Document z7zpDpzwvg0wNdQaBq5oVd2a

InternatioRneaslearcahndDevelopmenCtorporation SPONSOR: COMPOUND: SUBJECT: 3M Company Fluarad4oFluorochemical Surfactant FC-95 Ninety Day Subacute Rat Toxicity Study. Edwin 1. @oaaeuthal, Ph.D. Vice Presi@ent and Director of Research Collaborators: D. C. Jessup, Ph.D., Associate Director of Research R. G. Geil, D.V.M., Vice President and Director of Pathology N. D. Jefferson, B.S., Acting Director of Small Animal Toxicology R. J. Arceo, X.D Staff Pathologist Date: November 10, 1978 137-085 JL)N 3 1997 InternatioRneaslearcahndDevelopmenCtorporation TA BLE 0F C0N TE NTS 7nopsis . . . . . . . . . . )mpound . linical Studies Method . . . . . . . . . . . . . . . . . . . . . . . . . 1. General Procedure* 2. Compound Administration* 3. Observations . . . . a 0 4. Laboratory Tests . . 0 0 a. Hematology o . o . . o o . o 9 o o a o o b. Biochemistry . . . . . 0 0 0 0 0 0 0 0 0 c* Urinalysis o v o * o a o 9 a o o - - - o 9 9 d. Serum Samples . . . . . . . o . . . . . . . . o . 5. Statistical Analysis . . . o Results. . * o . . o . o . o o o 1. General Behavior, Appearance and Survival, o 2. Body Weights 0 3. Food Consumption o o 9 o a o o * 9 o o o o o 4. Laboratory Tests . . 0 0 0 . 0 0 0 0 0 0 a 0 a. Hematology . . . . . 0 . 0 0 . 0 0 . . 0 0 0 . b. Biochemistry . . 0 * t 0 a * . 0 * 0 0 0 0 0 * c. Urinalysis ......o . Athological Studies . o .o o ..o . . Methods . . . . . . . . . . . . . . . . . . . . . . . . . 1. Gross Pathology* 2. Histopathology . Results. . . . 9 9 1. Gross Pathology and Organ Weights. 2. Histopathology . . . . . . . . a. Liver. . o o o . o . o . . b. Stomach . . . . . . . . . . c. Bone Marrow. o . o ......... d. Thymus . . . . . . . . . . . e. Spleen . . . . . . Page 1 3 4 4 4 4 5 5 5 5 5 6 6 6 6 8 8 8 8 9 9 10 10 10 10 10 10 37-085 InternatioRneaslearcahndDevelopmenCtorporation TA BLE 0P C0NT ENTS (Continued) f. Meseuteric Lymph Nodes g. Small Intestine. . 0 0 h. Skeletal Muscle. . . . . . 0 0 . 0 0 . . 0 . io skin 0 * 0 0 0 & 0 0 0 0 0 0 a a 0 0 0 0 a 0 * a Table No. 1. Group Mean Body Weights; Weight Ranges; and Survival . . 2. Individual Weekly Body Weights . . . . . . . . . . . . . 3. T-test Comparison Between Average Food Consumption of Control and Treated Groups. * o o o . * 0 4. Mean Food Consumption* oooaoooo..o .o . 5. 6- 8. 9. 10-12. 13. 14-16. 17. 18. Means and Significance of Rematological Values 9 * o o o Individual Hematological Values . . . . . a 0 0 0 0 0 a . 1,4eansand Significance of Biochemical Values . o . o o . Individual Biochemical Values. 00 Means of Sigtificance of Urinalysis Values . . o a o . o Individual Urinalysis Values . . . o o . 0 0 0 0 a 0 0 6 Summary of Gross Necropsy Observations o o * * o o a * o Absolute and Relative Organ Weights. * 0 0 0 0 0 0 a 0 0 19. Individual Organ Weights . . . . . o . 6 . . 0 . . 0 * 0 20. Histomorphologic Observations . . . . . . . 0 . . . 0 0 0 Page 11 12 12 12 14 15-16 17 18 19-20 21-26 27-28 29-34 35 36-41 42-43 44 45 46-51 137-085 InternatiRoensaelarcahndDevelopmeCnotrporation Page 1 1. SYNOPSIS Fluorochemical Surfactaut was fed in the diet at levels of 30, 100, 300, 1,000 and 3,000-ppm to Charles River CD rats for 3 months. Five male and 5 female rats were initiated at each dosage level and in the control group. The rats were observed twice daily for overt signs of toxicity and mortality. Individual body weights and sex group food consumptions were recorded weekly. Rematological, biochemi- cal and urinalysis studies were conducted during the pretest period and at 1 and 3 months of study. At dosage levels of 300, 1,000 and 3,000 ppm, all rats died prior to scheduled termination of the study. Overt signs of toxicity including: emaciation, convulsions, altered posture, red material (right eye and/or mouth), yellow material (auogenital region), increased sensitivity to external stimuli and reduced Aotor activity were evident prior'to most deaths. Time of death was directly related to the dosage level with the earliest deaths seen in the 3,000-ppm treatment group. At the 30-ppm dosage level, neither male nor female rats showed any compound related changes in appearance or behavior. Mean body weights were slightly lower for the rats at the 30-ppm dosage level when compared to the controls. At the 30-ppm dosage level, one female rat showed a slightly elevated glucose level and one male rat showed a slightly elevated alkaline phosphatase value at 1 month of study. At 3 months of study, one male rat at the 30-ppm dosage level showed slight to moderate elevations in glucose, blood urea nitrogen and Y-glutamyl transpeptidase activity. At the 100-ppm dosage level at 3 months, body weight means when compared to the controls were lower for males and for females. Food consumption means when compared to the controls were lower (statistically significant) for males and females. At the 100-ppm 137-085 InternatioRneaslearcahndDevelopmenCtorporation Page 2 dosage level, slight increases in creatinine phosphokinase (CPK) and alkaline phosphatase activity, slight to moderate increases in glucose and blood urea nitrogen, and slight to marked increases in plasma glutamic oxalacetic and pyruvic transaminase activities (PGOT and PGPT) were seen at 1 month of study. At 3 months of study, slight to moderate decreases in hemoglobin, hematocrit and erythrocyte counts were seen for male and female rats and slight to moderate increases in PGOT and PGPT activities were seen for two.of the three surviving female rats at the 100-ppm dosage level. Compound-related gross changes such as emaciation, areas of discoloration involving ths stomach and liver were observed among treated rats that died prior to sacrifice. Similar changes were also observed in the liver of a few rats sacrificed at termination of study from the 30- and 100-ppm groups. Microscopically, compound-related lesions were observed among all test groups. Morphological changes consisting of centrilobular to midzonal cytoplasmic enlargement (hypertrophy) of hepatocytes were observed in the livers. LNecrosisof liver cells was also present among these rats. The incidence and relative severity of the above lesions were more evident among male test rats. In addition, especially among rats from the 300-, 1,000-, and 3,000ppm dosage level that died prior to sacrifice, changes involving the primary (thymus, bone marrow) and secondary (spleen, mesenteric lymph nodes) lymphoid organs, stomach, intestines, muscle and skin, were observed and were all considered as compound related. The other changes described in the tissues of these rats were lesions of naturally occurring diseases. 137-085 InternatiRoensaelarcahndDevelopmeCnotrporation Page 3 II. COMPOUND The compound was received from 3M Company, Saint Paul, Minnesota on October 24, 1977 as indicated below: - Label Fluorochemical Surfactant Description white powder 6 x 5 lb tins 137-085 InternatioRneaslearcahndDevelopmenCtorporation Page 4 CLINICAL STUDIES A. METHOD: 1. General Procedure: Thirty male (196 to 232 g each) and 30 female (165 to 206 g each) Charles River CD rats purchased as weaulings from The Charles River Breeding Laboratories, Inc., Portage, Michigan were used in this study. The rats were distributed among the groups, based upon a computer-generated table of random numbers. The rats were housed indi- vidually in suspended wire-mesh cages and maintained in a temperature-, humidity- and light-controlled room. During the pretest period, rats were provided ground Purina'*Laboratory ChowO and water ad libitum. During the actual study, the rats were provided the appropriate test diet and water ad libitum. This study was initiated November 2, 1977 and terminated by sacrifice of all remaining rats on January 31, 1978. 2. Compound Administration: The compound was mixed weekly with ground Purina(DLabora- tory ChowS (i.e., ground basal diet) on a weight/weight basis to provide dosage levels of 30, 100, 300, 1,000 and 3,000 ppm for feeding. Five male and five female rats were used at each dosage level and in a control group. Control rats received the ground basal diet. Diet samples (100 each) were taken immediately after preparation of each diet and after 7 days standing in weeks 1, 4 and 12. The samples were frozen and subsequently shipped to the sponsor. Diets were prepared in the following manner: before weighing, the compound was ground using a mortar with pestle. The appropriate quantity of ' Fluorochemical Surfactant -as mixed with 500 g of the ground basal diet in a Hobart blender to provide the premix. The proper quantity of premix was sub- sequently blended in a twin-shell blender, equipped with an intensifier bar, with the amount of ground Puriuae Laboratory Chowe necessary to 137-085 InternatioRneaslearcahndDevelopmenCtorporation Page 5 provide the appropriate quantity by group of prepared diet. Control rats received the ground basal diet. 3. Observations: The rats were observed twice daily for overt signs of toxicity and for mortality. Detailed observations were recorded weekly or daily when a specific condition existed. Individual body weights and food consumption were recorded weekly during the pretest and treatment periods. 