Document k6vveKGxMrXozvj34k0MLLd50

~ i ~ := i i i i i i i i = ii, , le iy i, Oo 2 Ly AR 226 -- 1017 &rm Zug o CENTRAL SCIENCE LABORATORY CENTRAL SCIENCE LABORATORY SCIENCE SERVING AGRICULTURE. FOOD AND THE ENVIRONMENT SCIENCE SERVING AGRICULTURE, FOOD AND THE ENVIRONMENT CONTRACT REPORT stopy UBER HTS STUDY NUMBER HT5602 PERFLUOROOCTANESULFONATE, PERFLUOROOCTANESULFONATE, PPOOTTAASSSSIIUUMM SSAALLTT ((PPFFOOSS):): AN ACUTE ORAL TOXICITY STUDY WITH THE HONEY BEE - . | w&r == : 22773344..00000011 Exhibit Sm s2r 734 000000000022 ver ones MET 0438364 CSL CSL Study Study NNuummbbeerr HKTT35660022 TITLE PAGE PPAEERNRFFALLCUUUOOTRREOOOOORCCTATALASSNNTTTEEOUUSSXDUDUIYLYCLFIFNNOTOUUNYNMMASBATBETTEERUE,R:,D:PPYHHOOTWTTTsI5AA66TSS00HSS22IITUUHMMESSHAAOLLNTTE((YPPFRBOOSES)%:): AN ACUTE ORAL TOXICITY STUDY WITH THE HONEY BEE REPORT REPORT AAMMEENNDDMMEENNTT NNOO 1 1 EPPO GUIDELINE 170 EPPO GUIDELINE 170 OECD OECD GGUUIIDDEELLIINNEE 221133 i AUTHOR : AUTHOR: Paul Wilkins Paul Wilkins STUDY STUDY INITIATION INITIATION DDAATTEE:: 1111 SSeepptteemmbbeerr 22000000 STUDY STUDY REPORT REPORT DATE: DATE: 2233 MMaarrcchh 22000011 SSUUBBMMIITTTTEEDD TO: TO: Envir33oMnMmeCCnootrrapploorrLaaattbiiooornnatory ESntvPi59ra33ol55nmBMBeuiunsnsthhnaelAAsLvvoaeenbnnouu$ree5a1to0r6y St Paul, Usa Minnesota 55106 USA : Performing Laboratory: Performing Laboratory: York YO41 117, UK 3 EnEEvnnivvrioirrnoomnnemmnetenantltaalBl iRRo&l&oDDgyTTGeeraaomump. Environmental Biology Group CCeennttrraall SScciieennccee IL_a,baobroarattoorryy SSaanndd HHuuttttoonn York YO41.1LZ, UK EEnnvviirroonnmmeennttaall LLaabobrsotroarytoorry PPrroojpjroeecctt NNuummbPbeaegrreUU212o77f22'33s Page 1 of 5 _ 22773344..00000022 000003 000003 ver orsssss MET 0438365 CSL CSL SSttuuddyy NNuummbbeerr HHTTS5660022 SSttuuddyyttiitlee TTeesstt acy facility SSttuuddyy nnuummbbeerr NNuumbmeroobffppaeaggeress:: CCEENNTTRRAALL SSCCIIEENNCCEE LLAABBOORRAATTOORRY El CCOONNFFIIDDEENNTTIIAALL REPORT REPORT AAMMEENNDDMMEENNTT 1 1 tPPoeercriffclliuutooyrroosotocuctdtaaynnwcesisutulhlfftoohnneaathteeo,,nePPyootbtaaessessiiuumm SSaalltt ((PPFROOSS)):: An An acute acute oonraall toxicity study with the honey bee EEnnvviirroonnmmeennttaall RR&&DD TTeeaamm,, CCSSLL,, SSaanndd HHuuttttoonn,, YYoorrkk,, UUKK HHTTS5660022 . 55 | i Signature, Alitheg Signature. /~~~ Dute5/B4t @dimmiyy) Date~ (dd/mm/yy) SSP tWtuuiddlyykiDDniirsreeccttoorr EEPnnWvviiirrlooknninmmseennttaall RR&&DD TTeeaamm,, CCSSLL --_--e AAuuthotrihsatoionroofifrreselleeaaasseetoofifrreoeppoonrrtt aammeennddmmeenntt sigue Signature [1 f1 he~ MMMaaHnnaaBggeeemmweenntt RReepprreesseennitattiivvee MHHeeaaHddBEEennwvviirroonnmmeennitaall BBiioollooggyy GGrroouupp,, CCSSLL DDaattee_~-'. ~0:.0o1-t t(did/mmmm/yyy)) -_-- 4 "This This amendment amendment has has been been nnootteedd bbyy QQuuaalliittyy AAssssuurraanncceeaass ccoorrrreeccttiioonn ttoo tthhee rreeppoorrtt 3 4o] signature LA ana LI. Date|CcShlol(dd/mmlyy) Datel~[t~[~l(dd/mm/yy) QMMrArss ACCudVVitWWoarallkkeerr CCQeeAnnttrAraaulldSSictcoiireennccee LLaabboorraattoorryy EEnnvviirroonnmmeennttaall LLaabboorraattoorryy PPrroojjeecctt NNuummbbeerr UU22772233 PPaaggee220o7f55 ee --_-- 22773344..00000033 000000000044 VET 0430385 MET 0438366 CSL Study Number HTS602 CSL Study Number HT5602 Reason or amendment There was typographical er om pages 7 and 8 of he final report = Reason for amendment: There was a typographical error on pages 7 and 8 of the final report This hsbee wwhheerree tthhee TTeesstt IItteemm hhaadd been mmisis--ssppeelltt as Peerrfuoorrooooccttaanneessuullffoonnaattee,, PPoottaassssiiuumm SSaalltt ((PPFFOOSS).). Tor had an comectd to Peflucroocamesalone, Posi So (FOO) A. for aios This has been corrected to Perfluorooctanesulfonate, Potassium Salt (PFOS). A formatting ccu on page 7. hich had ekeurd he length of expos Th ho error had occurred on page 7, which had obscured the length of exposure. This has been Coneted corrected. Distribution Distribution SSponsor's Representative MSRRobins 2copies Sponsor's Representative homors iomtor MiDPumer teem Sponsor's Monitor Stuy Director MIPWikne leon Study Director (CCSSLL MMaannaaggeemmeenntt RReepprreesseennttaattive 4 GAManager Mis RM Brookes 1 copy QA Manager Ms R Robideau Mr D Palmer Mr P Wilkins MMrr MM HH BBeeww Mrs R M Brookes 2 copies 1 copy 1 copy 11 ccooppyy 1 copy 3 Environmental Environmental LLaabboorraattoorryy Proje Project NNuummbbeerr UU22772233 Page sors Page 3 of 5 ---- 22773344..00000044 000000000055 ver ois MET 0438367 FFONGOR SPONSOR: [SPONSOR'S 3PONSOR' S RREEPPRREESSEENNTTAATITVIVEE:: [RLAAELONNPOCDDOCARAAATTT:CICIOOOONNPPYYOOFFOOSSFFTTTTUUHHDDEEYY,FF, IIRRNNAAAAWWLLDDAATTAA REPORT: omy CSL CSL Study Study NNuummbbeerr HHTTS5660022 SUMME33AnMMRviYCCrooornrpmpoeornratatatiiloonnLaborstory 9S9Et33nP55vaiBBurou,unssmhhMieAAnnvvnteaeenlnsuouLeteaabo5r5a1to0r6y Usa St Paul, LISA Minnesota 55106 Ms Ms Rochelle Rochelle RRaobbiiddeeaauu CSCeaennntdtraaHllutSStccoiinee,nnccYeeorLLkaa,bboYorrOaaAttoo1rryy112 UKSand Hutton, York, YO41 1LZ UK oY CSSLLSSTTUUDDYY NNUUMMBBEERR:: TSE i T TEST IITTEEMM:: 4 HT5602 PPFeerrOffllSuuoo)rrooooccttaanneessuullffoonnastte,, PPootaassssiiuumm SSaalltt (PFOS) STUDY: STUDY: P(PePerFrffOllSuu)oo:rrooAoonccttaaanneesscuuollfrfoauolnnaattttoe,x,icPPeioottytaassssstiiuuudmmywSSiaatltht the Honey bee (PFOS):.An honey bee acute oral toxicity study with the NNOOMMIINNAALL TTEESSTT CCOONNCCEENNTTRRAATITOIONNS:S: - 0NNe9eg9gaa1tt,iivv0ee4cc5oo0nn1trrnoodl,,0PP.oo2s0si5tiiuvege ccaoionmterroell, ,4.74.875,, 22..1177,, 0.991, 0.450 and 0.205 lag a.i./bee T TEST DDAATTEESS:: RRDeaafnniggnieettiteevssett::te1s1t22:--112551-SS2ee4pptSteeemmpbtbeeermrb22e00r00200000 Definitive test: 21-24 September 2000 LENGTHOF TEST. LENGTH OF TEST: [LENGTH LENGTH OF OF EEXXPPOOSSUURREE:: 7722 hhoouurrss 4 4 hhoouurrss 3 [ES -- Ado TESFTTOORRGGAANNIVSME: FONEY BEE (ips mellifera) HONEY BEE (Apis mellifera L.) 3i gSOOUURRCCEE OOFF TTEESSTT OORRGGAANNIISSMMSS:: NNcasatttiioonnaall BBeeee UUnniitt { CSaSnLd Hutton, York YO4I 1LZ Sand Hutton, York YO41 1LZ LIFE STAGE OF ORGANISMS: Adult ORAL ORAL Lis LDso ((7722 hryhrs): [SST(OTAAOTTBIISSSTETIRCICVAAELLDLLYYEFDDFEEETTCEETRRLMMEIINVNEEELDD OORRAALL BASED ON MEAN INTAKE): NO OBSERVED EFFECT LEVEL BASED ON MEAN INTAKE): 0O(.9f4500%plaCggLaa.i0i..s/b3ee3e- 048 pg aioe) (95% CL 0.33 - 0.48 I.tg a.i./bee) 021 0.21 ug aise btg a.i./bee EEnnvviirroonnmmeennttaall LLaboraatorbyPProroojjeerccttNNauummbtbeerreUU2277y2233 PPaaggee d4ooff 55 --_-- -- 22773344..00000055 660604 VET osasass MET 0438368 -- INTRODUCTION (CCSSLL SSttuuddyy NNuummbbeerr HHTT5566002 aq This study was carried out by the Environmental R&D Team, Central Science Laboratory nt (CSL), for 3M Corporation at the CSL facility in Sand Hutton, York, UK. The tests were ccoonndduucctteedd ffrroomm 1122*"h SSeepptteemmbbeerr too 2244t*h Seepptteemmbbeerr 22000000.. TThhee pprroottooccooll,, pprroottooccooll aammeennddmmenetnsts,, raw data, all notes to file and the final report associated with this study will be retained in the CSL GLP archives for a minimum of 20 years after submission of the final report. The test item will be disposed of by CSL within two months after issue of the final report. OBJECTIVE The objective of this study was to evaluate the acute oral toxicity of Perfluorooctanesulfonate, Potassium Salt (PFOS) administered to the honey bee (Apis mellifera). EXPERIMENTAL DESIGN i allows for precise exposure over the &hour dosing period. This flees ane man poencal The oral ingestion method with administration of the test item in aqueous sucrose solution allows for precise exposure over the 4-hour dosi:ng period. This reflects one main potential route of exposure for bees and other non-target insects: oral intake of contaminated food (pollen, nectar etc.). At the initiation of the test three batches of bees in groups of 10 bees at each dose level received a single dose of the test item dissolved in sucrose. This was removed after 4 hours and replaced with untreated sucrose. To determine an approximate toxicity a range-finding test was performed. Four doses of the test item separated by a factor of ten were administered to groups of 30 young adult worker honey bees. Five geometrically spaced doses of the test item were then administered to groups of 30 young adult worker honey bees to define the LDso. The toxicity of the test item could not be accurately determined in that trial due to high levels of mortality. Therefore another test (second main test) was carried out with using a lower dose of test item (issued as a protocol amendment). Only the results of the range test and second main test (hereafter referred to as the definitive toxicity test) are reported here. All data are retained on the study file. At initiation of the test, each group of 10 bees was offered 200 ~tl of the test item in 50% w/v sucrose. After 4 hrs the test feed was removed and the dose taken by each group of bees calculated. Mortality and sub-lethal effects were assessed at 4 hrs then a further 24 and 48 hrs after removal of feeders. In the range test and definitive test an additional assessment was made at a further 72 hrs after removal of the test feeders. All doses of test item are reported as ~tg a.i. (active ingredient)/bee and have been adjusted for analysed content. At the end of the tests any remaining live bees were killed by freezing and incinerated. - Environmental Laboratory Project Number U2723 PPaaggee 5Sooff 55 22773344..00000066 _-- 0000 07 MET 0438369 # CSLStudy Number HTS602 CSL Study Number HT5602 SSttuuddyy ttiittllee CENTRAL CENTRAL SSCCIIEENNCCEE LLAABBOORRAATTOORRYY AAMMEENNDDMMEENNTT TTOO PPRROOTTOOCOL O oral cniciy study with he honeybee PPeerrfflluuoorrooooccttaannee SSuullffoonniicc AAcciidd,, PPoottaassssiiuumm SSaallt (PFFOOSS):): An acuutte oral toxicity study with the honey bee `TTeesstt ffaacciilliittyy ECECennenvvntitirrrraaoollnnSmmSecceinineetntnaacclleeRRLL&&aabbDDoorrTTaateetooarrmmyy,,.. ESESnaanvnnviddiroHHnuutmmtteteoonnntn,ta.alYYl oBBoririkko,l.looYYggOOy44GG11rro11u1Lp.Z2 {Am= endment number: 2 `SSttuuddyy nnuummbbeerr HHTTS5660022 Amendment number: 2 . Number of pages: 2 PSSPattauuuudldl yyW WiDDliilikrkrieeinncctstsor . SignatuA re M. in, Signature Date /#~.