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@HAZLETON wie eONS LN Soonsor: st. pant, Himesota FAL RevoRT 13-Meek Dietary ToxSitciutdyyTStiutdlye:with 180, Ammonium hn Perfluorooctanoate (CAS No. 3825-26-1) Guideline 82-1 Author: Matthew J. 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SHR mre 0 EERT REE I Gm cn moran on sou vee jor. 82 g88 2 EY EE i Tu Tm ga g Emddne,g hn!SRH ESEdE keisR EErovEes oo BRIBERR ENE cm i more onson i on, 73 , <n BEET ill vo i ri n or n GEL ineunsBitnn EE AL, Shy im GE wi a io jae ES TONE evanrs ons RESE, no sire 8 g A arenes pesien. 1 -- rr A ae, in SERTRrE m LBC nL E EL it T roI oEm nalE HEE ae mer omsover, -3 8 or a Rr A ea og, geen SECT EL Whit eo) "gm im Pha NL da gi gE: Fi 88 & = | EEC EPlale gm 1d nag iH EEE Re i Sie R Fig E a E SHTEL mr BERR A,tmot ET ion, 3 88 g EglEm, 1 "fHiy imHo EguEe ge SdARRASRRnR n ar OBE Al rbsies onan omni TTT - a ! Cee RRge Se ART ote lle HEH EEE G SCL or BL aTinsewti!i SRT TR RR E neris Sah: "Bedale SORE ncieid chai Hines ici' Cc PF 2854 x QQ EOE WE ar iw Ge iil 1 i i iE | HEIRER RE EN mo wo mos im som eee sn. 3S a& LN a rr pie ERR a0i gp 00 SRL gow f EaAe ERNST SS RIA ofHg afHi Sr baii eiE n ay 33 & 8 | se fi 5 di SRR SE mo A gs } Er i er REI coves s matonnmone rones wr i BER Rh ni AEE Co ir mesa on. som ree or, &a3ss & HA HE ane HR ge aElET fm Ti EN Hi #8 gia i TER GEAE ot i mi ] Er ET 5 , ELT, rptbins a885 g ame SR GEL mR or CHET ETRE HA i se or 3 && " wT im am im Ga ee CHRIS SES a3 2gc: RE BANE cme wr mos on smaverte ar. EFlr Sg i Tfitr imHo gEEwE hy INIRL mieletems, ...... o 3 8 ~~ BBFIATEISS li aShe nn 7aSnSEEHREfoP ER i Ronee oT a HE EAI oti fiial Ga fi E HEiE H JiiEa SEAT BHHigEE8E } SEG ja rm ns 2,TENE Sf, reetotes LL ERR PH EE mo mone om soo ii 888 & HE Fl a Hi ha"a i WERE vi Sud Gi i ge ne in]SURES SHAE we or EIRBAR SST ERE, en. tt m,n i 3a & ~< HE EG pane. ERIESly gen wo, BoimnAF EEhEgRe, i amir osr ecor EL siilnrie,inns wSdERRESag iJm EgEgEa Lo - 883 s= 8 ie mo pn. ve gy gn a cfr epn SS i 1 SA wu Ra EE i Bi ani ggifig Eel, dw HEH REE SE nm cores SOLE1 mmoorro ootuic RL a3 au | HPI SB wan,SC oe BRsReSTOSTog S SE RG aFEl) WHaE SHEi Ege ViiHs FgiIgEiRi | CE] TM EER SA EN RPE, SERENITY me won. most (on. soma vest tv 38? s sm coildl ed, W{aEi rT CtHH EBa EEISEE Ey enSh en momen von o 5Ss # & HE dm pve. ve BEC i TH RTE RE LL 2 <8g g gg gre ame gan REESE en hdd GEE SEE ig Ti #1 E| Sieh is ] SE gy Eo: . SOIEINETCLCNNS / mstormeufoiwnmiosc ptiiie LL EESA RE RIN i moma mr,soivee or. 8g & ETRESUTg on 20d pm se ipeee Ae BET i hidPd, Hi CREE HtEH ham enh E Hi EBiE sh fie ] Ei gy rt. 210% THE ION coments | wistomnowroonmiac aon EI TiEA ERI ec sr im so wit son, 883a o&s & LN HB EG pyres ve ge: ggisern J FE Pe SHAR FalU l T +i ci wig ia ah 2g ] Ey i eT lan A teenie roe 3 <x hEllma LBE BWE@ BBEE BFC Bl HTS on,$7 ttn gy a ven BRE Lk iEle Hil gy eeinARAL ehfb ono ros 88 & 8 a C a Eee l, eLieiES ] #I @eoT om. mE o & fr fr otal i il dle GE Jai] WE Sai EE "Hiig SESigEiEn Ey AT RINE COnenrs7 mistommovonteimetnas ernrereeen Se ]g ll BEE BEE gE Ct iE gE LS eee ol a mns FettS oAAT T EEeaeLro ms heERR Rn ne ii UNO (LU). PROSTATE (PR). SNIMAL VESICLE (59) . 