4. Laboratory Tests: Once during the pretest period and at I mouth and 3 months of the study, blood (orbital sinus puncture technique) and overnight urine samples were obtained from all surviving rats for analysis. Food and water were withheld prior to the collection of samples. a. Hematology: Hematological studies included: hemoglobini hematocrit2, total erythrocytes3, reticulocytes4, and total3 and differential leucocyte counts. b. Biochemistry: Biochemical studies included: fasting glucose5, blood urea nitrogen5 (BUN), plasma glutamic pyruvic (PGPT) and plasma glutamic oxalacetic (PGOT) transaminase activities5, plasma alkaline phosphatase activitys, y-glutamyl transpeptidase6 (y-GTP), creatinine phosphokinase (CPK)7 and calcium8. Plasma alkaline phosphatase values were not obtained for the pretest (baseline) period because of interference from the anticoagulant. c. Urinalysis: Urinalysis included: measurement of volume, pH9, and specific gravity; description of color and appearance; qualitative tests for protain9, glucose9 . ketones9, bilirubin9, and occult blood9; and microscopic examination of sediment. 137-085 InternatioRneaslearcahndDevelopmenCtorporation Page 6 d. Serum Samples: Serum samples were obtained.for all surviving rats at 13 weeks of study. The samples were pooled by sex and group, frozen, and subsequently shipped to the sponsor. 5. Statistical Analysis: All statistical Analyses compared the treatment groups with the control group, by sex. Body weights (week 13), food consumption (1-13 weeks), hematological, biochemical and urinalysis parameters (1 and 3 months) and absolute and relative organ weights (terminal) were compared by analysis of variance (one-way classification) Bartlett's test for homogeneity of variances and the appropriate t-test (for equal or unequal variances) as described by Steel and Torrieio using Dunnett'sll multiple comparison tables to judge the significance of differences. B. RESULTS: 1. General Behavior, Appearance and Survival: Control rats exhibited no behavioral anomalies related to the testing procedure. Rats given the compound at 30 ppm of the diet exhibited no specific behavioral signs of toxicity and there was no mortality. Changes in general behavior and appearance for male rats at the 100-ppm dosage level included the following: increased sensitivity to external stimuli and red material around the mouth were observed for one rat; increased sensitivity to external stimuli, leans to left and convulsions following handling were noted for a second rat; and one rat was cold to the touch (one day after collection of blood) and died. No compound-related signs of overt toxicity were noted for any of the females at 100-ppm dosage level. 137-085 InternatioRneaslearcahndDevelopmenCtorporation Page 7 All rats in the 300-, 1,000- and 3,000-ppm dosage levels died prior to the scheduled sacrifice. Most exhibited one or more of the following signs of overt toxicity: emaciation, convulsions following handling, altered posture (hunched back), red material around right eye, yellow material around the anogenital region, increased sen- sitivity to external stimuli, reduced motor activity and moist red material around the mouth or uose. At 300 ppm, death for males occurred from day 13 through day 25 and for females from day 18 through day 28. At 1,000 ppm, death for males and females occurred from day 8 through day 14. At 3,000 ppm, death for males and females occurred from day 7 through day 8. Observation Number of Rats/Observation/Dosage Level 100 ppm 300 ppm 1,000 ppm 3,000 ppm Emaciation 5 10 6 Convulsion 1 10 3 1 Altered position (hunched back) 1 1 7 6 Red material (right eye) 2 1 Yellow material (anogenital region) 2 3 Increased sensitivity 2 2 Reduced motor activity 4 Moist red material (mouth) 1 1 1 Survival (prior to sacrifice)after 3 months of compound admin- istration was as follows: Dosage Level No. Surviving/N2. Initiated Male Fe-Ale Control 30 ppm 100 ppm 300 ppm 1,000 ppm 3,000 ppm 5/5 5/5 5/5 5/5 2/5 3/5 0/5 0/5 0/5 0/5 0/5 0/5 137-085 InternatiRoensaelarcahndDevelopmeCnotrporation Page 8 2. Body Weights (Tables 1-2): Comparison of mean body weights of the 30-ppm and 100-ppm groups indicate a lower body weight gain at both dosage levels when compared to the controls. Moderate to marked losses of body weight were seen prior to death for rats at the 300-, 1,000- and 3,000-ppm dosage level. Group mean body weights after 90 days of compound administra- tion were as follows: Dosage Level Control 30 ppm 100 ppm Group Mean Body eights, g Male Female 472 431 (- 8.7) 393 (-16.7) 264 243 (- 8.0) 221 (-16.3) 3. Food Consumption _(Tables 3-4): Group mein averages for food consumption declined consistently as the dosage level was increased. Dosage Level Control 30 ppm 100 ppm 300 ppm 1,000 ppm* 3,000 ppm* Average Food Consumption (grams/rat/day) Male Female 27.4 26.0 23.9 12.8 7.5 5.0 19.7 19.0 15.8 9.3 6.6 4.2 *values for 1 week only 4. Laboratory Tests (Tables 5-8): a. Hematology: At the 30-ppm dosage level (1 and 3 months) and at the 100-ppm dosage level (1 mouth) hematologic values were within the expected range. For the 100-ppm dosage level at 3 months, the two surviving male rats had lower erythrocyte, hemoglobin, hematocrit and leucocyte counts (differences when compared to controls were statistically significant, p<0.01). The three surviving female rats had slightly 137-085 InternatioRneaslearcahndDevelopmenCtorporation Page 9 lower hemoglobin and reticulocyte counts (statistically significant, p<0.05) as well as slightly lower erythrocyte, hematocrit and leucocyte counts. All other hematologic values of male and female rats at this dosage level were within the expected range. b. Biochemistr_v: One control rat showed moderately elevated PGOT and PGPT activity at 3 months of study. At the 30-ppm dosage level, one female rat showed a slightly elevated glucose level and one male rat showed a slightly elevated alkaline phosphatase value at 1 month of study. At 3 months of study, one male rat at the 30-ppm dosage level showed slight to moderate elevations in glucose, blood urea nitrogen and Y-glutamyl trauspeptidase activity. At the 100-ppm dosage level, slight to moderate increases in glucose (3 males',1 female) and blood urea nitrogen (5 males, 4 females) were seen at 1 month of study. Also noted at 1 mouth of study, for 3 males and 2 females at the 100-ppm dosage level, were slight to marked increases in PGOT and PGPT. The creatinine phosphokinase (CPK) levels generally were slightly increased for rats at the 100-ppm dosage level at I month of study. Alkaline phosphatase activity also was slightly elevated for a few rats at the 100-ppm dosage level at 1 mouth of study. The group mean glucose values for males, blood urea nitrogen values for males and females, and CPK and alkaline phosphatase values for females at the 100-ppm dosage level were significantly higher (p<0.05) than the control means at 1 month of study. At 3 months of study, 2 of the 3 surviving female rats at the 100-ppm dosage level showed slight to moderate increases in PGOT and PGPT activity. One of these females also showed a slight elevation in CPK activity. c. Urinalysis: No changes considered to be related to compound were seen in the urinalysis studies. 137-085 InternatioRneaslearcahndDevelopmenCtorporation Page 10 IV. PATHOLOGICAL STUDIES A. METHODS: 1. Gross Pathology: After 90 days of compound administration, five rats/sex in the control and 30-ppm groups, and two and three female rats in the 100ppm groups were sacrificed with carbon dioxide and necropsied. At necropsy, organs and tissues were examined for gross abnormalities and collected in 10% neutral buffered formalin (eyes in Russell's fixative). Liver samples, obtained from all of the rats at terminal sacrifice, were frozen and subsequently shipped to the sponsor@ All rats that died prior to termination, were also uecropsied, and tissues were collected as above. 