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% AR226-2750 Copies to: E. I. du Pont de Nemours and Company Haskell Laboratory for Toxicology and Industrial Medicine HASKELL LABORATORY REPORT NO. 253-79 ______ Material Tested______ Ammonium Perfluorooctanoate Haskell No. 12,037 Other Codes Sample Ready for Testing 1/5/78 Material Submitted by k lastic Products and Resins Dpart ent, Washington W c k s INHALATION SUBACUTE Introduction: Existing datal indicate that Ammonium perfluorooctanoat auses significant liver enlargement following acute exposure via inHSlation or oral routes. This experiment wa'lasi desiatied to Investigate a "no effect" level. Procedure: Houseline air-- (~20 IL/min) was passed through a cyclone-head dust generator connected to a particle agitator reservoir which c o n t a i n e d H p K l The resulting airborne particulate was passed into a 30 i battery jar exposure chamber. The atmospheric concentration o f ^ B ^ | . n the exposure chamber was monitored at 30-mln intervals with Celman glass fiber filters (type AE, 47 mm). The particulate concentration was determined by weight gain of the filters. Particle size was determined on a Brink cascade impactor. S lxlyChR-CD'"-1 albino male rats (20/group) were exposed to design level atmospheric concentrations of 0, 0.008 or 0.08 m g / R a t s were exposed, head only, for 5 consecutive days, 6 hr per day. After 5 days, the rats were given a 2-day recovery (weekend) which was followed by 5 daily, consecutive 6-hr exposures. All rats were weighed and observed daily (except weekends) during the exposure and recovery periods. Purina Laboratory Chow and water were available ad 1 ibltum at all times other than during actual exposure. Clinical laboratory examinations were performed on 10 rats from each group at 0, 14, and 28 days post-exposure (details in Appendix B ) . After a total of 10 exposures, 5 rats from each of the 3 groups were pathologically evaluated at 0, 14, 32 and 42 days post-exposure (details in Appendix C). Select organs were weighed; absolute and relative organ weights were calculated. On the 5th and 9th days of exposure, 10 rats were selected from each group for eye examinations. Company Sanitized. Does not contain T S C A CBI HLR #253-79 - 2- Results: Design Atmosplieric^BJ Concentrations Ab b /.*2______________ Atm.'spilerie S.D. 0.0 (control) 0.008 0.080 0.0 0.011 t 0.005 0.083 0.017 Mean Mat..3 Median Diameter . S.D. (u) 0.0 1.63 i 0.70 4.47 3.41 Weight Gain: Rats exposed t<^B ^ |showed a suppression of body weight throughout the test. The 0.083 mg/i. group was more severely affected during exposure (loss of body weight) and for 14 days of recovery (Appendix A). Clinical Observations: During each exposure, sporadic cases of blinking, pawing, chewing and red eye and nasai discharge were seen in all groups. Clinical Pathology: Some rats exposed tc^p^Jdust displayed elevated alkaline phosphatase activity. Effects were seen in glutamic-pyruvic transaminase and glutamic-oxalacetic transaminase after 0, 14 and 28 recovery days. The incidence was related to dose; higher activities were found at the higher exposure level. However, L * the latter activity at 0.083 rag/1 was statistically different from controls Appendix B.) Pathology: Pathologic evaluations showed cloudy swelling or granular degeneration of hepatocytes in the livers of rats (5 of 5 examined) exposed ti^BHQf'or 10 days with no recovery. This effecl longer recovery period. No other compound-related histologic changes were noted. or Appendix C.) Organ/Body Weights: Various organ weight effects were evident in rats exposed t seen after 32 or 48 recovery days and in the absence of related histologic abnor compound-related (Appendix D). The dose-related increase in liver weights correlates with existing data] described above. 