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AR226-2777 FOR DU PONT USE ONLY REVISED REPORT E. 1. du Pont de Nemours and Coapany Haskell Laboratory for Toxicology and Industrial Medicine Elkton Road, Newark, Delaware 19711 HASKELL LABORATORY REPORT NO. 589-80 Material Tested Octanolc Acid, Pentadecafluoro-, Ammonium Salt* Haskell No. 12,057 Other Codes Study Inltiated/Completed 10/17/79 - 12/14/79 Material Submitted J>1 'Polymer Products Washington Works RAT SKIN ABSORPTION SUBACUTE STUDY U I T H ~ B B B y I. Introduction: One o b j e c t o f t h i s subacute study was to determine any toxic effects related to a p p l y i n g f i H H H o n the shaved intact skin of rats at various concentrations. Another objective was to define the correlation between the effects seen and determined blood organofluorine concentrations. II. Protocol and Procedures: A. General Protocol: Male ChR-CD rats were exposed dermally t o C m B ^ a p p l i e d as an aqueous paste, at concentrations of either 20, 200 or 2,00$ m g Q B H K l k g body weight. A control group of rats was exposed simultaneously to distilled water only. Four groups of 15, 8 wk old rats (initially weighing from 210 to 245 gms) were exposed for 6 hrs/day (compound wiped off after 6 hrs), 5 days/wk for 2 wks. Doses were based on daily, individual body weights. Exposure period was tollowed by a 42-day recovery-observation period. Five rats per group were randomly selected for sacrifice on exposure day 10, and recovery days 14 and 42. Rats were collared oy securing 1-1/2" ID x 2-1/2" OD Teflon rings around their necks. Collars prevented ingestion of compound by rats when preening and grooming for 2 wk exposure period. After collaring, rats were housed individually and held for a 1 wk pretest period to ensure suitability. Compound was applied to the backs of each ret on a J" x 1-1/2" area shaved free of hair. Less than 0.5 ml of compound was applied in aqueous form to the shaved area. Daily 6 hr exposures' ended when compound was wiped from the rats backs with a gauze pad. Collars were removed after exposure day 10. Throughout the test period, Purina Rodent Chow and water were available ad libitum. All rats were weighed and observed daily (except weekends) throughout the exposure period and approximately every 3 days thereafter. - 1Company Sanitized- Does nol contain TSCA CBI B. Clinical Pathology Procedure: After exposure day 10, recovery day 14 and 42, blood waa taken from the tails of 5 rats from each group for heaatology measurements including: erythrocyte count, hemoglobin, heaatocrlt, leukocyte and relative number of neutrophils, lymphocytes, eosinophils, monocytes, basophils, mean cell volume, mean cell hemoglobin, mean corpuscular heaoglobin concentration Clinical chemistry measurements included alkaline phosphatase, glutamic-pyruvic transaminase, glutamic-oxaloacetic transaminase and total protein. C. Pathology Procedure; Gross autopsy and hlstopathological examinations were done on 5 rats/group after exposure 10, and on recovery days 14 and 42. Histopathology was done on the bone narrow, brain, cecum, colon, epididymides, esophagus, eyes, heart, kidneys, liver, lungs, skin, small intestines, spleen, stomach, testes, thymus, thyroid, trachea and lymph nodes. Mean absolute and relative organ to body weight analyses were done on the following: liver, spleen, testes, kidneys and thymus. D. Organofluorlne Analysis: After exposure 10 and on recovery days 14 and 42, blood was collected from 5 rats/group by cardiac puncture for organofluorine determinations. E. Ocular Examinations: Eye examinations were done on each rat after exposure 9, and on recovery days Id and 41. Procedure included gross observation of the eyes using a bright light and semimicroscopic observation using a hand magnifying lamp and a slit-lamp biomicroscope. IV. Results: There was no difference in body weight gain between the controls and the 0 mg/kg group during the test. The 2.0UU mg/kg and 2U0 mg/kg groups lost weight from exposure days 1 to 10. Both groups recovered the weight loss and exhibited normal weight gain after exposure day 10. A graph of the body weights is presented in Appendix