTS 2285 1Ts sSai
so 5 2 2 a
PROJECT NO 454-102
DChuayngSes. CThoaunle
796
213910
s&t
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1500
2To7n
is
3
0
2' 11000 325 113s2 E3es 91060
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0197
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34
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------------------------------------------
WILDLIFE INTERNATIONAL, 10 ae
APPENDIX VI In"dAicvuitdeuDalieBoadyyTWoexiicgihttysS(tp)dfy wirtahMaPolFlOamSrd
Pages
PROIECTNOS 4st102
GGnraom), Be Doo Dus DChwaongse bus bChuatngse cwTmoeulos
5
Com wm wm ees
26
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216
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_--mm-s
WILDLIFE INTERNATIONAL, LTD
a8.
PROJECT NO: 454-102
APPENDIX VI In`dAicvuitdeuaDlieBtoardyyTWoxeiicgithysS(tgu)dfyrwoimtha MPaFlOlaSrd
Page
I Experimental ---- pGrmou.p, Bid Dao Days DChnan0gSe Dw ChDaunsges CThoanugle
153
1 2 2 1s 355 9
m
2 " 27 156 a En
20
3 150 m 0 361 0
am
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2
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MseDn Tw TS1a mIee mis w 2 5 m2Es
13 TToeulw
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2
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2s
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0
MSepn w WiTs E35 mm2 w 3 5 m2o
Msenn Hie 22m ihtl m 2 m i r2s
Goin
_--_m--m-------
WILDLIFE INTERNATIONAL, LTD 9
PROJECT NO. 454-102
APPENDIX VI Individual Body Weights (g) from a Mallard Acute Dietary Toxicity Study with PFOS.
Page
_Ex-- perim mentam l (ppGrmosuip.. Bid Dwy0 DiyS ChDainygoes Duk ChDaanygses CThoanugle
366
1 3 261 130 mn
m
1
2 32 256 124 356 100
24
3 142 269 2 369 100
2
" 160 m Ho 353
El
193
s 1m 3s 136 3 9%
254
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m 2 Ta2m 0 w 3 10 am5
366 Gow TToeuwl
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185
2 136 Ed 151 02 ns,
266
3 197 263 ns 37
16
um
. 151 255 104 344
5
193
s 160 324 164 EY 103
267
MseDn W 12 Tu3am e 2 2) m 14 39 S MSaDn D W 16 s N 2 a28 y a3n7 e w2o 0 a3m0
[ERE
_--_--
"so- WILDLIFE INTERNATIONAL, LTD
APPENDIX VI In"diAvcidtuaDlicBtoadyyTWoexiicghhttysS9)tyfowmiath MPRaOlSr,d
Pag6e
PROETNO. 454102
Goin na
--
be wo Dus ChDaunoges bys oCwagies dTeomwe om om ow wm om os
dm wm wow oe ns sow ww ow os 2 Cos mm mw 2s som om om ow we son 0MEeAw ww T A oWwA E w IEnwm5
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dw am ms ow om n
som wm wm ww 0
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som am was we 250
Go
MM S en m e we o n am im w wWm a8 01i60
Tou! Ty omBe ww OT erww ee8 e
600
_--mmmm
WILDLIFE INTERNATIONAL, Lo si
PROJECT NO: 44102
APPENDIX VI In`dAicvuitdeuaDlieBtoardyyTWoexiicgihttysS8t)udfy wito ahMPaFlOmlSa.rd
Page?
= gGeromusp). Bid Duo Dus DChOangse Dyk ChDaunigse CTouuml
1
me a om @
128
Tom ow om aw 2
1
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26
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7%
165
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or
199
216
x
186
wi
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_--mmm--
WILDLIFE INTERNATIONAL, LTD
52. APPENDIX VI
PROJECTNO. 454102
In`dAicvuitdeuaDlieBtoardyyTWoexiigchttysS8t)udfy witoahMPaFlmOlaSrd Pages
--------- pmGsrlo)w met wo bus DChwanOgSe bus bChuatngse cTmouel
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166
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_--_--m--mm----
WILDLIFE INTERNATIONAL, LTD
5
APPENDIX VI In`dAicvuitdeuaDlicBtoardyyTWoexiicgihttysSgt)ufdryowmitah MPaFlOlaSrd
Paged
PROJECTNO: 454-102
-_Ex--perimental pGprmow Bid Diy0 Days ChDann0gSe Days ChDaunsges CThaonuge
sss
1 I} 10s a 16 2
4
2 1s 143
5 29 o
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3 12 160
8 pt 5
os
4 154 ne as
-
.