3 ~ i STR HrgS pp BEARD RA ot Ee tell HE WE BEE al i "Hi Hi gif ea poy Ey (SRO TRITON CONS | mistommoioatefumes a. RE EE HE rnin ot [I 3 ETEll 05 woEH RE F TEE bi] a EDR ee o all IBim "m Hi B"i HgEEE Egy SE otddi end maevm aneso "ERR AT HE EL a on sowr i, 2 F8A DY en ne meews pen pe Sa rr EaOlECCTOGJE] amiy Lol dH OgoE] GBishgit BEE | ay SME EL IN, ecnsns "BER SE rd nde wm on sia on.sou vice 3 S gy eta e Ser on SSs BRRLD SAA SeRmE re nE eve "Ei in ER ENE (mo ran on soon wc jo. i83 os a a oy eT gt IEEE HfFeelti dle iTae 4iF3E1 J "iisnesse GEi i EaaTEa gue nee,din?SERGE TSR ee SE | hiSani sm mc en 83- =z oo Eo i B FoEl ue CY djas FR RR fe GH BR CIE i HH iB fmi EEggisiig SOIL Lee TE... Mute poht ERI mm cn. moran cmsomver sr. 2 3<8Q 22 HAHEE pyre. ve ng fen c ame i S FEL ELS A ooeB EeAll f t KhS i] aH E ah p-- i igh fie 3 0:os SS iEdEC Cif mdei hHeo ESEEE } gn aE Si gysr e Sy idSR mr eens SOLINIICLIONNE Mmomnowonte iene EET BA SA EP EEE. oe io meat on son vere po, 35 z << ~~ BEC Hol eo WTiE i mn iiamn GdEae o ml n dli, bn!SRESAREAE si EEE pi Th isiy cl a3 BBg oe2 S Hm "i LoE li M o , I1E3H Trae ii] b al| gee Efia i con StdSRN SSA En rn ag88s 2 & a mg gen come RO ff a A ee EH im a HE S ee ldmove, SrnindSEEE ESRRERL oL EEE -38 & SPALL | mm lm REA a El ko ga fered i i El is A eee MELLONEICe LINMfNMDommo ie rome I SIRSE HR EI Lo om on son nce py, aJ 8 ]e ERSR BURIS Enad e, MiTg WHR WOECT le, ER amy Ge bea Hi BBE Er I EESM EY ER "vy otnd fd a, 3- <B g wi 6325-100 Aopenonx Quality surance Stitanent Individual Hormone Analysis Data 620 c02384" @ HwaAsZRLINEETTOONN Hanes at TRUE& Expygggy=== pe QuaLzey assumaxcs ozs Projecto. zaoomes Project Title:ELH: rapid Estradiol:rapidTestosterone Assay Run No.: 12.18.19.20.22.23.06.87,88.90.92.1266,1277.1201, 1294,2304 Tashcictsurarteeployrtrehfalsecbtesenthiensdpaetcatedgenbeyrattheed QSuaarliint)y dAusssiuSrianncceeUUnniitt aanndd 2329a/91m Inspection/Date 3 i Sassa s m Sis Ea2i/n2de8i/n9g1s Reported s[ii7iaAn sia Aubb.disftlaaonrna 555 YYjeeaaattneess 5a ui"cCeyhterireueeo"nutesJ)reuo<ann.ei DatCe-rFe7l-s9e,se 02356 HH 6329-100 IndividuaW]eeHkormone Data Animal Hunber cceerrdse?l CCeeT7d4sesh ce677a4s76 ccee77540s02 cec677550023 Estradiol Loa/nl) <2200..10 2200.00 220000 2200.00 220000 Testosterone --oo/al) 9opn T5180 10767,,000000 4258,,000000 24151,,000000 45851,000000 6139,,000000 Luteinizing --(na/al) 00.4 006s 00s3 0034 00s3 Loon 1:5180 ccee77ss1114 <2200.00 ce7515 200 355,5,000000 45,000 01.04 07 ccee775s2245 <2200.'70 cersed <20'0 2279,,000000, 289,000" 00ss 12 CC667755550 220000 ce7558 21.3 36,00064,000 0048 07 ce7559 200 16,000 04 1T0o5om180 CC6e7756628 20-.0 ce7s69 <20'0 63,00028,000 00.66 04 ccee775s7746 <<2000 cres91 200 16107,,000000 66,000 0006s CCee7759025 220000 ce7507 200 4875,,000000 16,000 0o6s 10 ce7601 20.0 82,000 06 a Ressay value. 622 C 0238 HHI 6329-100 IndividuaW)eeHko-rmone Data Animal Number ceC677662370 cer67663323 CeC677664306 ceCTo7e6t5s4 CC6677666539 Estradiol Loa/nl) 2-.8 221s7 2200.00 <220000 2320.00 --Te(sntao/sntelr)one 30oon T-5180 4256,,000000 3892,,000000 4126,,000000 7862,,000000 435,1,000000 --Lu(tneign/inziln)g 00.66 004s 0037 o1l0s 0046 100opm1-5180 ceco7r6e7s5] 22.517 ce7682 313 1010,,000000 45,000 0.43 06 cce677668970 33878 Ce7696 526 1319,,000000 24,000 03's 07 cceo77761938 3a615 cer715 258 5070,,000000 18,000 000446 cer7iy 2.7 85,000 06 0oon T-5180 ce6r7772373 <2200.00 ce7735 <20.0 478,,000000 104,000 00.63 0s cceerTraaz <220.50 ce7751 20.0 470,,000000 63,000 0073 07 cecr67776519 2200..00 cer762 20.1 262,2,000000 82,000 00.47 05 ce7769 200 22,000 03 b Atnhirnoaulgshouwteretrepaatimre-nfte.d with the high-dose animals 623 02358 fboomibelr cccorrredsess cceoc7snrduasrssz cords CCc6o67r74d48s34s8 IndividuaVleeHkor8mone Data tEsotroadlio)l 2200'.00 <22n0.'05o 2200'0 <222000.000 hTeostgosatelro)ne oonT5180 51024,8,.13700000 2394,,330000 30,4780200 36024.,,757000000 cors1z cc6oo77r55s33e19 cec7or354i0z ceCc76o73r55s47e5 C7360 Loon T-5180 20.0 12,200 <2 202000.00 32382/7,,147000000" 18,500 19.500 2<2220000.0000 1473,71.724003000 2000 cce6r7s57780 cersel 200.00 200 ccc7oor5rs0sea5 a230l0'o2 ccceoorr7ss5ee9s4 2n0o <n' 67614 <20.0 4 Ressay value. 