IV. No rats died during exposure or recovery periods. A. Clinical Observations: There were no differences in clinical signs displayed by the control and the 2l) mg/kg groups during the exposure or recovery periods. Rats exposed to concentrations of 200 mg/kg or the 2,000 mg/kg showed slight redness in the treatment areas throughout the test. Rats exposed to 2,000 mg/kg were salivating during the 2 wk exposure period and had wet stained perineal areas during the recovery period. Treated and control groups showed hair loss and cutaneous lesions in areas where collars rubbed the rats' necks. Treated and control rats also displayed red discharge from the nose and eyes while wearing collars. When collars were removed, discharge subsided. Neck irritation, red nose and eye discharge were probably not compound-related. B. Clinical Chemistry: After the last (10th) exposure the treated groups showed dose-related increases in serum enzyme1activities such as alkaline phosphatase, GPT and GOT. In the 2,000 mg/kg group the serum total protein was lower than controls after the last (10th) exposure. In all treated groups the erythrocyte counts, hemoglobin and hematocrits were lower chan controls on recovery day 14. These measurements from the created groups were similar to controls after exposure 1U. Erythrocyte counts, hemoglobin and hematocrit measurements in the 200 mg/kg group remained lower than the control at recovery day **2. The 20 mg/kg, and 2,000 mg/kg showed measurements similar to controls at recovery day 42. Clinical chemistry results are reported in detail in Appendix 1. 2- - Company Sanitized. Coes no? contain TSCACBi c. Pathology: Liver changes, basically characterised by one or sore foci of coagulatlve necrosis containing large vesicular nuclei with markedly Increased numbers, were seen in all treated groups of rata sacrificed following exposure 10. The Incidence and severity was dose-related although in each group there were rats which did not show this lesion. In the 2,000 mg/kg group, 3 rats sacrificed after 14 and 1 rat after 42 recovery days Bhowed this lesion. None of the recovery rats in the 200 mg/kg group and 1 rat from each recovery Interval in the 20 mg group displayed the change. The liver lesion, both in terms of Incidence and severity, appears to be reversible. Coagulatlve necrosis of the epidermis at the dose site of 2 rats exposed to 2,'>00 mg/kg and sacrificed immediately after the last dose was observed. No other skin lesions were observed microscopically and all other tissues and organa examined appeared normal. Details of pathologic results are presented in Appendix 11. D. Organ and Body `telgl t Analysis: There was a statistically significant Increase in the mean absolute and relative liver weights In all treated groups on exposure day 10. By recovery day 14, this increase was still significant in the 2,000 and 200 mg/kg groups. At recovery day 42, che mean absolute liver weights were significantly increased in the 2,000 mg/kg group only. On exposure day 10, the mean absolute spleen and kidney weights from the 2,000 ag/kg group were significantly less than controls and in che 20 mg/kg group the kidney weights were significantly greater than controls. The mean relative testicular weights from the 2,000 mg/kg group showed a statistically significant increase on exposure day 10. On recovery day 14, there was a statistically significant increase in the mean relative kidneys and testes weights of the 2,000 mg/kg and 200 mg/kg groups. Details of mean absolute and relative organ to body weight ratios are in Appendices V and VI. E. Organofluorine Analysis: Organofluorine measurements in each treated group were greater than controls after exposure 10, and recovery days 14 and 42. These increases were dose-related and appeared to be reversible with time. The controls had the lowest measurements of 10.2 ppm, 2.6 ppm and 0.6 ppm (after exposure 10, recovery days 14 and 42). The 2,000 mg/kg group averaged the highest values of 117.8 ppm, 4-4.8 ppm and 8.2 ppm. The 200 mg/kg group averaged 80.8 ppm, 26.2 ppm and i.7 ppm. The 20 mg/kg group had average values of 52.4 ppm, 9.5 ppm and 1.2 ppm. There were rats in