Document Ne2Lpp9v08nzZryQ463bE6opg

AR RR 6 1358 3M Strategic Toxicology Laboratory Study Title: Acute Inhalation Limit Toxicity Study of Perfluoroctanesulfonyl Fluoride (POSF) T-70983 `Test Number (sample): ~~ T-7098.3 (Perfluoroctanesulfony! Fluoride (POSF) Laboratory Project Identification: ST64 In-Life Initiation Date: ~~ 5/1/2001 In-Life Completion Date: 5/14/2001 Research Client: 3M Specialty Materials Division 3M Center Building 236 Saint Paul, MN 55133-3220 Testing Laboratory: ~ 3M Strategic Toxicology Laboratory 3M Center Building 270-SB-181 Saint Paul, MN 55133-3220 Study Director: ~~ Andrew M. Seacat, Ph.D. Toxicologist Specialist 3M Medical DepartmentToxicology Services 3M Center Building 220-2E-02 Saint Paul, MN 55133-3220 Study Toxicologist: Tommie Yvette Tumer, Ph.D. Post Doctoral Research Fellow 3M MedicalDepartmentToxicology Services 3M Center Building 220-2E-02 Saint Paul, MN 55133-3220 001525 7Fi0na9l8R.e3poArctute POSE lnhslaon Limit Test. Summary: An inhalation toxicity screen was conducted using 3 male rats for control (aironly) and 3 male rats exposed to perfluoroctanesulfonyl fluoride (POSF) at a nominal concentration of 1000ppm for 4 hours. Overnight urines were collected on day zero to analyze for perfluorooctane sulfonate (PFOS) in the urine. Animals were observed for any adverse effects for up to 14 days after cessationof exposure. At necropsy, serum was collecte"dThfeorcoPnFtrOoSl adneitemramlisnastiginoinf.icantly gained weight (p<0.05) during the recovery `period and showed no clinical pathology. The POSF treated animals tolerated the 4 hour inhalation exposure, had significant (p<0.05) body weight gain over the 14 day recovery period. Organ weights `were normal and histological evaluationofthe liver, lungs, bladder, kidney and brain revealed no significant findings. The amountofPFOS in urine collected overnight on the day ofdosing, and the amount of PFOS in serum on day 14 post-dose were 3.08 +- 1.60 ppm (N=3) and 4.57 +- 0.70 ppm (N=3), respectively. These data indicate that POSF `was absorbed from the lungs, metabolized to PFOS and found present in the serum then eurxicnree,teads PinFuOrSi,newaassP<F1OS%. The average percentage ofinhaled POSF in the serum or Conclusion: The 4-hour LC50 of POSF isgreaterthan 1000 ppm. L TShtiusdystOubdjyewctaisved:one to establish data on acute inhalation toxicity and to approximate the 4-hour LC50 valueofPOSF. The resultsofthis range finder study will `be used as guidance for a 13-week inhalation study. IA.n IdentTiefsitcaAtritoinc:les: T Name 7098.3 POSF Structure MW (/mole) Density (g/ml) CiFpSOF 502 1.69 B.Purityand Stability: Responsibilityofresearch client. The POS sample has been lab-fractionated and acid/water washed. Chemical characterization by 'H-NMR and A1nSaEly.tNiMcRalSLpaebcotrraotsocroyp!ySwMasDctoonddeutcetremdibnye t3hMe pSupreictiyoalfttyhAedhneosmiivneasl &proCdhuecmticaanldsto chhiagrhacptureirtiyzefaosrmmoafntyheimnpourmiitnyalcoPmOpSonFepnrtosduacstpo(1s)s.ible. The sample was found to be a C. Hteamnpdelriantguraen.d Storage: Upon receipt, the test article was stored tightly sealed at room D. Dispositionof Test Material: Any remaining unused portionofthe test article was. returned to the research client after completionofthe study. HL Test System: 001526 2 TFi-n7a0l98R.e3poArctute POSF Inhalation Limit Test Male rats were exposedto test compound in air for 4 hours for one day in 13-liter glass exposure chambers (3 rats/chamber/compound). The flow-through atmosphere was generated by mixing test compound from a syringe pump into a streamoffresh air flowing at 3.0 Liminintothe chamber and exhausted through a vent. The syringe flow rate was calculated based on using the Ideal Gas Law equation: (target concentration)(MW)/24.79) x (1g/10 ug) x (airflow rate) x (1/density) x (1000 wml) Gtairvgeetnconcentration (ppm) MW = g/mole) conversion