10 oom T5150 520,,860000 35841.,333000000 14521031,30100000 221800 35,900 MLutfeiRni0ziTMng o0l.s4 0074 3[%4 0% 000.6s5 0.4 0o0.s69 0087 o00.s74 3 @0.s3 0o'8s <o03l3d 0044 0.3 WN 6325-100 624 cozggq HI 6329-100 IndividuaVleeHko'rsmone Data Aenimral cocc7eer6r61e279 67639 cc6ee7rr68e44sr4e ccceerrreeessetel oEstmradi)ol 3203.4 zl 34.1 222004.004 220000 33 fTesotosaterolne 30 opm T5180 13225.2.98000000 252,000* 238256,.,382000000" 21210401423000000 lLuotegineiz)in"g 00.44 04 0.6 oo0'sl.d4 30o'ss 1000m 1-560 cceoC777676309039 2200.0 3204 7323116.,571000000 C7708 2003 251500 00.66 o0'7s cceerrr0o9 cern 220000 00 3522,6..806000000 0o3k ole ccceeennrrzzaee 382.74 500 1343743./008000000 0ols ols oon T-5160 cces71712298 carrie 2200.0 20 232112,5000000 0o.s4 03 ccceeerrrrioy 222000.08 cers0 200 411087..47000000 261300 3ot 00173 ccoorrssae 22000048 13180,880000 o0''ss Ce7768 <20.0 15,000 0.8 a ARtNheirsRosaualgyshovuwatelrueet.repaatimre-nfte.d with the high-dose animals 62s 02399 wi 6325-100 IndividuaetekHorsane Data fim] Number Estradiol og/ml) Testesterone --lna/al) Luteinizing --ing/al) Ecornioso Ga0o oon T5180 $12i.d200 cars Goo pret oo0dts cieirYnes GGGooooo Crise Goo 72f.1r52i08e0 tio 33a3 0 cECEerrrisiossre pia0]00 31560...586%80 3o033 me Teste Jccoartsros <GGao0.o0 cs G00 620.0.2500%0 1500 00a3ss 03 ccGerrieesses GGGoo0oo cre G00 f2140..i200000 forz00 o0ai3s 0 ccreeres &0@%9 i &e00 a033 0 con 1.5180 Ccnoorresnae:s <G2o00.o0 152$1200.0200000 cine G00 30 0o0.3a4 os cCCerrrieeeaosrs p2Tre) rent 00 5106e:.e5100]0 Yerian 0aa%33 1H cGorrees E85000 a 6.5% 0o3s 626 c0239g WW 6329-100 IndividusV]ekHar1s4ane Data . Animal Estradiol Testosterone Luteinizing Number Loa/ml) --lno/ml) --lna/ml) 30ppm 1-5180 cccraoerraeetls Ga2n00.000 511603,.102000000 000.334 cccaoarrr6eezzsys a27907 11367...047700000 0o0's3s Ceccaoerresess: GGGo00o00 i5171103s000 0023s cress 0 6.600 07 100ponT-5180 ccaerreers aa00 77.46200 00.3 Cccae6arr7ree6ess8nse6 2<Gi2800.00 2232973.,.1578560000000 o01's .s5 o's ceCcn7en6r3io7z aS2niloo 39"122.7020m00 0o'3s 03 nz 0 20.300 od oon Tosser ccceaoTrmremases aaaln0oo 1s95a.:.25800000 000.385 CCceo6rr7rr7ae9rs G02000.00 11321628.0040000 00a47 cccaea7rr7esss 2aol00a 26H.E600 1500 003s 0s csrrer 00 25.400 a3 b Atnhirmoaulgshouwteretrpeaatimre-nftad with the high-dose animals ar COR3GR Ww 6523-100 IndividueaeHormone Data mil Number Etragil pa/ml) Testosterone ng/ml) Luteinizing ng/ml) cCEoeernrieeessss 3p00r.0]0 0opmT-5180 71 10.:43050 30t.o fCoereed 300 5S.i2e% i3 cceerrssotse 29.0 opm 1-5180 21f.a3r0k0 <03. G6Cer7re5ce3sr4s 3a04.s9 2i 175,5%00 bo0.td3 - 10ppm 7-5180 cC6ro7rra5e6as7 55i29].8 162S31,i175s500k500o0 fie i Wis I00.345 os Cccoorrreeedsszse <a35 30 pon 71-5180 175i.58]0%0 0oisfi Ctrreeenr S306 f2i0.t10]0 ootd 00 oom 5180 CcCeeerrreeearnsss 303:08 cent i 225.315%03 1f-6a0 <o03e 03 Erman Hi 50.55% 8 a 02398 Ww 6325-100 soon Results of Individual Test Mataral Residue Analyses as cozaod Anmal Number const sre cores coress cme srs conse co7500 co7s02 corso WW 6328-100 Results of IndividuVaelekTe5s+t MMaatleersial Residue Analyses 1:5180 (opm) :. Group| ::: : : : :i ::: Mean(pom) . B . i Ri . . wrsn corse Group2 80e1.3i3% 0.2 corsis s6assr 6. ir) cor524 corszs SH&aE5l] 6. ra cores IrH7t3 se 4 Below the limit of detection 0 0239' jo Number rss css corse pr corse sen corer se css corsa corm cores conser corso corr ceo wi 6228100 Results of IndividuFalRTest materia Rosie alyses 1:5180(pom) ipu8Hrfnte Gr 2(o Contu inup ed) . Grou3p opGwitsi aaEFA3E] iaiie 2aa4i7s a23 ns . RsiHs Mean (ppm) . so . on . "no ss 5. a 50s as 65.2 a 5. . 