the 20 mg/kg group which had net values significantly greater than the controls. [At recovery day 14 one rat had organofluorine concentration of -l.l ppm and recovery day 42, two rats had values of 0.9 ppm.] Control values appear somewhat elevated at the exposure 10 and recovery day 14; however, all control rats received only distilled water, applied with ' uncontaminated, sterile syringes and were housed in separate rooms from treated groups from study onset to sacrifice. Results of the nonvolatile fluorine measurements are reported In Appendix III. IV. Summary: Three groups of 15 male ChR-CD rats were exposed dermally for 6 hrs/day, 5 days/wk for 2 wks. to concentrations of either 2,ODD mg body weight. A 4th group, exposed simultaneoulsly to distilled water, served^as a control. Groups of rats were sacrificed and examined immediately following exposure 10, after 14 recovery days (no additional exposures), and after 42 recovery days. Company Sani.u.d, Does no. con.a.n TSCA CM 9 During the 10-day exposure period, rats treated with either 200 or 2,000 mg/kg lout weight followed by a normal growth after exposure period. Slight redness ot the skin was seen in these 2 groups along with salivation in the 2,000 mg/kg group only. Rats treated with 20 mg/kg showed normal body weights and no unusunl clinical signs during the experiment. Clinical enzyme determinations monitoring liver function (alkaline phosphatase, GPT, GOT) showed dose-related increases in all treated groups after exposure 10. These values returned to normal at recovery days 14 and 42. Liver damage characterized by coagulative necrosis was seen in all treated groups following the 10th dose. The incidence and severity of liver damage was, in general, dose-related. Recovery was complete in the 20 mg/kg group 14 days following the 10th dose and was essentially complete in the 200 mg/kg group at the same time. On recovery day 42, reversal of liver damage was essentially complete in the 2,000 mg/kg group. Two rats exposed to 2,000 mg/kg had coagulative necrosis of the epidermis at the dose site following 10 exposures. Liver weights, both on an absolute and relative to body weight basis, showed a dose-related increase on exposure dey 10 with a return to normal weight seen in the 20 mg/kg group at 14 days and in the 200 mg/kg group at 42 days recovery. The increased liver weight persisted for 42 days in the 2,000 mg/kg group although a trend toward normal was seen. Blood organofluoride levels showed dose-related elevation on exposure day 10 followed by a decrease in the levels at recovery days 14 and again at 42. These values after the tenth exposure ranged from 52 ppm (20 mg/kg), dl ppm (200 mg/kg), and llo ppm (2,UUU ppm) to 1, 4, and 6 ppm (20, 200, and 2.U00 ppm, respectively), at recovery day `2. At a blood organofluoride concentration of approximately 10 ppm there were normal liver weight to body weight ratios, serum enzyme activity, no clinical signs or body weight differences from controls. One rat showed liver changes at this level. At a blood organofluoride level of approximately 50 ppm there were marked liver changes and significant increases In the mean absolute and relative liver weights. No other toxic effects were evident at this blood organofluoride concentration. r ______________ _ *v ha*. -4 notconW'T S C A C B ' Sanitized- Does Compaq Report by: 0 JW l Lynn S. Silber Technician Approved by: _ ' i y -- f n ___________ Gerald L. KennefBy Chief, Acute Investigations Section LSS:jrg Study Director: O. L. a s h i e l l ^ D a t e issued: Uctobei^lu , m u Report So. bri9-u Date Reissued: January 15, 1982 - 50oe3 noi contain TSC CB Company Sanitizs'i- HASKELL LABORATORY MO. 12037 CLINICAL PATHOLOGY REPORT PROCEDURE: Sixty male rats were divided into four groups of fifteen each. This* four groups were exposed dermally to 0 ng/kgt 20 mg/kg, 200 tag/kg and 2000 tag/kg Blood wits taken from the tails of five rats from each group after the tenth raposure, :hen these rats were sacrificed for pathology. Fourteen days late* (REC 1), blood samples were again collected from five rats from each group and these were also sacrificed for pathology. Forty-two days after