~l' (rid/ram/.) 5 Medvin Bew MMaannaaggeemmeenntt RReepprreesseennttaattiive Medwin Bew i Signature Signature _ \ ~ ~ fRP ou ,~n DDaattee_i~y-- ~3..~2-11 ((dddd//mmmm//yyyy)) Environmental Laboratory reject Number U2723 Environmental Laboratory Project Number U2723 --- ppaaggee i1 ooff22 22773344..0000007 000000000088 verona MET 0438370 CSL CSL SSttuuddyy NNuummbbeerr HHTTS5660022, CCEENNTPTRRRAAOLTLOSSCCCOIIEELNNACCMEEELLNAADBBMOOERRNAATTTOORRYY PROTOCOL AMENDMENT AAmmeennddmmeenntt NNoo 22 ttoo PPrroottooccooll HHTTS5660022 ccAAhhfftaateennrrggececlddlaarrffiirrffoioimccmaattPPiioeeornrnffblbluyuyoottrrhhooeeooccSSttppaaoonnnneessoSSoruur,l,lffiioottnnwwiiaccassAAcrrceieiqdqd,uu,eePPssoottteetaaddtsshtsshaiiuatumttmthhSSeeaalTlTttee(s(sPtPtFFItOtOeeSSmm)) nnaammee ttoo bbee cPPheerarffnllguueoodrroofoorccottmaa:nneessuullffoonnaattee,, PPoottaassssiiuumm SSaalltt ((PPFF'OOSS)) aanndd ccoonnsseeqquueennttllyy tthhee ssttuuddyy ttiitllee changed from: {i hPPoeernrffellyuuoobrreooeooccttaannee SSuullffoonniicc AAcciidd,, PPoottaassssiiuumm SSaaltt ((PPFFOOSS)):: AAnn saccuuttee oorraall ttooxxiicciittyy ssttuuddyy wwiitthh tthhee honey bee tO i PPheoernrfueloyuroboreooeco.tcatnaenseusluflfoonnaattee,, PPoottaassssiiuumm SSaalltt ((PPFFOOSS))::AAnnaaccuuttee ooraall ttooxxiicciittyy ssttuuddyy wwiitthh thhee honey bee. "TThhiiss hhaass nnoo iimmppaacctt oonn tthhee SSttuuddyy CCiirrccuullaattiioonn lliisstt ``SSSpppooonnnsssooorrr'''sss RMReoepnpriretesoseernnttaattiivvee SSSttpuuoddnyysoDDiri'rrseecMcttooorrnitor CCCSSSLLL,MMQaaAnnaaUggneeimmteenntt RReepprreesseennttaattiivvee CSL, QA Unit MMMrssRDR RPRaoolbbmiieddree,aauuW,i,l33dMMlife. Intemational M Mrr M Mrr PDP M W WPBialeillkwmki,inensrCs,,,SWCCLSSilLLdlEEifnnevvIiinrrtooennrmmneaentnitotaanllalRR&&DD M MMrrrssMRR BBBerrwoooo,kkCeesSs,,LCCSSLL Environmental Environmental LLaabboorraattoorryy PPrroojjeecctt NNuummbbeerr UU22772233 ppaaggee 22 ooff22 --_-- a 22773344..00000088 000~00000099 MET MET 0043388337711 Wi....iglldao lliiffeeo IInntstmeeo rrunnraaottiiToonneaasll,s,auLLctetdds.. EC;OTOXICOLOGY & ANALYTICAL TESTING SERVICES ANALYSIS FOR PFOSIN AQUEOUS AND SUCROSE SOLUSUEDBTYCIENTROAL NSCIESNCE ANALYSIS FOR PLFOABSOINRAATQOUREIOESUSFAORNDCOSNUDCURCOTSIENSGOLHUOTNIEOYNSBEUESETDESBTYINCGENTRAL SCIENCE LABORATORIES FOR CONDUCTING HONEY BEE TESTING Enclose piss find a ata summary fo ursre ox FOSin peo an sucroseoutons Enclosed please find a data summary for our screen for PFOS in aqueous and sucrose solutions employed by Central Science Laboratey in the honeybee studies (Sample IDs HTSG0L02 Water and employed by Central Science Laboratory in the honeybee studies (Sample IDs HT5601/02 Water and HISGOU02 Sucrose), Included are the ret able, calbration curve, ad sundardsample. fon HT5601/02 Sucrose). Included are the results table, calibration curve, and standard/sample ion chromatogra for theecens performed omour triple quadrupleLOMSIMS system.As cbaesnea fn chromatograms for the screens performed on our triple quadrupole LC/MS/MS system. As can b~ seen in hedata,the sampleswer es than the lit of quattaion, 0.0500mg a. (000500mg a1.l the data, the samples were less than the limit of quantitation, 0.0500 mg a.i./L (0.000500 mg a.i./L low standard x sample dilution factor (100). The cronwasconducted on October 26, 2000, Thee ow standard x sample dilution factor (100)). The scr~n was conducted on October 26, 2000. Thes~ results apply solutions rprsenoaf htosieuvseed nthe llowing sis els apply to solutions represemative of those used in the following studies. PPEERRFFLLUUOORROOOOCCTTAANNEESSUULLFFOONNAATTEE,, PPOOTTAASSSSIIUUMM SSAALT (PPFFOOSS):): An AAccuutte CCoonnttaacctt `TTooxxiicciittyy SSttuuddyy ~wviitthh tthhee HHoonney BBee:c ((CCSSL Stuudy NNuummbberer.: HHTTS5660011)) PERFLUOROOCTANESULFONATE, POTASSIUM SALT (FOS): An Ace Oral PERFLUOROOCTANESULFONATE, POTASSIUM SALT (PFOS): An Acute Oral Toxicity Study withthe Honey Bee (CSL Sucy amber T5602) Toxicity Study with the Honey Bee (CSL Study Ntunber: HT5602) [S=cien Scientist av Van Hoven, PhD. Van Haven, Ph.D. _DATEalslo DATE MANAGEMENT MANAGEMENT: sndTone WWDiiriellcllatroadrr,BBdA..mNNaiixxlooinne,, PPChhh.eI~m.isty Director, Analytical Chemistry Yi DATE DATE 5598 Commerce Ove Eton.Monyond21601 +Tol 410.6Fa74210-.22806 60im0o acolos 8598 Commerce Drive Easton, Maryland 21601 . Tel. 410-822-8600 Fax 410-822:-0632 E-mail: ecotox@wildlifeinfemafional.com eee Oe Behm 73 War To earaL311753756 S355 81 m@wciocmlsaontoomnacitoonmsmoiecnom European Office: Bergkampweg 1 . 7231 CL Wamsveld The Netherlands Tel. 31 (0) 575-573848 ~, Fax 31 (0J575-574813 E-mail: aleopold@worldonline.nl _ reese. ` Sosa IlVooET osasar2 MET 0438372 22773344..00000099 : PaPagge~ 11 ooff,44 gaz yes HEE Faiz 3382 SHEE fran ii fs oes seme i ressagggs i Ii iI1] il i flill Waill HH RiEi3Hi33idini ts lull i 22773344..00001100 OOCCFFEE. RRTTTHHTIIEEFFIIOEOERDRDIIGCGIOINNPAAYLL ll. Sy DR pr ss Se] 7 MET 0438373 Lo Micon wn 6 MacQuan, version 1.6 PPagae 22g0oeff 44 Fras To 03 30 a0 250 Pdnted: Thu, Oct 26, 2000 12:58 CCooirn PE0G84FES 0500 MO a: cos DA3P00PRUEGT OATAS4G.01 4541015000: Pe Cali'l:~"MJon Rle: Cal PFOS 102600 MD Path: Macintosh HD:API3000:PRO~ECT DATA;454C-t01:454C101_102600 MD: 0101 as Oo Sons os wi sr rea Comments: PFOS: 454C-101: Method Development - Screening fish flake, water, sucrose -(LC/MS/MS-MRM) T 705 199.0269.1 Ho ana ris E Tl SSs PFC6 499.0->99.1 No Internal Standard ns 7 Weighted (l/x) ovrcont < 323.17 Intercept = 323.17 Sone Paros Slope = 8193.32 Tio on. = 055040 Correlation Coeff. = 0.99949 owArea ano42000 sao36000. 200030000" 240024000- 100018000- 120012000- eno600(: ook Tia The zGhone gaeaene Take TE ike "OCFERTTHIEFIOERDICGIONPAYL eT ---- es anpe --- --_-- 22773344..00001111 000000001122 rowan MET 0438374 . : . MMacacQQuuaann,,vveers~ioonn 11.66. Pin:To, 0a30, 200 1269 Pdnt~d: Thu, Oct 26, 20~0 12:59 Page 3of4 Paasgee 3 of 4 CGubaanmFPiRaO.ui4tF6OS 102500 pa: MacoHAP PRUECTDATAS40-104SUHO1_28M0D0: C~llbra~on File: CaJ PFOS 102600 MD Pat~: Madntosh HD:API3000:PROJECT DATA:454Co101 ;454C101_102600 MD: 41,01 Maho Doran Sevonebsrcsot ros ie Commenls: PFOS: 454C-101: Method Development - Screening fish flake, water, sucrose -(LC/MStMS-MRM} P_--osy DOM uA Tok28 20 1030 O00 ---- PFOS_I STD 0.500 ug e.IJL 4675A-011D-6 49901patos 4.98 in 1 pedod jha ra -- ent 400cu No Internal Standard " Use Area :497 rw250sans . 1:4.97 MRM, 298 scans Lanne 409.0->99.1 Nat 20 Noise Three. 2.0 oS hm i - Quant Three. 1.0 F Min. Width 1 2 mh Mull Width 10 Base. Width 40 Abn 10 5 RT Win. (secs) Soo 3 : ,.Smooth Stim in Expected RT 10 2 3.70 anes 5 Area 4231.52 fox "i Height 303.9O Sh a0 2 3.50 Eteionwan 3b37o > = Integration Width Ew = 1121316s 3 7 Retentk~ "Rme login Toe 4-58 h te te e Bh Hh h Hh e Wl Te hm Integration Type 4.17 0.$7 3,70 A- 66 Thu, Oct 26, 2000 10:34 100" 9080" 70" 6O5O" 4030 20 10 0 intensity: 4000 cps 4'1 0.69 221 76 112 143165 193 A 270 1.36 2.03 2.70 3.37 4.04 4.71 ~me POSE Sotmugain 44P68F77O55SAA_-o20011110D--77STD 1.00 _ug e,IJL 13001prea 4.98 ~ 1 pedod =Sanasun No Internal Standard jkrUse Area 5 437 rw20sa 1:4.97 MRM, 298 scans Lae 498.0->99.1 rR zo Noise Three. 2.0 Sah 10 Quant Three. 1.0 wha Min. Width 12 mh Muff. Width 1o fli 4p Base. width 40 Wine 10 RT W~n. (secs) Tame 3 Smooth Epeasamr 3m Expected RT I 0 2 3.70 oaA~ea 8421.33 SSHutenaitrgthTTtiimmee: 746.57 Sogeatin via Integration Width RP~eintinon yTiimne mnTe Integration Type 33.4477 i4.14 3310.67 3 3.69 A753 A - BB Th,On a 00 "l'hu, Oct 26, 2000 10:40 0Oao>70 hs60 50 :40 a30 :220 10 . 14 sty: oto ox Intensity: 4000 cps 2=20 tteteT 1 OTe a 1a 1 38 4'1 e He Hh Wh WN 0.69 75 116 156 ,1,'~9"1~ r' 0'I ' 1:~1 ' 1(~1 1,36 2.03 2.70 201 ' 3.37 22~4.11r~ 4.04 Thm 281 Sean 4.71 Time "OCFERTTHIEFIOEERDDI"CGOIONPpAYyL OF THE ORIGINAL -- ---- HESEET). | nessmommamse. --_-- 22773344..0000112 o0o0o0o01_33 VET osasars MET 0438375 `MMa~ccQQuuaann,,vveerrssiioonn11..66. PPagae 44goofef44 Prva Tu,Oct28, 200 1250 Pdnted: Thu, Oct 26, 2000 12:59 CuFlf:GltPFOS1o250n 0MO Pa MacoHOAPBOOLPROJECT DATAASAC-10ASICIN. 10240Wo: Calibration File: Cal PFOS 102600 MD Palh: Macintosh HD:APi3000:PROJECT DATA:454C-101:454C101_102600 MD: ont POS 64G101 Mead evinced inTare, a, rt CHAS) Comments: PFOS: 454C-101: Method Development - Screening fish flake, water, sucrose -(LC/MS/MS.MRM) Prose PFO~_9 wn WATER 454C-101 - 4stintputea 4.98 in 1 pedod oeR:OS Ro otrasacar No Intemal Stan0ard fs Use Area, 47 ue,20 sears 1:4.97 MRM, 298 scans Lise 499.0->99.1 freA Noise Thres. 2.0 Ste 30 Quant Thres. 1.0 frat Min. Wldth 1 2 Wan 13 Muir. Width 1 0 Elm Base. Width 4 0 Ar S`RrsTmeoWoeitnh. = (secs) EgeciedRT 3221.070 Expected RT 3.70 ao Area 0.0 bo Co StHSatersitrgthTTtiimmee. 0.0 00.0000 ogi wan a0 in End Time Integration Width ree' Retention Time %0.00 O.00 0.00 Integration Type Tomou naz Thu, Oct 26, 2000 11:22 "100.90a80 -7 0 :60 5 aA40 b220 1 tintoennsyity::44000000ccppss the th Yh 3388_6666 110055 11: 38 Th1666 PHT 118809 221177eWo TE 220686 OSI whom PFOS_IO jaca 454C-101- SUCFIO~ stint parca ms 4.g8 in 1 pedod uma sans NO Internal Standard a Use Area : 47 250sans t: 4.97 MRM. 298 scans {onan 49g.0->gg.1 ost 20 Noise Thres. 2.0 san 12 Quant Thres. 1.5 fr Min. Width 1 2 wm 1 Mull Width 1 0 fa Vian 40 Base. Width 40 Rio 10 RT Win. (secs) 1 0 ,EESxxmppoeeocttchedtAeRdTT 332.7700 nanos Area 389.06 oan *Wi% StHSatearirg~thTTtiimmae. 