.
5 16s 195 0 us EK)
2
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se
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55
6
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.
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57
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.
.
-
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a Toul sD 23 2 2 3 a 36
Suift lerenTfi oom hel cone)poop1<p001 Dusmen's is)
--
Go
--_--m
WILDLIFE INTERNATIONAL, Lo
sa APPENDIX VI
In`dAicvuitdeuDailctBaordyyToWxeiicgihttysS(t8u)dfy wirtahMPoaFlOlSma.rd Page 10
PROJECT NO: 454.102
TGe roup w
pm
Bid
Dny0
Days DChwaynOgSe Des Chbaanygees
ChTaonugle
un
' ns B El
-
.
.
20
.
.
.
2
3 1s 96 "5
.
4. 150 ns 2
-
.
s 166
-
-
-
-
s --_-- sMsaoon THESiw s iW n o5 w . iTT:
nm
' us 0 a
.
.
2 no
-
.
"
.
.
3 14s m2 EY
.
4 157
-
.
s 20 154 a6 15s
5
as
Cow
Msepn. Tw EE3 m5E4 m 1s : a : Men 0 ine am we
o T"Nooamoraliiws acTcuSiDs.y nof yhe 2esam coi2l groups afc5Dy -
ee
Sica differs fom the control group sep S001 (urns ies)
Reet
_
se WILDLIFE INTERNATIONAL, LTD
PROJECT NO
APPENDIX VI
In"dAicviudluaDlicBtordyyTWoexiigcihttysS(t8)udfyrwoitmah MPaFlOlaSrd Page 11
Eorom Gomel)
Bi
Dus
Duis CDhawnigles Dyn DCuhaniges
>Laeow oome oomw m msw m
PSoooams ooeems oomm o semus
MeSnowsa9 eemn aaem mw I e
sh 2 26 a
454-102
cmTeoul o39m9 pwest CaH2
o
2Lowme aws oomw wmme awwm
Sooke oa om wom
3a5n7 ht
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3a866 Tie
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V SS e2m1 E5m1 I sw 56 rl u
w2i% 2
GRO;
WILDLIFE INTERNATIONAL, LD
-56APPENDIX VI
In`dAicvuitdeuaDlieBtoardyyTWoexiicgihttysS(tgu)dfyrwoiatmh MPaFlOlaSrd Page 12
PROJECTNO: 454-102
ETGroup. pai) 91
Bid Days
' 39 2 334 3 399 . an sm sMDe.n TEE3
Dayls ChDawnSgles Day
"0 i ol 9 as 70 oie ar 0 6 29 77 sts 2 "a s2m m20 o3m
-- Toul Cham Change
29
mn
15
39%
16
"0
208
asa
198
an
imsE
a3m
E[T 2 pm
Ch-- ange Change. Toul Bid Dis Deyls DayIS Dey Deyis2 Change
153
' 355 on 26 mow
a5
2 a 0 m os us
siz
3 61 Ed wr mos
3
` 390 a 2 0 ies
a0
s mm as 2 ww
an
MsaD.n T 2 2 a 2 W5T W m a a @ Seay Geen om he cl ro 1 <0 (Duane ie)
GROG
WILDLIFE INTERNATIONAL, LTD os
PROJECTNO: 454-102
APPENDIX VI Individual Body Weights (g) froam Mallard `Acute Dictary ToxPiacgiety13Study with PFOS
------
re
"pGrmow, Bid Dey$ Duis CDhuayn8g1eS Daya? ChDaanygies.2 CThoanugle
366
' mn or 305 sa es
50
2 356 ar 1 sr 20
6
5 36 on 4 wm
ed
4 35 59 26 [Et
30
s wm 2 29 soos
w
MseDn TTT3 e#w am2 am3 w 3 w a
---- gpGrmoup. Bid Days Davis CDhauniges. Dayar CDhawngie.:
es cThoualm
n2
' m w 0 CT
367
2 28 0 0 6616
ss
3 30 6 246 wus
a"
a am 99 2 we 97
a
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as
MsaDn Tm R0 S9 we2 w 1s 3 a3m6
coy
_-_----------------
WILDLIFE INTERNATIONAL, LTD
oss.