the tenth exposure(REC 2) blood was collected from the remaining five rats In each group. Hematology measurements on the blood included the following: erythrocyte count (RBC), hemoglobin (Hb), hematocrit (Ht), leucocyte (VBC), and relative number of neutrophils (Neut), lynghocytes (Lymph), eosinophils (Eosin), monocytes (Mono) and basophils (Baso). Mean cell colume (MCV), mean cell hemoglobin (MCH) and mean corpuscular hemoglobin concentration (MCHC) were calculated using the appropriate data. Clinical chemistry measurements included alkaline phosphatase (AP), glutamic-pyruvic transaminase (GPT), glutamic-oxaloacetic transaminase (GOT) and total protein (TPR0T). STATISTICS: The data were analyzed statistically by a one-way analysis of variance and least significant difference tests, conparing treated groups with the controls when the ratio of variance (F) indicated an effect due to treatment. Significance was judged at the 0.05 probability level. RESULTS: Results of the- clinical pathology measurements are summarized-in Table 1; statistical analysis in Table 2. A dose-related increase in the activity of the serum enzymes, GPT and GOT, in all groups occurred after the tenth treatment. The alkaline phosphatase activity in the serum of these groups was also higher than the controls at this time. The statistical analysis, however, indicated only the GOT in the group treated with 2000 mg/kg was significantly elevated. The serum total protein was also lower than the controls in this group. . 1 S j j i f ( j ; - 6 Qompany Sanitized. Does not contain TSCA CBI - 2- . Increased activit/oof the serum enzymes and a decrease in serum protein are associated with liver involvement.-Similar effectsh m v e been observed in the serum enzymes of rats i n h a l l n g M ^ M H B H V y ^ d8* ingesting Fourteen and 92 days liter the last treatment the serum enzymei^amRotar protein of the treated rats were not different than the controls. Erythrocyte counts, hemoglobins and hematocrits were lower than the controls in all treated groups 14 days after the last treatment. For the group treated with 2000 mg/kg these measurements were also lower 42 days after the last treatment. Report by: t- fobby L. Moore Clinical Chemist ^ 1 BLM:JRB:1jm Date: February 29, 1480 Approved by: fJohn R. Barnes Chief*'Clinical Pathology Company Sanitized. Does no* contain TSCA CBi 7 TABLE I AVERAGE CLINICAL LABORATORY VALUES ON MALE RATS EXPOSED (DERMAL) TO 4 RBC X lO^/mm1 Hb g7. Ht X MCV ^ MCH pg MCHC X WBC X 10 Vtran5 Neut X Lymph '% Eosln 'X Mono X Baso X AP XU GPT IU GOT 1U TPROT g7. Exposure _0_ 6. OS 14 8 45 75 25 33 16.3 23 72 1.6 3.6 0 250 27 62 5.86 TEN DAYS Concentrati on 20 200 5.34 5.23 14. 1 14.0 45 47 84 89 27 27 32 30 16.8 23.4 24 28 72 68 1.0 1.4 2.6 2.8 00 332 370 44 52 87 <6 .2 0 95 6.42 mg/kg 2000 6. 30 14.6 44 70 23 33 15. 1 23 72 1.0 4.8 0 330 67 155 5.16 FOURTEEN DAYS Exposure Concentration 0 20 200 6.38 6.01 6.02 15.2 14. 1 14.2 45 44 45 71 73 74 24 24 24 34 32 32 18.2 16.0 16.5 35 28 28 59 0.2 69 0.4 68 1.8 5.4 3.0 3.0 0 00 253 199 256 33 28 27 72 53 48 6.44 6.50 7.12 mg/kg 2000 5. 74 13. 1 39 69 23 33 14.9 29 67 0 4.0 0 307 36 65 6.38 FORTY-TWO DAYS Exposure Concentration _0_ 20 200 5.87 5.94 u. 02 15.1 15.4 15.5 45 48 47 76 81 77 26 26 26 34 32 33 14.6 18.5 17.7 25 17 17 71 80 79 0.6 0.4 1. 2 3.6 3.2 2.6 0 00 160 176 178 24 24 23 53 6.84 50 7.08 m 7.22 %/k* 2000 5. 14.5 42 76 26 34 14.8 23 72 1.6 3.4 0 187 23 45 7. Sanitized. Doss not contain tscacbj Company TABLE 2 SUMMARY OF STATISTICAL ANALYSIS FOR RATS EXPOSED (DERMAL) TO a RBC Hb Ht MCV MCH MCHC WBC Neut Lymph Eos in Mono Baso AP GPT GOT TPROT F 8.9 1.0 0 .8 28.4 25.9 7.1 2.5 0.6 0.6 0.6 1. 1 0.0 1.7 2.3 4.0 15.2 ______ TEN DAYS Expoiun Cone, mg/kg 2000 200 20 0 -* 0 00 0 00 - t 0 -0 0 00 0 00 0 00 0 00 0 00 0 00 0 0' 0 0 00 f 00 - +0 F 3.4 14.7 20.8 1.9 1.0 1.3 0.9 0.5 0.7 2.3 1.2 0.0 4.9 1.1 2.9 5.8 FOURTEEN DAYS_______ Exposure Cone, mg/kg 2000 200 20 --- - -- -0 - 000 0 00 000 0 00 000 000 000 0 00 0 00 + 00 000 000 0 +0 F__ 2.5 3.4 9.9 2.4 0.2 5.0 0.2 1.4 1.5 1.5 0.2 0.0 0.4 0.2 1.0 4.7 FORTY-TWO DAYS Exposure Cone. 2000 200 __20 000 000 - f + 000 0 00 00- 000 000 000 000 000 0 00 000 000 000 ++0 0 not signiflcantly different from control - significantly lower when cohered to controls + * significantly higher when compared to controls Company Sanitized, Does not contain TSCA CB! m central research and development department HASKELL LABORATORY FOR TOXICOLOGY AND INDUSTRIAL MEDICINE APPENDIX I I - ^ 5 -if Octanolc acid, pentadecafluoro-, a-- onlum salt H-12,037 Petrochemicals Depart-- nt Subacute Detaal Toxicity Study - C^w-CD Rata May 27. 