2.2 631 cozasf wi 6328-100 Results of IndividueaelkTe$s7t MMaatteerrial Residue Analyses animal tuner 1:5160 (opm) ean (ope) corezz G(rCouotn4tinued) or cores coress 15a%Ii wo no creo 2wl or cress iit m corese cores =108 1Iie I. I" cores 1rs . corers cre Ji1m3o Grow5 i. coreaz cores? 15% 3i ro re core 1iz ne coress =i or coresn cms a5 5= " ms Bi8 cm 183 2 for 632 0239 HHI 6329-100 NAnuimmbaelr cerr27 ce7733 67735 cerrer cer782 cer751 ce7759 c67761 cer762 cer769 Results of IndividuWaelekTe5st- MMaatleesrial Residue Analyses 1:5180 (oom Grow 6 a a a a i a - Mean_(opn| - . - a Below the limit of detection. 633 02398 HAI 6329-100 HAunmipmearl coress csr4s8 core e746 crn cera78 =r [= cs7435 22 Results of IndividuVaelekTe8s+t HMaalteesrial Residue Anslyses L:5160(opm) Srow : : ii ::: a: i:: : Hoan (opm) . i . . . - consi cor831 cors32 cr539 cr540 corse co7505 Grow2 6.4957 88l103s 55.3265 6i8s T7! 8z8 67.551 4 Below the limit of detection. 5.12 8.09 5.31 6.61 7.58 . 7.03 ou C0239 WW 6328-100 Results of IndividuVaelkT8est- MMaatleersial Residue Analyses - NAnaismealr 1:5180 (opm tan teas corsse Gr 02.1(o Contuinup ed) oo corssn 977506 a cors60 87:088 78 760 cers70 corsre corsa corse corsa cosas cosas corse corsse cei a.Grou3p os 6HH0]3 aus 2i we 230 oi iass Ps aoss oe n50e2 ser 2BHs "se 3888 we anlas a fins ere 3Group as cores IH53 oe coreae 107 1 or 87 635 0400 Animal Number C7638 cores cor647 co6ss cer661 cores co7667 cores0 cress ce1703 cer708 cer709 mo cen 67721 cena corre WWI 6325-100 Results of IndividuVaelek Te8s+t MMaalteesrial Residue Analyses 1G:r5o1u8p0(4pom()Continued) 110058 88.s59 27892812 8al3t 14o65 d 14 Group 1Is8l 11161243 f34e).e4d 137 11176502 113398 11155057 2l0o3l 218 Hean (oom) 106 8.5 0.2 0.7 6.9 0.9 15 150 13 10 Ex 15 1 13 164 168 m a coza0f wl 6325-106 Results of IndividuVaelekT8est MHaatteesrial Residue Analyses Antnal Number 1:5180 (opm) Mean (pom) cman cernas come cnr . corms cone cormso conse corsa cones . . Group : ::: . . : . ::: h . : . : . :: . + Below the limit of detection a 0240, Animal Nunber corso corer conn corns caress cone corse 0) cerson corsos wl 6325-150 Results of IndividWueaelk T1e4st Meatenrial Resicue Analyses 1:5180(ppm) ron . ::: ::: :: ::: ::: ::: Mean (opm) R . . i . Ri . . censor costs rsa corsa corsa cosas cosas Grou2p p77..e860 8I000]88 bGsrae] HHHHtH 773h8 a Below the limit of detection. 0.03 191 0s 5.95 S91 6.6 7.2 ` 638 c02408 wi e325. ico Rests of Individeuaml TTeEsL Material Reside Analyses . fie tunber 1:5180(oom) Hean(oom) ws ws Gr is2na(oConu tinuped) sn ob ras corse ass5a a ces cee 8isdzGroup3 it 0s %s cn corse FE2de 32 as ne css conor a8333 83 as 2 cece cin a&t33 23 9.1 ws cron ws i#13 EH ns ne core ron Parl 6s crete conan ajaoi:s 83 as wr Pa cozaof wl 6325-100 Results of IndividVuealx T1e6st Maatteersial Residue Analyses Ansan) Nunber 1:G5r1o8wn0(opm(C)ontinue Mean(pon) cores corezs 10I3 )i ne 108 crs corses 15o08si 58 ws se. ceresz corese sh3ied) 78 na a0. ceress 86a1.73 ar cern caren 113s Group 1s &.0 66. ceress coress 1106025 120 103 vs caren caress 111657 139 155 we cores camo i11s9t is 0 m comz wn 13i10% 15 10 18 i% 640 02408 1 6329-100 Results of IndividAual RTesTt MaAterial Residue Analyses pfniieall 1.5180 (oon ean (oon cnn Group :? . corres : . coms come ::3 . . cones comes I::: ; . cose conse :::: . . correo cone :::? . . : 3 Belo the init of detection. C2406 HN 6329-100 HuAnnibmealr co74s3 corse cores cs74%0 C7435 Results of IndividVueaelk T2e2st- MMaatleersial Residue Analyses 1:5160(pom) Gro|w aa aii ai a a: Hoan(opm) . . . car508 ce7516 cors3s co743 Grow z aa - 1.18 1.20 aa1.21 . 