PROJECT NO: 454-102
APPENDIX VI I`nAdicvuitdeuDailetBaordyyTWoexiigchittysS(t8)udfy wirtahMPoalOlmSard
Page 14
= Group.
ppm
Bid
16
'
2
3 +
s
-- sLDT.
Cha-- nge Toul Dus Dayis DaySIS Duy Change Change.
26 as 2 69 18
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30 569 29 7 mn
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6
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30 576 196 550 2
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2 16s as 250
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----eeeeeeeeee.
WILDLIFE INTERNATIONAL, LTD "59.
APPENDIVXI I"nAdciuvtieduDalitBoadyyTWoexiicgihttysS(u)dfyiwoitamh MPaFlOlaSrd
Page 14
PROJECT NO: 454.102
--T--eGETpne--r --------------C--hang--e ----C--hng--e --iTount Gemil) Be Dus Duyis Duis own omitn dee
536
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9
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4.
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sow ww se ae
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MS en TW w TT Cw m Hw a CB o
-- T TGrom E u--------Cha-- nge --Cha-- nge -- eTrorurl gpl) Bu Dy Dwi DoRls wn pein cee
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M>en TETA TT CoE Be eed Arey Bam
:
er
ET
nog
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--------
WILDLIFE INTERNATIONAL, LTD
PROJECT NO: 454-102
-60
APPENDIX VII
Feed Consumption (gbirdiday) by Pen from a Mallard Acute Dietary ToxiciPtaygSetu|dy with PFOS
Experimental Group. (oma) Pen
Exposure Period Dyos
PostExposure Period Tb T wes
Control
'
109
14s
2
2
190
3
8
nr
4
2
132
5
2
100
6
9%
ne
MSeDn. Tm 10
w13o
GRO oD
_--
WILDLIFE INTERNATIONAL, To "a
PROJECT NO: 454102
APPENDIX VI
Feed ConsuDmipeitoanry(Toixidciety)SabdylyPwnitrh PEGoSM.llmer Acc
oeEre
Page? Ep-- on Food FosTaper Perot
pi
ren Duos
Dues
2
2:
nn
110340
Mew
=
7
183
2:
5@
iuss
Men
0]
i
66
2:
0ss
11390
Men En
12s
na
2:
u
1509
Mean
7
Tor
1
Me2: m Toort
a0n2 5
=
2C5 TE a
Mew
a
@
sto
2:
0
@a
Mean
36
ss
un
2:
n2
22s
Mean
2
3
ano Ee
_--nm ms M_-- - m------
WILDLIFE INTERNATIONAL, LTD -a-
PROJECTNO 4se02
APPENDIX VII
Fecd ConsumDpiicotnar(ygTboixridcditayy)StbuydyPewnitfhrPoFmaOSMallard Acute Page
TEvpenmenal Grow (opmai)
FxpowrcPenod
PostExporare Period
Pe TDT mels bwna
Control
1
155
159
2
161
182
3
166
199
sMDen T 1 r 20
GOO TE
_--
WILDLIFE INTERNATIONAL, LTD
PROJECT NO: 454-102
-
APPENDIX VII
Feed ConsumpDtiieotnar(ygT/oixridc/idtayy)StbuydyPewnitfhroPmFOaS.Mallard Acute
Paged
-- Experimental Group.-- Expo-- sure Period P-- ost-Exposure Period
_-- (ppmai)
Pen Days1s | Daya
91
1
1m
198
183
1
186
204
366
1
165
79
n2
1
148
3
146
1
159
164
203
1
109
32
sso
'
na
143
nn
1
106
154
600-5: <
--_------
WILDLIFE INTERNATIONAL, LTD
ee APPENDIX VIII
PROJECTNO: 4sd-102
