Document EzNp5qj1b06M9Xp3KzoG35q0

AR226-2794 FOR DU PONT USE ONLY Approximate Lethal Concentration by Inhalation (ALC) of Haskell Laboratory Peport No. 293-85 MR No U E. I. du Pont de Nemours and Company Haskell Laboratory fo r Toxicology and Industrial Medicine P. 0. Box 50, Elkton Road Newark, Delaware 19714 Date Issued: June 3, 1985 Date Revised: December 31, 1985 mmniliSiilm Company Sanitized. Does not contain TSCA CBI mm m HLR 293-85 Approximate Lethal Concentration by Inhalation (ALC) of Summary To determine a^ALC, groups of 6 male Crl:C0($D)BR rats were exposed to dust atmospheres o f * f l f o r a single, 4-hour period, followed by a 14-day recovery period. Afffr determining an ALC, 2 groups of 10 male rats were exposed for 4 hours to ajn^ginally lethal and approximately l/10th of a lethal concentration o f M 0 For each test group, a control group of 10 male rats was exposed simultaneously to air only. Five rats per group were killed on the 5th day of recovery, and 5 rats per group were killed on the 12th day of recovery for a gross examination of the liver. Under the conditions of this test, the ALC f o r M W w a s 590 mg/m^. This material is considered moderately toxic by inhalatloTT. 3 Rats exposed to 67 mg/m had significantly elevated mean liver weights and 11ver-to-body weight^ratios on the 5th and 12th days after exposure. Rats exposed to 590 mg/nr had significantly elevated 11ver-to-body weight ratios on the 12th day after exposure. Liver weights for these rats were not significantly different than controls on the 5th day of recovery, and mean liver weights were significantly depressed on the 12th day of recovery. However, these seemingly inconsistent changes were due to severe body weight loss in rats exposed to 590 mg/m. 3 Gross pathologic examination of rats exposed to 590 mg/m revealed discolored livers with prominent lobular patterns in 4/5 rats killed on the 5th day of recovery, and similar gross liver lesions in 2/5 rats killed on the 12th day of recovery. Work b* : J f K t kxiu. Robert T. Tiurner Technician i- n Study Director: Laura A. Kinney Chemist A U LU LS LAK:smk:HLR9.14 Approved by: Nancy , Chromey, flh7u7 Section Supervisor Acute Investigations ^ (A 2- ,,,> '1 .; . . I "z ' tm wrnim aim Sim m ^^ Haskell Laboratory Report No. 293-85 Revised HR No. Material Tested: Haskell No. 15.437 Nonanoic acid, heptadecifluoro-, ammonium salt Sponsor: Polymer Products Department E. I. du Pont de Nemours and Company Wilmington, Delaware Material Submitted b y : S. C. Mai hotra Polymer Products Department E. I. du Pont de Nemours and Company Parkersburg, West Virginia Test Faci Iity: E. I. du Pont de Nemours and Company Haskell Laboratory for Toxicology and Industrial Medicine P. 0. Box 50, Elkton Road Newark, Delaware 19714 Study Initiated/Completed: 1/3/85 - 2/3/85 There are 15 pages in this report. Distribution; -3- not contain t s c a c b i Company Sanitized. Does INTRODUCTION HLR 293-85 The purpose of this study was to determine a 4-hour inhalation ALC for C-9 in male rats. The ALC was defined as the lowest atmospheric concentra tion tested that caused the death of 1 or more rats either on the day of exposure or within 14 days post exposure. In addition, following the determination of the ALC, additional exposures were conducted to monitor the effects of C-9 inhalation on the liver. Except as documented in the study records, this study was conducted according to the applicable Good Laboratory Practice Regulations. MATERIALS AND METHODS A. Animal Husbandry Young adult male Crl:CD(SD)BR rats were received from Charles River Breeding Laboratories, Kingston, New York. Each rat was assigned a unique 6-digit identification number which :orresponded to a numbered card affixed to the cage. Rats were quarantined for one week prior to testing, and were weighed and observed twice during the quarantine period. During the test, rats were housed in pairs in 8" x 