factor: (1 gram)/(1 x 10%) airflow rate: (Lmin) density: (mL/g) conversion factor: 1000 p/mL : `The target concentrationsofeach test compound, the syringe flow rate and the airflow are provided in the following table: Dose Group c Syringe Flow Rate Airflow Rate Treated: POSF (p1p00m0) (0min) (2U0m-i3n)0 Control: Air only 20-30 `Standard target concentrations for each compound were prepared and analyzed by Fourier Transform Infrared Spectrometry (FTIR). Chamber air was monitored by FTIR dfoaryczoenrcoeonfttrahteiosntsuodfy.teOsnt cdoamypzoeruon,daannadtomxosypgheenre(2)o.fTaphperodparyioaftdeocsoinncgenwtraastidoensiwgansated generated by adding a calculated amount of the test material to a test chamber ofknown volumecontainingthe animals. Airflow was increased when necessary to maintain a Level of 1000 ppm. Control animalswere exposed to room airunderequivalent chamber conditions as the POS dose group animals. Animals were handled in accordance with an animal usage protocol approved by the 3M LARC. Body weights were determined immediately prior to dosing and immediately prior to euthanasia. At the terminationofthe study, all animals were `humanely euthanized by CO; asphyxiation and necropsies were performed. Serum was collected at necropsy for POS analysis. Analysis ofPFOS levels by highp`wraessspuerrefloirqmueidd cahcrcoormdaitnoggrtoapphuyb/litsahneddemmeetlhecotdrsos(p3r)a.yUrmianseswsapsecctorlolseccotpeyd (ovHePrLniCg-hMtSo/nMdSa)y zero from ratsofthe limit test group. Urine PFOS levels were determined following a validated extraction procedure (4), witha slight modification in that no surrogate: standard (THPFOS) was added. 001527 3 7098.3 Acute POSE Inhalation Limit Test Final Report Lungs, liver, kidney, bladder and brain were collected at necropsy on day 14 from the limit test group. A portionofeach tissue was immediately fixed in a 10% formalin solution for subsequent histological evaluation. The remaining sections were placed directly into propylene tubes filled with liquid nitrogen for possiblefuture evaluation. IV. Results and Discussion: Test Atmosphere `The POSF atmosphere generated in the exposure chamber averaged 912 = 91 ppm ftoarrgtehte cloinmcietnttersattgiornoufpor(5t)h.eTlhimeitmeteasstugrreodupc.onFcaecnttorrastiwohnicihn tmhaeychaacmcobuenrtwfaors t9hi1s%oinfctluhdee the factthatthe calculated concentration based on the Ideal Gas Law and the actual concentration in the flow-through chambers when the system reaches equilibrium may notbethe same. The airflow was increased to 3 liters/ minutes to adjust chamber `concentration. ' Control Group: `The three control group male rats had no adverse clinical observations noted during the 4hrair-only exposure. The average body weight was 378 + 12 g at the initiation ofthe study. The control group gained an average of 22 g (5.5%)ofbody "weight during the 14 day study period. Liver weights averaged 14.1 + 1.0 g and the `averageliverto body weight ratio was 0.035 + 0.001 (Tablel). `Necropsy findings and the histopathology report by Dr. Elden Lamprecht, 3M Surgical and Histopathology Services, is found in Appendix 1. All rat lungs (both control andtreated groups) had mild to moderate congestion ofalveoli with occasional extravasationof blood around arterioles and bronchioles. This was possibly an agonal change associated with CO, euthanasia. It was noted that someofthe lungs were not. `well insufflated. Other than these artifacts, histological analysis revealed no significant changes in lungs and urinary bladder in all threeofthe controls. POSF Limit Group: `ThethreePOSF group malerats were exposed to 912 + 91 ppm POSF measured continuously for four hours. There were no biologically significant effects over a 14-day recovery period after exposure for the POSF treated group. As a gross observation, the POSF treatedanimalshad an increaseofurine and feces