CHANGES TO PROTOCOL
changesT:his study was conducted in accordance with the approved Protocol with the following
1. Theprotocol was amended 10 add an additional test concentrationof 10pprna. 2. pTrhoetopcrooltwoacoslclwaarisfiaemdeondiendditcoatitnhdicactoelltehcattiobnoief iwivlelrbferocmolbliercdtsedthaftrdoimedaldlursitnugdytbhircdosu.rseThoef
he study: 3. T64h0e parnodtoc1o2l80wapspmamein,detd0 t0o,c9h.a1,ng1e8t3h,e 3te6s6t,co7n3c.e2n,tr1a4t6i,on2s93f,ro5m806, a1n0d, 2101,7140,pp80m, a1i6.0, T3e2s0t,
acnoanlcyesnitsr.ations were changed io reflect the test substance purity given in the new certificate of 4. rIenqduiivriedduallelgybnaunmdbser1edbewuingsboaindcdesntwdiefyrethuesebdirdtso.uniquely idenify cach duckling. The protocol 5. Btelsototdubessa.mpTlhees pwreorteoccoollliencdtiecdatoend hDaayh8epaanridniDzaeydv2a2chilninnoenr-shewparoinuibzeeludsdedmi borosilicate lass 6. rTehceorfdienadl temperature from one brooder unit on Day 8 and all brooderunitson Day 22 were not 7. MThaeya8f,t1e0m9o9.on observations were inadvertently not recorded for birds in a single brooder unit on
8. OoOfbbsseetrrhevvaattteiisoot.nnss ffoToremccoopnneitrrraootllubbriierdsdssffforroommthotenwseeobbbirroooooodddeeerrrlleelvveeevlleslwsewrewreeerneaindavsdevroeirrtentnolttylynronetcoorrerdceeocdrodreoddnedontohniDsaDaydya2y26 ofthe est.
M05
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WILDLIFE INTERNATIONAL, LTD "6s
APPENDIX IX PERSONNEL INVOLVED IN THE STUDY
PROJECNTO 454-102
"The following key Wildife Intemational Lu. personne were involved in theconductor managemoeftnhits study: (1) Mark Jaber, Wilde Toxicoloist 2) Joann B. Beavers, Director, Avian Toricology ) Sean P. Gallgher, Snir Biologist (@) Courtney Casey, M.., Senior Biologist (5) WillrBd. Nixon, Ph.D. Manager Ansytca Chemisty (6) Tim Kendal Supervisor, Ansytca Chemistry (0) Raymond L. Van Hoven, PhD, Scientist (8) EllenMak,Chemist
COORG
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WILDLIFE INTERNATIONAL, LTD 66
APPENDIX X REVISED STUDY SUMMARY
PROJECT NO: 454-102
This study was reported using a test substance purity of 90.49%. The test substance characterization wasrevised on 9/7/00, follstuodycwomiplentiogn, and thepuritywas determined to be 86.9%. The Sponsor requested that the results of this study be revised to reflect the mst recent puriy analyses. The results previously presented were simply corrected by the ratio of the two purities or a factorof 0.9603. No attempt was madetorecalculate analytical standard concentrations or measured concentrations from original raw data. As such, minor errors due to rounding may be preset. tachedi the revised summary page from the inal report and the most recent certificate of analysis.