41.96 335.500 Er i End Time 3.77 ngtooWan 037 Integration Width 0,27 DeonionTies "358 Retention Time InIntteeggrraattiioonnTTyyppee 2 3.59 A* - BB 0a20,200 1128 Thu, Ocl 26, 2000 11:28 -10090o80.7060E5 0a40- 30- RB io 40 E2B0- 1 0- TAT 10 4_0 O 00.6699 ney: intensity: 14000000 1cps 00411 tea17y gue seeses TART 9.._~_01 I~I 13 203 dy 270 B12 337 404 2 sen 471Time 1.36 2.03 "OCFERTTHIEFIOERDICGOINPAYL -- E-- HES) SY PECEED. mm A ia AUDIT A 22773344..00001133 00O000001144 TT MMEET TT o0a4s3e8a3r7e6 [A 5=yd 7he 7 I EN i1 7 i king BEY J J a= i > i i = 7 1 A ii , fo Ay i, a5= aa mnERace TITLE PAGE CCSSLL SSttuuddyy NNuummbbeerr HHTTS5660022 GL PPAEERNRFFALLCUUUOOTRREOOOOOCRCTATALANSSNTTETEOUSUSXDUDUILYYCLFIFNNOTOUUNYNMMAASBTBTTEEEUER,R,D::PPYHHOOTWTTTSI5AA66TSS00HSS22IITUUHMMESSHAAOLLNTTE((YPPFRBOOSwS)):: AN ACUTE ORAL TOXICITY STUDY WITH THE HONEY BEE REPORT REPORT EPPO GUIDELINE 170 EPPO GUIDELINE 170 OOEECCDD GGUUIIDDEELLIINNEE 221133 AUTHOR: AUTHOR: Paul Wilkins Paul Wilkins STUDY INITIATION DATE: 11 September 2000 STUDY INITIATION DATE: 11 September 2000 STUDY STUDY RREEPPOORRTT DDAATTEE:: 2233 M Maarrcchh 22000011 SUBMITTED TO: SUBMITTED TO: 3MCorporation 3M Corporation EEnnvviirroonnmmeennttaall LLaabboorraattoorryy 935 Bush Avenue 935 Bush Avenue StPaul, Minnesota 55106 Usa St Paul, Minnesota 55106 USA Perting Lata Performing Laboratory: EnEEvnnivvriiorrnoomnnemmnetenantltaalBliRRo&l&oDDgyTTGeeraaommup EnCCevenintrtorranalmlSSSeaccnniitdeeannlHccBueetioLtLlaoaobnbgooyrraaGttoorroryyup YorkSYanOdSIHuIttLoZn UK York YO41 1LZ, UK Report Report NNoo 11 ooff 66 Environmental Environmental Laboratory Laboratory Project Project NNuummbbeerr UU22772233 Page Page 1 1 oorf 2255 -- --_-- 22773344..0000'1144 000000001155 VET osasar MET 0438377 re [iId - - -- 7 [ind |i | iElde i: iv i ind ri ie ing ie -- le A i dese (CCSSLL SSttuuddyy NNuummbbeerr HHTT5566002 SL GOOD GOOD LABORATORY LABORATORY PPRRAACCTTIICCEE CCOOMMPPLLIIAANNCCEE SSTTAATTEEMMEENNTT `SSttuuddyy nnuummbbeerr:: HHTT55660022 honey bee RReeppoorrtt ttiittllee:: PPeerrfflluuoorrooooccttaanneessuullffoonnaattee,, Poottassssiiuumm SSaalltt ((PPFOOSS):): AAnn aaccuuttee oorraall ttooxxiicciittyy ssttuuddyy wwiitthh tthhee honey bee cddI1,ahatetttahamheeiggceeuuannnnlededrereaaarnrtstaseeiliddgygsnniaaeesrrdd.ee,, vvTddaaleheliccieddl.laa.rrrAAeeepssotthrhssatatttatatfttteuehhdldeelyiinoonbabjttnjhheedeeccttpapiivrcvrooecetusstoorcllacaotoiilelddlnnyddoooorwwssoaannlmmepiipnlnsleettsshthehooeffpprdpdrooroottossoociiccnenoogdgllussrwwooeelsleururtteeuiiosoaanenccdsshhiiwwaeeenvevrdreeeeddtcchaaooennlllddlereccttwhttheaeadtdtdttfafhhooteerr eerste chemical generated inthe above study. analysis. The report in the above study. fully and accurately reflects the procedures used and the raw data `This This study study was was conducted conducted in in accordance accordance wwiitthh tthhee UUKK GGLLPP RReegguullaattiioonnss SSTI 11999999 ((NNoo.. 33001166).). TThheessee epultions are in accordance with the Organisation for Economie Cooperation and Development regulations are in accordance with the Organisation for Economic Co-operation and Development (OECD) Principe of Good Laboratory Practice (1997 ENVIMCICHEM 98) 170 bom (OECD) Principles of Good Laboratory Practice (11997 ENV/MC/CHEM (98) 17). It is my u`unnddeerrssttaannddiinngg tthhaatt tthhee UUKK aanndd OOEECCDD rreegguullaattiioonnss mmeeeett tthhee ffoolllloowwiinngg sttaanndaarrdd:: JJPaarppoaadunncMMtiinioninissBtturryryoeoaffuA,Agg(rTrciockuuyllotuu,rre,1,98FF4oo)r.reessttrryy aanndd FFiisshheerriieess,, 5599 NNoohhSSaann,, NNootitiffiiccaattiioonn NNoo.. 33885500,, AAggririccuullttuurraall Production Bureau, (Tokyo, 1984). acTcThhhhwaaeerraadcactttteeteessrtrtaiirzzieaaetettimimbooenniwnwwwgaaaasssmsappinneenoorrtttffaooirccrnmhhmeaaederrddaaiccanattcceehccrreooiizrzredde3didiMnnggiiannttroochaa33iccvMMcceoso.SrrSdtdtaaaaTnnnnhdcdceaeearrSddwwpiOOoitptnhpheseroraffartuutlilihllnnaggsGGPPaLLrrtooPPecceedccddoouutmmrrhpeetplssliitaaaahnnnneddcceteMMe;;setethhththoooeowwddmeses,ev,ireaar,nnb,dedittaanhhllgeell ccrraehhwceahmraidarcacaattcaleteraarirnieazzleebyddseiinsiin.ng accordance maintained accordance with in the with GLP. No samples 3M archives. The GLP. No samples of the dosing solations were Sponsor has stated that the test of the dosing solutions were collected fos item is being collected for chemical analysis. Air'. Signed: sss: Due_23030) (@armmiyy) (dd/mm/yy) Envionmental R&D Team, CSL Study Director MMrr PPWWililkkiinnss Study Director Environmental R&D Team, CSL Environmental Environmental Laboratory Laboratory PPrroojjeecctt NNuummbbeerr UU22772233 Page20f25 Page 2 of 25 --------------c TT 22773344..00001155 000000001166 MMEETT 00443388337788 -3 a i! 3 ? | h| d Id > |iJ i li e J DP i-- + i? i. Rl > i. *i1 e 3| . CSL Study Number HTS602 CSL Study Number HT5602 QUALITY ASSURANCE STATEMENTS QUALITY ASSURANCE STATEMENTS QQuuaalliittyy aassssuurraannccee iinnssppeeccttiioonnss ooff tthhiiss sstuuddyy wweerree mmaaddee oonn tthhee ffoolllloowwiinngg ddaatteess:: DaDtatee Activity Activity 0088/100991/0000 RR11aa/nn0gg9ee/0tt0eesstt 1209000 11/09/00 12/09/00 1144/00990/0000 VVeerriiffiiccaattiioonnofofststuuddyy ppllaann PDDriiesspppaeernnasstiiinnoggno((ftteedssittliuittteeimmoanansndd ttooxxiicc rreeffeerreennccee)) ADPDsoroessspeiiasnnrsggamtbbeioeeneentsssof dilutions AAsssseessssmmeennttss Assessments 1809/00 MMaaiinn tteesstt 18/09/00 00900 20/09100 DDoossiinngg bbeeeess Assessments Assessments 200900 2DDe0ef0fii9nn/iit0tii0vvee 20/09/00 tteesstt DPDriiesspppaeernnasstiiinnoggno((ttfeessdttiilittueeimmonaasnndd ttooxxiicc rreeffeerreennccee)) 21/09/00 PDADsoroessspeiiasnnrsggamtbbeieoeneentsssof dilutions 200900 AAWWsesseiseigegsshhssiimmnneggenntffteseseeddeerrss 22/09/00 Assessments DDaattee rreeppoorrtteedd 0088//0099//0000 111555///000999///000000 15/09/00 1155//0099//0000 2211/00990/0000 22111/0099/0000 222555///000999//000000 25/09/00 2255/00990/0000 walla Sek pie 28sale mm MMrrss CCVVWWalaklkeerr QCQeAAntAAauluddiSitctooirrence Laboratory Central Science Laboratory "TThhiiss rreeppoorrtt hhaass bbeeeenn aauuddiitteedd bbyy tthhee QQuuaalliittyy AAssssuurraannccee UUnniitt ooff thhee CCeennttrraall SScciieennccee LLaabboorarattoorryy.. SiSgined. Ca 2st Saker... DDautee....~2...3~..l..8C~3ch]]..S~L.1.... a(ddt/mmm/y)y) QA Auditor MMrrss CC VV WWaallkkeerr QA Auditor CCeennttrraall SScciieennccee LLaabboorraattoorryy avie Ly i-- Environmental Laboratory Project Number U2723 Environmental Laboratory Project Number U2723 PPaaggee 330of2255 AE--bo mem 22773344..00001166 0O0000001t77 VET 0430379 MET 0438379 ad sn i i i i : = i - = - ad a ----- B=) Co Tr TM Thre. Hiy = SSPPOONNSSOORR:: 33MM CCoorrppoorraattiioonn CSL CSL Study Study NNuummbbeerr HHTTS5660022 RREEPPOORRTT AAPPPPRROOVVAALL TITTITLLEE:: Honey bee. PPeerrfflluuoorrooooccttaanneessuullffoonnaattee,, PPoottaassssiiuumm SSaalltt ((PPFFOOSS)):: AAnn acuutte ooral ttooxxiicciittyy ssttuuddyy wwiitthh tthhee honey bee CCSSLL SSTTUUDDYY NNUUMMBBEERR:: HHTT55660022 --_--m------ SSTTUUDDYYDDIRIREECCTTOORR:: Pllthg DDAATTEE_~2"3'~|I Oo'B~IlOo~L (ammyy) (dd/mm/yy) MEMnrvriPPrWoW inlmiklekinintnsasl R&D Team, CSL Environmental R&D Team, CSL AApppprroovvaall ffoorr RReeppoorrtt iissssuuee a MMAANNAAGGEEMMEENNTT:: I -- DATE, DATE 2etot 7..~,, "~. erl @(dddi/mmmmi/yyyy)) Head Envonmenal Biology Gra, CSL M Mrr M M HHBBeeww Head Environmental Biology Group, CSL dg I 4 Environmental Environmental Laboratory Laboratory PPrroojjeecctt NNuummbbeerr UU22772233 -------------- PPaaggee4 odoff 2255 . ees -- 22773344..0000"1177 000000001188 MMEETT 00443388338800  3 +9 -= - - EJ im . ~~ i is a iy le = :TM ie As = CSL CSL Study Study NNuummbbeerr HHTTS5660022 `TTAABBLLEE OOFF CCOONNTTEENNTTSS TTIITTLLEE PPAAGEG.E........coco. -- ree - - crsre are wf GOOD LABORATORY PRACTICECOMPLIANCE STATEMENT... 2 GOOD LABORATORY PRACTICE COMPLIANCE STATEMENT ................................................. 2 QUALITY ASSURANCE STATEMENTS... 3 QUALITY ASSURANCE STATEMENTS ............................................................................................ 3 REPORT APPROVAL sss ie cmd REPORT APPROVAL............................................................................................................................ 4 TTABLAEOOFFBCCOONNL TTEENNTETSS ccs 5 SUMMARY SUMMARY . ST ............................................................................................................................................ INTRODUCTION ce SE P--. INTRODUCTION .................................................................................................................................... 8 OBJECTIVE ............................................................................................................................................. 8 EXPERIMENTAL DESIGN... cr EXPERIMENTAL DESIGN ................................................................................................................... 8 MMAATTEERRIIAALLSS AANNDD MMEETTHHODOSD..S.............................................._..--..................s..e..r...e..n..e...............se.r................ Condidurtinig tohenesst... pons wm cwmmsnmmnnn 10 TTeEst SitemS....r..r...r...s..s...s...s..s...m...s...s..s...e...m..e...r...m..m...e...t...................a.......................p.r.m ...................-- .......................... 10 ToTxoixiccrreeffeerreennccee....................s ...........-- ...................................s.....t................--...............................................1. 0 10 Conditions during the tests .................................................................................................................... 10 DDososeeprpereppararaattiioonn ............-- ...........................s...e..m...a...n...a............-- .....................................................vse............. 111! DDoossee aaddmmininisisttrraattiioonn ..........r...e..............