1 These effects were not es were not considered and hepatocyte changes .gye Examinations: No corneal, iritic or conjunctival effects were seen in any of the rats examined after 5 or 9 exposure days (Appendix E). TTom p n S 'S anUiicd. Poos n ^ g n t i n T S C A C B 1 - 3- Summary: Groups o f J ^ n a l e rats were exposed, head only, to particulate atmospheres of 0, 0.011 or 0.083 rag/e ammonium p e r f l u o r o o c t a n o a t e M ^ n 6 hr /day, 3 days/wk for 2 wks with 42 days recovery. Rats exposed to^ppj|showed a dose-related suppression of body weight. Rats exposed to[(flhad dose-related elevations of plasma enzyme activities for up to 28 days of recovery; liver injury or dysfunctioni w a r suggested. Liver degeneration was detected at the end of the 2-week exposure period in all examined rats exposed t c M B i l This liver effect was not detected after 14, 32 or 42 days of recovery. Eye examinations revealed no compound-related effects. CTHsscg Report No. 2S3-79 .Date Issued: Jul\ Report by: 'T- $ (Saferge T. Hall Toxicologist Approved by: ____ __________________________________ Gerald L. Kennedy Chief, Acute Investigations Section .DOCS not contain T S C A CB1 Sanitized C o m p 30* .-wy-t. 450. t<J#.61J| 14iTO 13-JU.-79 APPENDIX A GROUTH CURVE-AMMON!UH PERFLUOROOCTANOATE RATS n EAN GROUP BODY U E I G H K GRAMS) 400. 350. 300. 250. 200. 150. I ILE LEOEM>s i unni -- LOU IOh -- *- DAYS ON TEST \ 9 m m * ~ * * ~ '* ~ ~ 1 * * ~ APPENDIX B SUBACUTE TWHAT ATTON OF AMMONIOM PERFLUCROOCTANOATE Hagfcgll Laboratory No..12037 Clinical L fboretorv Report Saiwlas of urine were collected overnight (16 hours) from groups of ten rats that were exposed nine times for four hours per day, ?lve. days, per week over a two-week period to 0 mg/L, 0.008 mg/L or **"sL 'Jftlust. Blood was taken from the tail of these rats the tMth exposure. Fourteen and 28 days after the last exposure blood and urine (24 hour) samples were again collected from the animals. The examination of the blood and urine included the following: nomacoloev: Erythrocyte count (RBC), hemoglobin (Hb),hematocrit rtiM to"al leukocyte count (WBC), relative number of neutrophils O'*') * lymphocytes (Lymph), eosinophils (Eosin), monocytes (Mono) and basophils iSces. .a corpuscular *ol>je (KV), meeo ,,,, hesmgloSlu ocm and m e m corpuscular hemoglobin concentration (MCBC), calculated from these data. rhomiarry: Alkaline phoaphatase (API, glutmlt-pi ^ rtt ttma _-- ir5rriutaaiic-oxalacetic transaminase (GOT), gan*na-glutamyl S S T b S S S . C M ) , r e . u itro g m ) and to ta l protein (TPROT). . Urinalysis: A measure of the volume (VOL), osmolality (mOs) and pH; a semi-quantitative test for sugar, protein and urobilinogen (UROBL); a test fo? blood and bilirubin; a microscopic examination of the sed.me from pooled specimens. The data were analvsed by a one-way analysis of variance (AOV) to comp^^e t ^ m a i a -iti the controls at eech by a nested AOV for the three sampling intervals. Signi-icanc J 8* at the 0.05 probability level. Results: The results of the clinical laboratory measurements or ^ f " `" oas are sunmarized in Table 1, and for the statistical analyses in Table a . Measurements oh individual animals are listed in t e compu p attached to this report. contain TSC A C B I Company Sanitized. Does not t activity was found for the plasma enrymes AP, Higher than nor"*1 * U | a s t after the last .exposure and GPT and GOT in some rats exposed t w activity was re- 14 and 28 day* Utar. lated tbthe dose, occurrig Lc!y\ t h r ^ t . exposed to 0.08 jwalvgia however, indicated that only elev a ted . Increased activity of the,, SSy'SS or dysfunction. A similar eJe^ - dose o ^ l H k l ) - These results, that were given a single "on: ^ "1 J ts expoid $ inhalation t f l p Q I o :" S ' b r ! f e c t l 5 " y tie e x p .. r . * 'th at the e f f e c t - y p e r s is t fo r davs or longer. WBK:JRB:ljn Date : O c t o b e r 19- 1 9 7 8 Report by ,'j J / Z B . Walter B. Koniecki Clinical Chemist Approved by:. V*dm STJo Hd Re Barnes Chief,\CUnical Pathology Section ... . p rot contain TSC CBi HEHAIPLQfiX Erythrocytes X lOVnm' Hemoglobin g X Hematocrit X HCV MCH ng HCHC X Leukocytes X 10*/i*i* NeutrophlIs X Lymphocytes X EosInophlIs 1 Hoimcytes X Basophils % CHEMISTRY AP 1U GPT IU GOT 1U g-CTP 1U Bilirubin sgX Urea nitrogen agt Total Protein gl URINALYSIS Volume ml Osmolality nOs pH Blood, nusdier posit!