19S Introduction Male ChRS>-CD rats were arranged into four treatment groups as follows: Results Group I Group II Group III Group IV Distilled water control (0.05 ml) 2,000 mg/kg (as an aqueous paste) 200 mg/kg (as an aqueous paste) 20 mg/kg (as an aqueous paste) The morphologic changes observed at necropsy are reported in Table I. Representative sections were prepared for histomorphologic evaluation from bone marrow (sternum), brain, cecum, colon, epididymides*, esophagus, eyes, heart, kidneys*, liver*, lungs*, skin (control area), skin (treated area), small intestine (duodenum), spleen*, stomach, testes*, thymus*, thyroid, trachea, and lymph node (mediastinal). The results of histomorphologic evaluation of tissues j I Company Sanitized. Does not contain TSCA CBI Mk m fran the control and high-dosed rats are presented In Table 11 Upon finding a compound-related effect In the cutaneous and hepatic tissues of animals In the high dose group, similar tissues were examined microscopically from all animals in the intermediate (200 mg/kg) and low dose (20 mg/kg) groups. The results of these examinations are reported in Table III. Compound-related hlstomorphologlc changes are summarized for between group comparisons in Table IV. Discussion and Conclusions A compound-related hepatic effect was observed in rats from all three compound-treated groups (Table IV). Affected livers contained one or more foci of coagulative necrosis. The Kupffer cells, within the foci of hepatocellular necrosis, contain large vesicular nuclei and were markedly increased in number. Inflammatory cells were occasionally present within and at the periphery of the necrotizing lesions. Dermal lesions at the site of compound application were observed in two animals in the high dose (2,000 mg/kg) group (Tables II and IV). Affected areas of skin were characterized by coagulative necrosis of the epidermis and papillary and reticular dermis. A minimal lympho-histocytic inflammatory infiltrate was present in the subadjacent dermis bordering the necrotic zones. The remaining histomorphologic changes shown in Table I were changes that were observed with similar frequency in the control and compound-treated groups Summary The dermal application of octanolc add, pentadecafluoro- aamonium salt for 10 days resulted in hepatocellular necrosis and necrosis of cutaneous tissue at the site of application.* * " Tissue weighed Report by: Raymond M. Everett, D.V.H., Ph.D. Senior Research Pathologist Approved by: RHE:vlm . Date Issued: May 27, 1980 <7> 'U m v C William C. Knuss, D.V.H. Chief, Pathology Section - 12 - Company Sanitized- Does not contain TSCA CBI TABLE I MACROSCOPIC OBSERVATIONS RECORDED DURING NECROPSIES OF ChRR-CD CONTROL AND TEST MALE RATS TREATED BY < DERMAL APPLICATION WITH OCTAHOIC ACID. PENTADECAFLUORO- AMMONIUM SALT! Rat No. Amount of Autolysis Recorded During Necropsy Group 1 -- Control Males 12 Days on Test; 0 Recovery Days 263204 None 263205 None 263206 None 263207 None 263206 None Mode of Death* S S S s s Observations N.A.D.t Thymus - pink N.A.D. N.A.D. N.A.D. j* b Days on Test; 13 Recovery Days 263209 None S 263210 None S 263211 263212 263213 None None None S S S Thymus - right lobe - dark red mottling Lungs - all lobes - pin-head sized red foci Thymus - right lobe - dark red mottling N.A.D. Salivary Lymph Nodes - large, mottled red Thymus - pink * Mode of Death: E Sacrificed in extremis; FD - Found Dead; S - Sacrificed t N.A.D. - No abnormalities detected. .Company Sanitized.'0** n^vo...air. TSCA CBI TABLE I (Cont'd) MACROSCOPIC OBSERVATIONS RECORDED DURING NECROPSIES OF ChR-CD CONTROL AND TEST MALE RATS TREATED BY DERMAL APPLICATION WITH OCTANOIC ACID. PENTADECAFLUORO- AMMONIUM SALT Rat