-- .............................r..e..r..e.....................--...--..................-- ...................... 11 AIYSOFI08S. S-- 12 ObOsebrsevravtaitoionsn.s .........e.m.s..e.s.e.s.t.s.e.s.n.......e.s ............................-- ........-- .................e.n.e..r.a.n.n..n.e.n............I .....................s.wn.e.s.se.r.so.n.s.... 1J1 Analysis of data ...................................................................................................................................... 12 RESANDUDISLCUSTSIONS... wmmm---------- RESULTS AND DISCUSSION ............................................................................................................. 12 Range test.. . - SE wml Range test 2 Definitive test COCNOCNLCULUSSIIOONNSS w------ rer-- A. 2 ron 133 Table 1. Oral LD~o of PFOS from definitive test data ........................................................................... 13 `TTaabblel2e2.. OOrraall ttooxxiicciittyyooffddiimmeetthhooaattee ttooxxiicc rreeffeerreenncceeffrroommddeeffiniitinveitettesstit dvaeta ...da.t. ................................. 13 13 EEnnvviirroonnmmeennttaallLLababoorarattoorryyPPrroojjeecctt NNuummbbeerr UU227723 Page 5 of25 Page 5 of 25 ~ 0ll 60 00 00 11. 99 I -- 22773344..00001188 er MMEETT 00443388338811  !2 CSL Study Number HTS602 CSL Study Number HT5602 [ h REFERENCES... REFERENCES 13 iI I3 PReINCIoPALsWORKEiRS..n. -- --m118 TM PRINCIPAL WORKERS ...................................................................................................................... 4 DISTRIBUTION OF REPORT ................................................................................................... 14 APPENDIX 1 (TABLES AI-A8) ......................................................................................................... 15 DDEETTAAIILLEEDD OORRAALL TTOOXXIICCIITTYYTTEESSTTRREESSUULLTTSS............................................................".....................i. s 15 ib INATPEPREINMDIC2EXRTIFICAc TE OFANALYSIs S FOR TEST s SUBTANCE.. _-- 2x3 i APPENDIX 2 ......................................................................................................................................... 3 INTERIM CERTIFICATE OF ANALYSIS FOR TEST SUBSTANCE .............................................. 23 I>| . i i z7J -4 1 --m Ze Te == he ED A = 4 000020 Environmental Laboratory Project Number U2723 Environmental Laboratory Project Number U2723 -- -- : TT WETomens PPaaggee 66 ooff 2255 000020 MET 0438382 .......... 22773344..00001199 ' iii3 i; % i Rd iii i> i la i 1 -J Ii im -1 ~ - ad EJ = iz@ =m Te = ad = FONSOR: SPONSOR: SPON`SOR'S REPRESENTATIVE: SPONSOR'S REPRESENTATIVE: LOCATION LOCATION OF OF SSTTUUDDYY,, RRAAWWDDAATTAA RANENPDDORAAT.CCOOPPYY OOFF TTHHEE FFIINNAALL REPORT: CCSSLL SSttuuddyy NNuummbbeerr HHTT55660022 SUMMARY SUMMARY 3M Corporation 3M Corporation Environmental Laboratory Environmental Laboratory 935 Bush Avenue 935 Bush Avenue SSttPPaualu,l, MMiinnnneessoottaa 5555110066 UUSSAA Ms Ms Rochelle Rochelle RRoobbiiddeeaauu Central Central SScciieennccee LLaabboorraattoorryy SSUaaKnndd HHuuttttoonn,, YYoorrkk,, YYOO4411 11L2Z UK est mew: ISSSLLSSTTUUDDYY NNUUMMBBEERR:: rEST ITEM: bron STUDY: NOOMMIINNAALLTTEESSTT CCOONNCCEENNTTRRAATTIIOONNSS:: rEESSTT DDAATTEESS:: ENGTH LENGTH OOFF TTEESSTT:: HHTTS5660022 Pertuorcosaneslfonte, Potassium Slt Perfuorooctanesulfonate, Potassium Salt ((PPFFOOSS)) A Perfuorooctanesulfonate, Potassium Salt (PFOS): An acute ora toxicity study with the h(hPooFnnOeeyySb)b:eeeAe: n acute oral toxicity study with the NN0eegga9attiivv9ee cc1oonnattr,nroodll0,,0P.Po2os0.is5tiptiivv4geeacc5oionntbtr0eroocll,, 4.7487,8,2.12.717,, 0.991, 0.450 and 0.205 lag a.i./bee DRReaafnniggneieteitesvstet::te11s22t--121551-SS2ee4ppStteeepmmtbbeeemrrb22e00r00002000 Definitive test: 21-24 September 2000 72 hours 72 hours @ SL TEESSTT OORRGGAANNIISSMM:: [SOURCE OFTEST ORGANISMS: SOURCE OF TEST ORGANISMS: [LLIIFFEE SSTTAAGGEE OOFF OORRGGAANNIISSMMSS:: HHOONNEEYY BBEEEE ((AAppiiss mmeelllliiffeerraa LL..)) =NNoattiioonnaall BBeeee UUrniict `CSSaSannLdd HHuuttttoonn,, YYoorrkk YYOO4411 11LLZZ AdAudlutlt OORRAALLLLDDsso0((7722 hhrrss)):: N[SSTOTAAOTTBIISSSTETIRCICVAAELLDLLYYEFDDFEEETTCEETRRLM MEIINVNEEELDD OORRAALL N(O BOBASOENRSVMEEEDADENFFIENCTTAKLEE)VEL (BASED ON MEAN INTAKE): (95%CL0.30.348 ugai bee) 00..4400pbgtgaa..iiiffobeeee (95% CL 0.33 - 0.48 gg a.i./bee) 021 pgaisbes 0.21 btg a.i./bee TT Page Tol 5 Page 7 of 25 -- 22773344..00002200 00~000002211 MI~EETT 00443388338833 vy 9 - ct. id > - - J - - - -Co --- . cCsSLt sStuugdyyNNuummvbeerrHtTs5e6n0e2 Jie INTRODUCTION INTRODUCTION "ST33TeMhMhpisstCCessomottrubruppdedooyryrraaw1wtt0aiiao2sosn4ncc"aaaatrtSdrettehhpideemooCCubuSStteLLrbbyyff2a0attch0hci0eeli.itEElynTnvhiivnineirrSoSopannarmonnmteddoencntHHotaulault,ltttpRoRornn&&o.,tDDoYYcoooTTrlrekeka,aa,mmmUU,e,KKnCC.de.emnTnTetthrnhrateealsl tt,eSSescscrttiiasseewnnwwccedeeerareeLL,aaccbbaooeolnnrrddaaeututcoovctrrtseeyydd((tCCfforSSoolLLmme)),,a11ff2n2oo~d+rhr tm2St2hho00eeenpyyttffeeieihnamnasaarrlblssaerrfaaeretffpprtttooeorrrirt2st4sssaatuuushsebbssSmmoooicecfiispistaassthtetiieeoemoddnnibnwweooarififltt2httrh0hettee0phh0oiiffsrs.iitnTnssaatthlulueddrryyeeppprowwoorittrilol.tll.cbboTTeelhh,eerrpeerttttoaeeitissonnttceeiioddttleeinmnamtmthwwheeienilldlCCmSbSbeeeLLnddtGGsii,sLsLpprPPaoowssaaeerrddcdchhaooitiavfvf,eebbssayylfflooCCnrSoSLatL.emmswwiitinnoittiihhmfimiilnunuemstwawnoooodff months after issue of the final report. OBJECTIVE OBJECTIVE The abjctive of this study was to value the scut ral toxicity of Peruoronctncsufonae, PTPoohtteaassossibiujumemcStSiavaleltt ((oPPfFFOOthSSi)s) aasddtummdiinyniisswtteearrseeddtottooettvhhaeeluhhaootnneeeyythbbeeeeea((cAAuppteiiss oral toxicity mmeelllliiffeerraa)).. of Perfuorooctanesulfonate, EEXXPPEERRIIMMEENNTTAALL DDEESSIIGGNN prpTbTeerhheeceecsiiososraeeraanleeldxxiipnpnooggotsehesuseustrrtriieeoonnnooonvvmm-eeetreatthtrhhhgeooeeddt-4w-whihioniottsuhheurcrataddsdd:moomissnoiiinnirnasiggslttrppreaiaertntriitiioooaodnkdn.e.oofoTTffhhttihicesseorrtnteeeetffsatlltemeccititttneessammootneniiednen amaqqauuieneoopuuosstessnuutccirarolossreeosstooellouuttfiiooxnnpaaollsllooowrwsss mfoaoind p(opotelnletina,l nroecuttearoifceyx.posuArte ffffoooorrrr the dubdiinoeonisnesatesietaritaooiooneffnndtthfsoohueftchrttttehoheesresesett.ntteoeitstnteet-mmtttahhrrddegeiieeesstssbbooiaallntvtvscceeehhddceetsssii:nnoofofssrububaceclerereoosisnsseteiia.n.nkeggrTrTooohhuuifippsscssowwoonafaftsasm1100rrieenbbmmaeeotoeeevvssdeeaaddfttoeaeaoaafdltcceehhr(pdd44ooollshhseeoonuu,lerervsvsneeelalacnntrraddeercceerreeiietvcpvpe.ell)adda.cceaaeddAssitiwwnngtgiihltlhheee untreated sucrose. separated by a factor of en were administered to groups of 30 young adult worker honey bece. `TToo ddeetteerrmmiinnee aann aapppprrooxxiimmaattee ttooxxiicciittyy aa rraannggee--ffiinnddiinngg tteesstt wwaass ppeerrffoorrmmeedd.. FFoouurr ddoosseess ooff tthhee tteesstt separated by a factor of ten were administered to groups of 30 young adult worker honey bees. Fiveiitteemm Five dw`gwgeoeoetroorekmkmreeeemtrrtirrinihhcceoaoadlnnlllieeyynyyhssbbppaeeaaeeccsiseeddattoodddooddusseeeeefsfsii0nnooeefif gtttthhhhheeee lttLeLeevDssettDlss0ii.totefemTTmmhhoewewraeelrttriooeetxxyiitt.cchhiieettynnyThaaeooddfrfmmeifttnihonhierisestttteeaeerrsnsetetoddtithtetteeoormmtggerrcctoooouuu(upplslssddecooonnffnoodtt3300mbbaeeyyioonaauucc1nnccsgguur)raaaatddtweeuualllleyytt cctdtHhehaaereemrterei.irrreeamaAddnnliggnoloeeueudtdtteaetswwisatnitittataahhrhnneauuddtrsseitssinrteeigancaciolgonanndeddllduooeowmmwnaeteaorirtinnhhddeiootgeessshsestttldooe((yffvhehteefelirslteseet.aorfiffttAeeeemtmrmoirrre((netiffaisseelstriuerrtiyeeedod.ddna03Ttoosahfaaeptsrrhpeoettrfhototheotreoecsddc,oaeoelfenfliiaonnaactimmihhtteieievngvrnedretdmoetmteusoxntpxeti(nio)scctfe.i)ht.cyy1oOO0n1tnbed0nlse5ytm0)sthahaaiweerneaeersterereslaseuptpe)losotrrswrteeeoaodddsff 200 4 f the est tem in 505% wv sucrose. Afe4 hrs he test fed was removed and the dos taken here. All data are retained on the study file. At initiation of the test, each group of I0 bees was offered 200 lal of the test item in 50% w/v sucrose. After 4 hrs the test feed was removed and the dose taken further 24 and bbyy eeaacchh ggrroouupp further 24 and 48ooff 48 his ater removal of feeders. In the range test and defntve Let bbeeeess ccaallccuullaatteedd.. MMorotratalliittyy aanndd ssuubb--lleetthhaall eeffffeeccttss wweerree aasssseesssseedd aatt hrs after removal of feeders. In the range test and definitive test aa44nn additions) hhrrss tthheenn aa additional h`praeasoepssrseotetsresstesesmdmsdeeaanann5tstyu~wwtrgagaessmaa.a.mmii.aan((ddiaaeencctgtaaiivtltvieeaaveiifnfnubugergrrttseehhsddeeiwrrieene77rt22n)eb/hhbkerreisselelaaeaaffdnntteddebrryhhrrafeaevrmmeeeooevbbviaeeanleelgnnooafaafndddttjhjhuueiessnttttceeeeidsdsntteffrffooaeerrteeeaddadennerarassll.y.ysseAAedldlllccoddonoontssteeeensnts.t.ooffAAtteetssthttheeiitteeeemnnmddaarooreeff the tests any remaining live bees were killed by freezing and incinerated. i= i J c=. a de i a Environmental Laboratory Project Number U2723 Environmental Laboratory Project Number U2723 PaPgageeB8ooff2255 rrr re 22773344..00002211 TT 000022 000022 weroMET 0438384 - ' ii `TThhee nnoommiinnaall ddoosseess aaddmmininiisstteerreedd aarree sshhoowwnn bbeellooww.. `B. areBacainngorst te doses Range finding oral test dose levels ; r rrionse Dose group PFOS dy Number (CCSSLL SSttuuddyy NNuummbebrer HHTT55660022 Treienoe Dimethoate a) SL i CocoS naing 5| 3 one| oman [ 1 T (lag a1e.0is.3/bee1 ) (G ~g0a.0 6i/0bee) 2 1 Cs 3 6 e11.00s.7 33 o o00s..13i 50 s]| 4 0.103 0.075 Control I 50% w/v sucrose 50% w/v sucrose > -9 3 -3 _- - - - :J - ) CoCnotntrroall2z containing 5% acetone containing |s5o 0% wm /v suw crosei|vs TTrriittn w ooNn/AXX-a m -11000 e ] Dente est ds nt Definitive test dose levels Dose group PFOS Dimethoate Gaines O 