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FINAL PROJECT REPORT 50-514 Developmental Toxicity Study of Inhaled 1,1,1-Trichloroethane in New Zealand White Rabbits June 25, 1987 BUSHY RUN RESEARCH R. D. 4, Mellon Road, Export, Pennsylvania 15632 CENTER Telephone (412) 733-5200 PROJECT REPORT 50-514 TITLE: Developmental Toxicity Study of Inhaled 1,1,1-Trichloroethane in New Zealand White Rabbits AUTHOR: Rochelle W. Tyl SPONSOR: James A. Barter Halogenated Solvents Industry Alliance 2315 M Street NW, Third Floor Washington, DC 20037 DATE: June 25, 1987 SL 042566 Bushy Run Research Center A Joint Mellon Institute--Union Carbide Corporation Operation Report 50-514 TABLE OF CONTENTS Page Abstract...........................................................................................................................................................1 Objectives...................................................................................................................................................... 2 Materials and Methods . . ......................................... 2 Results \t ...... .................................................................... 6 Discussion................................................................................................................................................ 8 Conclusions ............................................................................................................................................ 8 References...................................................................................................................................................... 9 Good Laboratory Practices Compliance Statement................................................................... 35 Quality Assurance Summary Statement ....................................................................................... 36 Inhalation Chamber Operation and Analyses ....................................................... Appendix 1 Individual In-Life Maternal Data............................................................................. Appendix 2 Necropsy Data Including Individual Fetal Body Weights ............................ Appendix 3 Malformations and Variations by Fetuses and by Litters............................Appendix 4 Protocols and Amendments............................................................................................... Appendix 5 Developmental Toxicity Exposure Range-Finding Study of Inhaled 1,1,1-trichloroethane in New Zealand White Rabbits............................Appendix 6 SL 042567 ii Report 50-514 Table No. LIST OF TABLES Title__ Page 1 Exposure Parameters: Definitive Study 2 Distribution and Fate of New Zealand White Rabbits on Study 3 Mean Gestational Body Weights and WeightChanges 4 Maternal Clinical Observations by Gestation Day 5 Maternal Gross Necropsy Observations 6 Maternal Organ Weights at Sacrifice 7 Gestational Parameters at Sacrifice 8 Malformations Observed in Fetuses andLitters 9 Variations Observed in Fetuses and Litters 11 12 13' 15 16 17 18 20 26 SL 042568 iii BUSHY RUN RESEARCH R. D. 4, Mellon Road, Export, Pennsylvania 15632 CENTER Telephone (412) 733-5200 CONFIDENTIAL Project Report 50-514 37 Pages June 25, 1987 Developmental Toxicity Study of Inhaled 1,1,1-Trichloroethane in New Zealand White Rabbits Sponsor: Halogenated Solvents Industry Alliance ***** ABSTRACT Timed-pregnant New Zealand white rabbits were exposed to 1,1,1-trichloroethane vapor for six hours/day on gestational days (gd) 6 through 18 at target concentrations of 0, 1000, 3000, or 6000 ppm (mean analytical concentrations of 1017 17.7, 3122 38.6 and 5906 45.1 ppm for the 1000, 3000 and 6000 ppm groups, respectively). Maternal clinical signs were taken daily, and body weights were measured on gd 0, 6, 9, 12, 15, 19 and 29. At scheduled sacrifice on gd 29, maternal body weight, gravid uterine weight and liver weight were taken, with liver sections and nasal turbinates retained. Ovarian corpora lutea of pregnancy were counted and all uterine implantation sites were identified and recorded: resorptions (early or late), dead fetuses and live fetuses. All live fetuses were examined for external malformations including cleft palate, and variations, and for visceral malformations and variations (including craniofacial alterations); 50% of the fetuses in each litter (intact) were examined for skeletal malformations and variations. Maternal toxicity was observed at 3000 and 6000 ppm: significant weight gain reduction for the exposure period and gestation period and an apparent reduction in gestational weight gain (corrected for gravid uterine weight). Gestational parameters exhibited no significant changes, including number of corpora lutea, total, nonviable, or viable implantations per litter, sex ratio, pre- or postimplantation loss, and fetal body weights (total, males or females) per litter. There were no significant changes in the incidence of any individual malformations, malformations by category (external, visceral or skeletal), or of total malformations. There were also no treatment-related changes in the Incidence of individual variations, variations by category, or of total variations, except for an increased incidence at 6000 ppm of bilateral thirteenth rib (a common fetal variation). SL 042569 Bushy Run Research Center A Joint Mellon Institute--Union Carbide Corporation Operation Report 50-514 Page 2 In conclusion, exposure to 1,1,1-trichloroethane vapor by inhalation during organogenesis in New Zealand white rabbits resulted