output on day one post-exposure. `as compared to control. The average body weight was 404 + 31 g (N =3) fourteen days `post exposure. The limit group gained an average of 19g (4.9%) of body weight during the 14-day study period. Liver weights averaged 14.2 + 2.0g and the average liver to `body weight ratio was 0.035 + 0.002 (Table 2), similar to controls. Day zero, overnight urine samples were collected and extracted, to measure the levels of PFOS. Each urine sample was prepared in duplicate. Independent dilutions of the urine were done with each value calculated from the averageofthe duplicate determination. The average amount of PFOS in urine was 3.08 1.60 ppm and the average percentage of PFOS in urine was 0.05 0.03% (Table 3). The standard curve 001528 7F.i7al09R8e3poArtci POSF nblation Limi Test. used to calculate the levels of PFOSinextracted urine samples is shown in Figure 1, and thearea under the curve (AUC) for these extracted urine standards are in Table 4. Serum samples were collectedatnecropsy on day 13 post-dose to measure the `concentration of PFOS (Table 5). The average amountof PFOS in serum was 0.058 + 0.004 mg. The average percentage ofinhaled POSF in serum at necropsy was 0.101 0.007% (Table 5). `These findings are similar to the PFOS levels observed in an accompanying toxicokinetic study of POSF (T-7098.4) in which a small percentage of PFOS was also found in the serum and urine (4). Vv. Conclusion: The apparent inhalation 4-hour LCS0in rats for POSF is greater than 1000 ppm. 001529 s FTi-n7a0l98R.e3poArctute POSF Inhalation Limit Test. VL References: 1. SKepsetcnterro,scTo.pCy.he3mMicSaplecCihaalrtayctMeartiezraitailosn &ofMPaOnuSfFac(tFuMr-i3n2g7D0i)vibsyio"nHA-nNalMyRti&cal"F-NMR Laboratory. Request No. 6370. May 15, 2001. 2. Modified EPA Method 320. Pace Analytical. (2001). 3. SHpaencsiefnic,,KQ.uJa,ntCilteamtievne,CLh.arAa,ctEelrliezfastoino,nMo.f OE.r,gaJnoihcnsFolnu,oHro.c0h.e(m2i0c0a1l)s.iCnoBmipoloougnicda-l Matrices. Environ. Sci. and Tech..35, 766-770. 4. Method Number: ETS-8-96.0. Extractionofpotassium perfluorooctanesulfonate or eotlheecrtrfolsuporraoyc/hmeamsiscaslpeccotmropmoeutnrdys/mfarssomspuercitnreomfeortrayn.al3ysMisE.nuvsiinrgonHmPenLtCal Laboratory. 5. GInuct.z3koMw,LaTb.or(a2t0o0r1y) RPeOqSuFesAtnNiom.alE0E1x-p0o6s4u8r.e3TMestETRe&pSorSt.FiPealcdeAAnnaallyyttiiccaallSSeerrvviicceess, and Technologies. - 6. Seacat, A. and Tumer, T. (2001) Acute Inhalation Toxicokinetic Study of Peflucoctanesulfonyl Fluoride (POSF) T-7098.4. 6 001530 T7098. AciePOSEabalaion Limit Test. Final Report VL Signature Page Authors: onus He `Tommie Yve furner, Ph.D. Post Doctoral `Study Toxicologist Fellow Linde 7 nN Andrew Seacat, Ph.D. `Toxicologist Specialist Study Director Lea ef * Reviewed by: geben 3Fle John L. Butenhoff, Ph.D, CTH, DABT `Toxicologist Specialist Study Director 7 o/ ate Ios) ' Date w/i9fot Date. ov1s3t 7 rl] ih rh i fl1 [EH - cf. g 3| fe 3 | il g iiss i {Lote a fee ik Fe | E.5s 1258s k J 0 [e mall: 001532 g mo iT p L2 at TiTT 55 3 i[ITF] 5 aiiees [[[11] 1iz FaIs t|f al 12 | 3OitsRT i So 2 Mal 3 . iH La 2Bo% ln shldAEH } 001533 Hop Bopp i 32 i ily ots I iEE ol ETT 5 gas _ . . ls :tty4 = ir EE+1E 4 IN le 3HE$E3 i fi lo S le aE Eg jet s35s sobs esas) i "I 8al mes a5or ame | 7HRE } oe PH :: 8 Bgyl oaEgmes ess asles [1[5 2i8 2 Bliss lls mais sags | 32 IRE TT i:BilFfi s gg gg gg " hol fEga50]0 3i f 8 BE 2s5a88fes g 29 33383s og esefaiseniE 3H:3E104E3 . 