-', Sugar, number sbn:nl Protein, nunber ''normal Urobilinogen, Ehrlich units Bilirubin, number positive Microscopic, nunber abnormal TABLE 1 SUMMARY OP CLINICAL LABORATORY MEASUREME S CN RATS EXPOSED TO AMMONIUM PERFLUOROOCTAMOATE Test 1 iiik/L Recovery 14 28 3.86 13.1 46 78 26 33 15.15 18 81 0.8 1.0 0 6.30 14.8 46 72 23 32 17.76 22 75 1.2 2.1 0 6.25 14.9 47 75 24 32 15.73 18 76 2.7 3.4 0 201 23 62 1.7 0.3 16 6.6 226 23 68 0.9 0.4 21 6.7 270 25 68 1.0 0.3 IB 6.7 7 2019 7.2 0 0 0 1.0 0 0 18 1327 7.1 0 0 0 1.0 0 0 14 1706 6.7 0 0 1 1.0 0 0 Test 5.H7 14.7 43 77 25 33 10.77 19 79 . 0.6 1.2 0 242 28 72 1.2 0.3 19 6.3 6 2337 7.7 0 0 0 1.0 0 0 0.008 raa/L Recovery 14 28 6.12 14.3 45 73 23 32 13.17 19 78 1.3 1.6 0 6.24 14.7 47 75 24 32 12.92 15 81 1.3 2.6 0 296 25 74 0.7 0.4 22 6.7 . 291 26 67 0.8 0.3 19 6.8 16 1490 6.9 0 0 0 1.0 0 0 12 1818 7.3 1 0 0 1.0 0 1 Test 0.08 Jma/1. 1 Recovery i4 28 5.99 15.2 46 76 25 33 13.44 20 78 0.8 1.6 0 244 27 72 1.1 0.4 20 6.6 8 1624 7.9 0 0 0 1.0 0 0 6.17 14.5 46 75 23 31 17.00 17 81 0.9 1. 1 0 275 25 78 0.6 0.5 24 6.6 16 1596 7.7 0 0 0 1.0 0 0 6.37 15.2 4B 75 24 32 15.87 14 83 0.8 2.2 0 307 29 84 0.6 0.3 22 6.) 15 1991 0 0 0 1.0 0 0 net contain T S C A CB1 C o m p a q Sanitized. Does Erythrocytes Hemoglobin Hematocrit MCV MCI! MC IIC Leukocytes Neutr ;nhils Lym phocytes Eos inophils Monocytes Basophils AP GPT GOT gGTP Bilirubin Urea nitrogen Total Protein Urine VOL Urine mOs TABU-: 2 SUMMARY OF STATISTICAL DATA F: RATIO OF VARIANCE Tenth Exposure 0. 1.3 1. 1 0.8 1.7 0.1 5 7.4 0.4 0.7 0.2 0.7 0.0 4.4 1.4 1.5 1.3 3.0 4.8 1.9 2.0 7.4 14-Day Post Exposure 1.5 1.7 1.5 2.0 0.0 2.9 4.4 1.6 2.7 0.5 1.2 0.0 2.0 0.8 1.1 0.6 2.1 1.4 0.3 0.3 0.9 28-Day PostExpQBure 1.0 1.7 0.7 0.2 0.5 0.4 2.2 1.0 2.9 4.9 2.0 0.0 1.0 1.4 5.4 1.8 2.5 1.8 0.4 0.8 1.0 All Data 0.7 2.2 1.8 0.3 1.4 0.4 5.2 0.5 1.4 2.9 1.2 0.0 2.3 2.1 2.7 1.6 6.1 3.2 0.1 0.6 1.0 Company SanilizerL Ones nel contain TSCA CBI APPENDIX C AMMONIUM PERFLUOROOCTA.NATE H - 12037 PP&R Department Inhalation Subacute - ChR-CD Rats February 27. 1979 Summary and Conclusion Sixty young adult male rats were divided equally into three groups with Group I serving as a negative control and e x p o s e ^ n l | to air. Groups II and III were exposed to ammonium p e r i l u o r o o c t a n a t e M ^ ^ ^ J a t concentrations o 0.01 and 0.08 mg/1 respectively, 6 hrs/day, 5 days/week for 2 weeks^ On 0, 32, and 42 days post-exposure, 5 animals from each group were sacririced for pathological evaluation. At necropsy, the animals were examined grossly, ana the following tissues or organs saved for histopathologic evaluation: external ear, abdominal skin, trachea, lungs*, heart*, thyroid, adrenalsthymus , mediastinal tissue, spleen*, sternebrae with bone marrow, stomach, smal_ intes tine, large intestine, liver*, kidneys*, testes*, epididymis, brain and eyes. The results of pathologic findings are summarized in Table I. As the ^ table shows, an apparent compound-induced toxic effect consisting of a clouay swelling or granular degeneration of hepatocytes was detected in the livers of all five rats (5/5) in both Group II (0.01 mg/1) and Group _li (0.08 mg/1) at the end of the two-week exposure period. However, the hepatotoxic errect was not detected after a 14-day or longer post-exposure recovery p e n o a . all of the remaining pathologic findings listed in the cable are considered to be spontaneous or the result of intercurrent disease. * Organs weighed Report by:_ __________ Hans H. C. Chen, D.V.M. Senior Research Pathologist HHCC:WCK:mjfc Approved bv: William C. Rrauss, D.V.M. Chief, Pathology Section Company Sanitized. Does not contain TSCA CB/ D TAWJ; I BlIMMAnY OF rATlfOLOUIC FTNDIHnS IN MA1.E (Tlin-CP BATH EXPOSED TO AMMOMUM PEnnWROOCTAHATE r pwr-ii R R 2 ^ f*> R ?363f>7 R 23^358 R 236359 R 236360 R 2}636l R 2363*2 R 2Vi353 R P V tW i I0 T0 TO I0 I0 T0 I0 I0 I0 I0 I0 10 o 1 10 o 2 10 o 2 10 o 2 10 1 1 10 in 1 10 in 1 10 in 2 10 in 1 10 32 1 10 32 1 1 1 1 1 1 2 1 Code: -t no leelone lj very mlljd change 2: Slight change 3: Moderate change P: Present X: (Iroeely normal, the tlnnue not available for hlntologlc evaluation ' Company Sanitized. Does not contain TSC A CBt ,# 3 TART IT TARI.E I niJWARY OF PAT1I01XX1IC FTHDINOR IH MA1.E CIIR-CD RATO EXPOSED TO AHMWHJH W'.nFUJOROOCTAHAi'E R r 23*356 R ?3*?57 r 25*3118 R PV13I19 r ? T>fy$o R 23*3'51 n 23*352 R 2^*383 R ?3*3flh r 23*385 R 25*386 I0 10 32 l ~ * - - m '. I0 T0 I0 T0 I -0 10 T0 10 Y! l-1---- * _ 10 3? l - 1 - - - 1 . _ 10 te i - 1 - - - - - - . _ 10 ha l - - - - - - _ . - - . _ _ 10 ha l - 1 - - m . 