2. Histopathology: Microscopic examination of formalin-fixed, hematoxylin and eosin stained, paraffin sections was performed for all rats in the control, 30-, 100-, 300-, 1,000- and 3,000-ppm dosage levels. The following tissues were examined histologically: brain with cervical lumbar spinal cord peripheral nerve eyes pituitary thyroid parathyriod adrenals lung heart with coronary vessels cord B. RESULTS: aorta spleen mesenteric lymph thymus bone with marrow (sternum) salivary gland stomach small intestine (3 levels) colon nodes pancreas liver kidneys urinary bladder testes ovaries prostate uterus skin (mammary gland) and all tissues with gross lesions 1. Gross Pathology (Table 17) and Organ Weights (Tables 18-19): Compound-related gross necropsy lesions and/or observations were present among rats in all experimental groups. The livers of these rats showed varying degrees of discoloration"ancr/or enlargement, while in the stomach, foci of discoloration in glandular mucosa, 137-085 InternatioRneaslearcahndDevelopmenCtorporation Page 11 presumably hemorrhages were also observed. In addition to the above gross observation, emaciation was evident.especially among rats that died prior to sacrifice (300-, 1,000- and 3,000-ppm levels). 2. Hlstopathology (Table 20): Compound-related lesions were observed histologically in the following organs and tissues. a. Liver: Slight to marked, focal necrosis of liver cells. with no lobular distribution was seen among rats at the 100-ppm and higher treatment levels. The above lesion was accompanied by focal to multifocal, very slight to slight, cytoplasmic.enlargement (hypertrophy) of hepatocytes in the centrilobular to midzonal regions of the affected liver lobules in all test groups, especially amone male rats. There was a slight increase in cytoplasmic vaculoation of hepatocytes in 30-ppm treated rats, but this finding was not evident at the higher dose,levels; therefore, this lesion was considered insignificant. b. Stomach: Mucosal hyperkeratosis and/or acanthosis was observed in the forestomach (nonglandular portion) while mucosal hemorrhages c. and/or erosions were observed in the glandular portion. Bone Marrow: Hypocellularity was noted. d. Thymus: Depletion in number ans size of lympoid follicles was noted. a. Spleen: Decrease.in size of the organ (atrophy) with a corresponding decrease in size and number of lymphoid follicles and cells was observed. f. Mesenteric Lymph Nodes: Depletion in size and number of lymphoid follicles was noted. g. Small Intestine (duodenum, jejunum, ileum): Decreased atrophy in the height and thickness of the villi were noted. 137-085 InternatioRneaslearcahndDevelopmenCtorporation Page 12 h. Skeletal Muscle: Atrophy was evident. i. Skin: Epidermal hyperkeratosis and/or acanthosis was noted. The findings in the stomach, lymphoid organs, intestine, muscle and skin, were prominent among all rats (300-, 1,000- and 3,000-ppm levels) that died prior to sacrifice. 137-085 InternatioRneaslearcahndDevelopmenCtorporation Page 13 References 1. Coulter Remoglobinometer, Coulter Electronics, 590 W. 20th Street, Hialeah, Florida. 2. Microhematocrit, John B. Miale, 3rd Ed., 1967, The 0. V. Mosby Company, p. 1154. 3. Coulter Particle Size Counter, Model ZBI Coulter Electronics, 590 W. 20th Street, Hialeah, Florida. 4. Gradwholls Clinical Laboratory Methods and Diagnosis, Frankel and Reitman, Editors 6th Ed., 1963, The C. V. Mosby Company, p. 1132. 5. Technicon Auto Analyzer 6/60 Micro Methodology. 6. Sigma GGTP Procedure Bulletin #545. Sigma Chemical Co., St. Louis, Missouri. 7. Micro Auto Analyier 11, 6/60 Micro Methodology. 8. Photovolt PV4, Photovolt Corporation. 9. Multistix (-AmesReagent Strips). 10. Steel, R. G. D. and Torrie, J. E. (1960), Principles and Procedures of Statistics, %icGraw-Hill,New York, N. Y. 11. Dunnett, C. W., New Tables for Multiple Comparisons With a Control, Biometrics, Sept. 1964. 137-085 1,1*1ItI$&'ItI$!(!1I4":iIl ;till I (.'iliitrill sillily itioy Wt.. Well-.lot Siii*Klolil-,i!o viv;ii NiiieLY I)iky%IillllctlItteiltTtoxit;ity;Ltllly Body Wt. (:r4iiiii Itiellwyeiritte. (:rllms.W,L!IrlltkilligLIrn.o;ttlqtlrviviil. W4vll%iit SterNjiliges v IVill 100 lloltlyWeiglL WL. Rangtin sell-vivaki ltudy Wetglit Wt. Ralines StirVIVAIL Body Wt. (!OILJ!M Wo-,Iltltt Sisr gailiges viv., -1 (1 I 2 3 4 5 6 7 a 9 to IL 12 2 3 4 5 6 7 a 1) 12 Is 224) 245 'Sol 12'1 14@b 'it$8 'IRO 416 434 444 451 4b3 475 486 472 212-228 2.11-259 301-350 3OR-'176 346-405 352-406 380-444 400-466 414-48) 418-487 437-495 448-51H 458-526 438-S23 5/5 its 5/5 5/5 5/5 5/5 5/5 5/5 5/5 5/5 5/5 5/5 5/5 5/5 5/5 2is 2'19 294 303 330 3st 356 389 404 41.1 422 429 440 444 4]t 206-227 227-265 272-328 274-332 303-360 3tt-387 328-392 360-420 3. 70-440 176-448 386-455 399-458 410-476 418-466 408-464 5/5 5/5 j/S 5/5 5/5 S/5 5/5 5/5 5/5 5/5 5/5 5/5 5/5 S/S 5/5 IR6 16R-197 5/5 186 172-199 5/5 11)1 ]bR-207 !i/i 192 IBO-208 5/5 219 190-236 5/5 220 206-240 5/5 219 186-236 5/5 224 204-244 5/5 233 207-244 5/5 231 211-255 5/5 141 212-264 5/5 235 216-262 5/5 252 222-278 5/5 246 232-264 5/5 2h2 224-286 5/5 256 238-276 5/5 266 ZJO-21)0 5/1 256 238-2RO 5/5 271 2'16-297 5/5 254 234-28S 5/5 2b7 233-21)8 5/5 254 236-278 5/5 21". 2 14-:110 5/5 252 238-274 5/5 ?Bt, 142.--illi 5/5 2(,4 244-290 5/5 3116 2/i.l-'14)lo5/5 260 240-284 515 264 224-20)8 5/5 243 228-262 5/5 217 226 276 291 .301 107 308 324 351 373 :188 391 398 412 393 2t2-224 2(10-252 2ti6-306 244-330 258-350 262-359 278-362 275-363 280-390 2R4-42(i Zlil-442 292-457 :120-47h 344-480 ')20-466 5/5 5/5 5/5 5/5 5/5 5/5 3/5 3/5 3/5 3@5 3/5 3/5 2/5 2/5 2/5 188 170-206 5/5 198 182-222 5/5 212 190-234 5/5 204 182-226 5/5 202 192-214 5/5 210 195-235 5/5 217 208-222 3/5 272 212-234 3/5 22t 212-214 3/5 224 217-235 3/5 227 217-245 3/5 225 216-241 3/5 2:16 230-244 3/5 234 225-250 3/5 221 20(j-238 3/5 202 223 217 2(17 192 196-214 210-250 200-244 200-224 IR:1-200 5/5 5/5 5/5 4/5 3/5 0/5 187 165-202 5@5 197 175-215 5/5 176 164-188 5/5 166 t5l)-182 5/5 166 152-178 4/5 0/5 217 207-232 5/ 23t 21.2-251 S/ 106 164-204 5/ 178 172-igg 5 180 179-181 5 142 120-150 5 0 .. ... .. ............ I Fluarochosical Surfaccanc Sinat7 Day Subacut* Rac Taxieltv Scudy. Page 15 TABLE 1. Individual Weekly Body Weights, Grams. Group. Prarast F &at qo. Sax -1 0 Control: Week of Scudv 2 3 4 5 6 7 3 9 10 --Tl12-13 73886 r 73887 73888 73889 73890 x 222 249 314 330 354 372 396 424 438 452 !,53 464 494 4a2 466 x 225 248 286 302 328 346 352 380 @@oo 414 418 437 448 464 456 x zzs 159 330 358 376 405 406 444 466 483 487 495 513 526 5'3 x 212 '133 3CO 322 342 367 372 404 424 422 442 "l 448 45g8a 4&38 x 21-1 i35 -.76 302 308 352 374 428 440 451 456 476 476 4 477 73891 73892 73893 73894 73895 p 194 200 230 230 240 251 262 278 280 285 270 279 294 394 268 F 179 184 208 210 230 227 238 248 252 246 248 256 260 262 243 F 194 207 232 236 2" 264 278 286 290 297 298 310 310 320 298 r 168 168 L90 186 207 212 222 224 230 236 233 234 242 243 224 r 197 205 236 234 242 252 260 274 280 293 286 296 300 310 2SS 30 con: 73896 x 216 232 '@82 300 325 343 3" 382 394 408 425 424 434 "4 417 73897 m 227 265 328 332 360 387 392 420 440 "a 455 458 476 466 464 73898 x 217 240 306 316 344 366 368 410 4ZO 420 431 "4 "6 454 438 L 73899 x 208 229 280 292 318 346 350 372 394 401 413 420 434 436 428 73900 x 206 227 272 274 303 3U 328 360 370 376 386 399 410 418 408 73901 73902 73903 73904 73905 p 172 iso 208 204 213 220 232 238 238 234 236 238 248 241 228 p 199 208 240 242 254 262 264 274 280 285 278 274 290 284 262 r 195 191 210 216 216 216 236 250 244 245 244 243 260 253 235 F 193 201 238 2!64 255 255 264 276 276 273 272 265 276 280 259 F 172 181 206 212 215 221 232 244 240 2.35 240 240 2" 240 231 100 20m: 73906 73907 73908 73910 73946 73911 73912 '!3913 13914 73915 x Z13 200 270 m 224 252 306 x 212 224 272 m 221 245 284 x 213 208 246 F 206 217 232 F 205 222 234 F 170 182@ 190 F LBI i35 2C4 F 178 196 198 292 292 330 350 300 319 290 284 244 258 2'-4) 210 2z6 214 188 196 200 192 182 200 309 Died 359 362 363 390 305 284 335 382 302 Died 262 278 275 280 211 222 234 234 235 Died 195 208 212 212 204 Died 206 220 220 216 415 439 1.20 442 284 282 235 2145 217 218 221 217 424 Died 457 476 480 466 292 320 2!.l 244 344 320 230 238 217 230 228 Z06 216 234 225 219 137-085 Fluerachomical Surfactanc 2. C-.U. Group, Bac so. Sam Pretest -1 0 300 cam: 73916 73917 73918 73919 73920 x 199 210 x 200 213 x 196 212 x 203 230 x 214 230 73921 73922 73923 73924 73925 F 190 200 F 183 192 r 202 215 F 194 202 F 165 175 1,000 