14" x 8" suspended, stainless steel wire-mesh cages. The rat assigned the lower number in each cage was identified by a slash in the right ear. Prior to exposure, rats' tails and cage cards were color-coded with waterinsoluble markers so that individual rats could be identified after exposure. Except during exposure, Purina Certified Rodent Chow #5002 and water were svai.able ad libitum. B. Exposure Protocol To determine an ALC, groups of 6 rats, 8 weeks old and weighing between 234 and 298 grams, were restrained in perforated, stainless steel cylinders with conical nose pieces. Each group was exposed nose-only for a single, 4-hour period to a dust atmosphere of C-9 in air. Rats were weighed prior to exposure, and were observed for clinical signs of toxicity during exposure. Surviving rats were weighed and observed daily for 14 days post exposure, weekends excluded except when deemed necessary by the rats' condition. To monitor the effects of C-9 inhalation on the liver, 2 groups of ip rats, 8 weeks old and weighing between 237 and 277 grams, were restrained and exposed to a marginally lethal and approximately l/10th of a lethal exposure concentration of C-9, respectively. Two groups of 10 rats, 8 weeks old and weighing between 231 and 267 grams, were restrained and exposed to air only. Each control group was exposed concurrently with HEikYiiniinia*iiItiMi -4Company Sanitized. Does rot contain TSCA CRI Revised HLR 293-85 one of the test groups. Five rats per group were killed 5 days after exposure, and 5 rats per group were killed 12 days after exposure for pathologic examination of the liver. C. Test Material Other Codes: Stabi 1ity: the test. assumed to be stable throughout D. Atmosphere Generat <on For most exposures, dust atmospheres of C-9 were generated with a K-Tron Bin Feeder equipped with twin feed screws. The feed rate was regulated with a K-Tron Volumetric Feed Controller. The bin feeder metered test material into a glass transfer tube. Air introduced at the tube swept the test material through a size-reducing cyclone and into the exposure chamber. The cyclone removed large particles by inertial impaction, while aerodynamic particles passed through the cyclone and into the exposure chamber. The atmospheric concentration of C-9 was controlled by varying the feed rate. For the lowest exposure concentration, the atmosphere was generated by passing pressurized air through a 2-stage glass generator. A round flask at the bottom of the generator served as a dust reservoir. A cyclone elutriator was inserted above the reservior. A motorized stirring rod wit! plastic paddles agitated dust in the generator. Air introduced at the bottom of the reservoir and at the cyclone e'utriator swept dust particles into the exposure chamber. The atmospheric concentration of C-9 was controlled by varying the 2 airflows. E. Analytical The atmospheric concentration of C-9 was determined at approximately 30-minute Intervals by drawing calibrated volumes of chamber atmosphere through preweighed. Gelman glass fiber filters. Filters were weighed on a Cahn model 26 Automatic Electrobalance*. The atmospheric concentration of particulate wa$ determined from the filter weight differential before and after sampling. Particle size (efass median aerodynamic diameter and percent respirable) was determined with a Sierra model 210 cascade impactor during each exposure. During most exposures, chamber temperature mas - 5 - Company Sanitized. Does not contain TSCA P ! HLR 293-85 measured with a mercury thermometer, relative humidity was measured with a Bendix model 566 psychrometer, and chamber oxygen content was measured with a BioMarine model 225 oxygen analyzer. F. Pathology Two groups of 10 rats were exposed for 4 hours to 67 or 590 mg/m of C-9 in air, and 2 groups of 10 rats were exposed to air only. Five rats per group were arbitrarily selected and killed by chloroform anesthesia and exsanguination 