10 ha l - - - - - ' - a m 10 ha a - 1 - - - 1 - - - - r - IT 0.01 in 0 2 - 1 1 - ,, ma II 0.01 10 0 i - 1 2 - - - m . II 0.01 10 0 2 - 1 2 * - - . _ _ II 0.01 10 0 2 - 1 2 - - - - - - - - - roAe! - j no lenlnna Is Very mild cliange 2 j S lig h t change J : Moderate change Ps l* a io n p re sen t X Oroaely normal, the rlnnnp nol nvnllnble for hlntnlngtc evaluation Company Sanitized. Does not contain TSCA CBS rAjrr m TABLE I miwuvnr o f pathologic rmmiiGn tn male cim-cn rats EXPOSED TO AWOHIUH rEHFUK)mX)CTAHATE Heart: Chronic focal tvocarditis Anlmnl No.. 1*1 m fi IV* 3 g I? to Q R IT 0.01 10 0 R pV-VfR IT 0.01 10 ii* R 2*M70 r 236100 236101 III 0.01 30 ii TI O.Ol 10 1 U TI . O.Ol 10 1 U r 216302 R 23637' n 23637'* R P3617' IT 0 .0 1 10 11 II 0 .0 1 10 32 II 0-01 10 32 IT 0.01 10 32 R 216376 R 236577 R 236360 R 236369 R 236370 R 236371 R 236372 II 0.01 10 3? II 0.01 10 3? II 0.01 10 1*2 IT 0.01 10 US II 0.01 10 us II 0.01 10 us II 0.01 10 us Code: -5 Ho lesione 1: Very mild hange 3 Pi I f 2 a&B j m f1 *4 |i +14* 12- - 1 -1 --- 2 -- - -- P-1 --- 2 - 1- ~ - - 2 -2 --- 1 - 1L - - - 1 -1-- - 1-1 3 -1 2-1 1 -1 1 -1 1 -1 1 -1 1 -1 2! Slight change -"- -- -- --------- 3! Moderate cliange t; i 1 1 - 1 1 1 1 1 1 .- 1 pt Lesion present Xi (Uoimly nonwl. not contain T S C A CB1 Sanitized- Does Company PART IV TABLE I OIIMMARY OF PATIIOUXJIC FTNimiOfl TN MA1.K Cim-CD RATS EXPOSED TO AWOMIW PERFLUOUOOCTANATE # Animal Ho, R PVJ1O3 n ?y>bob n 231^05 n 23r.iior> R 23W107 r 23M9B R 23*399. R 23*bOO R 23*1101 R 23*b02 iOX- ITT HI III TIT III III III III III III ,Qo . o.on O.Ofl o.on 0.00 o.on 0 .0 8 0.08 0 .0 8 0 .0 8 0 .0 8 84 <p I i? JL 10 0 1 10 0 1 10 0 ? 10 0 2 10 0 2 10 lb 1 10 lb 1 10 lb 1 10 lb 1 10 lb 2 3 I tu 11 -- -1 -1 -1 21 1 -1 1 h If 1~S-- H r-- ----T-- S P)8JUS q la hu(04) P.5 H0H4 S. ( c'a -TT<u401 >S1-4 If cfl in<4 u O Oe *h901 4 SiU* I -PPSH0I 4PI33A .. t? VU01I u4G101 UO031 s e 2_ _ a <h n 'a UOPU 0) -VP) lb fl#] rtftluO 3hur00M41tH>OoPVuH>I. a-t0oviI>1tr.I4Tu0UoQCM:9) coP 0) *PP pPu] O1J o> -UX. g .q 31 . 1 3- - 21 2 , mm . < MU 1 m m2 m - -- .. ,, .m 1 m 2 .. .. rode: No lesions Is Very mild change 2s S lig h t change 3: Moderate ctiange Ps le sio n present X: Orossly normnl, the tissue not nvnllnblfc for lilntnloglr evnluntlon Company Sanitized. Does not contain T S C A CBI. PftHT V inDiiCi & SWMARY OF PATHOLOGIC FINDINUS IN MAI cim -cn RATS EXFOBEP TO AWOHIW rERFLUOROOCTAMATE vOTvo4i h 0 MliflB I I Animal Wo. ! P 8 fc I IV 8 PI 3 l e S R 236393 III 0 .0 8 10 32 2 1 1 1P R 23639'* III 0.08 10 32 2 - 1 - - 1 m H 236399 I II 0.08 10 32 P 1 1 - - - - _ . 2 R 236396 . III 0.08 IO 32 1 1 _2 R ?36397 I I I O.O8 10 32 2 - 1 - - . 1 . .. R 236308 I I I 0.0 8 10 1? 1 - 1 . - m R 236309 I I I 0 .0 8 10 1*2 3 - 1 - 1 _ R 23639O II 0,08 10 1*2 1 - 1 . . 1 m m R 236391 I I I O.08 10 *2 X - 1 - _ _' _ R 236392 III 0.08 10 >2 1 - 1 - - - 1 - - l_ - - - Code: - No lesions l: Very mild change 2j Slight change 3t Moderate change Ts Lee Ion present X: Oroealy norml, the tlnnue ott nvnllnh&t for hi8inlogic evohint ton Company Sanitized. Does not contain T S C A CBI APPENDIX D i_ MEAN ABSOLUTE ORGAN AND BODY WEIGHTS OF MALE RATS. &jiP AMMONIUM PERFLUOROOCTANOATE VIA INHAL. (SUBACUTE-0 RECEDE* GRUE'____________EINAI BUI-------HEAEI---;--- LUNGS--- ~-- LI! I CONTROL II LOW LEVEL III HIGH LEVEL 273.6000 258.8000 239.0000+ 0.9720 0.8340+ 0.7760+ 12.3 F RATIO<1> LSD <2) DUNNETK3) WMS (4) 6,125* 21.6130 24.7992 246.0000 11.280* 0.0924 0.1060 0.0045 ____ GEQUE_____________ KIDNEY......._._IESIIS----- IHYMUS I CONTROL II LOW LEVEL III HIGH LEVEL 2.5560 2.2840+ 2.2 ISO-* 2.8060 2.5860 2.6520 0.6600 0.4660+ 0.3420+ F R A T I O <1) LS D (2) D U N N E T T (3) WMS(4) 5.920* 0.2269 0.2603 0 0271 1.540 0.2803 0.3216 0,0414 13.861* 0.1327 0.1522 0.0093 (j) RATIO OF AMONG- TO WITH IN-GROUP VARIATION-- ONE-FACTOR ANALYSIS OF "" . |J^ | # 'gM (2) LEAST SIGNIFICANT DIFFERENCE-- GIVEN A SIGNIFICANT (ALPHA=0.05) F ANY TWO MEANS DIFFERING BY MORE THAN THE I..SD ARE SIGNIFICATLY_BIFFERENT|^||a ;gg WITH A- VARIABLE-WISE FALSE POSITIVE (ALPHA) ERROR RATE OF 0.05. '"li (3) DUNNETT TEST-- ANY TREATMENT MEAN DIFFERING PROM THAN THE DUNNETT STATISTIC IS SIGNIFICANTLY DIFFERENT FROM THE CONTROL MEA^Lj WITH A VARIABLE-WISE FALSE POSITIVE (ALPHA) ERROR RATE OF 0.05. 