001534 o g g l I1i 3 33 3: 3: td 5 i 31 4 gHE 3 i . nn 8 afzlg i a8 8l2(a2lg lols 001535 hin szoafss Big rege| fer) 1 ds f . fees hes i FR & i ' 3. g fie pager i|iidf Bin) od| 2 Hl: 88 gis i : gir | geen BFIR ote ie SHeields eo F[ti sriihe ik : uiy 1d fill : ||| B55 ds ad sity fp || FB Bul BLL ud bbl, | HEE = 001536 Ee ole & ERNE oe el Tf | 1 5 8 [EEEECCEUIE OCNNp b Sten 2 s 232 ifEm onyislaeblAN ce Ne fe - :| g : gE Belo hal ee, Lt | 28 8 5 i :8 A2 : iiz 2 BE EE aN 3 ES a ee EhE PRRe s aN Na] gi aia N= f Sa EiEe a Eaos Ee Ed LT aE e Ee LN ERE is: i= biy iF FF fq Fa 23 (iw) ony 001537 z 3{ :2 2i%c z: Is 3 53 8 : iif i: 1 yyy g iEH i 3%S:O 2: iz 8 $i 2 3H ts 1 i= i;i :83 Hl i 321 10% 12 id 2 B BLL oobfr eien 2%ld22 2B2r _5gf3f 8% $i5f 222 fii;t 2% i$3 ERHEIRIH EL 2 885 25 288 i: - } 001538 SAeprpuemnPdFiO2xS To: Andrew Seacal/US-Corporate/3MUS@3M-Corporate `Tommie Tumer/US-Corporate/3MIUS ox John L. ButenhoftlUS-Corporate/3M/US@3M-Corporate. Paul Lieder/US-Corporate/3W/US@3M-Corporate `Sue Tanaka/US-Corporate/3M/US@3M-Corporate Kathy Thompson/US-Corporate/3MIUS. Deanna J. Luebker/US-Corporate/3M/US@3M-Corporate Subject PFOS Levels from rats exposed to POSF via inhalation T-7098.3. Andrew and Tommie: `The 1 to 100 estimate for the serum levelsof PFOS was right on target. The following results are basedonthe rat serum being diluted 1:10priorto extraction. Theextractionwas carried out at pH 3.0 using a singleshakeout in Ethyl Acetate, TheEAwas transferred to a clean polypropylene tube and blown to dryness using nitrogen gas. Theresultant residue was redissolved in 100 uL of 25% acetonitrile and 75% 10 mm ammonium acetate. Ofthis solution 20 ul were injected onto a Betasil C18 reversed phase HPLC column (50 x 2 mm, 5. `micron particle size). The flow rate was maintained at 0.5 mi/min. A gradient flow of `acetonitrile was used to elute the compounds of interest starting with 15% acetonitrile and ending with 65% acetonitrile over 5.5 minutes. Return to initial operational parameters and catlimn equilibration requires and additional 2.5 minutes for a run time cycle of 8.0 minutes per run. `The MS was operated in the Electro Spray negative ionization mode with a constant 3.0 kV `potential applied to the source. The method used was an MS/MS method involving the transition of the following ions: Compound Q:2 Offset Voltage Base Peak MS/MS lon Internal Standard (PFHS) 60 volts 299 amu 80amu --60 volts PFOS 499 amu 80amu `The capillary heater was held ata constant 300 degrees C. All quaniitative calculations were `based on the MS/MS ion rations between the compound of interest (PFOS) and the intemal standard (PFHS). The standard curve ranged from 5 ng/mL (pptob25)0 ng/mL (ppb). A `aqnuRadrsatqiucacreduvraflivutweoaefs0.u9s9e3d5w.iththe standard value weighted 1/X. This cufirtprovduceed `The first tubeoftwo available for control animal number 1R01464wasdiluted and run with the initial extractions. This tube (1:100 dilution) had a PFOS level of 2.3 ppm. Repeat of this tube dilution produced comparable results (2.1 ppm). The second tube labeled 1R01464was diluted 1:100 and extracted. The second tube on this control animal had aresultof less than the lowest standard (5.0 ppb on the diluted serum). 001539 : T-7098.3 AcutePOSF Inhalation Limit Test. Final Report. ORnOl4y6o5nweistehtrhuem ctoubmeecwtasdaatveaainabdlTe f#orrecanoirmdaeld.1R|0d1i46t5e, uannfdoertuxnattelry ttaihssctuutbbeeewaandsdlabeled recorded resuts under the animal number 1RO1465. ~~ "RTOheTs4a7mpZlaeppfeorar1eRd0t1b4e6swiagsninfoitceadnatslohweemartlyazned1(R20+14he6m8olaynsids)1,ROT1hATe.niTahlirsessaulmtpolfewas aelxstorarcet-ieoxnt.racted andthe analysis repeated. No significant diffewraesnnocteed on re- Resuls: Ratt Control PFOS Level 1RR0O14E62 yyeess ROMES yes RROO14M6E5 nnoo iRROO11446678 nnoo ROW no 1R01472 no <s05o.0 ppb (on .:diluted serum) 5413 ppppmm 38s8 ppoomm 93 pom 50 ppm (5.1 ppmonduplicate) --- die. 3DaMvCiodrJp.oErharteesTmoaxnicology BPlhoogn.e23665-11.87:3232.5070 FAX 651.737.4754 diehresman@mmm com UUL540 1 : = F`Ti-n7a0l9R8e.p3oArctutePOSF Inhalation LimitTest. es ae Ta mL 3 e -- Re s a Ta --am t-- ea |]--aam-- minyateiom-- n 1TH iEE =a[mn Ba reRo m Wedb e a waa e lk . Imm oooA m m b en Teete: noodr ie on 1 i rr * I. i. 2. I; CIR: ACA oremp] nm JOEJ JENr EEA WR UU1541 "