5 days after exposure. The remaining 5 rats/group were killed 12 days after exposure. Rats were necropsied and the livers were weighed. Mean liver weights and 1iver-to-body weight ratios for test rats were compared to their respective controls by Least Significant Difference and Dunnett's tests. Significance was judged at the 0.05 probability level. G. Records Retention All raw data and the final reports will be stored in the archives of Haskell Laboratory for Toxicology and Industrial Medicine, Newark, Delaware, or in the DuPont Hall of Records, E. I. du Pont de Nemours and fompany, Wilmington, Delaware. Characterization of C-9 Atmospheres and Associated Rat Mortality HL 293-85 Concentrt ion (mg/m^) Mean S.D. Range % Respirable3 620 910 16CJ 4600 180 490 - 930 760 10 - 1700 420 920 - 1900 600 4100 - 5900 91 94 93 90 59607cc 35 290 .12 - 120 300 - 1200 95 75 b MMD(jm) 3.4 3.2 3.4 4.2 3.5 3.7 Mortality i* deaths/# exposed) 0/6 4/6 6/6 6/6 l/5d C7 tit 3 Percent by weight of particles with aerodynamic diameter less than 10 um Mass median aerodynamic diameter. c Exposures for subsequent pathologic examination, d Although this exposure was conducted for subsequent pathologic examination of the rats, 1/5 rats designated to be killed 12 days after exposure died on the 12th day. B. Clinical Observations During or immediately following exposure, some rats exposed to the 5 lowest exposure concentrations had red or brown facial discharges. At 620 mg/m3 and 1600 mg/m3, rats had test material on their heads. Rats exposed to 4600 mg/m3 had labored breathing, profuse clear nasal and oral discharges, a diminished startle response and test3materiaT on their heads. All rats died during exposure to 4600 mg/m3. 3During the postexposure period, 1/5 remaining rats exposed to 590 mg/nr died 12 days post exposure, 4/6 rats exposed to3910 mg/m3 died 9-11 days post exposure, and 6/6 rats exposed to 1600 mg/nr died 4-8 days post exposure. Rats exposed to 67 mg/nr lost 1-9% of initial body weight 1 day post exposure, fallowed by normal weight gain. At concentrations greater than 67 mg/m3, rats lost approximately 6-15% of initial body weight 1 day post exposure, and continued to lose weight either throughout the* recovery period or until they died. Most surviving rats exposed to 590, 620 or 910 mg/m3 weighed only 54-71% of initial body weight when they were sacrificed 12 days post exposure or at the end of the recovery period, respectively. No adverse clinical signs were observed in rats exposed to 67 mg/m throughout the recovery period. Common clinical signs at higher concentrations included hunched posture, ruffled or discolored fur, red Coinpany SanKlaad. Does no*contain T S C A OB* H S HLR 293-85 or brown facial discharges, wet or stairtid perineum, pallor, lung noise or labored breathing, lethargy, limpness and hair loss. Clinical signs were observed throughout the recovery period. C. Pathology 3 Gross pathologic examination of rats exposed to 67 mg/m revealed large livers in 5/5 rats killed on the 5th day of recovery. After 12 days,of recovery, none of the rats' livers were noted as large. At 590 mg/m , 4/5 rats killed on the 5th day of recovery had discolored livers with prominent lobular patterns or markings. These patterns were probably attributable to diffuse fatty change with a zonal distribution. On the 12th day of recovery, gross liver lesions were noted in 2/5 rats. The Pathology report is attached as Appendix I. Statistical analyses showed significantly elevated mean liver, weights and liver-to-body weight ratios in rats exposed to 62 mg/nr on both the 5th and the 12th days of recovery. In the 600 mg/nr exposure group, mean liver weights and 1iver-to-body weight ratios were not significantly different from controls on the 5th day of recovery. On the 12th day of recovery, mean liver weights were significantly lower than controls; however, liver-to-body weight ratios were significantly