No. Amount of Autolysls Recorded During Necropsy 53 Days on Test; 41 Recovery Days 263215 None Mode of Death* S 263217 26321 263219 263277 None None None None S S s s Observations Lungs - few dark red foci 0.5 to 1.0 on in diameter, few grey streaks Nose - red discharge Skin - neck - Irritation Lungs - grey aottiing 1 to 3 on in diameter Skin - neck - necrotic area 1.5 z 0.2 cm; skull - necrotic area 3 on in diameter Thymus - left lobe - dark red Liver - lobular markings prominent N.A.D. - 14 Sanitized. Does not contain TSCACBi ~ Company TABLE I (Coat'd) MACROSCOPIC OBSERVATIONS RECORDED DURING NECROPSIES OF ChR9-CD CONTROL AND TEST MALE RATS TREATED BY DERMAL APPLICATION WITH OCTANOIC ACID. PENTADECAFLUORO- AMMONIUM SH.t Rat No. Group 11 Amount of Autolysis Recorded During _____ Necropsy______ 2000 mg/kg Males Mode of Death* Observations >J 12 Paya on Test; 0 Recovery Days 263220 263221 None None 263222 None 2o3223 None 263224 None Animal - thin Liver - heavy Nose ~ red discharge Perineal Area - vet and stained Skin ~ chin - stained and moist Eyes - red discharge Liver - heavy Nose - red discharge Perineal Area - stained, wet Skin - Irritation Spleen - small Urine - bright yellow S Animal - very thin Liver heavy Nose - red discharge Perineal Area - stained, wet Skin - irritation Thymus - very small Urine - stained red S Liver - heavy Nose - red discharge Perineal Area - stained, vet Skin - test area - dark red crusty area 0.8 cm in diameter Spleen - small Thymes - small Urine - bright yellow S Eyes - red discharge Liver - heavy Nose - red discharge Perineal Area - stained, vet Skin - irritation - 1-5 Sanitized. Docs ;cQ'ein i-- Compaq MACROSCOPIC OBSERVATIONS RECORDED DURING NECROPSIES ni.Bull 0F ChR-CD CONTROL AND TEST MALE RATS TREATED BY I DERMAL APPLICATION WITH OCTANOIC ACID. PEMTADECAFLUORO- Atamviirw cM T | Rat No. Amount of Autolysis Recorded During Necropsy 26 Days on Test; 13 Recovery Days 263225 None 263226 None Mode of Death* S S 263227 None S 203228 203229 None None S S Observations Liver - heavy Liver ~ left lobe - numerous pale brown foci up to 1 1q diameter, heavy Liver - heavy Thymus - right lobe - dark red mottling Liver - heavy Liver heavy 53 Days on Test; 41 Recovery Days 203231 None S 203232 263234 263235 None None None S s s 263236 None s N.A.D. N.A.D. N A# D* N.A.D. N.A.D. 5 TABLE I (Cont'd) MACROSCOPIC OBSERVATIONS RECORDED DURINC NECROPSIES ChR-CD CONTROL AND TEST MALE RATS TREATvn nv PMWL applicatioh m m octakoic acid. , Rat No. Amount of Autolysis Recorded During Necropsy Group in - 200 mg/kg Males 12 Days on Test; Recovery Days 2oJ23 None 263239 None Mode of Death* S s 263240 263241 None None s s 263242 None s Observations Liver - heavy Liver - cystic lobe - grey a 0.8 cm in diameter, heavy Liver - heavy Liver * heavy Nose red discharge Perineal Area - stained, wet Skin - neck - Irritation Eyes - red discharge Liver * heavy Nose -- red discharge Thymus - pink 26 Days on Test; 13 Recovery Davs 263243 None 263244 None S S tJ i.3 None S 2b32*6 263246 None None S s Liver - heavy Liver - heavy Thymus - pink Liver - slightly heavy Testes - right - yellowiah-white subcapsular streaks Kidneys - right - hydronephrosis Liver - heavy Liver - heavy Lungs - few pin-head size red foci scattered throughout - 17 oc anv S a * n ^ a ' Cawy ,,,t S C A c S TABLE I (Coat'd) MACROSCOPIC OBSERVATIONS RECORDED DURING NECROPSIES OF ChR9-CD CONTROL AND TEST MALE RATS TREATED BY Rat No. Amount of Autolysia Recorded During Necropsy Mode of Death* 53 Days on Test; 41 Recovery Days 263249 None S 26325v None S 263251 263252 None None S S 263253 None S Observations Skin - test area - thick Lungs - fine grey nettling scattered throughout N.A.D. Skin - test area - slightly thick Thynus - right lobe - pink Testes - slightly snail i 3 MACROSCOPIC OBSERVATIONS RECORDED DURING NECROPSIES OF ChRe-CD CONTROL AND TEST MALE RATS TREATED BY DERMAL APPLICATION WITH OCTANOIC ACID. PEMTAPECAPLUORO- AtMONIUM SALT Rat No. Amount of Autolysis Recorded During Necropsy Group IV - 20 ag/kg Males 12 Days on Test; 0 Recovery Days 263254 None 263256 None Mode of Death* S s 263257 263258 None None s s 263259 None s __ _____Observation Liver - heavy Liver - heavy Skin - neck - scaly, crusty Liver - heavy Liver - heavy Thynus - left lobe - pink Liver - heavy 1/ 26 Days on Test; 13 Recovery Days 263260 None s 263261 2o3262 263265 253268 None None None None s s s s Liver - slightly heavy Lungs - grey aottling scattered throughout Skin - neck - necrotic area N.A.D. hA* D Skin - neck - necrotic area 8 mm in diameter . 