1 T ((a ~ptgg 4aa..w .7ii.8f/bbeeeeT )) oe (llgg0aa.i6i//0bbee)) F F 23 T oos02a.9e.1971 4 0.30 oi0.1s5 oos | co Fs CCoonntt5rrooT ll 11 o 5500%% wws 00i./.24vv05T ss50uuccrroossee. --50-- % w0i.0v7s5ucrose 50% w/v sucrose ming ne | omen containing 5% acetone containing (CC ontrol 2o |n So50f%wwi/svwsuocrroosee | TTrriittoon XX--1100 -- w N/Aa] Emitsof Da vhs were clutter st, The herd ct els we trol? Estimates of LDs0 values were calculated for the main oral test. The no observed effect levels were e ddeetteerrmmiinneeddbbyy aa ssttatissttiiccaall evvaalluuaattiioonn ooff tthhee mmoortratalliittyy ddaattaa,. oremacltianig ononrmrailyvoatlhecteas iemnd4ur0in thewso nh pms E NNeeggaattiivve ccoonnttrroollss,, ssoollvveenntt ccoonnttroollss ((55%% aaccetoonnee)) aandd ppoossiittiivvee coonnttrrooll g`grroouuppss ((ddiimmeetthhooaattee aass BBAASSFF 4400 bode 1 BASE40 formulation nominally containing 400 g/l) were maintained concurrently to check that the bees were tad reacting normally to the test item during the studies. ed - TB 1 TO *2 Le MMAATTEERRIIAALLSS AANNDD M MEETTHHOODDSS cof lonhPtec n EO Goin 1 n TThhee mmeetthhooddss,, ssppeecciieess uusedd aandd rroouuttee off addmmininisisttrraattiioonn ddeessccrriibbeedd iinn tthhiiss pprroottooccooll aarree bbaasseedd uuppoonn pprroo-C on rods on Hoes ec soo oeni rol bs gonp cedures specified in EPPO Guideline 170, Guideline on Test Methods for Evaluating the Side-Effects rouprs.oNno atTesr p nner ome ew snhlo retin he Sik fr of Plant Protection Products on Honey Bees; and the OECD Test Guideline 213, Honeybees, Acute otential soures of bias ae expected 1 n 21%Hore, t c Oral Toxicity Test. In order to control bias, bees were impartially distributed to treatment and control 0 ae the ress of t e groups. No other potential sources of bias are expected to affect the results of the study. The study ewanhe OFC nt 1 108 se ne study. The ny wwaass ccoonnssiideerreedd vvaallid wwiitthh:: aa) ccoonnttrrooll mmoortratalliittyy ooff 1100%%, oorr lleessss;; aanndd bb)) tthhee ccaallccuullaatteedd ddiimmeetthhooaattee LLDDs0 fell within the OECD standard, i.e. 0.10 - 0.35 ~g ai/bee. | .ilx= e a EEnnvviirroonnmmeennttaall LLaabboorraattoorryy PPrroojjeecctt NNuummbbeerr UU22772233 --_-- Pagesors Page 9 of 25 2277344..0002222 ~ 000000002233 veronss MET 0438385 :. i i `le ii | = | li e 1 i i || RE| J 11 . i iim |i -i -ig = _-- = - C- m C- c. =. ) = ee le ay ER TTeesstt iitteemm (CSL CSL SSttuuddyy NNuummbbeerr HHTTS5660022 gl sTTahlhlee, CPPAFFSOOSSNu((mIIUbUePPrAACC219NN5a-a3mm9e-e3,11--OwOchctitaatnnecessupulolffwoodnneiicrc,aaccliioddt,, Isanltte, rCnaAtiSonNalumLibd.er o2n79b5e-h3a%lf3,ofw3hiMteCopropwordaetri,onl.ot 221111,177,),)2,12uu,ss32ee,3dd.,4ii,2n4n,53tt,h5hi3i,6ss,46ss,t7t4uu,7dd,5yy8,8.ww8,8aa5-hsshee,ppstsautu6adppdepp6ecllcaiifa,eelfddluu1oorbbm,oyy--p7poWWoti8talailsdsd.slsiliiui8fufmeem Each sample of the material tested in this study was utnInhnietieqgsrunpeeoalnltyisoollnraa,abbleaeLltllete1ddd6. :wwo2ni0ithbCtethhhhpeearlTTfieeososfttr03eIMtuesmem.CnnouurpmmobbreaertrioNNnBB.UU1E10a05c5h.. The tet er sample of the The test item ws ord,as material tested was stored, as rrieencctoohmimsmmseetnundddeyeddwbbayys the sponsor, at 16-20 C prior to use. (PPaFFpOOprSSoxwwiamaassteddliiysspp5ee0rr0sseemddg/lii)nn. aaAcceedttoiosnnpeeer((saainnoaanllyyttetiiscctaawllasggrrcaaaddreer))iebbdeeoffuootrr,eebeuufssoeeredduutehee ttoo aa llooww ssoolluubbiilliittyy iinn wwaatteerr toxicity tests were performed. (iinanpwwphrhoiixcchhimPPaFtFeOOlySS5ww0a0assmddgii/1ssp)p.eeArrsseedddispiinneraasccioeenttoonnteeestaawtt atthsheecaaarppripperrdooxxoiiummta,attbeeefccooornneccetehnntetrrtaaottxiioiocnnityrreetqqeuusiitrsreewdd ettrooe performed, ddeelliivveerr tthhee uhuPhissiFggeeOhhddeSesttsototwedidinornosccesellesucus.ddh. eeeTTcmmhhkaeeeatdethheroorviimimaaslolougawwgelhenhlineyicecidihhttuyyrmmioaoanffyygtthbdbhieeeesmmspseuiuirxssxstpptiuueeornrnneeddteewweddasasotosrraabdsdseissifsseeoppssreessereressddteehddaae.f.fdttHeHearroyom22moofhthogoogehuuenerrnessei.t.iettIsyynt of such solutions of the this reppoorrtt ""soolluuttioonn"" iiss of such solutions of the and immediately before PuupsusFeer.O.pSoAAsllwellseosorfoleluauctdthimiooienncniskssewtwdreaevtreiiesounrr.aeel--lmTymhiidxexueedrdtienpptgrriiidootirrespmt0oefruusosisreoe.mn.edteSSosaotllscuulttbeiieoaofrnnosssreooolftfuhttteihhoeednaetyoessntot fmdditoohxsseieensstgeswwtaeiearrnheedehhioomhmmimgoohgegeedsenitnaootdueuolsssyeff,booerraffottehhrreee (pt4wtuh0orepohaHopsouepurrrsosofxttiahhmdeearrmteeeinwwciaasostsnrcaaaetinssotllnriig.aghhtTtithossneeeddrteiiemsqmtueeininrttteemdaatttfoorooporormmmeepdattereamempctpehleereraaahtrtiuusgrroheeelusaattttiodaainlccuoootnnneccdemendnitoxtrsriaanettgiiiooanntth of 86 ig &.i/l PFOS the highest dose, after of 86 pg a.i./pl PFOS tes). (the approximate concentration required to prepare the highest diluted dose in the test). TTooxxiiccrreeffeerreennccee DDiimmeetthhooaattee 37.45% wiw ttooxxiicc ssttaannddaarrdd ((NNBBUUS9O0)),, BBAASSFF 4400 lloott ai, 400 g/l nominal concentration), was 3377MM 99002200110066 ((aa bblluuee purchased from UAP lliiqquuiidd EECC ffoorrmmuullaattiioonn,, York on 14/06/00 and tww3h7aae.ss4d%ssottooorwrree/tddwoaattathi00e, --41r1e011f0rCCigg/e((loroannntoooomrnniheenaaoodlcccccbaaoessneiincooennonptttrhehaeneteimmodna)a)xx,siiumwimtuauasmbmlpettueermtmcopphemeararsaiaetntuudtrraeeifrnrroeemscctoaobrrUiddleAeiddtPywwfaYaosrsor211k44yoeCCanr--s1tthha4iic1ssc0oww6ra/ad0ssi0nafgafitnee0drr ticihnonefnfocodrermonmtaoartrtaiiottooinnotnsshuuebppapprsleleiifdeerdidgoenbbryydaatttothhraeeshmmuaapdanpnulbfuieafeecadtncubtruoyerpreBe.rAn. SedFDD).oosssueeitaaannbddleLLtDoD50mccaaailnlcctauulilnaattiisootnanbssilaairrteye fffooorrr cc2oorryrreeeaccrttseeddacddciiommredettihhnoogaattteoe concentration based on data supplied by BASF. "TtTohheenohhnootannreegyyetbbeeieens((eAAcptpsiisssmmienelcllliififeterirasa)ainiss uiusmsepefofuurltlaiinnnteepvvaaolluulaattliinngigotntfhheeavapptrooitoteoenunrtstiiaaallgrhhiaaczzuaalrrtddussraoolffcaaroggprrsii.ccuulltTtuuhrreaalrlecchhiseemmailicscaoallssa tssouuwbbanssrtotdaansnntt.atiiraaglletddaainttaasebbcaatssseesioonncetthihteeiseeffaffneecctitmoopffoaratggarrnootccphhoeemlmliiinccaaaltlossr uuoppfoovnnarbbieeoeeusss With Which agricultural with which ct0oroccpaastt.eeggToorhriiezzreee pote is also a potential hazards. WWUnoiort.rkkeeBrreehhsoonnfeeryyombbeeteehsse((cAAoppliiossnmryaele3ll2liiffweeerrraa)e) wwuesererede ootbhbtrtaaoiiunngeehddoufftrrotomhmecctooellsoosn.niieeBsseebbseellowonenggriienngegxttaoomttihhneeedCCSSpLL.riNNaoatortiiootnnhaaellsBBtaeercet otUoaffnkttiehthn.eeBhtteeaessedttsaanfnnordtdomssbhheoteohwnwenntcrttoeoolaotbbneeeydffrw3rei2eethwooffearaaevccaaaurrrsirineondea,,ctinnhdooreosseuewmgimhtaahoiaaunnntddtthhaaeemmotloeeaesbsbttasa..4. BTTwheheeeeeskccswoo.lleoorneWnyyoerxffrkaroemomrminwwbehehdeiisccphhrwiottehhrreeetobbceetoeehlsselewwscettearerreedt ttffarhrokoememmntt(hhh4eea0d-hh5iinv0voeeptebbrbyyecuueanssgieint)grneagiaantsseommdacalywlllliitaanhmmdroaoiucuvnanatltrrmooofefassschmmidoocekakegew,e,si.ggtheeninnIttnllytythhsseehhaallakkasiibtnnogg4ratttwhhoeeremeymktsffhr.reoommWmettoshhrhekccecoraomgmbebebssesswaaenwnrddeertteprralancanocssfelfeldeerrcrniitntengdog ttthhheeemiinnc(c4uu0bba-a5tto0orrp((e22r55c+1a2g2e)CC,i,n66to55 3% relative humidity) cylindrical mesh cages. + 5% relative humidity) oIn to s:tshttaearrvvlaeebttohhreeatbboeereeyss tffhooerr 1.5 10 2 hs mesh cages 1.5 to 2 hrs befor the tet. were placed into before the test. C.oCnodnidtitiioonnssdduurriinngg tthheetteessttss TmTeeesaststubbreeieenssg wwaeperprereoxhhioomuuassteeeddlyiinn9ttmeesstt iccnhhaadimmabmbeeertrsesr.wwhhiiAcchhsaamrraeelccllieenaavnne,,rewwdeelllpl evveeenidtiillsaehtedd(,,spiinpnvrveeorrsttieedmdippneeyttrrii3 ddmiisshh5eess,, `mcbcoeoeennasttsaauaiinnrnidntthghweeaassspuupiccrdrroeooxnsstieemifssaiooetlelduulyttbiiyoo9nnsctwwmuadasysinnaauffdffmiiibaxxemeerdde,tiiednnro.eesaaaccAghhe sccgmhhraaoammullbpbeiearnrn.v.derEErteeaapdcclihhcpaettteteerss.itt dccAihhsfaahemmrb(baeedprorpsccriooonxnngti,tmaatiianensteeetddlyca11g003ewwsmoorwlr)kekereteror bkrKeeeelppeastttiiiavnnnedddhaauwrrmkkainnsdeeissitdssyeaanwtttii22tf5i5heid+n 2bt2hyeCCsetnuaavdnniyddro66nn5u5m%m%enbt+#earl,55%d%cohshhaauumgmmbeiedidgriirtwtoyyeu,,rpeeexxarccneeedcppottrredddpueulrdirciincanotggen.toobibsAnseuerfortvuveasratlitdiyooonndss.usin.rgiTnT, geetmemtsphteperec(araeatgstue.ursreewaaennrdde relative humidity within the environmental chamber were recorded continuously during the test. Environmental Laboratory Project Number U2723 Environmental Laboratory Project Number U2723 PPaaggee 1100.o0ff2255 TT TT 22773344..00002233 000000002244 Eros ~E~'-0438386 - 9 ; CCSSLL SSttuuddyy NNuummbbeerr HHTTS5660022 Gi i Brim i. i2I PJM 3r ce m m E owitmh 0m 55ma e wisirm oe.em To er eT wiecm aia ed ci ut e eam srilwithT e v aforps roo nLy ly DoDsosee pprreeppaarraattiioonn 2 Doses of the test item were made by preparing a solution of 103 ~tg a.i./~tl (range-finding test) and 47.8 ~tg a.i./pl (definitive test) PFOS and making a series of dilutions from this. Doses were prepared in acetone and then further diluted in 50% w/v sucrose for dosing. In the range test and definitive test 0.5 ml of each acetone solution was then diluted to I0 :ml with 50% w/v sucrose giving a final acetone `ccoonncceennttrraattiioonn ofof 5%. Althoouusgh this concentration of acoetsone was grer ater tha an recommeoneded in the OECD guidelines (1%) it was required to ensure the highest concentrations remained in solution for the oral test doses. One set of control bees was treated with the same level of acetone in 50% w/v sucrose (0.5 ml acetone in 10 ml sucrose) and a second set of control bees was treated with 50% w/v sucrose. Stock solutions of the positive control, dimethoate, were prepared in deionised water containing 1 g/1 T Triton n X-100. BStoeckasolutionnss oof f 2.80 ~utgg aaii//ppll ((rraanngge-ffiinnddiinngg tteest) and 3..000 ~ptgg aii/~/tll (ddeeffiiniti ive ty est) dimethoate in deionised water containing 1 g/1 Triton X-100 were prepared and a series of dilutions made in 1 g/1 Triton X-100. This series of dilutions (0.5 ml) were then further diluted to 10 ml with 50% w/v sucrose. Control bees were dosed with 0.5 ml 1 g/1 Triton X-100 in deionised water diluted to 10 ml with 50% w/v sucrose. The tests were carried out in parallel with those for PFOS. SSaammpplleess ooff tthhee ssuuccrroosse solluutition usedd in the study were analysed by Wildlife International \Ltd. and no PFOS background levels were found. 