3 ? 4 (4) WITHIN-GROUP MEAN SQUARE. Pi + SIGNIFICANTLY DIFFERENT <P<0.05> FROM CONTROL GROUP BY LSD. BeTM t SIGN IF 1CANTLY DIFFERENT (p<0.05 > FROM CONTROL GROUP BY DUNNETT TEST. f^ji _ * SIGNIFICANT AT THE 0,05 PROBABILITY LEVEL, ISfii YyI i M p ;si ii* 'M m Company Sanitized. Does not contain TSCA CBI 111 'lyi|i|i ZXS7S rS B K `X X M a ia !UaStbt * FOR MALE RATS EXPOSED TO MEAN RELATIVE ORGAN fJJTSIAUfuSi[S AMMONIUM PERFLUOROOCTANOATE VIA INHAL. (SUBACUTE -0 RECOVERY DAYS) :n "4 !| __ UEABI------ .LUNGS----- _LIVER--- SPLEEN---- __UIDNElJ| ' I CONTROL II LOU LEVEL III HIGH LEVEL F RATIO*1) LSD*2) DUNNETT(3) UMS *4) 0.3548 0.3243 0.3254 1.725 0.0406 0.0466 0.0009 0.6206 0.5447+ 0.6083 4.569* 0.0587 0.0674 0.0018 4.7259 4.9997 5.9226+ 4.316* 0.9299 1.0670 0.4554 0.2075 0.1665+ 0.1455+ 17,638* 0,0232 0.0266 0.0003 " 0.93481 0.8920 1 0.9292 ''v-1 0.273 | 0.1372 If 0.1574 il %0.0099;| ' -1 ` "T? ____ GEOUE---------- IESIIS - I CONTROL. II LOW LEVEL H I HIGH LEVEL 1.0253 1,0059 1.1105 0.2418 0.1808+ 0.1438+ F RATIO*i) LSD <2) III INNETT * 3 ) ' inS*4) 1.836 0.1266 0.1452 0.0084 7.713* 0.0549 0.0629 0.0016 U, RATIO OF AMONG- TO UITHIN-GROUP OARIATION r) ONE-FACTOR ANALYSIS OF VARIANCE (2, LEAST SIGNIFICANT DIFFERENCE-GIVEN W I C A N T ^ H A = 0 . 0 5 > F ^ ,TI' Z X J S Z S E f f S A ^ T i r (I i H ^ R S^ FOF ..OS. (A ) WITHIN-GROUP MEAN SQUARE. + SIGNIFICANTLY DIFFERENT (P<0.05) FROM CONTROL GROUP BY LSD. . SIGNIFICANTLY DIFFERENT <P<0.05) FROM CONTROL GROUP SY DUNNETT TEST. # SIGNIFICANT AT THE 0.05 PROBABILITY LEVEL. QW Vm Sanitfced-Do notcontainTSCACBl of MALE RATS EXPOSED TO_ ; MFAN ASSOLUTE ORGAN AND BODY "EIGHTS (SUBACUTE-14 RECOVERY DAYS) ammonium p e r f l u o r o o c t a n o a t e via inhal. ____ GROUP--- I CONTROL I I LOU L E V E L I I I HIGH LEVEL F RATIO<1) L S D (2) DUNNETT(3) UhS(4 ) ElblAL-BUI-- ___ HEARI---- 341.2000 341.0000 303.8000 1.1660 1.1720 1.0480 2.604 41.1241 47.1867 890.6333 1.810 0.1601 0.1838 0.0135 LUNGS-- _ 1.8480 1.6900 1.6640 1.226 0.2771 0.3180 0.0405 LIVER-- 14.4980 16.0560 15.0360 0.684 2.9474 3.3819 4.5750 ..,,SELEEN-;! ' 0.7500f| 0.6260 i 0*5860 ' " ' -i''r? 3.029f 0.1514 ? 0,1737 " 0.0121 ____ GROUP. I CONTROL I I LOW L E V E L I I I HIGH LEV EL F RATIO(1) LSD(2> DUNNETT(3 ) WHS<4) __ KIDNEY-- ___ IESIIS-- .,,.__IHY0US.-...___________ 2.8520 3.0060 2.6360 3.2760 3.1580 2.5840+ 0.6040 0.6520 0.6020 1.793 0.4278 0.4908 0.0964 3.949* 0.5740 0.6586 0.1735 0.250 0.1743 0.2000 0.0160 (1) PATTG o"f AMONG- TO UITHIN-GROUP VARIATION ONE--FAST OR ANALVSIS OF VAp A N C E < ,, LEAST SIGNIFICANT ^ F E R E N C E - G I V E N S nT S K VESTS# -- - OF o.os. <3, DUNNETT TEST-ANV T R I i ^ F ^ ^ T T ^ o S ^ r ^ ^ ^ L M E A N S TM IHU A R I A B L ^ E TFa" e POSITIVE (ALPHA) ERROR RATE OF 0.05. (4) WITHIN-GROUP MEAN SQUARE. : + SIGNIFICANTLY DIFFERENT <P<0.05> FROM CONTROL GROUP BY LSD. _V ,, ,, r m n o T (f 'O 05' FROM CONTROL GROUP BY DUNNETT TES.. ., * SIG N IFIC A N TLY DIFFERENT (I -0 .0 .J, /i * SIGNIFICANT AT THE ,,0.05 FpRpOnr^<BABiILLi1T1YY LLEtVEtL. Comosnv Sanitized-Does MEAN RELATIVE *Sa ?EB5 Ia INHAriS <SUBACUTE-14 RECOVERY DAYS);. TO AMMONIUM PERFLUOROOCTAwOATE VIA INHAL. 'M ill I CONTROL II LOU LEVEL III HIGH LEVEL F RATIO1> L S D (2) DUNNETT<3) UMS(4 > .HEART_______ LLINCS- 0.3420 0.3438 0.3458 0.5392 0.4965 0.5514 0.029 0.0339 0.0389 0.0006 1.952 0.0637 0.0731 0,0021. LIVER______ SELEEN------.tSiDNET- 4.2487 4,7164 4,9095* 3.350 0.5720 0.6563 0.1723 0.2180 0.1838+ 0.1925+ 5.090* 0,0243 0.0279 0,0003 0,8368 0*8837 : 0.8647 ' .,v!l 0.624; 6).0920. 0,1055. 