elevated. The lack of significant liver weight changes on the 5th day of recovery, and the depressed mean liver weights on the 12th day of recovery were due to a large loss of body weight in rats exposed to 590 m g / m . Mean body weights for rats exposed to 590 mg/m11 were only 70* and 48* of controls on the 5th and 12th days of recovery, respectively. However, mean liver weights were 83* and 72* of controls, respectively. Statistical analyses of mean liver weights and 1iver-to-body weight ratios are presented in Appendix IT. CONCLUSION 3 Under the conditions of this study, the ALC for C-9 was 590 mg/m . This mater1al,1s considered moderately toxic by inhalation (ALC between 200 and 800 mg/nr). However, C-9 caused elevated liver weights in rats exposed to 67 mg/nr on the 5th and 12th days after exposure; and gross liver lesions in rats exposed to 590 mg/m. 1 Calculation described in Sierra Instruments, Inc., Bulletin 7-79-219IM, Instruction Manual: Series 210 Ambient Cascade Impactors and Cyclone Preseparators. - 8- Company Sanitized. Do ,s not contain SCA CW m iSSa HLR 293-85 Appendix I - 9 ,,,,co o ttln T S C A C B t C o P an* Samtized`DoeS > 08* ir*4*s-is<ox E. I. ou Pont oe N emours & C ompany CO*iCMMVrD Haskell L aboratory for Toxicology amo Industrial Meo ic.ne P O Box SO. Elk to n R o a d NEWARK Delaw are 197(1 C E N T R A L R E S E A R C H a n d D E V/ELO P'.U n r D E P A R T M E N t MEDICAL RESEARCH PROJECT NO.! HASKELL LABORATORY MO 15437 NONANOIC ACID, 2,2,3,3.4,4,5,5,6,6,7,7,8,8,9,9 -HEXADECAFLUORO-, _________ AMMONIUM SALT C-9 APPROXIMATE LETHAL CONCENTRATION (ALC) TOXICITY STUDY _____ _________ IN MALE Crl;CP*(SD)BR RATS______________ CROSS PATHOLOGY POLYMER PRODUCTS DEPARTMENT DATE ISSUED: MAY 16, 1985 Q - ALC TOXICITY STUDY IN MALE RATS WITH C-9 Introduction and Results Male Crl:CD(SD)BR rats exposed to C-9 for A hours via Inhalation were necropsled and livers were weighed and examined at 5 and 12 days post exposure. Method of euthanasia was chloroform anesthesia and exsangulnatlon. Table I identifies the exposure groups and contains the gross observations made at necropsy. The liver of only a few exposed animals was noted, as large at necropsy, however, liver weights (weights and statistics will not be presented in this report) indicate that the liver of all exposed animals was large. The prominent lobular pattern noted in A of 5 high dose rats exa<"'..ed on day 5 post-exposure Is most likely attributable to diffuse fatty cnange with a zonal distribution (centrilobular or periportal). A portion of the Increase in liver size may be due to fatty change, however, it is more likely that the liver functions as the primary site o f metabolise of the compound and that the increase in size is due to a proliferation of smooth endoplasmic reticulum. Acknowledgement Joan A. Wolfe was pathology supervisor for this study. Report by: Theodore W. Slone, D.V.M Diplomate A.C.V.P. Staff Pathologist TWS/VCK/wfd SLONE 3.13 Approved by: Manager, Pathology Division -.11 - not contain t s c a c b * c o m P any Sanitized- DeS 3 H - 15437 TABLE I Animal Test Recovery Number Days Days Group IA - Control 386616 6 5 386618 386620 386622 386624 386617 386619 386621 386623 386625 6 6 6 6 13 13 13 13 13 5 5 5 5 12 12 12 12 12 GROSS PATHOLOGY C-9 MALE RATS - INHALATION Mode of* Death Observti SD Liver - discoloration, dai mm), left lobe SD No abnormalities detected SD No abnormalities detected SD No abnormalities detected SD No abnormalities detected SD No abnormalities detected SD No abnormalities detected SD No abnormalities detected SD No abnormalities detected SD No abnormalities detected Group IB - Control 386988 6 5 386990 6 5 386992 6 5 386994 6 5 386996 6 5 386989 13 12 386991 13 12 386993 13 12 SD No abnormalities detected SD No abnormalities detected SD No abnormalities detected SD No abnormalities detected SD No abnormalities detected SD No abnormalities detected SD No