2i i I3- a 2 le Dose administration te re ri i cet ie The bees were anaesthetised with carbon dioxide irnrnediately before dosing and gently tipped out onto filter paper and counted into the petri dish cage (drones were discarded). Each group of 10 bees a te wasis ooffffeerreedd 00..22 mmll ooff aa ggiivveenn ccoonncceennttrraattiioonn ((oorr cccontrols as above),a the dose bmeimng pmroeuapsuarfed10inbtuossa small, pre-weighed, glass feeder within the cage using a variable volume pipette. This is equivalent to memLomeerre ism sstriet tMwinene 20 lul/bee. At each dose for the test item and dimethoate and for each control there were 3 replicate sme cages of 10 bees. Doses were administered within 2 hours of preparation. After 4 hours the glass feeders were removed and weighed and the sucrose feeders filled with approximately 3 ml 50% wlv aqueous sucrose so that bees had continuous access to sucrose for the Srenmgairndieruodfithae nstudyJy.. TThheeddoossee ccoonnssuummeedd wwaass detteerrmminneedd by comm parison off ththee weight of thD e di ose remaining in tl~e glass feeders with the weight of a known volume of the test solutions. = Observations MMorotratalliittyy aanndd aannyy bbeeeess kknnoocckkeedd ddoowwnn,, ii..ee.. aalliivvee bbuutt imrniommbobiille, or stumbling were assessed when the feeders were removed and 24, 48 and 72 hours after removal of the test feeders (i.e. 4, 28, 52 and 76 hours after the start of the test). = Te = Te RY -- == ---- Environ abor je Environmental Laboratory Project Number U2723 PPaaggee 1111 ooff 2255 Jr300028 2734.0024 TT 000025 Soevzs MMEETT 00443388338877 Ed ii iiI iVio i iI i1s i ii i - - i - i -1 i-g J i: e "= a zm BE Ry " Te ~~. Z=oe --- CCSSLLStSuttduuyddyy NNuuNmumbmebbreerr HHTTS56600:2HTS602 AAnnaalyslisyoofsfddiaatstaa Hogarth of dos (CSL Probit | package) A fest - quars regression (Fny 1971was fed to AAnnaallyyssiiss ooff tthhee ddaattaa ccoommpprriisseedd pplloottttiinngg pprroobbiitt mmoortrtaalliittyy rreeccoorrddeedd aafftteerr 2244,, 4488 aanndd 7722 hhoouurrss aaggaaiinnsstt tthhee tstlhohleegosspaeeer..iothfTTmotoxhxioeicfcriidtetyyosspwweoaa(nssCseeSxxcLppurrrPeeerss.ossbeeTdditoaa1ss eeslope of the response curve. The package). A least - squares Lu in kg 4.1. per bee with LDso in lag a.i. per bee with NOELS (1 cbse effet NOELs (no observed effect regression (Finney 1971) was fitted to the 95% confidence limits, NOEL and the 95% confidence limits, NOEL and enh) wee cima wig Sen's levels) were estimated using Student's t-test (p<0.05). cTTahhteeegLoLrDiDse0s uwwsedaass uuwessreeedd: ttoo ccllaassssiiffyy tthhee tii toxicity ooff tthhee tteestt itteemm aaccccoorrddiinngg 0to tthhee IICCBBBE ((11998855).). TThhee categories used were: MHHoiidggehhrllyayttteoolrxiyicc:t:oxic SMStliioigngdthhuetalrllyayltyeotolnnyxonietc-o:txoixci:c: Virtually non-toxic: ll1ee1ssss0tthphagannsi11bbgtegsaaiif/bbeeee g111r00-e1--a011t00e~00rgtilaahggia/nbaaiie/1erb0ee0pse g aes greater than 100 lag ai/bee DpDraaottagarffaoomrr (ttPhhreeobddiiitmmeIe,ivthheoorsasttieeon22444-.),. 4488-- aanndd 7722--hhoouurr mmoroarltailtiyty' rreessuulltts wweerree aannaallyyysseeeddduuuissiinnngeg tthhee CCSSLL poorporobbiitt program (Probit 1, version 4). RREESSUULLTTSS AANNDD DDIISSCCUUSSSSIIOONN TATphhpeeenrredesisuxultss1. ooFffuttthhheeeroordraealtlaietlessstsasreaarrgeeivsseuunmmbmmealraoirwis.seeddThiienn oTTraaalbbllteeossxic11itaaynnoddf 22theaanntddesddteetetaamiillereddesrrueleseaudlltsisn aaarre7e2llihisrstteeLdd Diinn value for the honeybeeof 0.40 ig a. bee. Appendix 1. Further details are given below. value for the honeybee of 0.40 ~tg a.i./bee. The oral toxicity of the test item resulted in a 72 hr LDs0 RaRnanggeetteesstt bMMoerottwratealeliinttyya wwmaeassanoobbissneetrravkveeeddofaatt1a0allllandddooss1ee0ss uffrgroomam.i/mmbeeeaea.nn Siinunbttalakeketeha11l..00efi/afggect&as..i/.wibebereeee owwbiisttehhrvaaedsstteaeeseppknddoooscseekdrroeewsspnpooannnssdee stumbliatng4 hrs at the twohighest dose levels with these individualsdying by 24 between a mean intake of 1.0 and 10 p.g a.i./bee. Sublethal effects were observed stumbling at 4 hrs at the two highest dose levels with these individuals dying by 24 hs.as knockdown hrs. and DeDfeifniintitiivveetteesstt 0`Trr.eTeh4hss0eepprrouoeenngswswaeeaaissbbJeessttiiowggwenneeiieeff(nini9cca5aaann%ttmmcmmeoeoanoarnfntrtaialdiilineinttnttyyacakkeaaeettliaaoolmlflflitdd00soo..04s4s.e55e5ss3aaaann-bbddo0ov.224ve..82a2eauplmmaiggegeaaaanna..bi..i/ie/nbbnectetea)a.e.kk.eeTohoTfTefhh00eNe..22O7711E22ulL.tghhgrrwaaaLL..isiD.D/b0bss.e0eee2ww1wwaaiisistthhgccaaaallbccssuttueeleleaaeeptpteeaddddtoo7ssaa2eess K0hHon.o4uuo0rrcsskp,dbbgaoaassw.eein.dd/baooetnneth(mme9e5ehaa%innghceiinosntttnaafdkkidoeeseddenaacltteeaavle((ilmSSattiuutsdd4ee0nnh.t3rt'3s'ss w-tLi-i0ttehe.4sttt8h/ppe,tsg<<e00a.i.0.0ni5.d5)/ib.)v.eieSSd)uuu.babTllleshettedhhyaNaillnOeegaEfffffLetececwtrtssa2sww4e0er.hreoe2u1rssseeb,eetgnn observed a.i./bee at observed as72 as knockdown at the highest dose level at 4 hrs with these individuals dying after 24 hours. TTTahhbeele22442,., M44o88rtaaannlddity7722of--hhbooeuuersr cecoxonpntotaasccettdLLtDDos~tyohvvaatloluuxeiecss rffeoofrrerddeiinmmceeetthihnooattteeh iitnnesttshheealddleeoffiwinneiidttiivcvaeelcttueelssattiaaorrnee ossfhhoaownwnnoraiinln LhTLTDhaeoDbses0slswoeeeof2rrefe.e00ssrMu..e11lloat22scrttu~assihlgghinotogywaawiinoodaadrfimitaommboleelxeyteiithschioiioetayaxtntpetellfohe/bbvvsieeeesleedltewwaatsotittitt22hhth4i4inen--hhtttooohhuuxeerrisscrraaaarngenngfeddeessr00err..n1e1ec1p1peoorlpir.tntgeegddtaahiibbeyyddttiietmhmhseeteesithOOhaoolEEalaotCCtweeDD//ebbdeegieeudcieadaallttecilu44inln88aeetssiaaonnsnsddhhooo77wvf22i-ai-hnnhnoggouourttsrrhha.seel. bees were reacting normally in this test. Environmental Laboratory Project Number U2723 Environmental Laboratory Project Number U2723 TTT PPaaggee 1122o0f2255 TTT 22773344..00002255 000000002266 oa MET 0438388 vy i . iiI e i ii| i ii g i i i i - | 1> i? tT -i i i iEaY i Z-e = .. Te Te Te =. 2=. --_ Exposure of honeybees 1 PFOS by oral dosing resulted in a stp dose response cure with CSL CSL SSttuuddyy NNuummbbeerr HHTTS5660022 CONCLUSIONS CONCLUSIONS al Exposure of honeybees to PFOS by oral dosing resulted in a steep dose response curve with significant mortality above a mean intake of 021 ig ai/bec and por significant mortality above a mean intake of 0.21 p.g a.i./bee and a 72 hr LDso calculated as 040 ug abe. The NOEL based on siaisical anslyss of the data a 72 hr Lg caloninc uy 0.40 ~tg a.i./bee. The NOEL based on statistical az~alysis of the data (Student's t test p<0.05) was 021 ugaifabt7c2hes. (Sudent' t ot pe0.08) wag 0.21 ~g a.i./bee at 72 hrs. (Th1he9es8s5ee)irreessscuulllttasssddiefememdonoassntrhasitgetthltrhhyeeaotootrxraieacllttbooyxxiiiccniigtteyysotoifofnPP.FFOOSS aass 00..4400 up.gg a.i.bFeoeeeaanndd aacccorcdiongrttoodtthhieIInCCBBgBB (1985) is classified as highly toxic by ingestion. ab1l.Oeral LD ofPFOS Table 1. Oral LD~o TiHrmse Time 0f P[ 'OS frofrommLdedfDeifninititiivveetetesstt ddata (ug LaDiss0bee) I99n55t%e%rvCCaolonfnfofiriddLeennDccee | EEssttirimemsaapttoeeonosffesslllioonppeee ooff H24rT s op (/.tg 0a..7iJ2bee) 7 Inte00r.6v6a00l--f00o.8r[ 855LDs0 sreo spo5n.s0e line 7] L 4782 T o00w ..4406 Two 0.32 s - 0.55 0.33 - 0.48 a44..87 x] Table 2 Ora toxicity of dimethoste toxic referees from definitive est Table 2 Oral toxicity, ot dimethoate toxic referer e from definitive test data "Time. Do 95% Confidence | Estiomfastpe of HTrHimsrse ug LaDdJsboee) (~tg a.iJbee) I9n5t%ervCalonfofirdLenDc,e F 2 4284 1 s oo0.nn 12 1 1 Iontooe0r.vIm-0al-eofo0r.mn1L4D 0.11 0.096 - 0.13 Toit teen 72 0.11 0.096 - 0.13 Estirmespaontsee of sllionpee of ee] response line 6.4 5s 7.3 ] 8.1 REFERENCES REFERENCES ans European for European aanndd M Meeddiitteerrrraanneeaann PPllaanntt Evaluating the Side-Effcts PPrrootteeccttiioonn OOrrggaannizizaattiioonn.. of Plant Protection Produc 11999922.. GGuuiiddeelliinnee ts on Honey Bes. oonn TTeesstt EPPO MMeetthhooddss Bulletin, for Evaluating the Side-Effects of Plant Protection Products on Honey Bees. EPPO Bulletin, 22, 203-215. Fioney, Finney, DD...JJ.. 1199771.1. SSttaatiisstiiccaall MMeetthhooddss iinn BBiioollooggiiccaall AAssssaayy,, 220ndd eedd.,, GGrriiffffiinn PPrreessss,, LLoonnddoonn,. International International Commission Commission ffoorrBBeeee BBoottaannyy ((IICCBBBB)).. 11998855.. methodsfo esting hazardof pesticides to honeybees. RReeccoommmmenenddataitioonnssffoorrhhaarrmmonoinzizaattiioonn ooff Third Symposium on harmonization of methods for methodsfor methods for testing testing testing hazard of pesticides 0 bees, England. hazard ofpesticides to honeybees. Third hazard of pesticides to bees, England. Symposium on harmonization of Organi.zation Organization ffoorr Economiic Economic CCooooppeerraattiioonn aanndd DDeevveellooppmmeenntt.. 1997. Guideline 213 Honeybecs, AAccuuttee OOrraall TTooxxiicciittyy TTeesstt.. 