AMENDED 000756
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WILDLIFE INTERNATIONAL, LTD
_61-
PROJECT NO: 454-102
REVISED STUDY SUMMARY BASED ON A PURITY OF 86.9%
SPONSOR. SPONSOR'S REPRESENTATIVE:
LDAOTCAAATNIDOAFNCSOTPUYDOY,FTRHAEV FINAL REPORT:
SM Corporation Ms. Susan A. Beach
WEiasltdoinf,eMIanrtyelmaantidon2a1l6L0i1d
LWTIDL.DLPIRFOETEICNTTENRUNMABTEIRO:NAL TEST SUBSTANCE stupy: CNOONMCIENNATLRATTEISOTNS: TEST DATES:
LENGTH OF TEST:
asa102 PFOS (Perfluorooctane Sulfonic AcidPotassium Salt) PFOS: A Dietary LCS0 Study with the Mallard 0,88,17.6,35.1,703, 141, 281,562 and 1125 ppam.
EBixopleorgiimceanltTalerSmtianta-tiApor-nilM2a,y 1149,991999 Experimental Terminatio-n May 14, 1999 24
TEST ORGANISM: SOURCE OFTEST ORGANISMS:
AGE OF TEST ORGANISMS:
Mallard (insplagyrhynchos) `Whistling Wings, Inc. HanoveIr,L 61041-3512 10 days at cst niiaion
To 95% CONFIDENCE LIMITS: CONNOCMEONRTTRAALTIITOYN: CONON-COEBNSTERRAVTEID-OENF:FECT-
503 poma 430and 919ppm
141 ppmai
35 ppmai
AMENDED CRUST
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INTERIM CERTIFOIFACNAALYTSIES ConeAr abuso CONRea: 250180 DueorianAmi: 180 [e----_-- SoCo: Fenn 10
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AMENDED
60059
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ne APPENDIX XI
PROJECT NO e4.ter
Report Amendment
IL AOrmgeinndaeldReRpepoorrtt: TTihleeaPmaegneded report de was added. The total numberof pages
Reason:
wTaosicnhdaincgaetdtfhart ot6hm5e 0rep7o2r.t was amended and note change in
pagination
2 OArmgeinndaeldReReppoorrt:: Resson:
TPhaegea2mended report dat was added nd the exceptions 0the c"Toomsplhioawnctehesaiamteenmdeentdwreeproertredvaiste,d.provide nwsignatures and dSaattesmefnotr.the amended report nd to update th compliance
3 OArmiegnindaeldRReeppoorrtt: TPhaegseusit datsfothe amended report were add.
Reason
Tsiognsahtourwetahnedadmaetnedfoedthreepaomrteanuddeitddaptoesra.nd 0 provaindeew
4 OArmgeinndaeldReRpepoorrti: Reson
PNeawgesignatores and dats were affixed and a tite change was mTaodperovide signatures and desofspproval fo the amended veport and 0 updatea manager' le
5. ONregmiennadledReRpepoorr:t: PApapgeensdixX, Revised Study Summary, and Appendix XL, Report
Resson
TAmaevndimsenhte, wTearbeeaodfdCeodntteonthtesT0abilnecloufdCeotnthenrte.visd summary
and report amendmen.
6 OAdmgennadleRdeRpeoporr:t: Reason
PAafgoet?note and th amended report dat were ded tthe S"Tuomdmiraerctytphaegreeadeor Appendix X, which contin the revised Study summary atno shdow the amended report dt.
7 OArmieginndaeldRReeppoorrt: Resson:
APagfeosotnote was added to the Tst Substance ex. "rTeocednitecCtertihfeicrateeofatAonAadptpyieesn.drix X, which contain the most
ACME0ND0ED5_2
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I WIe LRDLIEFE IINNTTEERRNNAATNIOOINAALLL,e LTLDo a e FromCT NNO: o456o1 ween APPENDIX XI
Report Amendmens continued)
5 ARemaessodnedRopor: 5. ReAmseomndedRepori
PTaogpesro6v6i7d0,aRdedviosend SitnufdoymaStuimomnary, were added the report. "PTaogperso?v)idaendde72a,lReopfocrhtanAgmeesndmmdesnto,th weeoraidgdneadl troetpohretreport.
AMENDMENT SIGNATURES `
SoyDiecior 7)
BaAenloy
WhsittesrerYmLaWtiaonlz,tLid Maragemert
B2ue3a 200d,
RevEWED BY.
Susan Kf Colaman
Quality Assurance
3-23-04
Date.
AMENDED