3i . a Below the limit of detection. ` 2 0240 HWI 6329-100 Animal Huber 67567 67575 67579 67589 67593 Results of IndividWueaelk T2e2st- MMaatleersia) Residue Analyses 1:5180(oom) Grou3p 0.0.541612 00..128212 a a 22..5599 a a Hean(oom) 0.487 0.207 2.59 - 67626 C67637 C6764 Group 1.582 1.53 1.53 22..6509 2.60 0a.771 0.771 67683 C6769 c67701 Grows aa - 2.43 2.58 2.51 2.41 2.36 2.39 a Below the limit of detection. 643 02408 HI 6329-100 APPENDIX MetMheotdhoVdalViadlaitdiaotniofnorfo1r-5FC1-014i3n iRnatFeSeedrum su 2469 RoP1aEA0TPG-ELE4:A8C51Eo0Sl.G!1Fj0o5'ds1/1,9001 SECTION: 50] ASSAY TITLE: FE-143 In Rat Feed IN 6329-100 sAcRoErAe:OF APPLICABILITY: ReMsazdlueteonCwhiesaciosnnsriyn, Inc. TTevOe1ES EofR1 to5 1108apbole to the cuterination of Fe-143 0 rat feed sangre PRINCIPLE: CCCaorpdtPtubrreotndsheitEaeoucptalonyra141P0I. End1sGhaLterraiinecd btyatgeusSsCochtraenmeatetohpearyaotstnurengeerrtsce1t0s sewstmvin: PTRhEeCIsSeInOsNi:tivity of this method 15 1 ppa of FC-143 in rat feed. Accuracy: aTthe30replpamtiv(ene6s)t,andanadrd3d.e4v%iaattio1n00 1sPom21%(neagt).| Ppa (ne5), 5.4% at 10 ppm (ne6), 7.7% REFERENCES: 3T0heppamver(ange6e),recanodver9y6.9i%s 85.1% at | at 100 Ppa p(npemg)(.ee) 99.4% at 10 ppa (n=6), 100% at Fiethod provided by IW and modified at Wacteton Visconsin, toc. (wu). HWI Laboratory Notebook Mos. 3418 and 3450, as 02410 OHPAAoTG.EE:4p20E1G0/FH-0A4's/51 RSEECPTLIAOCNE:S! O60r0i]ginal WI 6325-100 APPROVED BY: Hind X)TahanaTata, #10 MSettahfofdsScDieevnetliosptaent REVIEVED BY: DKinSteogrearbC.oKenllanr Lo Kedln Quality Assurance Unit (14780) ote: [2/20/50 ores 2.24 LT 646 coz414 OHPAAPTG-EE:A3P0F1OO/F1-0A46/51 RSEECPTLIAOCNE:S! Origina1 6001 SAFETY PRECAUTIONS: HMI 6329-100 OTrbasienrivneg aMllanusatla.ndard laboratory safety procedures as outlined in the HWI Safety FUNDAMENTAL EQUATIONS: FC-143 (Ammonium perfluorooctanate) formula: CF,(CF,),C00-Ni, Nonadecafluorodecanoic acid (NOFDA) formula: CF,(CF,),C00H INTERFERENCES: - There may be When it does oacncuorc,castihoantalsamipnlteermfuesrtenbceerepaeasksatyheadt. is observed at the 1 ppm level. QUALITY ASSURANCE: orasarus: Two standard curves, low and are used for calculation. high, each consisting of a minimus of four points, 9 Epp FVoonleummaettrriiccMpaispketst,es,varviaoruisoussasiozes " endorf digital pipettes, various sizes Doposible Horizontal Sinatra vials, 20 al shaker, Eberbach Corporation Ack Sis chromatograph with electron capture detector, HP S890A, Hewlett o Autoinjector, WP 7673, Hewlett Packard Integrator, HP 3396, Hewlett Packard Note: Equivalent equipment may be substituted. REAGENTS: Mota: N-Q\ water IEstohoycltaenteh,er,Mahg-hp-upruirtiytysolsvoelnvte,nt,848 J & J HPyedrrcohclholroircicaciadc,id,cocnocnecnetnrtartaetde,d,ACGSR,reEagNenStciegnracdee,Aldrich Diazomethane described in solution Aldrich in ether, Technical prepared according Information Bulletin toprocedure No. AL-1 13. Equivalent reagents may be substituted. a C2412 TesT marcas: SRHPAEAeTPG-ELE:A1C49E`rSoo:01lr./sodr/gsina1 WSIEC6T32:9-100goof PROCEDURE: FC-143, Internal