01.0045 n o m iC' -- I CONTROL II LOU LEVEL III HIGH LEVEL F RAT 10 (T) LSD <2 > D U N N E T T (3) WMS<4) IESIXS-- ___ IHYMUS-- 0.9658 0.9263 0.8528 0.1773 0.1916 0.203 1.004 0.1763 0.2023 0.0164 0.408 0,0560 0;0642 0,0017 (1) RATIO OF AMONG- TO UITHIN-C < ,, LEAST SIGNIFICANT DIFFERENCE-GIVEN LSD^ARE^SIGNIFICATLT~DIFFERENT a SI t w o m e a n s d i f f e r i n g b y " r e t TM r n , ^ r h a > e r r o r RATE OF 0.05. UITH A VARIABLE-WISE FALSE POSITIVE . CONTROL EAN BY ORE <3. DUNNETT T E S T - A N T TREATMENT J ^ F E R E N T FROM THE CONTROL HEAN THAN THE DUNNETT POSITIVE (ALPHA) ERROR RATE OF 0.05. WITH A VARIABLE-WISE FALSE POSITIVE , (4) WITHIN-GROUP MEAN SQUARE. + SIGNIFICANTLY DIFFERENT <P<0.05) FROM CONTROL GROUF B . , SIGNIFICANTLT DIFFERENT (P<0.05) FROM CONTROL GROUP BT DUNNETT TEST. MVMk.lTC"l CAMI MB . t-nc. WBVW I TTV I P U E L . --- .jeaaiaiS-TM m _______ GBQUE___________________ Eibll-- BU- I CONTROL I I LOU L E V E L I I I HIGH LEV EL F RATIO(1) LSDC2) DUNNETTO) UMS<4) 3 6 7 .2 0 0 0 3 6 1 .6 0 0 0 3 5 1 .6 0 0 0 0 .4 9 4 3 4 .6 5 8 7 3 9 .7 6 8 1 6 3 2 .6 0 0 0 _HE6BI- 1.1100 1 .1 2 6 0 1 .1 3 4 0 0 .0 5 4 0 .1 6 2 8 0 .1 8 6 8 0 .0 1 4 0 .LUNGS______ LIUEfi 1 .7 9 4 0 1 .6 8 4 0 1 .7 6 0 0 1 2 .2 4 4 0 1 2 .4 2 4 0 1 2 .5 7 6 0 0 .5 7 5 0 .2 2 8 9 0 .2 6 2 6 0 .0 2 7 6 0 .0 9 9 1 .6 2 4 0 1.8634 1 .3 8 8 9 10 53g i^lj$O4S|| .GRQUE_______________________ KID N E*- I CONTROL I I LOU L E V E L I I I HIGH LEV EL 2 .6 7 4 0 2.8200 2 .6 6 6 0 F R A TIO (l) LSIK 2) DUNNETT(3) UMS(4) 0 .2 7 9 0 .3 0 5 8 0 .5 8 0 3 0 .1 3 4 7 _IESIIS_____ IHSUUS. 3 .0 5 2 0 3 .2 4 0 0 3 .1 0 6 0 0 .5 9 0 0 0 .5 2 2 0 0 .6 3 4 0 0 .8 9 1 0 .3 1 5 9 0 .3 6 2 5 0 .0 5 2 6 1 .8 1 6 0 .1 2 9 0 0 .1 4 8 0 0 .0 0 8 8 i|?( ( 1 , RA TIO o r AMONG- TO WITH IN-GROUP VARIATION-- ONE-FACTOR A N ALYSIS O P | A r t A N C E | (2 ) LEAST SIG N IFICA N T D IFFER EN C E-G IV EN ^ SNIPICANTpiALPHA O .O ^ F ^ ^ ,. y I-rHTA^VARI A B L E -W IS E "F A L S E ^ t S T < U a ERROR RATE OF 0 . 0 * . ;; ^ " <3, DUNNETT T E S T - A N Y TREATMENT FAN D I F F E R I N G J ir z r v A R ^ r * ** * of - ( 4 ) UITHIN-GROUP MEAN SQUARE. + S IG N IF IC A N T L Y D IFFER EN T < P < 0 .05 ) FRON CONTROL GROUP BY L S D . S IG N IF IC A N T L Y D IF F E R E N T <P<O.OS> FROH CONTROL GROUP BY DUNNETT T E S T , ^ j ;; , not contain TSCACBJ Company Sanitized. Does f I RATS E X P S s i& iiS ffil TO AMMMOENAIUNMRREELRAFTLIUVOERO^O^CTNAONOAATTEE ^VIIAA IINNHHAALL. ^ <SUBACUTE -3 2 RECOVERY ____ GRQUE------------------- I CONTROL II LOW LEVEL. III HIGH LEVEL F RATIO<1) LSD(2) DUNNETT(3) UMS <4) 0.3028 0.3104 0.3221 1.379 0.0255 0.0293 0.0003 LUNGS- 0.4884 0.4662 0.5004 1.449 0.0443 0.0509 0.0010 . H U E S _____ SELEBi---;if^lE5ii " 'iiMI MS 3.3384 3,4328 3.5735 0.1789 0.1661 0.1704 -0.7760% ^757ifj -;S| 1.472 0.3004 0.3447 0.0475 0.765 0.0231 0.0265 0.0003 --- JH29SS .;0'| p 0 6 7 f | IESIIS-- IHYUS---- I CONTROL II LOW LEVEL III HIGH LEVEL F RATIO (1> LSD <2) DUNNETT(3) UMS(4 ) 0.8336 0.8991 0.8849 1.067 0.1029 0.1180 0,0056 0.1627 0,1439 0.1810 1,913 0.0413 0.0474 0.0009 .I\ RATIO OF AMONG- TO WITHIN- g roup v a r i a t i o n ICANT (ALFHA=O.Oo> r **'**?' , , , LEAST SIGNIFICANT * W W ^ 5 LSD*ARE SIGN1FICATLY DIFFERENT ; UANT IWO MEANS DITM 1^, ^ POSITIVE (ALPHA) ERROR RATE OF 0.05. UI1H A VARIABLE - f f e RING FRO THE CONTROL HEAN BY MORE (A> DUNNHTT T E S T - A N Y TREATMENT ^ ^ [ r i . I F F E R E N T FROM THE CONTROL MEAN t han THE DUNNETT L g i T I V E (ALPHA) ERROR RATE OF O.Oo. WITH A VARIABLE-WISE FALSE FOSITIVt ^ . (4> UITHlN-GROUP MEAN SQUARE, ; , SIG N IFICA N TLY DIFFEREN T < P < 0 . > FROH CONTROL GROUP BY LS D . ^ . SIFNTF.CANTLY DIFFERENT CP<0.05> FRON CONTROL DROOP b y DUNNE.. .EST. ,.^ not contain TSCACBi tized. Doss Company Sam MTfiN ABSOLUTE ORGAN AND BODY WEIGHTS OF HALE RATS S S j AHNONIUH PERFLUOROOCTANOATE VIA INHAL. CSUDACUT-.2 RECOVERY ..GBQUE______________ E I N 6 I-- BUI. I CONTROL II LOW LEVEL III HIGH LEVEL 427.8000 409.8000 372.8000# F RATIO(1) LSD(2) DUNNETT(3) WM3<4> 3.449 46.3909 53.22*8 1133.3667 -UE6BI- 1.3460 1.2420 1,1640 3.078 0.1604 0.1840 0.0135 _LUUGS______ LIVER 1.9080 1.8920 1.8480 0,085 0,3275 0.3758 0.0565 i3.7:i|^a^:ia : 2.8192 "fS#*1 `*$1 3'..234%s^ ^ , . l r f 1 4.1855 ^-3^.009- ___ GEOUE______ -_____ KIDNEY---- .-IESIIS----- IBXI1US-- _ _L I CONTROL II LOW LEVEL III HIGH LEVEL 3.2100 3.2220 2.8500 3.3000 3.2560 3.2100 F RATIO(1) L.SD ( 2 ) DUNNETT(3 > WMS <4) 1.241 0.5846 0.6708 0.1800 0.235 0.2862 0.3284 0.0431 (Ii PATIO OF AMONG- TO WITH IN-GROUP VARIATION 0.6040 0,5920 0.5440 0.540 0.1331 0.1527 0,0093 ~'V "?