1997. Guideline 213 Honeybees, Environmental Environmental LLaabboorraattoorryy PPrroojjeecctt NNuummbbeerr UU22772233 i one -- PPaaggee 11330off2255 ee 22773344..00002266 0000 27 000027 VET 0430389 MET 0438389 ' ! i3. In | i i lie i ii i i I| ~ ' i -: -C: a - - Ze = Ce =] Te =. T ~e "a mm CC"SSLSLL SSttuuddyy NNuummbbeerr HHTT55660022 Sh PRINCIPAL WORKERS PRINCIPAL WORKERS STUDY MANAGEMENT STUDY MANAGEMENT Study Director VP ikin Study Director (CCSSLL MMaannaaggeemmeenntt RReepprreesseennttaattiivvee Mr P Wilkins MMrrMMHH BBeeww OPERATIONAL SUPERVISORS OPERATIONAL SUPERVISORS Principal Scien Dr HMThompson Principal Scientist Sciontat & Tot Hom Controller Miss UB Piewch Scientist& Deputy Test liem Controller Scientist & Test Item Controller Scientist & Deputy Test Item Controller ScSiceinetnitisstt Semin DI3SinRks Scientist SScciieennttiisstt Dr H M Thompson Miss C Green Miss U B Pietsch Miss S C Green MMrr AA HH BBaatttteerrssbbyy Dr R J Spinks MMrr MM JJ KKeellllyy QUALITY ASSURANCE QUALITY ASSURANCE OA Audi Q`QAA M Maannaaggeerr QA Auditor Ves CV Waker M Mrrss RR MM BBrrooookkeess Mrs C V Walker DISTRIBUTION OF REPORT DISTRIBUTION OF REPORT `SSppoonnssoorr''ss RReepprreesseennttaattiivvee Shorenr's onto SSSttpuuoddnyysoDDiri'rsreecMcttooorrnitor CO Mamsemen Repmamatve CSL Management Representative EoQA Manager MMss RR RRoobbiiddeeaauu MiDbmer MMMrrr PDP WWPialillkmkiiennrss MMHBew Mr M H Bew MRi Mbecskes Mrs R M Brookes 22ccooppiieess leony 11 ccooppyy I copy leony 1 copy copy 1 copy Environmental Laboratory Project Number U2723 Environmental Laboratory Project Number U2723 -- Page 140125 Page 14 of 25 -------- sr ---- 22773344..00002277 000000002288 --er oxsesso MET 0438390 vr 9 i[ [ng ii 1I 1s 1 i'> iim i eldn ii a| i] e 17 Ji = =J= Ce oy == Te ~~ = 9 Soa CCSSLLSStuttududydyy NNuuNummmbberebrerHHTTS5660022 Table AL Table A1. Nominal Nominal ple dosel (ng a (~g a.iJbee) DETAILAAEPPDPPOEERNNADDILIXXT1O1 X((TITACABBILLTEEYSSTAAEL1S--ATAS8R))ESULTS DETAILED ORAL TOXICITY TEST RESULTS Result Results ooff rraannggee--ffiinnddiinngg oorraall ddoossiinngg tteessttss wwiitthh PPFFOOSS - - MMorotratalilittyy AAbccttauuaasll ddioossnee'2 CCRaaeggee (~tg a.i./bee) No. Rm E h CCuummulualattiivvee nnuummbbeerr ddeeaadd ((nn==1100)) a4 bhrss 2244bhrrss 4488hhrs 7722hherss 103 103 96 96 646 s 10 10 10 110100333 9194041 4a417 5755 11110000 01100 10 11110000 103 101 48 7 10 10 10 1110003.33 111000 10310.3 10.3 1010 10 woe 1111..00003333 1111..0000 1.03 1.0 445909 0 00 111000 111000 111000 5150 51 00 0 1010 10 1010 10 1010 10 smo 0 2s 5555432 0000 00|1 02 1 1 2432 54 0 0 0 4 0.103 0.10 51 0 0 0 0 00..110033 0.103 00.1100 E5l5 010 56 0o 0 00 0 0o 0 0 0 0 0.103 0.10 56 o 0 o 0 0.103 0.10 57 0 0 o 0 oo03 00 0 355988 0 00 000 o 00 000 o03 03 000 k56s90 00 0 00 0 0 0 0 0o0 04 0 61 0 0 o 0 o 0 2 0 0 0 0 04 0 62 0 0 o 0 o 0 6 0 0 0 0 04 0 63 0 0 0 0 *bbbaeasseeddd ooonnnannmoomomiuinnnatallofccsoonntssuummspplttiaiooinnnoofcf otteensssttussmooelluudttii(oovnneoorfafg22s00p4o.1fb/be1eee0s) based on amount of test solution consumed (average of 10 bees) *undosed acetone dosed undosed Sl Environmental Laboratory Project Number U2723 Environmental Laboratory Project Number U2723 PPaaggee 1155ooff2255 erm cere-------- -- 22773344..00002288 0000000072.99 A VET 0438391 MET 0438391 . 3iI> ii |i i `eo Ii i 1 ilo iin 1 i = | i -| d>n 1i* i = J = . = ce = -- ~~. -- = CSL Study Number HTS602 CSL Study Number HT5602 y SL "Table Table AA22.. RReessuullttss ooff rraannggee--ffiinnddiinngg oorraall ddoossiinngg tteessttss wwiitthh PPFFOOSS - -- SSuubb--eletthhaall eeffffeeccttss i pia cor Nominal le, Gris a dose1 (1g 21 beo) (lag a.iJbee) Actual dosez (lag a.iJbee) Cage No. NNuummbbeerr kknnoocckkddoowwnn ((KK)) oorr ssttuummbblliinngg ((SS)) T4 hhrss ihe 24 hrs @hs 48 hrs The 72 hrs 103 103 9% 96 45 ax 0 0 0 110t00333 919%041 8a46 47 sk 145K4K 1S,4K o000o 000 0 00o o 103 101 48 1S,2K 0 0 o 111000.333 111000 949 a22s%Kk 00o 000 000 10310.3 10.3 10 st 2% 0 10 50 IS,3K o 10 51 00 00 2K o 0 0 111.000333 111.000 5552 000 00 0 000 1I0KK 1031.03 1.03 101.0 1.0 553 54 00 0 0o o 10 K 00 1K 0 ol00.1o100s33 000.111000 s35s5 0 00 00 000 000 0103 0.103 0.103 0100.10 0.10 556 57 00 0 00 0 00 0 00 0 o03 0 0 0 558 00 0 0 00 0o 0 59 0 0 0 0 003 03 00 0 &59 60 00 0 00 0 0 0 0o o o 0 6 0 0 0 0 04 0 61 0 0 0 0 o 0 0 0 0 0 04 0 62 0 0 0 0 o 0 & 0 0 0 0 04 0 63 0 0 0 o bas on ominal consumption of tes solution of 20plec based on nominal consumption of test solution of 20glfaee sed onmoun fest slo comune (verge of 105s) based on amount of test solution consumed (average of 10 bees) acetone dosed *uuanncdedotoossneeedddosed Environmental Laboratory Project Number U2723 Environmental Laboratory Project Number U2723 -- Page 160f25 Page 16 of 25 -- --- 22773344..00002299 000000003300 TTET oxez -- MET 0438392 3I bP [id i i > TM 71 iie i =f:~ -~1~ ~=i1 |~I~1~ TM TM TM ~i~ de -1 - TTaabbllee AA33.. NNoodmomiseinn'aall (ug abe) dose~ (~g a.iJbee) 000.666000 000...666000 030030030 0.30 0.30 0.30 01s000.1115s5 0.15 0.15 [rH 00.007755 000..000777555 000 00 0 CSL Stdudyy NuNmubmebrer HTS602 CSL Study Number HT5602 RReessuullttss ooff rraannggee--ffiinnddiinngg oorraall ddoossiinngg tteessttss wwiitthh ddiimmeetthhooaattee -- M Morotratalliittyy A(Aucctgtuuaaailldldoeosesee')2 CCNaaogg.ee (ttg a.iJbee) No. CCuummulualattiivvee nnuummbbeerr ddeeaadd ((nn==1lO0)) 4ahhrss 224ahmrs bs 48 hrs 7T2hhers 000.535466 23311 033 111000 111000 111000 000..665004 332 33 000 111000 1100 1100 10 10 02s 2 2 10 10 10 030 36 1 10 0 10 030 0.25 0.30 33s4 35 0 02 111000 1100 1100 10 10 0.30 36 1 10 10 10 015 o 5 3 7 000.111ss5 3377 000 488 588 588 0.15 38 0 4 5 5 0.15 39 0 5 7 7 0075 0 0 0 0 00.007745 a4l0 00 0.074 41 0 0 0 0 000 00 0 00..007744 a422 0 0 11 1 1 2 2 0 00 5a43 000 000 000 000 00 0 45 0 44 0 45 0 0 0 0 0 0 0 0 0 0 a + = ze 1~ i ? based *based ~ based 2 based oooonnnn naanmoommmooiuuinnnnatatllooccffoottnneessssuttumssmooppllituuitotoiinnoonnoofcfcooetennssststuussmmooeleluuddttii((ooaannvveeoorrffaagg22ee00!opo.tflflfoc11e00ee bbeeeess)) 2 = ==. = Ze == Environmental Laboratory Project Number U2723 Environmental Laboratory Project Number U2723 res PPaaggee 11770o1f2255 -- 22773344..00003300 TTT 000000003311 TT erossss MET 0438393 ` 2 |ba ii | i i i a ii| > iTi Te i ii - 1| i1Ed 1i = .- Te . Te . = ea Table Ad. Table A4. (vg aiJbe0) nea , NNoommiinnaall dose~ (l~g a.iJbee) CSL Study Number HT5602 Results ofrange.finding oral dosing tess wiCthSdLiSmetuthdcyaNteum--Sbube-rlHeTth5a6l0e2ffect~ Results of range-finding oral dosing tests with dimethoate - Sub-lethal effec sib) AAccttuuaall ddoossee'2 (/lg a.i./bee) No. CCaaggee No. [2 ST 060 056 3 0.60 0.56 31 0.60 0.54 32 060 060 0.60 0.60 33 030 02s 3 0.30 0.25 34 [ES 0.30 0.30 35 030 03 36 0.30 0.30 36 01s 01s 0.15 0.15 37 01s ols EY 0150.15 0.15 0150.15 0.15 338 39 00.007755 00.007755 400 0075 0074 a 0.075 0.074 41 007s oon 2 0.075 0.074 42 0 0 a 000 00 p4r3 0 44 o is 0 0 45 NNuummbbeerr kknnoocckkddoowwnn ((KK)) oorr ssttuummbblliinngg ((SS)) Tie sms @hs Tee 4 hrs 24 hrs 48 hrs 72 hrs x 0 0 0 2K 0 0 0 2K 0 0 0 o 7K 0 0 0 1K 0 o 1K 0 0 0 0 0 0 0 o 0 0 0 0 0 00 o00 000 000 000 000 000 000 0 0 00 o00 0 o o00 0 00 0 0 0 0 0 0 0000 0 000 0 000 0 0 000 0 0 0 0 0 : Based `bbaasseedd based onoonn on amount of est sation consumed (average of10bees) nnoommiinnaall ccoonnssuummppttiioonnooftfetesstt sooluutiioonn of 20u~ltl//bee amount of test solution consumed (average of 10 bees) Page 150125 : `Environmental Environmental Laboratory Laboratory PPrroojjeecctt NNuummbbeerr UU22772233 Page 18 of 25 22773344..00003311 000082 000032 er onsen MET 0438394 3E} J [i? {i i i `a I i ie ile ii i m = ly i| II 1i a ibp> i 7 F ind Ce | "Bilned = = -- === mp mer om Si TTaabbllee AAS5.. Nominal dose' dose1 (ug abe) (lag a.i./bee) (CSL CSL Study Study NNuummbbeerr HHTTS5660022 Results Results ooff ddeeffiinniittiivvee oorraall ddoossiinngg tteesstss wwiitthh PPFEOOSS -- M Morotratalliittyy (ugailee) No. Actual dose2 (ttg a.i~bee) Cage No. Cumulative Cumulative nnuummbbeerr ddeeaadd ((nn==1100)) As be @hs The 4 hrs 24 hrs 48 hrs 72 hrs 478 43 316 3 10 10 10 444..777888 484.8 4.8 333111776 33 3 1100 1100 1100 44.7788 448.8 331188 33 1100 10 1100 10 111000 21 22 En 1 10 10 I 217 22 320 0 10 10 10 2.17 2.17 2.17 2.2 319 1 10 10 10 2.2 320 0 10 10 10 2.2 321 1 10 10 10 oso os ms 0 1 4s 000.999999111 00.999911 000.999666 3E22r2 000 00.9966 32243 00 775 68 1090 I11000 59 910 0.991 0.96 324 0 6 9 9 0450 04s EY 0 2 + 6 0450 045 326 0 3 7 7 0.450 0.45 325 0 1 4 5 0.450 0.45 326 0 3 7 7 0.450 0.45 327 0 2 4 6 000.222000555 00.220055 000.222111 333222898 000 00.2211 30329 0 0 000 0 0 00002 011 0 2 0.205 0.21 330 0 0 2 2 00@3 003 00o 3333x1 000 01 1 01 1 01 | 0 0 3m332 00 00 o 0 o 0 o 333 0 0 o o o 0 33s 0 0 0 o 0o4 0 0 333344 0 0 0 0 00 00 00o44 00 0 333356 336 00 0 000 00 0 0o o abbbbcaaaaessssteeeeoddddnooooennnndonanasommoemomoiduuinnnnatatllooccffoottnneesssstuut mmsspooptlltuititooiinononnooftccfooteenntssstuussmmooeleluuddttii((ooaannvveeoorrffaag22ge0e1ooaffcl/b11e00ebbecees)) "ndosed acetone dosed undosed Environmental Laboratory Project Number U2723 Environmental Laboratory Project Number U2723 S-- PPaaggee 1199.o0f2255 rm --_-- 22773344.00003322 000000003333 Werotens MET 0438395  RB? I i` I I i il.e iI i i| 1 | i lslSi Te ie _/ ~: l[i e~ ing _1 ~21~-?.', = ' ~ . i - - ~ = == Te ==. --= -- CSL Study Number HTS602 CSL Study Number HT5602 SL "Table AG. Table A6. Results Results of of definitive definitive oral oral ddoossiinng tteessttss wwiitthh PPFFOOSS ~- SSuulbb-lleetthhaall eeffffeeccttss er mi Nominal Ge Gein Ne dose~ (1 be) (lag a.iJbee) Actual dosez (lag a.i./bee) Cage No. Number Number kknnoocckkddoowwnn ((KK)) oorr ssttuummbblliinngg ((S5)) Th 2h dhe 72h 4 hrs 24 hrs 48 hrs 72 hrs am 4s a1 IK 0 0 0 4.78 4.8 316 1K 0 0 0 478 is 3 IK 0 0 0 444..777888 444..888 333111878 11IKKK 000 00 0 000 222.111777 222.222 33321199 000 000 000 000 2172.17 2.17 222.2 2.2 321 0 320 0 321 0 00 00 0 0 0 0 om oe m0 0 oo 0.991 0.96 322 0 0 0 0 0991 