Lot No. 115, purity standard, (NOFDA), 98%, IM Lot No. 1120LX, purity 98%, Aldrich To00-aL volumfeot.ric Flask' Sii,sollveecutrhaetalsyoluwteiiognh wSaotleurt.ionStvoarrey it maotntrroeorm temperature in a Po swmbeoustr2i0smeg1eoferF1a63oneto 84 lyethylene bottle. Prepare fresh : Ein 4 volumetric pipatte, transfor 20 st of s 100-uL water. volusetric Store it flask. Accurately weigh Dissolve the solution and about 200 dilute it Bg to of FC-143 into volume with a solution every 6 at room months. temperature in a polyethylene bottle. Prepare fresh . Solution solution temperat 8 To a S0-mL volumetric thoroughly and transfer flask. ft 0 2 Dilute it to polyethylene volume with water. bottle. Store it Mix the ure. Prepare fresh solution asneeded. at room "ether. Stora it 20-mL volumetric ML of perchloric at race temperature and prepare resn sepon tos seine yl acifldaska. ndDdiislsou-ltveeAtchcieutrsaiotnle9ult0y:1w0eivg/hvab1o3u0t0ct3a0n5e-atohfy]NOeFtDhAeri.ntoAdad 100 ion to voluse with 1s00ctane-ethyl Solution et D'Eo 8 1.600-aL volumetUrsiic:nga3skvo,lum~eDtIrTuitce'pi15p:etttoe,votturmannsrfaetrh 10Seateosf . hyl ether. Prepare fresh Nsioxluttihoen saoslunteieodnedt.horoughly and store 1t at room temperature. Solutions thoroughly ons vereek mice. 2ppropriate control rat stock feed, s4 oallu. tioofFnoswlaltoteowri,nsgepatrhaetescshceimentibellloawt,iontravniaslfsercoanltiaqiunoitnsg of1 and 2 L of hydrochloric acid. Mix the the g of oe 0241% Phat Ear MP-APFO-MA DATE: 01/04/91 REPLACES: Original SECTION: 600] HWI 6329-100 A 25 A 50 i i A 100 0.5 1.0 2.0 A io 5i0 A 600 c 2% c so 0 c 75 12.0 20.0 40.0 60.0 80:0 c 150 120 fog mtr pete, rust 110s000catLanofe-eSolution Eto LL volumirietiask. Biuve ors co t'thtn solution ther. every 6 Store the months. solution at room temperature and prepare fresh SamlePreparation nr Weigh ater ai and 2 feed sample into a mi of hydrochloric 20-al acid. disposable scintillation Mix the sample thorou vial. Add 4 mL of ghly on a vortex SameExtraction To each of the vials standard on a h extractant. containing the Cap the vials standards and samples, tightly and shake for add al of internal Tinatyoereatoorizsoenptaarlateshaakuetro,injCeecnttorrifviuagles,andAdtdra0n.s5fe=r 1o-faLthaelidqiu2aozthsourosf at high speed the organic 10 to 1c5hsoecfontdhse. viaIlnsj.ectCa2pBLthoefveiaaclhs sanadmpmliexintthoe sthaemp6lCessyonstoesase.tvhoarnteexsmoixuetrionfor Column: 5% im phenyl methyl fil, Hewlett $1licone, 0.31 Packard, Co jm 1.d. x 50m, 1.03 PIunrjgeecttiiosne,pomritnultiense:r: Carrier gas: Splitlessglass Tiner ON 1; OFF = 38 lumn No. 42-85-04 Flow rate: Make-up gas: 5% methane/95% argon 1 al/minute Flow rate: Injection port temperature: 5% mathane/95% argon 35 al/minute 250 Detector tesperaturs: C 250C a cozadf T"emTpeEraItureSvenfempmrogerars:ature: RFianmIapnliAttitaelmpetriamteure: Ramp 8: FFIiinnnjaaellctittoiennmepsesrizaet:ure: 40C 101005:mCi/naiuntuetse 25C/minute 23820Csainutes wPA-GAEpr6 0O-Fn's RSDEEACPTTLEIA:GCNE0:S1:/04/Od9ro1iogyiny) WI 6325-100 . Bata: ATY conditions may be dusted to optitze chromstography. CALCULATIONS: Sitio sete tel actos Sac irtek ENE ites ofne Compute the linear regression #quation relating the peak height ratios of the . Peak height ratio tquation, compute othfetdhoesesacmopnlceenftereadtiionn thine pappmpraosprfioaltleowlsi:nearregression Ppa FC-143 = (y -b)/m x F u: oss where y b # = = = ipnetaekrcheepitghtfrormatithoe opfptrhoeprsiaamtpele lifneeedar slope regression equation F = calculaftrioomn