you: 73926 73927 73928 73929 73930 x 207 212 m 207 212 s 220 230 x 232 251 m 217 248 73931 73932 73933 73934 73935 F leg ISO F 175 ISO 7 176 Lai F ISO ISO r 172 179 3,000 vam: 73936 x 218 242 73937 %1 217 237 73938 x 223 252 73939 m 216 245 '-73940 x 223 248 73941 73942 73943 73944 73943 r 187 197 F 169 178 192 198 F 177 183 F 170 177 Ninety Day Subacute Rat Toxicity Study. Individual Weekly Body Wtights, Grams. Week of Studv 1 2 3 4 5 6 7 a 212 200 210 220 2!64 iss lis 178 184 164 200 Died 204 200 224 174 162 182 164 150 183 192 200 Died 165 168 178 L52 Died Died Died Died Died Died Died Died 164 Died 170 Died 190 Died 202 Died 204 Died 150 Died 120 Died 148 Died 150 Died 142 Died 164 158 Died Died 156 11.0 Died i-io 126 D!.ed Died Died Died Died Died Died 9 10 Page 16 12 13 137-085 L Fluorochemical Surfactant Ninety Day Subacute Rat Toxicity Study. TABLE 3. T-test Comparison Between Average Food Consumption of Control and Treated Groups. week of Study Sex Control 30 ppm 1-13 27.4 19.7 26.0* 19.0 100 ppm 23.9** 15.8** 137-085 *Significantly different than Control group mean, p<0.05. **Sigaificantly different than Control group mean, p<0.01. FI SIIF t illit 'I'Aill4..l-' 114.mI t.411 Week .11 Sillily rist/ dity (illy 2 .1 4 5 6 7 8 9 10 It 12 I'l FFMAI.ES: 1 2.1 4 5 6 7 a 9 10 It 12 13 21.() @I/I.9 26.7 24.7 28.7 28.5 29.5 26.9 28.0 2B.5 28.3 26.5 28.2 19.9 16.9 20.3 18.7 20.3 20.5 21.0 24).3 21.1 21.1 21).j 11.2 18.3 H9.6 71.11 18.1 67.0 75.6 68.5 67.9 61).6 62.0 61.5 59.5 54.6 59.7 90.8 77.2 87.3 77.5 80.7 78.1 79.1 74.7 78.9 76.Y 71.7 56.1 69.2 30 ppo it/ R/ rat/ kg/ day day Hiiit-ty1);iy RiitTiixic!ltyStittly. tit!ilVlOlI)D(,ollotlmllipstit. 100 ppo rat/ tl;ly it/ kit/ day 300.1ilm riot kg/ filly day 27.6 21.0 25.6 23.9 25.2 27.5 27.6 26.0 27.9 26.1 25.5 23.8 27.8 2t.2 16.9 19.1 16.9 t7.9 19.4 20.5 19.9 20.0 18.4 19.8 17.1 20.3 93.9 75.8 77.7 ba.i 70.8 70.7 68.3 63.3 66.1 60.9 Se.t 53.6 64.4 96.4 75.5 82.9 71.7 72.9 75.7 80.1 78.5 78.9 73.0 75.t 65.8 83.5 25.S 21.2 21.0 16.9 t9.2 21.1 26.0 27.5 26.2 26.1 28.1 25.9 26.5 18.9 13.9 15.7 t3.2 14.9 17.1 16.9 t5.9 15.9 16.3 16.0 14.7 tS.6 92.3 72.9 69.7 54.9 62.5 65.1 74.1 73.7 4?.5 66.9 70.7 62.8 67.4 88.9 68.3 77.9 62.9 68.5 77.2 76.3 7t.0 70.1 72.4 67.8 62.9 70.6 18.2 7.9 12.2 $4.0 38.0 63.7 11.0 7.0 9.8 62.7 42.3 59.2 1,000 I!ilm A rat Lil day till 7.5 40 6.6 46 . . .............. Fluorachamical Surfaccaut TABLE S. Hamtology Erythrocvcas, Rematocrit, % Hmaglabin. 2/100 al 10 /cm Neutrapbils, z Lymphocytes, 2 Zosinophils, 0 .4onocytes, Basophil-s, z Raticulocyces. 4inety Day Subacute Rat Toxicity Study. 4&LZS: m-jknsand Signiflemcea of Rematological Values. Scud3.4on-.b Control 30 ppa Precosc 1 3 Pratast 1 3 Pretest 1 3 Pretest 1 3 Protest 3 Protest 1 3 Precast 1 3 Precast 1 3 Orecest 1 3 Protest 1 3 5.96 6.50 7.57 45 45 50 15.5 16.0 16.6 U.32 L3.14 12.50 7 Li 14 93 as 94 0 1 0 0 1 0 0 0 5.3 3.9 2.8 6.42 6.28 7.43 47 48 46** 15.5 14.9 15.6 10.60 L2.69 9.63 11 12 15 as 86 83 1 2 2 0 0 0 0 0 0 5.2 3.2 2.9 100 ppo 6.40 6.30 5.52** 48 43 3 15.9 15.3 10.76 9.97 6.21** 17 17 30 82 81 69* 1 1 0 0 1 0 0 0 0 5.4 2.7 3.2 137-085 *Significantly different than Control group mean, p<0.05. *a*Significantly different chan Control group mean, p-cO.01. S-.acisticalanalyses were not conclude--o*n -.heprotest valu@s. Fluorochomical Surfaczauc iiamazology E-,vtgroc-!tes. 10 /cm Hamacocrit, Hawglobia, g/100 al Loucicytes, 10 /emm Sautrophils, Lympboe-ytes. tosinophils, fonoc7tas, Basoph:Us, Reciculocytes, Ninety Day Subscuce Rat Toxicity Study. r-MLU-rS: %leans and a Signi!icance of Rematological I:aLluts. Study Month Control 30 ppm Orttest 1 3 Protest 1 3 Precast 1 3 Pretest 1 3 Precast 1 3 protest 1 3 P.-acost 1 3 Iretost 1 3 ?retest 1 3 ?retest 1 3 6.74 6.54 6.39 48 4;11 45 16.2 13.9 15.4 9.23 7.23 7.29 17 15 21 82 al 76 1 3 2 0 0 0 0 4.4 3.1 3.2 6.69 6.58 7.28** 48 47 47 16.0 15.7 16.0 9.77 7.07 5.68 a 8* 19 91 gj** 80 0 0 0 0 0 0 4.1 2.3* 2.8 Page 20 lot pz- 6.36 6.69 6.03 45 48 41 15.0 16.1 13.8* 9.39 10.23 6.26 11 8* 25 sa 91** 75 1 Jt* 0 0 0 0 0 0 0 4.1. 2.4 1.8* 13,--085 *Significantl? different chau Control group u"n. P40.05. "Sigmificantly differon: than Control group M"C, p-cO.01. 'Scaziszical analyses were not perform*d an cio protest value. v Itoflat-litI-4m-.Il Stirl';ii6i-ttit 'I'Aill.61.1 I:rtiiol). RI$ t Sux Pryilirti- 4*yti.n I,)/com 114!0L1O1c I-It z Hitiety llityStilineute RBL T4ixii-ILy Stittly. liidivitisiIslaelmatolorii-iiVialtieti- llreteiiii. llemoi;lubiii m I.ittlet)tcilito 0 /cm met$ttt)J!Itkai ser.. Nuit-Seg. 1 2 l.yophucytes z 71886 11 6.25 47 16.5 10.20 a 0 92 0 73087 @l 5.95 44 15.2 12.38 9 0 9t 0 7 3H$R 5.77 4'1 14.9 11.55 6 (1 94 0 7'1889 ti 5.56 4S 15.4 11 .()1 6 94 0 71M90 ti 6.26 48 15.7 11.45 6 94 0 pit-all 5.96 45 15.5 11.32 7 0 93 0 7'1891 F 6.42 5() 16.4 8.65 9 (1 9t 0 7'JU92 F 6.16 41 15.5 7.83 11 0 89 0 73893 F 6.67 4R 15.5 10.05 12 86 2 7'1894 p 7.)6 st 17.4 9-52 34 0 66 0 73895 F 6.89 49 16.3 10.1)8 17 82 1 K!lllk 6.74 48 16.2 9.23 17 0 82 1 .IQill,!!: 7,IN96 6.77 49 16.4 1().1$ 12 87 1 l'IN97 pi 5.89 44 14.7 9.()S 1() 0 69 1 7'JU98 6.57 47 ts. 1 12.52 17 1) 82 1 71899 m 6.41 48 15.4 12.68 6 0 94 0 7 19#Xl N 6.48 49 15.7 8.61 a 91 1 F14!1611 6.42 47 15.5 11).60 it 0 as I 739111 r 7.16 5(1 16.9 9.49 6 0 93 1 73,102 v 6.2R 44 15.5 9.17 ti 0 89 0 Y'19111 F 7.()1 sto 16.2 9.57 11 41 89 0 719t$4 p 6.2*1 47 tS.4 6.St 6 0 94 0 7'19(1)' p 6.73 48 16.1 13.80 5 93 2 6.69 48 16.0 9.77 a 91 1 711)416 ti 6.bK 49 15.9 l(I.19 1 lp 0 ol 2 1'19111 11 10.99 45 15.2 14.31 It) 11 89 t 7 @i9@18 ti 6.25 47 15.6 8.01) Is 11 at 1 731)11) m 6.57 so 16.4 13.42 11 1) a/ 0 7'11)41P Pi 6.52* 49 16.2 7.88 1() 1 69 4) Fk-.Ill 6.411 48 15.9 11).76 17 11 B2 1 71911 r 6.(In 42 14.K 11.8% 11 11 87 0 7'111)2 v 6.51 46 15.3 l(P.7'1 11 0 By 2 7'1911 r 6.41 45 14.R 1.67 11 87 2 7'11)14 v 6.19 to] 14.2 lil.'14 41 11 ql 0 1'19js F 6.lol 41 1li.9 8.36 12 88 6.'16 ............. ... RR I @All I'l? fill,, F Ioisiriti-IlciitsI-o NicietyI)jiyStiliasetiRtaet 'roxii-ityStiidy. 'I'Aill.6V. t inilivititiltlielmitoti)glt-Y.ailities llrvto-Ni. (:riitili. Pryttirti- llemtto- lii-mo- Netitri)plills l.ymipllu- PAIGIIUI- Moll Nilt I. RIOI)III c SCR. HI)n-SeR. cytes pl%Ltpi cyt Nex 0 /4-00 z 5/100 mi I /com z 2 z 711)16 H 7.30 5() 15.8 1().11 16 8.3 1 0 731)17 ?1 6.111 42 7),)1n ti 6.(Il 49 14.5 8.85 6 0 Its.1 11.93 10 93 1 0 9() 0 11 '73911) m 5.!Ih 43 14.9 11).32 7 (1 92 1 41 731)2(1 m 6.31 47 15.1 13.36 1() 0 90 0 0 tic-ills 6.47 46 ts.3 10.91 10 0 ag I (I 731)21 F 6.69 49 15.8 7.76 14 41 85 7'1922 v 7.12 49 t7.1) 11.18 7 0 93 73923 6.66 48 15.9 9.30 .10 0 90 73924 fi.48 48 16.5 1t .49 12 0 as 73925 6.45 48 16.2 10.78 5 0 95 1 (1 0 0 0 0 0 0 0 0 fleiiii 6.68 48 16.3 to.10 10 0 90 0 0 1,(NIOJilti@: 71926 Fl 7.08 49 16.5 12.76 9 91 73927 m 6.14 48 16.1 13.67 12 as 71928 N 6.()9 49 16.0 9.75 17 0 83 7'1929 ti 6.R4 49 16.4 13.02 a 0 91 7391(1 6.53 47 IS.t ts.59 a 91 Ho--alt 6.54 48 16.0 12.9fi 11 11 89 73931 r 6.66 50 16.7 10.50 17 0 82 73932 F 7.60 51 t7.8 12.79 Is 0 84 73911 p 7.*13 49 17.6 14.12 17 0 83 71934 r 7.tZ 47 16.8 11.74 is 0 83 73915 v 6.:16 46 15.4 17.48 7 0 92 7.01 49 16.9 t3.33 14 0 as 0 0 0 0 0 0 1 0 1 0 0 0 1 0 1 0 0 0 2 0 1 0 1 0 73916 m 7.12 49 16.4 13.52 9 11 91 73917 7.11 47 15.5 17.97 8 0 92 7'1938 6.35 44 15.0 14.79 9 1) 91 739:19 ?1 6.12 46 15.1 17.20 6 1) 94 73940 H 6.4R 49 16.4 15.29 14 41 86 0 0 0 0 0 1) 0 0 0 1) "A-411 6.(18 47 15.7 15.77 9 0 91 0 0 71941 r 6.00 44 15.5 13.li 13 0 85 2 0 7:1942 r 7.15 49 17.3 12.25 14 85 1 7'1941 F 6.51 49 17.4 9.49 14 05 1 71944 p 6.59 4" 16.2 ti.87 16 1) "I 1 41 711145 v 6.il 46 15.6 lt.72 to 11 9() 0 11 6.56 47 16.4 11.69 13 1) 86 137-1111% t-r It IIH86 m 6.76 47 7')Htl7 m 6.35 - 7 JdHd ti 6.45 42 73"89 6.48 46 13890 ti.46 45 tjo.;Ios (b.50 45 73891 F 6.51 44 7'1892 6.68 51) 7.'1891 6.6t, 46 7'IH94 p 6.68 41 73895 r 6.19 46 6.54 47 7389h 11 6.87 51 7381)7 6.46 46 73898 N 6.59 49 738911 6.44 46 73900 m 5.04* 49 liezill 6.28 48 739(11 r 6.64 49 7'1902 v 6.50 51 7J903 p b.58 48 7:19(14 v 6.SS 45 7:190'i p 6.61 44 6.58 47 1!n i-iln* 71906 tl,. 7.t)R 50 73907 6.30 48 739OR ti 5.71 43 7'itI()I N 5.9(1 'IH 7'1946 H fo.'19 46 tleail 6.30 45 73911 v 6.6H 48 73912 r 7.018 48 73913 r 6. 1 47 73914 r 6.!io 47 7'19I!i F 6.48 /is lit,.III 6. 6'1 48 ..................................... *I I I...I V.-III...