# ONE-FACTOR ANALYSIS OF y ^ I A N Cl| A (2) LEAST SIGNIFICANT DIFFERENCE--GIVEN A STGNIFIChNT CALPHA O . O ^ F ^ R g ^ or o.oo. I (3) DUNNETT TEST-- ANY TREATMENT MEAN DiF P & ^ THAN THE DUNNETT STATISTIC IS SIGNIFICm NTu.'i D - ' W i t u WITH A VARIABLE-WISE FALSE POSITIVE (ALPHA) ERROR RATE 01- 0*0.. (4) WITHIN-GROUP MEAN SQUARE. ' + SIGNIFICANTLY DIFFERENT (PC0.05) FROM L'JNI i-.OL GROUF BY LSI. t SIGNIFICANTLY DIFFERENT <P-0,OS> FROM iNTNOI. GROUP BY DUNNETI TEST.. : J .3 not containTSCACB! Company S a n ltM * TM MEAN R E LA T IV E ORGAN A W BODY TO AMMONIUM PERFLUOROOCTANOATE V IA lNrlHL. % RATS EXPOSED ^ -42 RECOVERY DAYS) .GOUE- I CONTROL II LOW LEVEL h i h i g h Le v e l F RATIO(1) LSD(2) DUNNETT(3) WMS(4) .UERI- 0.3162 0.3029 0.3125 0.405 0.0333 0.0382 0.0006 .LUNGS-- 0.4451 0.4629 0.4950 1.530 0.0629 0.0722 0.0021 .LIVES-- 3.8684 3.7516 3.6835 0.573 0.3807 0.4368 0.0763 .SELEEN-- 0.1847 0.1613 0.1875 2.089 0.307 0.0352 0.0005 JEDUEX - . '- 0 .7 W G' 0.7855, 0.762S 0 .3 8 7 0.0904! 0.1037 0.0043 ____ SROUE______________ IESIIS------IHXttUS- I CONTROL II LOW LEVEL III HIGH LEVEL 0.7745 0.7982 0.8630 0.1422 0.1451 0.1456 F RATIO(1 ) LSD(2) DUNNETT(3) WMS(4) 2.687 0.0861 0.0988 0.0039 0.027 0.0345 0.0396 0.0006 u , RATIO OP a h o n g - t c u i t h i n - g r o u p m a r I a t I on o n e - f a c t o r a n a l y s t s OF OARIANLET <2 > LE A ST S I G N I F I C A N T D I F F E R E N C E - -G I M E N f S iu T < ^ > i g n IFICATLY DIFFERENT W R -- of 0 .0 5 . : ; , 3 , DUNNETT T E S T - A N Y TREATMENT MEAN D IFFER IN G FROM T H E ^ O N T R O D M E ^ B Y ^ O R E I - r r J S it Ssir^ -- of .. (4) WITH1N-GRGUP MEAN SQUARE. + S IG N IF IC A N T L Y D IF F E R E N T <P<0.05> FROM CONTROL GROUP BY L S D . . S IG N IF IC A N T L Y D IF F E R E N T <P<0.05> FROM CONTROL GROUP BY DUNNETT T E S T . APPENDIX E INHALATION TOXICITY OF OCTANOIC ACID, PERFLUORO-, AMMONIUM SALT HASKELL NO. 12,037 OCULAR TOXICITY REPORT :: iM *m r Hissites S.i`SfSiIf Procedure: Sixty ChR-CD young adult male rats were divided^nt|> . three croups of 20 rats each and exposed (head-only) six hours p^r day, five days per week for a total of 10 exposures over a "": week period to 0.0C, 0.01 or 0.08 mg/1 octanoic acid, perfluoro-, ammonium salt particulate. Ten rats were selected from each gropp for eye examination on the 5th and 9th days of exposure. Observe- ; tions of the cornea, iris and conjunctiva were made with a hand slit lamp and a biomicroscope after using Fluor-i-s^np stain, , Results: The results of the eye examinations for each rat are presented in Tables I, II and III. When rats were ^ o s e d b y ^ halation to octanoic acid, perfluoro-, ammonium salt particulate at 0.01 mg/1, two of ten rats at five days and five of ten rats at < nine days had dried reddish brown discharge around their eyes. When exposed to 0.08 mg/1 of test material, two of ten rats had dried reddish brown discharge at five days and five of ten had brown dis charge and one had dried reddish brown discharge at nine days. This discharge (chromodacryorrhea) is often seen in animals under stress. No corneal or iritic effects were seen m these *ats*. No corneal, iritic or conjunctival effects were seen m the control rats at any time. Summary: No significant ocular effects were seen in rat eyes when SEamlned after the fifth and ninth exposures to octanoic acid, perfluoro-- , ammonium salt. Report by: Doris F; Edwards Toxicologist Approved by : f. K Gerallld^^XL-. Kennedy Chief, Acute Investigations Section 1979 ,s not contain TSCACBI sd. Do; Company Sanffl* ua C:m spaeiry STM Iris Conjunctiva redness eheaosis ' discharge yiaor-i-strip Sisaie ef esrnss Carnes opacity area Iris Conjunctive radnass cseous discharge ?luor-i-stri?5 Bicsic ci caraea Cars** opacity tni Iris Conjunctiva redness eheaosis discharge ?luer-i-strips Bicnic -i cornea .6351 Caraea opacity &ru Iris Conjunctiva redness chaBOSia discharge 71uor-i-stria Slcsic ci saraea 36352 Cesses opacity STM Iris Conjunctiva redness chamesis discharge . ?luor-i-strip! Blcals of caraea a/4^ 9 ay* V9/ * 30 00 30 00 03 00 00 90 99 90 n9 ? 00 00 00 00 neg aeg " 3--- 5 03 00 9 o 0 00 00 09 aeg aeg 00 03 00 o 00 00 ns? 03 00 09 90 03 03 03 r.eg aeg La 00 00 00 30 00 00 -0 - aeg La 00 00 09 0S 00 00 -0 - aeg a a . 