tfhaectaoprpr(o1p/r6i)ate Tinear regression equation (1/ppm) = weight of sample feed Beasured out for extraction (g) Example: For a 1 ppm 3: le, y = 0.159 wl b = 0.08895 6 = 1.0222 Ppa FC-143 = (0.159 - 0.08285)/0.0846 x TE = 0.810 Average % recovery (Average amount found/amount claimed) x 100 Example: For a 1-ppa sample, average 5 recovery = $8511.00) x 100 650 02418 HPAPG-E A1P0FHFSA7 ROETPEL:ACES0:3/08O/r5i2ginal SECTION: 6011 HI 6325-100 ASSAY TITLE: FC-143 in Rat Serun AREA OF APPLICABILITY: RHeaszildeuteonCWhiesicsotnrsiyn, Inc. score: TTheivselsmeotfhod1.0fs4ga/pmpLlictoabl1e2.7tougth/eL.determination of FC-143 fn rat serum samples at PRINCIPLE: FaeClc-ei1cd4t3rcoonnidnictatiphotenusrs,eermdueesttehscyatlmoaprtl.eed,isanedxtdreatcetredminiendtobyisgoaosctcanhersoemtahtyo]graepthhyerusmiinxgturaen under SENSITIVITY: 0S.e7nshigt/iavlityofdFeCs-c1r4i3bedinasrattheserluomw.est fortification level that was validated, is PRECISION: s6To.h/3eo5lw,higt/h2ni.ln7,%da32ty.9%s1.t5aat6ndua7gr./d9m4ld,pev/i1ma.t7,i%on0at.9(%3n+.3a1)t8 pfoo/ralF,C-014.375 9.53 ja/aL, iniatndr4a.0t7._64s%ehrgua/tnaLi1,s3.700..u39y%%/aaoLtt. 0.794 TS1Eh:e567.bp9e8t/wnpege,fnnld1,a0y.s'8S%.s5at%tanatd3a.r91.d653pdoe/pvaail/a,atli,o1n0i.n2(d%ne68a).t4%4f.o7rat6FpC1g-2/1.m47l3,joi/n1a0lr..a2t% aster6u.n35isyg1/0a.l1,% a1t0.1% ACCURACY: Tahned 9av7e.r3a%geatre1c45ovsegr/yal.fs(9n9e.)9.% at 0.794 ug/ml (ned), 99.9% at 7.99 pg/ml (ne5), REFERENCES: Method provided by 3M and modified at Hazleton Wisconsin, Inc. (Wil). HN Laboratory Notebook Nos. 3418 and 3450. 51 coz416 worm DPSPAAhTGEeE:rs2:0O3F/087o/5n2e HHI 6329-100 APPROVED BY: RORTaiTnanane i fr--g SS d PanBsastetr) Assurance uate ote: 3- OR-92 oe: __3/sfre 652 coparg PHAPG-EAP3ESO-FHA7 RDAETPEL:ACE0S3:/05/O5ri2ginal SECTION: 6011 HI 6329-100 SAFETY PRECAUTIONS: WObissecrovnesinallSafsettayndaTrrdainlianbgorMaatnourayl.safety procedures as outlined in the Hazleton INTERFERENCES: tTihneerefmoaryPCb-e14a3n. enTdhoigsenomuasy with control blank serum. b{eIntoevrefrecroemnecebypeaakpprthoaptriaetleutedsiluatrioounndorthebe rdeatmepnetnieodn AQ'UsAtLaInTdYarAdSSUcRuArNvCeE:consisting of a minimum of five points is used for calculation. APPARATUS: ++VoluVEopmlpeuemntedrtorriifccdipfgilipatesaktlst,esp,ivpaervtiatoreuisso,ussviazserisizoeuss sizes ++ ++ GDHaoisrsipzcoohsnratobamllaetos1gh2raakxperh7,5wmiEtbherpbeoallceyhcptrroCoponryploecrnaaepttiutorenestdettuebceto(rw,ithHPca5p6)304, Hewlett Packard Autoinjector, HP 7673, Hewlett Packard Integrator, HP 3396A, Hewlett Packard Mote: Equivalent equipment may be substituted. REAGENTS: + + EHItsIhoyo-lctQaentehw,eart,ehrihgihg-hp-upruirtiytysolsvoelnvte,nt,BuBrudridcikck& J&acJkascoknson ++ + DHPiyeadrzrcoohmclehortlihocarniecacisadco,ildu,tcioocnnocnecnietnnrtaerttaehtdee,rd,,A`CpSGrRe,rpeaEarMgedenStcaicegcnrocareddei,ngAltdoripcrhocedure described in Aldrich Technical Information Bulletin No. AL-113. Mote: Equivalent reagents may be substituted. 653 c024 18 o Tecsteome, a: s:purity sen, 0 . Internal standard, (NOFDA), purity 98%, Aldrich pPhoatpesr. [Sier DATE: 03/05/92 WE 6325-100 SHPolaurktionet0iFooCef-1SaTt0o3crkiS(t2a0anoddtaarsa,di)S.olouMVteitnoeanhs1169039gooff FE-0114030 inton8atsoS0on-tet vvoa)tumeirc polypropylene centrifuge tube. Prepare fresh solution every 6 months S Hao k lpil iuNsOoFOhtA r(i 1T.7o maO/anLL). eWe1igh8403 m,g OoFf OKOTFOAAIninttoo5a2255--naLLvvoolluumseetric eptehrecrh.loriSctoraeciadt anrdoomdiltuetmepertahteurseoluantdionpretpoarveolufmreeswhitsholu{tsioooncteavnee:reyth6y]monetthhse.r. Prepoafr