096 32 0 0 0 0 0.991 0.96 323 0 0 0 0 0991 036 324 0 0 0 0 0.991 0.96 324 0 0 0 0 0450 04s 32s 0 1s 0 0 000..444555000 000..444555 333222656 00 0 100 S 0 0 0 000 00..445500 00.4455 332277 00 00 11KX I1KX ,000.222000585 000.2211 33222s98 000 000 o00 000 0205 0.205 0.205 ox 3% 0.21 329 0 0.21 330 o0 0o 0o 0 0 o o 00@3 0 00 3W33n11 00 00 00 00 03 03 0 0 EY 332 0 333 0 o 0 0 0 0 0 0 335 0 0 0 0 034 0 0 333344 00 00 00 0o 0o4 0 0 0 333356 00 336 0 o0o 00 0 0o o 04 b?bbaaassseeeddd ooonnn annmoomomiuinnnatalloccfootnnessstuummspoptltuiitooinnonoofcfotteensssttussmooelluudttii(ooannveoorffa20g0e~odfp/b1e0ebees) *ubaanccadeesteotoodsnneoeednddooasmseeoddunt of test solution consumed (average of 10 bees) undosed EnvirronomenntalmentLaboraretory Projiect NumberU2723 Environmental Laboratory Project Number U2723 ee PPaaggee 2200 o0ff 2255 --_-- 22773344.00003333 000034 000034 ier osssaos MET 0438396 = ha i! Jo | i i `e 1i i1 ilo i 1i nig i ii= 1 i i A7 e =] - = = Te = ~. --= --e " able AT. Table A7. SCSL L StSutudydyNNuurmber HTS602 RReseultssooffddueefifinlniittiivivee oosrraall ddoossiinngg tteessttss wwiitthh CSL Study Number HT5602 ddimiemtheoatte h-- MMooroatrattalltietyy NNoommiinnaall ling dose~ gas (pg a.i./bee) AAccttuuasllddoosseez' (pg a.iJbee) CCaaggee No. CCuummulualattiivvee nnuummbbeerr dead ((nn=1lO0) ET Ta Ta 4 hrs 24 hrs 48 hrs 72 hrs 060 057 01 8 10 10 10 000..666000 000..555887 03300212 2 28 1100 1100 1100 060 060 10 10 10 0 5 0 10 10 0.60 0.60 303 5 10 10 10 030 030 0 4 10 10 10 00.3300 03%0.30 00.3300 0.30 30350.4 305 4 2 2 111000 1100 1100 030 10 10 206 5 10 0 10 0.30 0.30 306 5 10 10 10 01500..1155 00.1155 330077 11 9 9 1100 1I00 als 00115s 308 0 7 ' ' 0.15 0.15 308 0 7 9 9 309 2 5 6 7 0.15 0.15 309 2 6 6 7 0000775s ooosn 30 0 2 2 2 0.075 0.074 3 I0 0 2 2 2 007s au o 1 1 i 0.075 0.075 311 0 1 1 1 om an 1 | i 0.075 0.074 312 0 1 1 1 0. oomwooo0 ooo o 0 o as i 1 i 0 0 313 0 0 0 1 0 0 314 0 0 0 0 0 0 315 0 1 1 1 uscd on amount of test ston consumed (erage of 10bees) by~ bbaasseedd oonn nnoommiinnaall coonsuummppttiioonn of test solutiioonn ooff2200uDblfeoee:e 2 based on amount of test solution consumed (average of 10 bees) Environmental Environmental LLaabboorraattoorryy PPrroojjeecctt NNuummbbeerr UU22772233 --_-- PPaaggee 2211ooff 2255 ---- 22773344..00003344 00000003355 -------- vgronn MET 0438397 i ie1 d ile 1 nd i Ii 1 | li e iSInmd i le+ | i >| 1<= i -ia 1 i > Te = = 5 = = = (CCSSLL SSttuuddyy NNuummbbeerr HHTTS5660022 S Nomi TTaabbllee AAS8.. Nominal dose' dose= (aioe) (lag a.idbee) Acta Cope RReessuullttss ooff ddeeffiinniittiivvee oorraall ddoossiinngg tteessttss wwiitthh ddiimmeetthhooaattee -- SSuubb--lleetthhaall eeffffeeccttss (ugaided) Actual dose2 (lag a.iJbee) No.Cage No. Number knockdown (K) or stumbling (S) 44 hhrs 2244hhrs a48hhsrs T72hhers 00.6600 060 00.5577 303011 2xK 00 058 302 6K 0 00 0 0 0 0 0600.60 0.60 0600.58 0.60 333000332 655KKK 000 000 000 030 00.3300 030 00.3300 304 303054 3K 33KK 0 0 0 0 0 0 0 00 030 0.30 0.30 0300.30 0.30 306 x 305 3K 306 2K 00 0 00 0 00 0 01s 00.1155 01s 00.11s5 307 330087 0 00 0 00 000 0 00 0150.15 0.15 01s Ee 0.15 308 0.15 309 00 0 00 0 10 X 1K 00 0 ms om ww 0 000..000777555 000..000777554 33311110 100K 00 0 00.007755 00..007744 331122 00 00 oo 000 00 0 00 0 0 00 0 0 0 0 331133 00 314 0 0 0 0 0 0 0 00 0 00 0 00 0 31s 0 314 0 315 0 00 0 00 0 000 b!babasaseseedddooonnnannmooommuininntaallofccootennssstuusmmoplptutitioioonnnoocffottneesssttumssooellduutt(iioaonvnoerofafg22e00poglflb/be1e0e.ebees) based on amount of test solution consumed (average of 10 bees) Environmental Laboratory Project Number U2723 Environmental Laboratory Project Number U2723 PPaaggee 22220o1f2255 -- DE 22773344..00003355 000036 000036 TMT ETo4M3E8T 0343083898 ...... 1d 9 !* id ini J :i`> i lTMa-! ud -7 :- = ! CT -- 2 dm --p = - id Te = Le =4 ' oT CCS-SLLSSttuuddyy NNuummbbeerr HHTTS5660022 APPENDI2X INTERIM INTERIM CCEERRTTIIFFIICCAATTEE OOAFFPPAAENNNAADLLIYYXSS2IISS FFOORR TTEESSTT SSUUBBSSTTAANNCCEE `CIIeNnNteTTAEEnaRRlyIItMiMcaClCELEaRbRRRoeTrevTaIvitiIssoFFiirooIiInneCCs11(A0CA97O/TT70A/EE000)R)eOOfeFFreAAncNNeAA#:LLY0Y2S3S-II0SS184 Centre Analytical Laboratories COA Reference 3M PRreofdeurce:nc#Pe:FOSSD,.L0o1t8 217 3M Product: PFOS, Lot 217 #: 023-018A [mT Reference #: SD-018 Puri: S$pe6c3t%r Purity: 86.9% Tear _ Test Name specifications Purityt 86.9% if'Appearance Identification VlE eon Lo ine NMR CIA) = die Metals (ICP/MS) 1 Gem |b sme 1. Calcium 2 Nag i 01 wise 2. Magnesium 3_ Sodium 4 Pont t | 4 eve 4. Potassium2 5 Na i Sk S05 Doms 5. Nickel ia 6 msi 6. Iron 5 Nimgues 5 Ooorwenath 7. Manszne~e [Teemp,(iy |-- 33a Wot~ % Impurity (NMR) Total % Impurity Ee ee] ~C/MS! Total % Impurity (GC/MS) ReoCorpo = Related Compounds - White Crystalline Powder Conforms Positive 1. 0.005 wtJwt.% 2. 0.001 wtJwt.% 3. 1.439 wt./wt.% 4. 6.849 wt./wt.% 5. <0.001 wt./wt.% 6, 0.005 wt./wt.% 7. <0.001 wt./wt.% 1.93 wt./wt.% 8.41 v~./wt.% None Detected [erst POAA Residual Solvent~ 0%) (TGA) | Neos Dec 0.33 wt./wt.% None icAen Purity by DSC Inorganic Anions (IC) 1. Chloride Tes 2. Fluoride 3 Bie 3. Bromide i Nome 4. Nitra~ PE. 5. Nitrite 6. Phosphate 7._Suliaet 7. Sulfate4 Organic Acids s (IC) EtmT 1. TFA 2. PFPA A 4_3. 4. NFPA HFBA NFPA Elemental Analysis6: TC I. Carbon 2 lydogn 2. Hydrogen 3 oNumm 3. Nitrogen + Sir 4. Sulfur S Fae 5. Fluorine FSGRA f[LHdreLeE nlHESesamaEtaBE E smEEaR eeE ]a)E nSCyl D17g33 aCS3arleeteoeB vosoeswm3wk BET RRR | TorsoVie- 178% | netwme 1. Theoretical Value = 17.8% 1 rcs Vi0e% 2 oak 2. Theoretical Value = 0% 3 Teomihenon 3 i 3. Theoretical Value = 0% [8 Teele | sums 4. Theoretical Value = 5.95% | 5 ThVeiensny 5 Sian 5. Theor~cal Value = 60"/, Not Applie-ahle 1. <0.015 wt./wt.% 2. 0.59 wtJwt.~/. 3. <0.040 wt./wt.% 4. <0.009 wt./wt.% 5. <0.006 wt./wt.% 6. <0.007 wt./~t.% 7. 8.76 wtJwt.% 1. <0.1 wtdwt.% 2. <0.1 wt./wt.% 43. 4. 028 wn 0.10 wt.lwt.% 0.28 wt./wt.% 1. 12.48 wtJ'wt.% 2. 0.244 wtJwt.% 3. 1.74 wt.lwt.% 4. 8.84 wt./wt.% 5. 54.1 wt.lwt.% common COA0234) 18A tors al fess Lder0Ta x Pa covronmenat Lorton ret Somber v2 Environmental Laboratory Project Number U2723 PPaaggee 22330o1f2255 TT -- 22773344..00003366 ~600003377 CT wETosssw MET 0438399 ...... ; v i i ii J ! I| ne Is Hii I iTM I -le i 11i i + To iidg I = L} o =>a,d =p pe ~-- = = CSL CSL Study Study NNuummbbeerr HHTTS5660022 Sl IINNTTEERRIIMM CCEERRTTIIFFIICCAATTEE OOFF AANNAALLYYSSIISS Centre Analytical Laborsorics COA Referenc#e: 023-0184 Centre Analytical Laboratories COA Reference #: 023-018A Date ofLast Ansys: 0931100 Date of Last Analysis: 08/31/00 E`Exxppiirraattiioonn DDaatete:: 0088//3311//0011 SSttoorraaggee CCoonndidtiitioonnss::FFrroozzeenn 10C _<-10C Reason De 301 Re-assessment Date: 08/31/01 uit 0s mod44o SLO pri. Aric Flore, 1Purity = Fluoride, 05%#NMR impuris, | 93% organic acid impuis, 0 8% POA, 100% - (sum of metal impurities, 1.45% +LC/MS impurities, 8.41%+Inorganic 0.59%+NMR impurities, 1.93%+organic acid impurities, 0.38%+POAA, 033%) 0.33%) Le ---- Total impurity from all tests = 13.09% Parity = 100% 13.09% ~ 6% Purity = 100%- 13.09% = 86.9% :"PPoottaassssiiuummiiss eexxppeecciteeddiinntthhiissssaalttffoorrm annddiiss tthheerreeffoorreennoottccoonsnidsereidadanneiimmprupreuirtidyty.. pby yDSisrlot pcoof os w pk, No darnas aPudty by DSC is generally not applicable to materials oflow purity. No endotherm was abervdfor tis spe. observed for this sample. Sulfa the sampleappear ob converted 0 S0u ad enc detected using the 4Sulfur in the sample appears to be converted to S04 and hence detected using the Prandin ith pr inorganic anion method conditions. The anion result agrees well with the sulfur deerminadonin he demetalsnlys, ending concto i tepretaion. Based determimtion in the elemental analysis, lending confidence to this interpretation. Based ontherot,the SO snotconser an mur. on the results, the SO4 is not considered an impurity. S~TTFFAA HBA HFBA NPA PPA NFPA PFPA TTrriifflluuoorrooaacceettiiec aacciidd Hepuooronuyarced Hepuduorobutyrie add Nonouoropemnesiisd PNPeeonnnttoaaffilluuuooorrroooppprreoonpptau~nnncooiiicce aaaccciiiddd "~Trhheeoorreettiiccaall vvaaluuse ccaallccuullattiioonnss bbaasseedd oonntflh'zeeeemmpipriiriccaall ffoorrmmuullaa,, CCI~FF~vSSOOV~K."+((MMWW-5=3583)8) Thiswor This work wes was oconnddouccetedduunnddeerrEEPPAAGGooooddLIa.~boorraattoarryyPPrra~ctti~SSttaannddaarrddss ((4400 CCFFRK 116600)).. : conmsain COA023-018A EnvironmentalLaboratoryProject Number U2723 Environmental Laboratory Project Number U2723 PaPaggee 2244ooff2255 oT -- 22773344..00003377 pas ZEON TCEOER F aR d oS \= 6Co0aS EMETT 00443388440000 | el ii inlI e r Ie | |. lt n I i lili sw + iI i| i ii i i ii| sg Jii e Je +1 fhe ` = = C8 = == CCSSLL. Nom Study Number Study Number T5602 HT5602 a Ste CeIInNtNreTTAEEnaRRlyIItM iMcaClCLEEaRbRoTrTaItIoFFriIIeCsCACAOTTAEERO OefFeFreAAncNNeAA#:LL0YY2S3S-I0ISS18A Centre Analytical Laboratories COA Reference #: 023-018A LC/MS Purity Profile: CC mImppurei~ g TT C4 a C5 Te C6 C7 a Total wot.w/wet. w 1.22 1w.3m3 a4.e 72 1.14 a8.a41 Not: Ths Ct C8 lswit ted igh O 0 6 sunt lon Note: The C4 and C6 values were calculated using the C4 and C6 standard calibration Ss oi, Th iwo ep curves, respectively. The C5 value was calculated using the average response factors EE ee red from the C4 and C6 standard curves. Likewise, the C7 value was calculated using the average respome factors from the C6 and C8 standard curves. Donat = Prepared By: Seas ET = David S. Bell Date SEEe nic abr RReevviieewweeddBByy:: _ Scien_ tist,_ Cen_ tre A_ naly_ tical/_ Labo_ratories a-- Ta = John Flaherty Date Emi Sanus,CentsAnal Libris Laboratory Manager, Centre Analytical Laboratories on COA023-018A SEEsNesSi - "" " page 3 of 3 CorXe o Enviommenal Laboratory Project Number 02723 Environmental Laboratory Project Number U2723 PPaaggee 2255ooff 2255 To -- 22773344..00003388 o00c0a0d3s9y MMEETT 00443388440011