*uI - ltif1.1 so,,1 ltv;i I III r/10 16.9 14.8 15.9 16.2 16.2 16.0 16.2 16.6 J6.0 15.8 15.1 IS.9 16.5 t5.3 15.4 15.0 12.5 14.9 16.2 17.0 15.2 16.3 13.9 t5.7 16.7 16.1 13.7 14.6 15.4 15.3 tS.7 16.4 If).1 15.9 16.1) 16.1 I- 13.63 6 9.95 9 16.14 11 12.44 12 13.52 17 13.14 11 ?.89 23 7.37 17 9.5() a 6.15 It 5.22 Is 7.23 15 9.11 16 11.15 12 12.12 16 15.74 a 15.3t 6 12.69 12 5.22 7 6.36 a 7.30 13 7.92 9 8.56 5 7.07 a 10.42 21 8.96 13 9.05 29 1:1.,12 R 7.6(1 16 9.87 17 12.65 to 13.08 a 7.26 5 9.82 a 8.4r, B 10.25 8 cyl elk z PIIJIB z 0 93 1 (1 89 2 0 as 0 0 87 1 0 at t 0 on 1 0 73 3 0 a[ 2 0 89 3 0 83 3 0 79 3 0 81 3 0 83 1 0 at, 4 0 82 2 0 90 2 0 94 0 0 86 2 0 92 1 0 92 0 0 86 1 0 90 1 93 1 0 91 1 77 1 0 85 2 0 70 1 0 92 It 0 79 2 0 81 1 (I an 2 0 92 0 11 9s 0 90 1 0 92 0 91 1 TAIII.V7. Criiiiii Niember Sex Pik", 73916 m 73917 N 73918 H 7:1919 71921 v 73922 v 71923 F 71924 v 73925 F is()00-PEN!: 73926 m 73927 m 73928 it 73929 m 73930 m 7393t p 73932 F 73933 F 73914 v 73935 F 3 a0(!q-m@: 73936 m 73937 m 73938 m 73939 m 73940 11 73941 F 73942 p 71943 r 71944 V 73945 r Frytiero- t:yiltif 1116/i.@ liesustocrit z HilieLy Day %tilidt-4stHoat Taxii-ity Stio4ly. loodividisist Vitloses I Ph)sitio. llemislobill 8/10(1 ot I.Pitc-stcli@ten I /cvm Notitre ilia seg. Noll-Seg. I.yoploocyten 2 PostaoPlitt* I)Iegl Died llled Died lflo4l Died Died Died I)Ied I)Iesl Died I)le4i Died Died Died Died Died Died Died tiled Die4i Illed Died I)Ieti I)ted Died Died Died Died I)Iefl 'I'Aill.i1.1 kilt %OX (:ootiro I 7'1886 m 7'1887 ti 7'1088 Pi 73MM9 ,71891) K.-;Iil I]HI)I F 7'IH92 F 7IH9] F 13894 F 73895 F ticiiii !(IJ.ilin: 738911 m 73897 m 71HqB m 718419 H 131)()Il Olt-Iies Bli(Ii p 119(12 p 7341(1) F 73@1(14 r 7,19fis F Fi@an l(PO 7*194)6 739(07 ti 739()K ti I'll)lil h 711146 H Pkilli 71911 p 73912 F 73913 1? 7*1914 F 7:1915 r I 17-i)HS Fry Ilireb- It /1'.M@ liewitto- crit "Inety I)ay%tibiieuteltatTtixicity Stootly. flematoloigletilViiluo--u I t4oiitlisi. liew)- GII)IIIII 2/100 NJ l.elicoI()I/e@ Metitrailt!tf.lif "Oil-.7q;-i. I I.ymllllocyted z H. 1() 52 7. '19 so 7.16 47 7.62 $1) 7.511 so 7.51 sil fi.an 4H ft.85 47 fi./7 47 5.9(1 44 fi.4 7 4f) 6.59 45 7.iR 46 7.18 48 11.21 46 7.00 46 7.111 45 7.43 46 7.71) 49 7.15 41, 7.36 48 7.10 48 6.91 47 7.28 47 17.3 16.4 15.1 17.0 16.4 16.6 16.1 15.5 15.5 16.0 13.6 15.4 16.1 15.2 15.7 15.2 15.7 15.6 16.5 IS.fi 15.8 16.3 16.0 36.0 10.35 14 13.89 17 16.20 8 10.25 12 11.80 20 12.511 t4 7.02 17 4.95 t2 8.71 is 9.60 36 6.117 23 7.29 21 8.60 11 10.65 13 9.9(1 22 10.35 a 8.66 20 9.63 15 4.8t) 23 6.)1. It 5.17 22 S.Ifi 27 6.77 It 5.68 19 0 82 at 0 91 0 85 1 77 (1 84 0 81 0 06 83 0 56 (1 75 0 76 0 88 0 85 0 77 0 90 0 77 (1 83 75 87 0 78 1 70 0 86 0 RO I)Ied 5.104 18 1111-41 6.114) 39 5.52 39 fi.14 40 I)Ietl 5.91 41 llled 6.(14 19 6.03 41 11.11 13.2 12.1 14.1 11.8 11.6 13.8 5.711 24 6.71 17 6.21 31 7.58 2R 6.*17 26 4.04 21 6.26 :!s 76 0 62 4) 68 1 71 74 (1 79 75 plifIs 2 2 1 1 2 0 1 2 1 1 6 2 2 1 2 1 1 3 2 1 1 0 2 3 I 0 1 1 0 0 0 0 TABI.F'.M. Rat HieWier Sex Erytllrogytes 10 /c@ lienatocrit z Hillety Day Subactite Rat Toxicity %tudy. loidividtinifenalological Volisom - 3 Mt)uitlio. llemoglobin a/too mi Ieucocites to- /em Neutroplille Sea. Noil-Sag. 2 2 Lymptiocytes 2 gosinoplitle 2 71916 m 73917 it 7391R ti 7'1919 7192(1 H 71921 v 73922 v 73921 r 73924 r 73925 p t-Lqqq-JL,l@n: 73926 71927 71929 m 73929 71930 H 73931 F 71932 r 73913 F 71934 p 71915 v '11(pillf. 7393fi 71917 m 73938 771939 H 73940 73941 73942 73943 v 73944 @F 71945 p Died Died 1)1 etl llittd Died "loci Died Died Died Died Died illed Died Died Died Died Died Died [)led Died Died Died Died I)Iegl Died I)IU41 Died Iliad 1)[ell flied nuorochanical Surfactant TABLE 9. Biachauiscry Glucose, mg/100 al B.U.N., ag/100 al 7-C.T.P. Siva units/al Craatimine Phosphakinmm. SLgma units/=I P.G.O.T., LzLt'iUnLts/i P.C.P.T., Lnt'l units/I Calcium, maq/1 AII,Alfna Phosphatame, Lnt'l =Lts/i N:Lnaty Day Subacute Rat Tox:Lclty Study. MALLS: "'Aaasand Si$niflcan=8 of Biachdaleal Values. Study Ilcnth Cantrol 30 ppa ftatest 1 3 Protest 1 3 Protest 1 3 Precast I 3 Protest I 3 Precast 1 3 Protest 1 3 Pracastb 1 3 84 120 L12 11.2 15.4 IS.3 2 1 1 13 is L2 95 LU 101 93 47 33 9.5 8.4 10.4 166 89 120 iu LID 15.4 15.2 28.9 1 1 3 15 10 1.1 LL2 106 as* 47 54 43 8.4 8.5 10.2 190 127 Page 27 100 pps 112 158* 112 15.3 29.5* Z3.9 1 1 2 12 10 101 539 90 33 485 37 10.4 7.6 10.1 229 103 L37-085 *SLgaL:-Icanelydlffaranc chan l.'ontroglroup a"n. p 0<03. *&s ignifir-ant.dLiyfferent than Concrol Iroup a"n, p 0<01. Stac:LstLcalanalyses were not conducced an the pracesc values. b .oc data--mlnedin protest period because of ancicoagulan."!acarfarance. Surfaccant TABLE 9. Biochemistry Ninety Day Subacute ]tatTozicity SeWy. FL4ALES: .4*a= and Significance a of Biochemical Values. Study Month Control 30 ppa Glucose. as/loo al B.U.N.. at/100 al @-G.T.P. Sigma units/21 Cr"t!tnl-* Phospbokinasa, Sigma units/ml P.G.O.T.. incllunits/I P.G. P. 'r., incliunits/i Calcium, maq/1 Alkaline Phosphacase. int'lunits/I Precast I 3 Precast 1 3 Protest 1 3 Protest 1 3 Pretest 1 3 Precast 1 3 Protest 1 3 Protasr-b 1 3 103 L19 U2 11.3 17.0 14.6 2 2 0 10 8 9 82 103 203 79 56 136 10.3 10.1 10.2 99 47 U9 125 Lio 17.0 20.0 20.2 2 1 1 8 Li 9 103 107 91 56 61 34 10.1 8.1 10.4 125 75 Page 28 100 ppa LL2 149 107 14.6 40.0* 20.6* 0 1 1 9 32* 25 205 290 397 136 179 504 10.2 8.7 10.4 182** 142 *Significantlydifferent than Control group mazu, p OCOS. '3; :S.4gal-licancldyl-Iferi=cchan Control group a"=, p O<OL. bScatl-szi:a'a-nalysas not conductedon the practst values. !.'ctiaca--zLL-&i*n-4pratesc ptr!.odbecause of ancicoagulant Lncar-jare-tca. Fluarochamical Surfactant TABLE 13. Group. Rac Glucose Number S*x mg/100 al Control: 73886 x 96 73887 m 93 73888 x 69 73889 x 81 73890 m 81 mean 84 73891 7 75 73892 r 87 73893 F 132 73894 F in 73895 r Ul 103 30 you: 73896 m 84 73897 x 93 73998 x 87 73899 x 75 73900 x 66 N.team $I 73901 F 99 73902 F 90 73903 F 102 73904 F 81 73905 F 93 Xaan 93 100 Dom: 73906 x 96 73907 x 93 73908 x 81 73910 x 93 .3946 x 34 *@lean d3 73911 F 90 i-3912 F 81 73913 F 87 r 72 /3v!3 F 78 Iaan 82 licaty Day Subacuca Rat Toxicity Study. Individual liachami=a Values - Pretest. B.U.N. mg/100 ol y-G.T.P. Sigma units/al C.P.K.* Six= units/al P.G.O.T. int'l units/i Page 29 e.G.P.T. int'l units/I Calcium seq/1 Li.7 1 9.9 2 10.5 2 LI.7 2 12.0 2 11.2 2 Li.1 2 12.0 2 11.7 2 10.2 2 U.7 2 11.3 2 12.9 2 9.7 2 U.4 1 10.8 1 9.0 1 10.6 1 15.3 1 9.0 1 12..0 1 11.7 1 6.6 - 1 10.9 1 9.0 1 12.3 1 9.0 1 9.9 1 12.0 1 10.4 1 10.3 1 11.7 1 6.6 1 6.9 2I 9.3 I 9.0 12 96 13 108 L5 87 L2 96 14 90 L3 95 14 87 13 93 12 66 a 73 5 87 10 82 23 84 42 96 38 93 15 90 12 93 26 91 13 al 5 81 12 102 24 87 9 78 13 86 9 1128 10 U4 11 90 10 81 37 96 15 302 11 75 20 U4 10 78 23 132 24 66 18 93 102 105 84 84 90 93 96 66 72 69 90 79 84 al 78 78 69 78 99 93 U7 96 90 99 1104 120 ill 87 ISO 314 al 96 72 90 66 81 9.5 9.6 9.0 9.4 9.9 9.5 9.3 9.3 9.4 10.2 12.8 10.3 9.7 9.5 8.5 8.9 9.0 9.1 9.7 9.8 9.2 9.5 9.1 9.5 9.1 9.7 9.9 10.1 9.8 9.7 IO.-@ 10.3 '-0.6 10.8 10.8 10.6 L37-085 *Creac!=ina phosphokinast. Fluorochomical surfactant TABLE 10- L@... Gcoup. &at Niumber Sax Glucose as/100 ml 3GO pum: 73916 H 73917 x 73918 x 73919 x 73920 x %loan 73921 r 73922 r 73923 r 73924 F 73925 r %I"U 1,000 P=: 73926 m 73927 x 73928 x 73929 73930 X"U 73931 F 73932 F 73933 F 73934 F 73935 r M.,@ 3,000 o=: 73936 73937 7393ts 73939 m 739.@O x @lean 73941 r 7394,' F 73943 F 7,394i! F 73945 F Xean 84 102 Los 87 93 94 Lil 90 108 93 108 102 108 108 96 105 90 101 Ul 90 al 96 99 95 87 93 78 75 81 83 102 87 102 96 96 97 Ninety Day Subacute Rat Toxicity Study. twividual Biochemical Values Pratest, B.U.N. mg/100 al Y-G.T.P. Sigma units/ml C.P.Z.* Sigma units/ml P.G.O.T. inc*l unics/I 8.7 2 7.5 2 9..3 1 9.0 1 9.3 2 8.8 2 Li.7 1 9.6 1 9.0 1 12.0 2 12.0 1 10.9 1 14.1 1 U.4 2 9.3 2 1.1.1 2 8.7 2 10.9 2 1-).6 1 10.3 1 8.7 2 9.3 2 10.8 2 10.4 2 9.3 2 14.4 2 11.7 2 12.6 2 23.7 22 14.3 6 11.4 2 U.4 2 11.7 1 17.7 2 12.3 1 12.9 2 19 B7 21 84 10 84 27 72 1.1 69 la 79 6 60 10 87 13 72 Li 63 10 60 10 68 14 84 8 60 is 87 9 75 41 129 is $1- 16 117 13 87 12 72 12 90 12 93 13 92 11 84 14 84 12 $1 10 108 14 105 12 92 32 93 16 78 9 72 21 90 11 90 IB as Page 30 P.G.P.T Lnt'l units/', Calcium maq/1 102 11.3 87 11.8 93 11.5 84 11.1 87 11.7 91 11.5 78 U.4 90 Ll.4 84 12.1 96 11.0 87 10.3 97 11.2 94 Ll.7 90 11.6 66 11.3 66 10.2 90 11.6 79 11.3 129 10.7 75 10.9 60 11.3 96 12.0 90 11.4 90 11.3 LOS 11.2 132 11.6 99 11.1 126 11.5 93 11.6 L12 11.4 99 10.9 ioa 11.1 117 10.8 102 11.3 126 9.2 110 10.7 L37-085 kCreaz4-nIrd phosphokl@nase. Fluarochomical SUr' &rrAnt TABLE Ll. Group, &at Glucose .4umber Sax mg/100 ILL Control: 73886 x 73887 x 73888 x 73889 x 73890 x Mean 73891 F 73892 r 73893 p 73894 F 73895 F K"n 30 ppa: 73896 H 73897 x 73898 m 73899 x 73900 m Mean 73901 r 73902 r 73903 r 73904 r 73905 r Mean 100 Dom: 73906 x 73907 x 73908 x 73910 x 73946 n 1.tean 7391'- F 73912 F 73913 p 73914 T 73915 r 1Mean L38 Los 117 120 L17 120 120 126 im 117 Los U9 Los U7 108 lu ill III 120 147 120 123 114 125 135 171 183 171 129 158 153 225 120 123 126 149 llmty Day Subacute &at Toxicity Study. Udividual Biochemical Values - 1 Month. 