Worakit ngSi olo utin on I1n.t0eraLnaloSftSaonlduatridoEn xHtrtaoctaan1t-L(1v.o6luumge/tmrli)c. Using flask." a volumetric Dilute it to pipette, transfer volume with isooctane:ethyl ether. Store solution every 6 months. the solution at room temperature and prepare fresh EC-143 Standard Solution. appropriate stock solution Following the scheme below, transfer to separate 25-m_ volumetric aliquots of the with Mi111-Q water. Store at room Prepare fresh solution as needed. temperature in flasks. polypropylene Dilute to centrifuge volume tubes. Start usher jPo-}1 =5P-3 CStoomcck eSnotluitoinon _Go/sl) 320 ==i320 Alot volume (a) s10 :7 often volume lm) 525 8525 conFcihnalhon oo feray 1%128 Hj89ia.d6 654. cozadq sundae Cgooncoenisroalttiioonn Aliuot Hymper vvr3ae #320 --luo/ml) 33i Yolume (ml) + i as PrepofaMatrriaxSttandiardoCurnve ,tution ii52 Yolume(ml) OWPAAoTG-EE9:FSs0o.3rH/a0857 RSEePLtAoCEnS! o5ol3d2% WI 6325-100 CanceFmineanlation --luo/nl) 12f0.se0 th aPitpeettGeertah 1c0a0s-)4.L cFonotrrtoilty sWerRum 10into a'o1F2stxan7d5armdm sdoitsopsoisaanbleSopboolRypTropOyplenesnedetsestt Mi111-Q water and 50 ul perchloric acid and mix. Sample Precaration Sime: " Pipette a 100-4L tube (with cap). serum sample into a'12 Add 110 ul of Mi111-Q x 75 mm, disposable polypropylene water and 50 uL Perchloric test acid. Mix the Mote: For samples containing prepare a dilution levels of FC-163 which excesd the standard curve, volume of the of the original serum with matrix diluted sample taken for control serum. analysis should The total be 100 ut. S1ample3ITEonxetteuerxraTcnch1atio8ofstnVatonhrdetaertsdeesnet)xttruFabocretsa3ncteo,inntcaciiaapnsi!ntghetheeststtanodsaerdsandandSosmaamnpless,onadsd t2 atho of 2. 3. Place Shake the tubes In a hortzontal position at high speed for 30 minutes, in a horizontal shaker. 4 Return the tubes until the layers to a test separate. tube rack and allow to stand on a lab bench 5. Transfer 1-nl aliauets of the organic layer (top Tayer) to separate autoinjector vials. 655 cozazd HPAES DRSPAeAETcGEtPE:iRo6nE0:3O!/F057/cr5o2rg]inat I 6129-100 6. AdS 0.25 aL of diazomethane solution fnto each of the vials 7. Cap the vials and mix the samples fn 4 vortex mixer for to 10 seconss 6. Inject 2 AL of exch sample fnto the gas chromatography (cc) systen Chronstooraphic Conditions Column: 5% pheny] methyl silicone, 0.31 um i.d. x 50 m, CPIaunrrjgteeicetriiongnea:p:omritnultiense:r: 1.03 gm film, Hewlett $SoNp5lin=telt1e3hsasn0gFeFl/sa3+sssToarlgionner Packard MakFeluopw oruast:e: $15mBle/tmhianunteesosx argon DIentFfelecoctwtoirornatteep:mopretratteumrpee:rature: $3315500-mCCl/minute Temperature progras: IInniittiiaal tie: oven temperature: 7130mCinutes Ramp A: 5*C/minute Final temperature: 95C Final tine: Ramp 8: Final temperature 25C/minute 12030miCnutes Iniection size: 24 Note: A11 conditions CALCULATIONS: may be adjusted to optimize chromatography. CGaolmpfubtreatitohne s1itnaenadrardrsegrtoestshieoinreqrueastpieocntivreelaStoinncgenttrhaetipoenask Jhneitgphst epqeuaaktihoeni,ghtcomrpauttieo otfhetdhoesesacmopnlceensterrautmioinn tInhepaappsreoprTioartreol.inear aravtiiso:s oCfvstihneg regression the 656 cozarg WPAoG-EAP7ES0.e7 DRSAEETCPETL:IAOCNE0:S3!/0560/0r91i21gina) I 6325-100 R ppm FC-143 = (y - b)/(a x F) where: 2y5 22= SIPlneotapekerchefepritognhtftrhorenataitpohperooSfpprptirhaaetpersiasmTtpielneea|risneerrauermgrersesgiroenssisoanuateqiuonation VF 2= CVoallucmuelatoifon`safmapcletorser(u1a00/mVe)asured out for extraction (ut) 57 coz