53 Days on Test; 41 Recovery Days 263269 None S 263270 None S 263271 None S 263276 None S 263278 None S N.A.D. Skin - test area - slightly thick N.A.D. N.A.D. Skin - test area - slightly thick 19"- mm San' usta c c ^ a',ri am , > J d w tahi.k h Pa*r II HMJH HALT. KAfS THKATEU BY HIST(JHOHPHI)U;|(. IIBSKNVATIONS lF TISSUES lT" Aci^^TADecAFuiot^ A f w m m salt. Hode of Ornili* Haya on Teat Arcovery Day T la a u e * * .md Ohnervat(on*** Aina 1 Nunbrr 261- one Harrow (aternnn)i rain: Cecun: Colon: Rpldldynldea: Per Ivan, .liar l.ynphocyt I, Infiltration, Focal/null Ifocal Mlateral Croup I - Controla S s "5 s T^ s T s s s s~ u0 TT 00 A > u T n u T u T~1 i) 13 i) 13 " T """T~"2T" 13 2 0 0 i1 1 *i 2 1 i 2 1 i l Al 41 iT 6 / 8 9 0 i 2 3 > i 8 9 / N N N H N N N N H 0 N N N N N N 0 N N N N N u NN II H N H N N N N N N N H If N h II II N N N N N If N N N N N N N II 1. N H 1. N 1. N N N N 1. N i. N i. n 12 0 2 2 0 e N-l r 2 2 1 i Croup II ^.000 ne/fce "s a s T ~ s s 8 S S 12 12 12 26 26 26 26 26 51 S3 S 53 S 53 S 3 u0 22 0 2 13 n 22 1} 2 Il 2 11 uaAZlS._3AlAJAA.Ja-U .. 1 222222 2 2 2 3 222 A S6 22 /8 2 9 3 1 3 2 333 A3 6 N N HH If H NN N NN NN NN Nn NN 1. N R N M H n RR RH HH HR 1. R R R H R R H 0 N N L RRR RHR RHH If R R N 1. R H R R N R R R R R R Prrlrpldldymal Fat, Neutrophilic InfIannatIon, Unilateral Perlvaacular l.ynphocytlc Infiltration, Focal/nult(focal, Unilateral Perlepldldynnl Fat, Pyogranulonatoon Inflaanatton, Focal/nultI focal, Btlatrral uopMAui: Kjfn: Retinal Roaettr, Unilateral Henrt: KJ O NNNN N NNH nnnnnnkn NN NN NN HN N ............. p R R fl R ** T^eaue'Account tna n " ! ' " ' ' 1 * " Fo,m,, S " crl fired. a*. "h i t<*Xoir0Phho*|fI`ca'lTevaPlruiayt5inotn'.*Uc ^ ^ Pr*-luailonr!'.l-tL M oi l,^Seerrvredd. 'RN*--lN^oTlua"a,lo'o'*!o!b"a'erve`d1i" 0" -Tlaa-neionnotaode^a-JaltorM;aDfo-rFr.^entatlo. Severity of bealon Code: I MininaI chaneei i . Uiij i u "' P -rn t. -e C X ^ . L ` ' d C' " e ' ! H<KUr" t ' * > - C h a n .e p r e a . n l . v e r i t y n o t g r a d e d ; Company Sanitized. Does not contain TSCA CB1 TARLK II (Cont> Pege 17 II!STUMOR pilot.<i|C OBSERVATIONS (IF TISSUES fiOH HAI.r. ChR-CD RATS TREATED -RY_gERHAL AfPLlCATIjjNJjITH OCTANOIC ACID, TEKTADECATLUORO- UtWRUM SALT Mode of Death* Osya on Tost Recovery Day Tissue* and ObservotIan*** AnInal Nunber 76 1- Kidneys: ProaIasi Tubular Eplthellua, ReRenerstton, Focal/suitIforal I - Controls s 12 12 0 2* - 0 z 00 s 12 0 2 0 sss 12 12 26 uo n 222 00 26 1) 2 l 3 I T T " ~3 I T T T " "g~ 26 26 26 51 51 31 51 31 1) 11 1) 41 41 41 41 41 2 2 2 2 2 r T~T iill1 17 4 46 / R9 0 i 2 i 4 / 8 9 / N N N N N N L N L i. N N N N N ------ Croup II - 2,000 M /ka S 8 j S *8 J V | 8 H ! ; ; } '}i1}|f r y I 2 l 2 ! 2 12 26 26 26 26 26 i l 33 S3 51 *3 22222 2222213J1j -g-L-i-L * > 6 I B 0 . 7 4 l l LNI.NNNNLN L N L L N L Perivascular Lyaphocytlc Infiltration/ Interstitial Nephritis, Subacute, Focal Hcdulla, Cyst/a, Tubular 1.1vert Eat raar,lul lary Hesiatopolesls Perivascular, l.ysiplin-hlstlocyt Ic Inf IItratIon/a NLNNNNNN L ri I. N N N P L I. N I, L N L B N I. L Focus of Cellular Alteration, Basophilic Centrolobular Vacuolar Cytoplaaalc Change Hepatocellular Coagulatlve Necrosis With Kupffer Cell Frollfaratlon Flbroala, Focal Periportal Lyaphocytlc >nflltratlon/a I Periportal Plbroala and Haaoaldertn Deposition, Multifocal M Centrolobular Sinusoidal Ectasia Ling: Perivascular Lynphocytlc Inf IItratlon/a N NN N N NNN CranuloMtous Inflannatlon. Focal/Mult Moral Hint lorytosla, Focal/Hul1 1foca I Lynph Nodes (HedlastInal): Cortes, lleaorrhage ONNUNONONNtINOO I2 I II 22 I NNOLONON ) 2 I L L L L NN 2 2 11 2 II I N O K O OO Company Sanitized. Does not contain TSCA CBJ i -i; ____ -------------- ~ ~ L - * - rS TAHI.E I I ( C i i n l ' dJ HISTUMURFHOMJIi: OBSERVATIONS OF t i c c h i c O fL H jT H W jT A W |C _ A C lP . PENTADECAFLUORO- AFWtmiUW SALTj P a * * 11 Mode o f D e a th * Days on Tent R ecovery Days T i s s u e * * mid Observ t Ion*** A n lo a l Nunher 261- S kln (U n tre a te d S ite ) ! Head and H eck, P a ra k e ra to s is , I n f ilt r a t io n by N e u tro p h ils S kin (T re a te d S it e ) : D e rm a t|tla , A cute M e c ro tlftln n S m all In te n t In c (D uodenum ): S p le e n : Stom ach: H y p e rk e ra to s is . G la n d iila r/N o n R la n d u la r. Ln olna P ro p ria / S nbsnii'osa, E o s in o p h ilic I n i I I t r a t lo n /n G la n d u la r, N e c ro s is w ith Y e llo w P lg o o n t D e p o s itio n , F o c a l/M u It I fo c a l s s s Ss Ss s sS SS S 12 12 12 12 12 26 26 26 26 26 S I T~0 >. 0 A 0 i 0 1 0 i it A 0 2 0 7 0 2 0 8 11 1 ) 11 11 11 4 | 41 41 41 41 2 2 2 i i 2 T~~ T 0 11111 Ii 90 12 I S 89 H N N N H N N N N N N N I. N N N N N HN N N N N N N NN NNNNN NNNN NN NHNNN NNNN L NNNNNN NNNN NN C ro u p I I - 2,000 -/fc. s S it t i s i s i S i s 12 12 12 12 12 26 26 0 0 0 0 0 11 11 2 2 222 22 2 2222 22 0 12 14 56 HN SSS L P NN HN NL NNNN NNNN NHNN Te"""j|,,ndul"r p>T` *o n , U w tn a P r o p r ia Edeoa P S p e ra a tlc C e ll A tro p h y , U n ila te ra l Thymua: hnnnnnnnnhhn to C o rte x , Lym phoid A tro p h y NN NN NN N HN NN N to T t COri ! * ' " ' " I * !" U e p o a lllo n , H n lt lf o c n l T h y ro id : C y s t, S<|uasua C e ll l.ln e d 0 MN NN N HN N T ra c h e a : ie p r!" * e M -- M - tlo e y tlr In filtr a tio n L a a ln a P ro p ria , L y a p h o c y tlc In f I ! t r a t lo n / s NNN NN N HN NNNNH 2 NHHNHHNNRNNNHNN ^^NNNNNNRNNNN I NNNNLOHHHIINLNI.N * PP "HNNHNHNNNNLLHN 4 2 n otcnta'n SawVit^- eS Compaq 'V -I ...I M A '3 <n ;-.>u4(5! ...isJSBSbSS TABLE I11 FROM HALE ChRO-CD RATS TREATED IIISTf)HORPtlllM;|(: OBSERVATIONS (if TISSUES Kode of Death* taya on TtRt Itcovery D a y Tlaaue** and ObservtIon*** AnIna1 Number 261- Llveri Nepatoccllulai Neeroala Kith Kupffer Cell Proliferation, Focal/Huit Ifocal Periportal rtbroats. Multifocal Periportal Lymphocytic InfIItrarlon/n Cent rolohular Sinusoidal Ectasia Estrnuedullary Hemtopolesia Skin (Untreated Site): Ulceration with Acute crmatItla Keck, Ulcerative, Chronic-ActIve tarant It la. Focal Skin (Treated Site): s s fi Group h i _ 20U g/kR S s s S S s s fi ~}T 17 12 00 T~ 7 )l 89 12 12 12 26 26 26 0 0 0 n 11 1) 2 ? 1 7 T~T 4 44 4 44 0 1 2 14 5 26 26 11 1 ) r 7" 44 68 51 51 53 51 51 41 41 41 41 41 Ti } i 2 4 9 5 0 5 1 23 5 I 1. 1, 1. 1. H N N L N N H N N N NNNLN P NNNNNNNNN NNNHNNNNNNNHNN Pane I* n i * . T w r r f r . . 12 12 12 12 12 26 26 26 26 26 )) 53 53 53 53 ? 0 J 3 t ) || U 4! 61 61 *1 Al 222222222222222 333556666667277 * 6 J 0 I 2 5 9016 MNNLLN II III a tT 1* ot Death: E - Sacrificed In ealrenla, F - Found Dead, S: - Sacrificed. I Cod' ! * " prevented conplete evaluations K - Autolysla present, tlaaue evaluation adeunatai D - Pra natation nr to >- - .... . , e,--5:Tor D08S noi contain TSCA O', Company. - t a b u IV OP SZZ.'SZSLZ SPSSST'Kr WITH OCTANOIC ACID. PEHTADECAFLUaiQ. Group Designation Dose Recovery Days Nuaber in _________ Treatment Group Tissue/Observations Liver: Hepatocellular Necrosis with Kupffer Cell Proliferation, Focal/Multifocal Skin (Treated Site): Dermatitis, Necrotizing 555 --- 5-- -S JC 5c 5 5 5 5 0 0 0 33 1 5j cJ Je 5e 5 5 00 0 20 0 pSs f 5353 S s 555s35 30 0 2 11 555555 0 00 0 00 - 24 nol contain TSCA r Company Sanilized. Does APPENDIX H I AVERAGE ORGANOFLUORINE VALUES FOR GROUPS I. II, III AND IV Group No. I II III IV Dose mg/kg j 2 #0 0 0 200 Total Nonvolatile Fluorine (ppm) Exposure Recovery Recovery Day 10 Day 14 Day 42 10.2 117.8 80.8 52.4 2.6 44.8 26.2 4.5 0.6 8.2 3.7 1.2 - 25 Company Sanitized. Does not contain TSCA CBI INDIVIDUAL ORGANOFLUORIDE VALUES (PPM) FOR GROUPS I, II. Ill AND IV Group T Group II Group III Group IV Exposure Day 10 14.0 8.0 16.0 11.0 2.0 104.0 133.0 144.0 116.0 92.0 91.0 41.0 72.0 122.0 70.0 52.0 58.0 53.0 51.0 48.0 Recovery Day 14 3.4 3.3 3.1 2.3 1.2 44.0 41.9 36.7 67.2 34.2 26.8 25.9 18.2 29.1 30.8 17.6 6.5 3.1 10.8 9.5 Recovery Day 42 0.3 0.8 0.4 0.4 0.8 5.0 4.2 7.0 13.9 10.7 2.8 2.1 5.9 5.9 1.8 0.9 1.0 0.9 1.9 * * Insufficient blood sample - 26 Company Sanitized Poes not contain TSCA CSS