3.U.S. agiloo al Y-G.T.P. Sigma units/aLl C.P.K.* Sigma units/21 P.G.O.T. Lnt'l unics/I 17.4 14.4 14.7 15.3 15.0 15.4 18.0 16.5 LS.3 L5.3 19.8 17.0 3 16 Los 1 a 93 1 a 90 1 30 150 0 11 117 1 15 1.12 2 6 L17 2 10 Los 2 6 93 1 a 105 1 12 93 2 a 103 L3.5 1 9 93 L5.0 1 10 90 18.6 1 15 123 LS.3 1 6 99 1.3.5 0 a L23 15.2 1 10 106 22.2 0 6 102 16.8 2 9 Ul 22.5 1 a 93 17.7 1 9 120 21.0 1 23 108 20.0 1 Li 107 29.7 30.0 30.0 34.5 23.4 29.3 29.7 42.0 38.4 60.3 29.7 40.0 1 36 363 1 29 243 2 96 1527 2 14 396 L 16 168 1 44 539 0 42 432 2 56 657 1 24 123 1 19 120 1 17 U7 1 32 290 Page 31 P.C.P.T. incll units/I -Calcium maq/1 Alk. Phos. int'L unitsij 54 39 33 66 45 47 60 60 51 54 54 56 54 42 75 48 51 54 45 72 51 66 72 61 345 180 1248 480 171 485 282 393 75 78 69 179 8.1 201 8.0 147 7.7 147 9.3 138 3.7 198 8.4 166 9.3 93 8.8 120 9.3 105 10.8 87 13.4 90 10.1 99 7.7 207 8.4 267 8.7 162 8.8 163 8.9 147 5.5 190 7.5 132 8.0 144 8.2 90 8.1 120 8.8 138 5.1 125 8.0 219 8.0 198 7.0 ISO 7.2 258 7.6 288 7.6 229 8.3 162 8.2 132 9.0 ISO 8.8 231 9.2 204 8.7 182 137-085 *Creacinine phosphokimas&. Fluorach=ical Surfaccant TABLE II. C Group. Rat NuzLber Sam Glucose mg/100 al 300 ova: 73916 x 73917 m 73918 x 73919 m 73920 m 73921 F 73922 r 739Z3 F 73924 p 73925 F 1,000 ova- 73926 m 73927 m 73928 x 73929 x 73930 m 73931 F, 73932 p 73933 F 73934 F 73935 F 3,000 cam: 73936 x 73937 m 73938 m 73939 x 739,LO %I -.1941 F i3942 F 73943 F #3944 F 714945 F Died Died Died Died Died Died Died Died Died Died Died Died Died Died Died Died Died Died Died Died Died Died Died Died Died Died Died Died Died Died Ninety Day Subacute Rat Toxicity Study. Tndiv:Ldual Biochemical Values - 1 Mouth. B.U.N. mg/LOO ml y-G.T.P. Sigma units/al C.P.K.* SLgm units/al P.O.O.T. intli units/1 Page 32 P.G.P.T. int'l units/i Calcium asq/1 Alk. Phos. int'l units/I 137- 085 *Creatictime phosphokinasa Fluccochodeal Surfactant TABLE L2. Group, &at .4unbor Sax Glucose mg/100 al Minaty D&7 Subacuto &ac To TACit.v 97tudy. Individual Bioch@ical Values - 3 Ionths. S.U.Ii. as/100 ol y-G.T.P. Sig= UtLits/ml C.P.K.* Sigma =its/ml P.G.O.T. intl units:l Control: 73886 x U6 14.1 0 73887 x 105 14.2 L 73888 x uo 15.7 1 73889 m ill 17.6 1 73890 x lis 14.9 0 %L"n uz 15.3 1 73a9l r 105 16.0 1 73892 F 102 14.0 1 73893 F 128 L3.9 0 73894 F ill 12.0 0 73895 p 112 17.0 0 V.46au 112 14.6 0 330 D9=: 73896 m 143 74.4 13 73897 x 91 16.5 1 73898 x 116 19.1 0 73899 x 94 18.3 1 73900 x 105 16.1 0 4ean uo 29.9 3 73901 F 105 23.6 0 73902 p 125 17.6 0 73903 F 121 22.1 1 73904 F 103 17.4 1 73905 r 95 20.1 1 %.lean 110 20.2 1 100 73906 x 73907 %1 Died Died 7a9O8 %1 100 21.4 1 73910 x Died ,-3946 h 123 26.3 2 @ican 112 23.9 2 73911 7 117 19.0 0 73912 p Died 73913 F 121 19.0 1 73914 F Died 73913 F 94 Z3.8 1 %lean 107 20.6 1 Li L09 L3 97 12 95 a 102 14 102 12 101 a 87 9 86 9 as 9 663 1.1 100 9 203 11 36 10 86 12 100 L3 79 8 90 11 as 6 71 9 90 8 90 9 106 u 96 9 91 7 87 13 93 10 90 7 95. 42 786 27 309 25 397 Page P.G.P.T. int'l =ics/l CiLlcium maq/1 Alk. Phos. int'l uuits/I 42 10.4 120 31 10.0 79 30 10.8 99 32 10.3 59 30 LO-3 86 33 10.4 89 29 U.0 46 27 10.3 43 24 9.7 47 560 9.8 47 41 10.2 53 136 10.2 47 39 10.5 175 48 10.1 139 38 9.9 107 42 9.8 105 49 10.5 lu 43 10.2 127 19 10.5 70 23 10.5 95 38 10.3 49 53 10.4 60 35 10.5 99 34 10.4 75 46 10.3 79 27 9.9 127 37 10.1 .103 60 10.8 95 13" 10.0 176 108 10.4 154 504 10,4 142 *Craaciaina phosphakinasa. Fluorachanical Surfactant TABLE 12. Gcoup, &at Glucose Number S*x mg/LOC ml 300 Pam: 73916 m 73917 14 73918 x 73919 x 73920 x 739ZI p 73922 F 73923 F 739Z4 F 73925 F 1,000 Pon. 73926 m 73917 x 73928 x 73929 x 73930 x 73931 r 73932 F 13933 F 73934 F 73935 F 3,000 pan: 73936 %1 73937 x 73938 x 73939 x 7394,0 x 73941 F 7391&2 F 73943 r 73944 F 73945 Died Died Died Died Died Died Died Died Died Died Di*d Died Died Died Died Died Died Died Died Died Died Died Died Died Died Died Died Died Died Died Nimcy Day Subacute Rat Toxicity Study. Udividual Alochomiua Values - 3 voutbs. S.U.N. as/ 100 SL Y-G.T.P. SIIM& UUiC3/Ml C.P.K.* Sign& UnitSiMl P.G.O.T. Lnt'l =its/1 Page 34 P.G.P.T. intli units/I calcium maq/1 Alk. Phos. intli units/1 L37-085 *CreatiaLno phosphokinase. Fluarochamical sttrfactant L@k= 13. Urinalysis 4ALES: Valme, al Specific Gravity. FL4ALES: Volum, PH. Specific Gravity, Ninacy Day Subacute Rat Toxicit7 Study. 4cans and Sigmificmca a of Crinalysis Values. S:udy .4*mt!l Control 30 ppm ?recast 1 3 pr4ktast 1 3 ?recast 1 3 Protest 1 3 Pretest 1 3 ?-retest 1 3 2.1 2.8 3.1 7.6 7.4 7.2 1.050 1.067 1.068 1.3 1.8 0.8 6.8 7.0 6.7 1.079 1.038 1.071 3.5 3.1 2.3 7.0 6.5 6.3 1.054 1.057 1.067 1.7 2.1 2.1 7.0 7.3 7.2 1.063 1.048 1.055 Page 35 100 ppa 3.2 3.3 3.5 7.0 6.8 7.4 1.047 1.042 1.042 1.6 2.9 0.0 6.8 6.7 Qh'S 1.069 1.054 137-085 OKS - Quanity not sufficient a@carisrical analyses were not zonducted on precast values. Sao f;tcL.Iiit TAIII.P*.14. I:r(iiiii, Alit Hismiler .4t-x YtiItime imi Callst I-iIs 73RS6 7 IHS7 7 JUBS 73889 ,71890 73891 73892 7389'J 7:1894 7389S ti 2.5 ti 0.5 It 3.5 ti 2.5 11 1.5 F -O.S F 1.0 p 1.0 F 0.5 F 2.0 ('41tur 111141 Aplitttir. III[ Spec. Prav. 1)8-6!1 IIN-6-1 lis-t:1 lts-C ll!;-c1 I)N-(: S-C S-C 8-(: B-c 1 7.5 1.070 7.6 1.010 8.2 1.035 6.9 1.065 7.9 1.070 7.9 1.075 6.3 1.080 6.0 1.095 6.0 1.090 8.0 1.055 Niiiel:y1).iySoiliiietiRtaet Tt@xicity Htiidy. leidlividtiaIlfriiiiilyuisValues 1'rete#iL. A] bu"III ('Iucfiue fitilrul)lii Occol It Dioud Ke- Erytioro- ttliielt cytes Ept. Calls 2+ N N 1+ N N N N N N N N N N N N N N N N N tr N N 1+ 2+ occ 2@i tr 1-3 1+ 1+ N I)CC It 1-3 act! 4+ 1.t 1-3 oce 1+ N oce N N N, N 3+ tr 73896 H 2.S 73897 ti 5.5 7'1898 ft S.0 73899 11 3.5 71ql)o 11 1.0 8-(: 6.8 1.065 tr I)S-el 6.9 I.OSO N lis-(: 7.2 1.040 N 8-C 6.9 J.050 N DS-cl 7.1 1.065 tr N n N N N N N N N N m 1+ 1+ tr tr N N 2+ occ 3+ N occ 7391)t p 73902 p 739(13 v 4.0 I.() I.S $-(: 7.0 1.037 N IIS-(: 6.1 1.090 tr 1)3-ct 8.4 1.039 N N N m N N N N N tr 1+ occ 2+ tr occ 73904 r t.5 S-C 6.0 1.080 tr N N N N ace 73905 F 0.5 to%-cl 7.5 1.070 tr N 4+ 2+ !yq. pl@R: 73906 ?1 71907 H 7390H m 7'11)10 H 7'194ti 11 3.5 4.o 2.5 4.0 2.4) S-C 8-C 8-C I..%-C 7.0 1.045 N 6.9 1.050 n 6.0 1.065 N 8.3 1.038 N 6.9 1.036 N N N 1+ tr N N N N N N tr L-3 N N 3@ N oce N N N 1*1911 V 2.0 8-(: 5.9 1.065 N N N N N 73912 v 2.5 118-1-1 7.8 1.070 N N N 4+ N 73913 r t.0 1.8-C 6.0 1.075 N N m N N 7:1914 p 1.0 ll!;-c1 6.2 1.085 tr N N 2+ N occ 73915 r 1.5 S-el 7.9 I.OSO N N N tr N Amur. llritten p occ r I)CC r r ace ot!C fice occ oce acc acc acc occ I? tiec ace acc tice oce oce p p lick, F uce 0 lice 137-085 (.,m.a L!. tr Trace 1-t Trace Let at friel 21- gligial tiewiderjite 31 tliiclerillt40+ H.iiketl 8 - Strtw 1.8 - 1.11-liSttraw IJS - lkork Stritw lAm - I.IlgliAtmiter I)Am - I)iirAkol)er el - C14)4141Y (:- (:14-;er N - Negiit Iviq p - Few 1. - I.tia4led H - thiliy k - Riore lice - fk!(!Ilp#lilliill VI41i)$lI.tl!la.Iian!III Seerfiti'altilt I*Alll.F1.4. R;it V.1Iliou Al)lie.*ir. Jill Crov. Hitit-ty1).iy%iiii.%4-tiRtIeOL Tt)Xlclty SLootly. ltialividiiUiriiiinlystoViiiiies- 1'rett@tit. will (.Iti- niii- (14,1!iiltKeclose rtil)in n I()did totter Erytitrocytes pliI Celle Amir. Uriate ti 7 191 ti 73918 ti 13919 11 71920 73921 IJ922 r 73923 v 1.1924 r 7J92'j F I-,Iloo.lillou: 711)26 7)927 ti )3928 2 J929 73930 ti 73931 F 739)2 F 7j9jj F 71934 F 73915 v I)NS 1.5 fo.5 4.0 7.0 2.() '1.5 4.0 2.0 2.0 8.0 4.5 5.5 5.0 4.0 -0.5 1.5 <0. 5 5.0 QNS 7)9*lfi 7'1937 ti 719'18 ti l39.j9 7*194(1 m 7*1941 p 13942 F 1'19163 F 7'1944 r 73945 F 2.5 4.5 5.0 3.5 6.0 4.o 4.ti 2.5 1.0 3.5 115-4:1 s-1: I)S-C1 S-(: s-1: 8.0 1.045 N 8.2 1.070 tr 5.3 1.042 N 8.0 1.036 N 7.9 1.085 N 5.9 1.040 N 7.8 1.045 N 6.9 t.065 N 6.1 1.071) N 8-c 1 1.8-c 1 I.N-cl 8-C S-el 8.9 1.032 N 8.0 1.016 N 7.9 i.Olfi N 6.3 t.042 N S.t 1@040 I)S-ct 6.5 1.090 tr 1)8-(: 7.0 1.080 N I)S-(: 8.9 1.070 tr I.S-C 7.1 1.036 I)S-cl 8.2 1.050 tr 8-4:1 8.4 1.038 N-c 1 8.4 1.037 N $-(: fi.1 1.045 N S-el 6.4 t.048 N 1)8-4!1 a. t 1.035 N 1)%-C 8.4 t.016 tr 1)8-cl 8.6 1.042 1.