0w 00 00 00 00 00 -0 aeg as a 9* 03 09 00 00 00 _0 Be 00 c0 00 00 00 00 -0 aeg ldried reddish brown 2brown 5 day* 9/4 _ 9 ays 3/10 236393 Caraea apacrry araa Iris Ccnjunetiva reds* elMBOSiS discharge riaor-i-strip* atffgjc at eoraea 23639 Csrnea opacity area Iris Canjuneniva radnass cheaiosls discharge riaor-i-str-?5 oi esraee 236399 Caraea opacity area eesjuaetiva radnass eheaosis d isch a rg e ? luor-i-stri?s Isaac ad esrsea 2363S6 Cssaea opacity area Iris Csnjuaetiva redness eheaosis . discharge rlucr-i-strip* 3iaaic of cemee 23639? Cornea ep*eiS7 area Iris Conjunctiva redness. eheaosis discharge 7lucr-i-sarip ionic of cornea Qo 30 00 00 00 - 0 nag d* A Qo 00 00 00 -0 aeg 1 l 00 C0 39 oQ 03 0 0 neg ga 33 0C 0o 0 Q *0 * neg a 00 00 ft e 00 0I 00 -0 - aeg da A oo 0 0 99 . 0 00 s0 -0 neg - .3 9.0 00 a0 00 00 00 -0 aeg -3 09 00 30 00 00 00 -0 neg 1 00 00 00 90 00 00 -0 nag neg 0c 0c 00 s9 09 00 -0 * neg Sanitized. Does not contain Compaq Bar 23=363 opacity area Irla Can]ascriva rtdsus htttii discharge Finer "i-srrirs Sicmic ci carnai 236369 Canta opacity arsa Iris Caajunceiva redness cbaaosis discharge Fluor-i-srris9 Biscia ci carnai Corsea opacity ares Cria Cnnguseriva cecca]] cnemosij UJCiiCCt Flucr-i-srrip9 3iasie ei cara 236371 Cornea =P*ciry area Iris Cnngcnerivs radaui chamesis disear? Flucr-i-serip icsic c carras 236372 Comea opacitv arsa Conjunctive rtetiss abasosas discharr ?iaer--sm?s Sieaic ei ccraas TABLE II -(0.01 mg/L) H-12,037 S sv* 3/4 9 avi 9/18 I" TT 00 00 33 00 00 ol 0* 00 369 3S9 4* ~S~ 30 s0 30 00 00 0A 0* c0 aeg I A 0 00 00 3 00 3 0 0 0 00 02 0` -3 - 360 I a 30 00 0l o-0 - nag -a 00 c0 00 33 00 0* -0 - a? ** R 00 0 03 03 3 *w a3 -0 nec -s c3 30 *W 00 2<% 0 3 -C 3? 00 C0 00 03 30 33 03 3 at; `m ** 20 - 00 00 w 00 -0 nag - 33 3mw 3 _ 03 3C 3 0' -c " nag s a v i 9 y* clac 235373 c:n> osaeiry ITU Iris Cshjaacsiva radnass cianosis diseiarg Fiuer=i-srrip* Siesis ai corsea 236374 Comea oeaeiay area ' Conjunctiva redness Chemosis discharge r is e r -e-seri?3 Simio of cornea Cernea opacity arsa Ccnguscriva redness cnasesis discharge Fisar-i-smy5 Siesic ei cornea Corroa cpaciav arsa Iris ~ ag ineriva rodnes* chasesis iischarga "iacr-i-srrip* 3ieaie ci esraat Cornea opacity area Ir;i Conjunctiva redness erase sis issharga ?iocr-i-ftripr Sicaic oi cam* ao oo oo 00 0 0 0 0 0 -a os oo oo 9s 3 0^ w 0 rag P a 0 00 00 00 00 00 -0 - nag aa 00 c0 c0 3c 0- 0* 0 * sag 1a e0 3 0c o2 20 C0 0Q 00 30 -0 - nag m* A 2 3 0* c1 5 ' - 69 2, a 00 33 3z .3 o 3 Q 0 - nag 4s a 0 w 33 3* 0z 3 .C - 69 0 3 w 2n m w 3 * 2 - 69 -s ,, ** ; . 2r 2 - 69 Company Sani! Oo"3 not contsin TSCA Kttrtfii TABLE III - (008 mq/L) H-12,037 5 days 3 day* ?.i- 8/4 8/iO TT TT 36333 canw opacity ITM iria Cesjuaetiva rcinese ebanosis discharge Pluor-i-s-- i?` 3icnie of e e m e * 138389 coma* epici:? rii zeta Conjunctiva rainaai chanosis discSarga piuor-i-strip* aicare of esr^** 236390 Carnot opacity area Zi. Conjunctiva rodaosa ehosoaia d iacbarge rluor-i-*-- -?S a to n ie o e s s a # * neg neg La 0 3 0 0 0 l* 0 = neg te 3 0o 00 00 Q o 00 -0 aag La 00 00 00 b0 0 0 0 a9 la o c> 00 09 00 09 09 00 _0 _ aeg H 0 Aw 00 09 00 00 -0 - nag La Carata opacie*/ ITU iris Conjunctiva rodaasa ehanoaia disebarga Pluor-i-rtri?9 Siate of cacasi - 2363S2 Coma* opacity Iras Conjunctiva rodaoaa e s an o s i s di aerarla atonie of ceraia 1redd ish brown o ft Q 00 ft o 0Al 0 w' -9 nag te a 00 99 9 00 0a 00 0 * aag 90 99 00 90 09 1 l1 . 9 sag te R 6e gc a0 0c 0p 1 -0 - sag 236393 Cazaoa opaciey tra* Zzi. Conjunctiva rodaass ehaaosis discharge . rlcor-i-a-- i?s Bionic o osait 236394 23 Cornea opacity ITU Iris Conjunctiva redness ehanosio discharge riuer-i-serip State ef corsa* - Cssaaa cpaciey area iris Conjunctiva rd&ats ebanoois discharge riaor-i-strip Sionie ef cerna* 236396 Cornea opacity asss iris- Conjunctiva rodaass ebanoais disebarga riuor-i-serip Sienie ef come* 236393 Corsa* opacity cre irta Conjunctiva rainas* ebascais disebarga riaor-i-steip9 atonie of cerna* 5 days mM 9 ays fa 1 " 00 00 90 00 90 00 .0 - n*g ia c0 cc 00 00 0 J .0s 0 ne Ta 90 00 00 03 C9 00 -0 - sag ta 00 00 00 c0 00 0` 0 -0 a te a 00 ss 00 9g 00 00 -0 - amg La 0ft a 0 g0 03 00 00 c - ne te ? 00 00 go 0( 0( 9 * 00 go o o1 -0 - aag La 0C 0* 0a -c - SM *te ? 00 oe 03 00 99 00 -g - ne S9 cg e9 09 gW 00 -c am Sanitized. Docs contain TSCA-CBI not . .