abnormalities detected SD No abnormalities detected * SD - Sacrificed by design; FD - Found dead 12 - Company Sanitized, Does noi confato TSCA CB! Mai MALE RATS - INHALATION s s s s c s m s s s s s s assaasssssas sasssisss3333S9aaBSBss>sa3ssa:ais3B3XBBacB*susxa Animal Test Recovery Mode of Number Days Davs Death Observations 386995 386997 13 13 12 12 SD No abnormalities detected SD No abnormalities detected Group II - High Dose (0.59 mg/L) 386545 6 5 SD 386546 6 5 SD 386550 6 5 SD 386552 6 5 SD 386554 6 5 SD 386547 13 12 SD 386549 386551 386553 13 13 13 12 12 12 FD SD SD 386555 13 12 SD Liver - discoloration, lobular pattern prominent Liver - discoloration, lobular pattern promlnent No abnormalities detected Liver - discoloration, lobular pattern prominent Liver - discoloration, lobular markings prominent, tan streaks scattered Liver - foci, white, scattered, (< 2 mm in diameter), all lobes No abnormalities detected No abnormalities detected Liver - discoloration, small, white area, left lobe, (< 2 tma in diameter) Perineum - alopecia, moderate Dorsum - alopecia, right, moderate Group III - Low Dose (0.067 mg/L) 387033 6 5 SD 387035 6 5 SD Liver - large Liver - large - 13 Company SanUliad. Doe. not.conl.In TSCA CBI [-15437 TABLE I ri ssasasBS S3SSSS Animal Test Number Days GROSS PATHOLOGY C-9 MALE RATS - INHALATION ssasssasmssss3am*iscsssssstaBaBflBsaBasaBsaBaBMsBSBKssscKmaratmas Recovery Mode of Days Death Observations 387037 387039 387041 387034 387036 387038 387040 387042 6 6 6 13 13 13 13 13 5 5 5 12 12 12 12 12 SD Liver - large SD Liver - large SD Liver - large SD No abnormalities detected SD No abnormalities detected SD No abnormalities detested SD No abnormalities detected SD No abnormalities detected Part 3 - 14 Crapn^ tsca APPENDIX II C9 - MEAN LIVER WEIGHTS AND LIVER-TO-BODY WEIGHT RATIOS RATS SACRIFICED ON THE 5TH DAY OF RECOVERY GROUP CONTROLAI 67 MG/M3 CONTROL II 590 MG/M3 FINAL WT. 275.6 (O.OO) 276.6 (0.912) 293.4 (0.000) 204.0 (0.000)# LIVER 11.306 (O.OCO) 14.501 (0.002)# 14.511 (0.000) 12.010 (0.164) LIVER-TO-BODY WEIGHT RATI# 4.090 (0.000) 5.243 (0.000)# 4.946 (0.000) 5.803 (0.053) Values in parentheses - P value of Student's t test conpari son of treatment mean to control mean. + - Significantly different (p<0.05) from control group by LSD # - Significantly different (p<0.05) from control group by LSD and Dunnett's test i a ;i| %"I C9 - MEAN LIVER WEIGHTS AND LIVER-TO-BODY WEIGHT RATIOS RATS SACRIFICED ON THE 12TH DAY OF RECOVERY GROUP FINAL WT. LIVER LIVER-TO-BODY WEIGHT RATIO f CONTROL-I 315.4 (0.000) 14.806 (0.000) 4.678 (0.000) 67 MG/M3 325.0 (0.374) 20.371 (0.001)# 6 .2 6 8 (O.OOOf# CONTROL II 335.4 (0.000) 15.557 (0.000) 4.649 (0.000! 6i 590 MG/M3 160.0 (0.000)# 11.198 (0.002)# 7.003 (O.OOO)# >$ Values in parentheses - P value of Student's t test comparison & of treatment mean to control mean. I + - Significantly different (p<0.05) from control group by LSD # - Significantly different (p<0.05) from control group by LSD and Dunnett's test - 15 Compaq S> DO! . si# . iAip iilfl : a m $$(p] -i ,-..;tag.:39iS: CENTRAL RESEARCH AND DEVELOPMENT DEPARTMENT HASKELL LABORATORY FOR TOXICOLOGY AND INDUSTRIAL MEDICINE December 31, 1985 k Amendment to Haskell Laboratory Report No. 293-85 The chemical abstracts name for ammonium perfluorononanoate was incorrect' listed on page 3 of the report as Nonanoic acid, 2 , 2 , 3 , 3 , 4 , 4 , 5 , 5 , 6 , 6 , 7 , ji 8,9,9-hexadecafluoro-, ammonium salt. The correct chemical abstracts nfSes'1 is Nonanoic acid, heptaaecafluoro-, ammonium salt. Page 3 of HLR 293-IB5^^5)| been revised to include this correction. Hie other code for rfluoroponanoate was incorrectly page 5 q f the report as" The correct code name i '1 Page 5 of HLR 293-85 has been revised to include this Study Director JUQ u a a Laura A. Kinney Chemist 12 / * / / 9 ^ Approved by: 4 4 &!. Chromey^Vi. D . M titin Supervisor Acute Investigations Section L ' .' ',`if Vr-