@ 118-c 1 6.7 t.070 N I)S-c1 7.0 1.038 u N N N N N N N N N N N N N N N N N N N N N N N N N N N N N N N N H N N N N N N N N N N N N N N 4+ N 1+ N tr N N N oce 4+ N N N N t)CC N N Lr oce N N N N N m N N 1+ N 2+ tr 1-3 4+ 4)CC N N li. tr 1-3 li. N oce tr N N N 4+ oce 4+ N 4+ N 1-3 4+ 3#- N ace Dec v ace r occ ace oce p lice oce acc lice acc tdcc Dec oce oce p oce oce (14:c oce H r ace ficc oce occ p oce r oce lice It - tfosligilt 2# - Sligill to W)JOI.;Itu 41- FL-irketi S1.8 IIS I.AmI)Am 4-1 - I' - St1*11W 1.11-ti!t&Lriw I)arkStriiw 1.11;leAtwlies, 1);irAktelit-r Citicilly ('I.-jor N - mooiIiIva F - Few 1.- IAI;vde(l N - @L-isoy k - Kiirti $see - 4k-4:,IIcHiliIa$ 4)Ns (pi,inity,titatirrit.i4@t Iitl$-Iit! keelIt!ii 'I'AIIII.SE. HilletyI)iiyStil)actitReiotToxicity Stucly. lit(liviciUtritaolnlysinValues - t Hisittit. R.4t Hooolier %ex Coiltro I 71886 73087 H 7388" ti 730119 11 I']ti6)o to ?3091 p 73892 F 73893 F 73894 F 73895 F .3 A j-L-s@: 73896 ti 7'JB91 ti 73898 ti 73899 m 739(10 71901 r 7'J94)2 r 73903 F 73904 p 73905 p lpkaol!n: 73906 m 73907 Pi 1391)0 731)111 to Y3946 ti 73911 p 73912 r 7'J913 r 73914 r 13915 p vi)Itime pt asid Alilseoir. pit Spec. Al bu- ('raiv. mill 1.5 (1.5 5.0 11.5 41.5 1.0 ONS 1.() I.o 3.0 5.5 5.0 1.5 1.5 2.0 2.5 1.0 3.5 2.5 1.0 2.5 2.5 7.5 2.0 2.0 1.5 'J.5 (INS 1.5 1.0 S-4-1 6.9 S-C 6.8 S-4-1 6.8 0.3 1)8-4*1 8.2 lis-C1 8.0 1)8-c1 7.6 8-(.1 6.2 LS-t. 6.0 1.040 1.045 m 1.080 N 1 .(1"4) N 1.090 N 1.045 N 1.045 N 1.040 1.020 N 6.3 1.040 N I)S-ct 6.2 t.045 N DS-et 8.0 1.040 N s-t: 6.2 1.070 s-1: 5.8 t.090 N S-el 6.9 1.050 N S-4-1 7.0 1.040 N S-el 6.8 1.055 N lis-el 8.0 1.045 u I)S-cl 8.0 t.050 N S-C 6.8 1.045 N S-C 7.0 1.045 N S-C 7.0 1.035 N $-(: 6.5 1.051) N 8-6.1 6.5 1.(117 N S-(: 5.9 1.040 I)S-ct 7.9 1.050 N S-1-1 6.5 1.045 N 8-t-1 6.3 t.080 N I:lueivue ltill- Occult rubin aloold N N tr N N tr N N N N Ii N 0 1@ N m 2+ N N 14. N N tr, N N N N 0 N N N tr N N N m m N N N N N N N N N 1+ N 1+ N N m N N N N N N u N N N N N N N 1+ N N tr N 0 N Ke- Erytllro- tailleff cytes Cpl. Celle Aimir. Uraten N Dec N N Ir 1-3 to, N "cc N N 1-3 N N 1-3 N 1-3 N N acc N occ N N 1-3 N 1-3 N occ N 1+ 1-3 1+ tr 1+ 1+ occ N N N N occ occ lice did-C Dec 1-3 occ Dec Dec occ occ oce oce occ oce ace occ oce occ ace acc F r Dec liec oce v r oce Gee r p v r oce r oce v v ucc occ p occ oce p p p cu.de: tr - 1'ractIt - rrit!tt-o aliplit 21- - .1;111-ItidLirxitierite Ik - maillersite 41 - FL-irkt!ii S - Strisw I..% - I.Il-,ISittriiw I)S- llioe*Sktriew I.Am - f.iilitAmlier IlAm - 1);irkAwfour 4-1 - (*14011(IY (: - 1:1 t-tor N - Mi-pjkt Ive p - Few 1. - lasiggleti H - Hility It - Itgroii gk!t, (INS litiallitylicitnliff titrstit-liIttt:miIi Solr IIII:illIs ........ Mlisety ljiiyStilaiieutNuot To)xleity Study. loidividianltlriiiikiyalVsatiies - I Husitit. R:lt yetI isoke itself Spec. Atbio- (.III- hill- (k-cuI t Ke- Prytitro- Bpi. %e x ot Alviiejir. pil (.I*nv. mitt cisou ruilill llIlead toose:4 cytits Cetla Amr. Uraten 7191(1 11 7J91 7 73918 73911) 7)920 73921 r 73922 r 7392:1 F 13924 v 73925 F !.,!Lo(!1.,1@n: 731)2(1 @l 7*1927 11 1:1928 ti 7:11)21) 11 739'io 11 7'1911 r 1 J9,12 p 7 )9'1*1 F 7'14)'14 p I"il)'Ii F 739:16 ti 7:19'17 @l 7,j93ll 11 7,191ll 7'it)4(i 11 7'1941 F 7)1)42 F 7*194'1 F 7391&4 F 7 J945 1. Died ltic4i 1)1eal ltiell 1)1 it4i llled 111Ld I)Ieal llled llical I)Iell Died Died Died Died 1)1egi 1)1u(i I)ICJ Died Illeal 0 1L-41 I)Ieil lituil 111ell llicii ljit!gl 111otd 1)1etl 1)1441 I r 'riit4*e li Itst;lllIiL 2t Hi ll-.Ilt 1#6 1*641i.l,461t. )I tiehit.l,;Ita. 44 tiiit,k4-ol 8 - SLriiw I.N - 1)!. - 1);ork %ti".iw lAw - 1.11-liAtmlit-i* I)Aim - ll;ii'ALmiti,r 4-1 i:16.liily No-l-;tiIve- V Va-w 1. ti tially P K;ird- 1-1t'i1l-lil)lid.!1m..I;l Startiit:titiit 'I'Alli.@. I:Ffltiib, milt Naimiser Sex ValI time wi Coicir toll4l Aplieiir. pit Ninety I)iiySul).ictitReiit*I*t)xli-iSttyijdy. littlividtial Uritinlysis Vnition - I Fli)ittlos. silec. Alboi('.riiv. milt itcalve KillrfibL.1 Oceti It ll1U4 lot Ketiviiam Erytierocyten Ept . Celle (:4itriotiI Fl 11.5 1 7.0 I.ITO tr m 7*1887 11 1.5 1)8-4.1 8.0 1.070 N Y'looo ti 4.0 $-a! 1 7.2 1.059 N N /.IHHII ti 4.() it 8-t-I 6.8 1.045 N 7'1891) 3. (1 H-t: 1 7.11 1.057 N N N 2,@ N h 1+ N 1-3 N I+ N oce lice N N N occ N 2-@ N occ I in'I) r 2.5 1.8-4: 6.2 1.042 N 0 N N 11892 r <0. 5 8.0 1.060 N N N 44- N 1-1 7189'1 r (INS 11894 F 0.7 1..%-c1 6.0 1 ItO N N N N t-I 71895 F (INS 10 illim: 73896 m 1.0 I.S-c 1 6.0 1.080 N N Y'le9i 5.0 I.S-t-i 6.2 1.042 N N 7 *IH98 m ].S S-4.1 6.0 1.062 N N 71899 ti 2.5 S-cl 6.3 1.080 N 719()0 ti 2.0 S-c 1 7.2 1.070 7191)1 p 2.1) S-el 7.8 1.022 tr 73902 p 0.7 S-c 1 5.0 1.120 tr N 139()'l p 1.0 8-a- 1 8.2 1.030 N N 73904 r 5.0 I.S-c 1 6.8 1.040 N 73905 p 0.8 1)8-t-l 8.0 1.062 N N 1-11).Oj)lti@: N N N N N N N 0 N occ N N 1-3 1+ 1-3 N oce N 3+ N oce N N N N 3+ N N N ill:c acc N I.#- N 1-3 73906 m llle4i 73907 ti I)ICJ 71908 5.() Ls-(* 6.0 1.042 N N N N N 7 194 (1 m 2.11 lls-4.1 8.8 1.042 N N N 1-k N 73911 13912 71911 73914 73915 F (INS F I)lid r (INS F 1)1ett r ims T Am(ir. 11 ur4ates p p p p v N ace oce r p v N v p p N (*4)dL.: tr 21 It 41 I*e*jii,L- Sligiot to mmli!ritte tll)41eriil.e tkirkl-d NI.S - 1)9 lAn I)Am - Striiw I.IglitSte-itw [liirk%t riew I.Il-,IAiwtlier liierkAmlier N - NegaI Ive V - Few 1. - laiodi!d N - Hasty it- Riare lisilIil I)NS Quistity @lastaliffi F I lilbl,401.114.emlil I .;IiIris@l!astI Riit Hiimljt@r Sex Vu I lamt@ al (:iilor and Spec. Alilicar. Jill Cray- Ninety Day StsliitciiRtret Toxicity Stildy. liidlvidtiatllrliialysiVsislues- 3 Maiiitlim. Alloumin ijucome liiiI- tk!ciIet riolitit 8 1o4)d Ke- Erytltro- t4iiieti cytes Bpi. Cells Am)r. Urates Trl llim Poot 73916 to 73917 it 73918 73919 ti 73921) N 13921 r 73922 r 7'1923 p 73914 v 73925 p I,Plo-iopl!: 73926 H 73927 73928 ti 73929 7193(1 73931 73932 F 73933 r 73934 v 739:16, v @.JAO-O lip.@!- 73936 N 73937 H 73938 73939 73940 H 73941 p 11942 r 73943 p 73944 p 73945 F 1)1 ell 1)1 L-ii 1)1 a--d I)Iej 1)1 a-d litotil 1)1 atil Died Died I)ie(i Died Died I)Ied Died Died 1)1 ell Died Died Died 1)1 L-ii Dicil Died Died Died Died .1)14.41 111ell Died llleti Ibleti 137-085 (:ode: tr 1+ 21 It 4-t - Trisce Trace to aliglit Sllrlit rip mideritte tioldiel'ilte Fl;orki!41 S1.5 1)8 lAm I)Am ri Straw I.IKlit%Lriow I)iorkStr4ow 1.1plilAmiss.-i, ItiarkAwlicir 1 till4iV 1; 11-11r N V 1. N R tici, Negis t Ivu Vew loilolpti mility Rare k*.Ii*s" II)II.I0 I'AIII.F1.7. miiiety i)iisyaiipist:ikijlietroxicity sttitiy. Sionmary of (:ratioNnt-i-roliBy Ntiml)ersiet!riijimied M41 groan It-silloill KxLuriiiii pwif-IfitIfill liiiir14ism. f4trellWo ruildials matee-tiol isrouiid f4iri!iiWs yellilwliqli/ri!tl muterliti, iiii4il-.i-oeltal reRf4sit Cotot ro 1 F 55 43 TermistalSacrifice 30 plin 100 plim 300 ppo tl V to V ti p 1 .000 lipm 3,000 I)Iim ti r mF 55 23 2 Deaths. lhiseliedu (Uioirtitt 30 ppo 100 pl)m '100 v N v 11 r m 32 5 12 Vyea corneistoloocity 1.101igB cibl'Rest1011 witite/tliii-Ik)it)wii/rA.-(I/yellf)twict 2 t iiteleettiesto I.yolilNei)tluo realdioc!ttloriitl-oiciervical red/dark blai!kdiscolaratton liylierentio/ct)itiogietst dark briowit/redfoci - giiindielmmrucoun liiie4rIoL-worrita-gegi.anditioratic-.ova $will filleatille diorkred - groosilaiis:i)itteiita fic-minitliicili.irk.lirtotwaiits.t!uitteitto ltisockm.et.-ijidwilttrliel ver eoiliorl-.ed cilloge-fntiill a4-ceoitoi:ltelotlilitoli%Lfciii yetli)winli/brotwit/wiftoictie Lrregoilierare;iu r,reenial-i yellow extidate atilietilttoitrtiglitkicitiey laritwliettc4sltir Kidsoeys loyarlesneptiromIm I J7-olti 22 22 2111 22 3 I 2 3 2 2 IlIiI'ilt'lClILIO!181 !;%Iffillt.;slit 'I'Alli.1F7. Utertim enlarged/loydrometrii "I alleoile si!illtty 8116414$011101 fat HilletY Oily StilincliteRitt T(ixicity Stiedy. Sainumary c)f (:rose Neer4jpoy Observattotin. CA)IitrOt 30 PPO Hv v TermitialSacrifice 100 PPO F 3()0PI)M 1,000 PI)N 3.000 PIIM Np HV HV DL-atits.Ultlil:lledu (:4vntrtol30 ppo 100 pibm 3(X) mr H FN F H 1 2 137-085 F I ijoir4i4-lit-Imca I 18. Criblip. Sex atitly WL. SitIcall g Niviely IL-lyI:tibni!tiktieitTaixicity Sttiily. AlisolitLe((;rams)atidRelative (2 lititWleyiglit)firgitWieolitlits. Liver K tdtieys brain Adrenal* -2 p zxlo H 4%4 v 253 I)POT 11 414 p 2*13 10()-.LIP!! m 373 206 1).1/ 0.48 (1.17 (1.19 13.80 7.74 1.05 3.0? 4.01 0.89 2.27 0.50 74 L. 64 A 2.21 0.88 2.06 0.82 76 2.99 0.61 0.45 0.15 11.11) 17.19 9.69* 4. lb 4.176* 3.60 2.05 0.87 0.88 2. 3U 0.56 5!5** 1.34* 3 2.13 0.92 82 3.56 1 0.56 0.14 29.51 7.88* 3.73 1.00-t 2.28 0.64 70 1.9fi 3 0.38 0.18 13.54** 6.57** 2.09 t.Olk 1.98 0.96 79 3.02 2 117-085 (:rtsowiljviereelative argaiiwelaistoalitiwIint titletable were coicislittest *Silteitficaiidtilfyferent from cistitroiitlrai liyavertiploiLglieliiii[vidiiiictalyculated relative 4irgiiwiti.-Igiets. **Slh-itific-atdiitflfyerent from coistraiglru totr)ot:lielImt-aI "Iii(ttIy)aySubacute Not Toxli-.1tSytimly. IiidividtiOarlgan Weigitte- TermlitatSacrifice. Hat Nit. CoittrioI 73086 73887 7]HSB 73889 '7189(1 73"91 73892 73893 73894 73895 30 ppa: 73896 73897 73119H 73899 739()1) i.)901 719(12 7*190.1 7394)4 1390,'i 7:19(L4 71946 71911 73911 1'19ts Wi . Sjileeis I.lvr KId Leys Brain Adrenal* pa Sex 9 9 450 H 4 *19 ti 5(wo It 423 ti 451 262 232 284 213 r 273 m 404 n 442 m 423 m 416 H 387 v 216 p 251 p 224 F 251 221 H 446 lixi F 221 r 192 206 0.77 0.63 0.76 0.84 0.85 0.40 0.42 0.54 0.60 0.45 0.58 0.77 0.59 0.40 0.69 0.40 0.51 0.45 0.48 0.39 0.77 0.35 0.38 0.35 0.40 13.44 15.24 13.92 12.83 13.58 7.02 7.05 7.97 7.05 9.59 18.48 19.75 19.63 9.46 18.65 9.46 10.55 8.98 Q.9n 9.51) 35.75 23.26 13.83 12.45 14.34 3.95 4.10 4.14 3.74 4.12 2.28 1.92 2.32 2.10 2.@44 3.70 4.75 3.96 1.80 3.80 1.80 2.22 2.13 2.16 1.93 4.50 2.96 2.38 1.78 2.1(1 2.31 2.19 2.47 2.23 2.13 2.19 1.07 1.87 2.11 2.27 2.35 2.34 2.01 t.R9 2.90 1.89 2.19 2..30 2.13 2.14 2.10 2.45 2.15 1.75 2.04 73 74 89 66 70 88 31 79 72 112 62 54 55 49 56 64 70 96 63 lifi.. 65 74 70 70 96 ii,riiselia-mIi-iIo 'I'AIIIl.(Fl. liriiiii(Wlili oil Periplieral Nerve Pitiiiisiry ftl4.aclyiuplaaimlc vocticollit[killlifI)OSOPIOI]A NiiittlyDay SuloiiciitRti-ot'roxic!ityStaitly. IlistomorpleotoRicOlitiervatioiis. O)ntrot 30 ppo :r :7' 9-. W W W 94 .co0 1 Gcoo ch 0 -0 C-4 on -s in m a@ m a@ a- a. ch 0 MMMM=MM in cn on en C-% en in in m P4 I% w 94 o 1, do m -1 C-4 in o@ 0- Ch m a 00 M A A A M d@ Ch Ob m I I I tI I II III IIIII ILI II 22 2 2 3 222 II II Tityrosial repos@tiotft ultfmobranciiialrest fot!ilflsiteratillallyaqiltiibnifdiltrates Paratityro[41 Allrt-siIn f(icalcytoplasmicvu4:tjolatio(foicoorticalcells ilifftisceongestion t.iii)B fiscalperlbronchiallymplioldItyperplasts fikeitplerivascularlympim)ldlntillrates focal &iggregatesof alvatilaruncrophages clingentio(lalgaigni) focailinterstitialInflammatorcyall infiltrates alveolar cilema Eye x xxx x x xx x 2 It II tI I 2 2 2 322 22 22 34 22 2 2323 I 22 2 2 2 2 2 3 3 2 3 2 2 3 2 2 2 3 2 2 2 2 2 22222 22 22 22 2 3 2 222 2 3 2 2 3 22 22 2 IItII I ftit.-iii r4yot-ardial dellooteratitpio/mineralization focal listermtitfallyw4)liuililsfiltratetsn myocardioim f4ecal mys)(-arolisol filorivi4to foof-iii ii4!(-rainin wilit muiii)isiit-ii-nr cell toofittratdta lit myiit:r.iolItoo Aiirtii 2 2 2 2 2 1 I II tII I I litrillilly tli.-pilceiitlitiijsutdiie)srtollvseo(lywislfitilldfi!IPR loid-r4,iitit-ii 1)lrmm-oiilot ri@411111111 2 *1-2 2 '1 4 2 Ct)tlex: pri@qpeil If '1 2 3 F Itioru4-ii(!Imca I Slirfai:si ltl ,rAill.2X1). Tinsise Ninety I)ayStiliiactiRtaet Toxicity Sttitly. llistamorploolaigiOcliservatioian. 300 ul &j @D r% 00 M C), r-I C4 cn a v 0-4% -0%4 ral. crh4 cm4 c's 4rm4 a". rm4 a, 10CIA r% 94 0C-0# OCm'bs 0in on4 44 4n en eq in on Ch o@ at m m a- Ch m x ri-imilloot sif tilt lmil@riiiii-Iiiiii rePL looi4-rmtltijil lvmiilitil4i laitillraitego foic-al i-ytcpliliaomlc vuvtot)lietfaioi of t-girLI4:al ceits 4lllfllti4.4-tiliguaticill x xx x x It I I t I I 2 2 2 2 2 2 32 23 lymi)lttolil liyl)erplaola foi4-ii1i)t!rlvans-iiiarlymiolitillitlifiltrateo Jut-al iogl-rekates it( ielvettilnrswicroii)liiigep 4-11111-elitI fill (BRIIIIIII) tID4'11f1ilitsrHtiLlilllllfiiinKitibrYCt-'Il 22 22 2 2 2 22 2 22 3222 32 2 1 222 3 2 2 2 222 2 2 2 !1 2 2 2 2 24 3 3 3 2. Fye lid-art fita.ill fiii-al litt&-rutitiml [yoqtlidtld Itifiltrates fast-iol mytio-iirillist filir#iolo ftif-sel iii-4-raitile wlili opittititio-ii-iir &-ell In my4ii-mrstitim lsifiltrati-a lei I I tI I t I I I I III I 2 Aiirt;i I sit ri-l-liy all-lils-iffiiltloitiomllll-irtilillly.4e-lf lymlliilllfiillitit-lep iiii-,wo-oIitlo ri-oftoulp x x x xx 4 '1 5 Is 4 xxxxx 14 4 3 1 xxxxxxxxx x 4 14 5 4 4 4 5 1 4 x - contlitit)isprent4it I - itiotreonrksolite - vfiry all.plit 4 - vx)ilt-riite 5 - ovorke-al 6 - (-xir(-wk- stirfno "Mile Ninety Ilay Subacute Rat I'Oxlcfty Study. llistoiwrpliologiOcbservations. Control -30 ppo 44 )r 94 ;k 14 94 94 x: :r )r )r. X: 14 14 S. 64 .0 CO a- 0 # C4 n @l n co cc 00 00 a, ION (h 4h ch Go a* 00 en w w 0 In w In 0i'n0 w0 el M m a. (folilutlt)tifit size slid ititwour if( foilliclea dolilettuoiIn size slidiiijwteorf lymptioldfollicles btioietl;irriow %'.%terstaim) liyl)uca-llgjiar t)utt-ocinaticresorption of bttiie So IIva ry (:Isooott stomaell fociilaticonalerosion lotglandular portioii focal mucoust ItemorritaKeItsglandular portion aticiiiiallaylvarkeraLoSto itsforestumacit (nonglandiular) %ul;il I lilli-tilIII$, .14-.Ililllim,I lq!tlm) &ott-cal)(l4ityecreiiseIn etze of mtici)nitvtilli) 1,111-ittl.#li4.til111(- (1:6114111) t I I I iI I I I t 2 2 I I t t I II fat-atInterstitial tyalilievliedifiltrates fa)cikl eytollinsele VBCINIIULII)gl I)( neinar cells febt-.oilstrulvlioyil iit!liiscrells 2 2 2 2 Ij 2 I I I II I I I t I 1 11tI III I 2 2 2 f-14-iIl,l&1--i1storr1i/l,41 ',inftiollIs lymislinitliitfiltrates i-;elii;iol;vr sliffil-Al.f.illitintii(le:(iolligi@stlean a-xtri&mL-tloillalrwymatopotents iia-c-ri&mlt4 fdlt-;Il 4-yi$DIDl;l&imlc Vill-filIlIIt Ifalloil firpatocyten fter;ol loylia-rtriolilay OUltil(k'iil. lit-liatoseellular ltyist!rtrujiley liscri-itytei-tl-iiatiwlal"orulwiiligmk,nllotcyt(sioluomiml 22222 2 2 3 2 3 3 2 2 22 2 2 2 22222 3 22 32 2 2144 2 2 2 22 2 2 x 1,t)it4lliioilitr(-n(@itt It j@-wileraill- I tocitra-marktibla- i sone-kf-tl 2 ve-ry s-xiri-um- r Iitort)cIlciatItm Stir f.,eLti,teit I*ABI.V 20. Les Ioil Heneiitt.-ric dolilelloti I-yol)ls N4&clu In size asigi isi&rAi4@r of folifeltin Nincity Day SeebactiteltatToxicity Sttidy. Iliatomorpliologic I)Iiservatiotin. 300 ppm 1,000 ppe in 1: 1-. Ir 1. 9. t. f- A. t3 ig m0 14 " n A m 0, "N" 0. Ch n Cohn n on o@ en 0% 4)k m m m on en en 4n r, r% r, %a r, A & 0 H N C4 C4 at a"t nat m a. n m 0m11, Cr't, r@ 11 r@ 0, nm r, t% rt on en r, r, I- r% 2 3 3 4 4 4 34 4 4 3 3 3 5 4 4 4 34 tl,,-Itleticiiifit size and otiewit.-rof lympimid follicles Done Hierr4iw llypticullotor outei)t.-Inaticrevorptioin of tione .qaI Iva ry ('Iand stoma4-11 focal mtortionteromicin In glandittar portion focal sui-opal itemorrionge in glandular portion was-cisalltyporkerativals fit fairesitoomels(nonklandiiiar) Smusl I lilit,ullit(@ (I)IJI4tillelloin. .1vitillosin.I It-im) atropliy (decrease In size of succual villt) litit-i4flile llancreas 14sent interstitial lymphold Infiltrates fiscal t!ytoplasmic vocuntaticin of aclnar focal atropity of scivior calls cells Liver focal per Ipdvrtal/sfnumoidnl lympitoid Infiltrates clopetilar fibiusto diffiess, sitittooldolcongestion focal loortiti/minumoidal uxtraowdullary liewtopoleefs fiscal feepatocelliater necrosis focal CyLaiiiammic vocuointitito cif lsepatocytes focal lieputticellular hypertropley ntiltlfik!iil,lsepatocelliolar leypertropisy inerenottd yellowtvii-lirowoipinuent In cytoplarm of liel)atticytes 2 34 444 33 I I 2 2 3 3 .2 3 4 3 4 455444 4 343433 3333 IIII II II II I 2 2 2 22 32 2 I I I IIII III 3 2 2 3 3 2 2 22 3 322 32 3 3323 2 333 1 1 1 tI I I I I I 2 32 2 32233344 33 I I I I I I I I II IItII III 2 3 2 332 4A 14 5 2 22 3 2 2 34 I 23 2 23 5 Code: x etipiditinitprearist 4 t r4-mnrkiilvle Yt-ry t;lll-,Iit IIIIl@iIs siotilerale ouirkatel I-xi If.-to PI titrsciclol-mriiel stirf;li1!.*illt I'Alll,F21). ------------- Ninety Uiy StobacutoKat TaixicitySttedy. 1411111. Control 30 ppo tA'$ 1011 6A 0 al Z Kidney liydrtio%Lplir4imis (tot-allistttr"tltliliyliplilitliiltdfiltratte italsieliioiri!lviorunin liyuliiitd!ropletra liivytciplitemof prioxiewl convolieted toiliulas tttl)iiiasroec!ruvfiwiltli olioursoIlziotlost ji;trestellycwyaslt protttiiisceottkeiil)telacrastis ittropliy(olecreaveIn size) tiriiiiilrlyijitlil(-r x: :r r. sk $4 w 94 1" W Cl@ 0 4 w w o@ m Mon 4 m In m w w w m w m w w en on m on on on m en 2 2 2 2 3 3 2 22 2 2 2 4 2 x z x x x P. N w IN oft %o r@ 0 m o@ 0 m 000 a m m m w 0 o@ m 0. m m m m m n m m n n m m r@ el r, r@ r@ f, r@ r@ 3 2 2 1 3 2 2 2 2 22 2 2 2 2 t)vjreIas ProseLate local lii[itratitiallymplioidliifiltraten lsydr43metra atr&bplly Skin focal t-I)IIietou&tI-alitilosto difft&Re epidl-r.stalltyperkeratuals k I tIII ItI 2 3 2 III 4 2 tI I 4 4 I I II t I I I I 2 I tIII I tII I (@ttle: x I 2 efilittlijiItIiIl-i-selitIs istitremirknivie v4!rytiligit 6 witli-i-nie it .2 -9 2 7 z 3r 0 to C %C @C IC Z M r- MM 64 .4 6J Li w 6J La 60 t.D to to w go ot x x 0-- x - X X x 0- x x n x x 9 x n 09 u x x .- "N X xx Group, Rat No. Sex 73916 73917 M* 73918 M* 73919 %1* 73920 -1* 73921 F* 73922 F* 73923 F* 73924 F* a .c 73925 F* 73926 M* 73927 .4* 73928 .4* 73929 Xb 73930 1.0 73931 F* 73932 F* 73933 P* 73934 F* 73935 F* 73936 %1* 73937 HO '73938 .4* 73939 't- -3940 Mk 73941 F* 73942 F* 73943 F* I 7394-'& F* 73945 F*