in maternal toxicity at 3000 and 6000 ppm and slight fetotoxicity at 6000 ppm. The "no observable effect level" (NOEL) for maternal toxicity was 1000 ppm and the NOEL for developmental toxicity was 3000 ppm. No embryotoxicity or' teratogenicity was observed at any exposure concentration employed. OBJECTIVES The objective of this study was to evaluate the developmental toxicity (including teratogenicity) of maternally inhaled 1,1,1-trichloroethane vapor in the pregnant New Zealand white rabbit. The potential of the test chemical to produce maternal toxicity was also determined. MATERIALS AND METHODS Generation of Atmospheres and Chamber Parameters Three 55-gallon containers of 1,1,1-trichloroethane test material (methylchloroform; TCEA; CAS No. 71-55-6; Lot No. TA8 606182, BRRC Sample No. 49-228 A, B and C) was received from the Dow Chemical Company (Freeport, TX) on June 25, 1986. Test material from the original containers was placed in one-gallon glass bottles and blanketed with nitrogen. Mr. W. D. Tasto, Dow Chemical USA, provided purity and stability analyses on the test chemical, conducted both before and after the exposure period. No significant compositional changes occurred while the study progressed; the test material remained 99.9% 1,1,1-trichloroethane throughout the study. The gases for the chromatograph were obtained from Linde Corporation. The nitrogen and hydrogen were Ultra High Purity grade and the air was breathing quality. A Perkin-Elmer Model 3920B gas chromatograph (GC) equipped with a flame ionization detector was used to monitor the 1,1,1-trichloroethane vapor concentrations in the chambers. The GC column was a 10-feet x 1/8 inch (i.d.) stainless steel column packed with 10% SP-2100 on 80/100 mesh Supelcoport (Supelco, Inc., Bellefonte, PA). A Spectra-Physics series 4000 central processor, a data interface, and a Perkin-Elmer automatic gas sampling system (station and valve programmer units and a gas sampling valve) were used for the analyses. Calibration of the gas chromatograph was done with dynamically generated gas standards of 1,1,1-trichloroethane prepared by syringe injection of test material into Tedlar (DuPont) gas bags. The series of standards encompassed the entire range of vapor concentrations generated in the exposure chambers. A linear calibration curve was obtained when areas (integration counts) were plotted versus the gas equivalent concentrations of the standards. The calibration factor (KF) was calculated as: KF = ______ABM___ CONCENTRATION The mean KF for all valid injections of standards was used in subsequent microprocessor calculations. This calibration factor was checked at least once each week during the exposure period and did not require adjustment. SL 042570 Report 50-514 Page 3 Each chamber atmosphere was analyzed for 1,1,1-trichloroethane approximately once every 25 minutes during each 6-hour exposure. Daily nominal concentrations (an estimated concentration calculated from the amount of test material delivered and the chamber airflow during the exposure period) were also calculated for each chamber. Liquid 1,1,1-trichloroethane was metered from a piston pump into a heated glass evaporator similar in design as that described by Carpenter et sX. (1975). The temperature in the evaporator was maintained at the lowest level sufficient to vaporize the liquid (35-64C). The resultant vapor was carried into the chamber by passage of conditioned air through the evaporator. Chamber atmospheres containing 1,1,1-trichloroethane were filtered before leaving an exhaust stack. The four chambers in Room 139 employed in this study were rectangular in shape, constructed of glass and stainless steel (Wahmann Manufacturing Company, Timonium, MD), and measured approximately 2.1mxlmx2.1m (height). Total volume in each chamber was approximately 4320 L. An orifice plate was positioned in the exhaust duct of the chamber and was connected to a Dwyer Magnehelic Pressure Gauge. Airflow in each chamber was approximately 1000 L/minute (14 air changes per hour) with a t99 (theoretically-derived time required for the chamber to reach 99% of the equilibrium concentration) of approximately 20 minutes. The chambers were illuminated with artificial room light. Temperature (Minimum-Maximum Thermometer, Abbeon Cal., Inc.) and relative humidity (Airguide Instrument Company) gauges were placed inside each chamber during exposures. Chamber temperature, relative humidity, and airflow rate were recorded eleven times during each six-hour exposure. Within each chamber, the animal cages were rotated daily to compensate for any possible, but undetected, variation in chamber exposure conditions (l.e., concentration, temperature, relative humidity). A detailed description of analytical procedures and results is found in Appendix 1A and B (definitive study). Target concentrations were 0, 1000, 3000, and 6000 ppm. These exposure concentrations were determined by the Sponsor based on an exposure range-finding study also performed on pregnant New Zealand white rabbits (see Appendix 6). Rabbits One hundred eight (108) virgin female New Zealand white rabbits (at least 2.5 kg, approximately 5 to 5 1/2 months old) arrived from Hazleton Dutchland Laboratories, Inc., Denver, PA on September 23, 1986. Each animal was assigned a unique number and received a stainless steel Monel ear tag (Gey Band and Tag Co., Morristown, PA). They were quarantined for two weeks, during which time they were examined by a veterinarian and fecal samples taken for detection of intestinal parasites. Quality control data indicated that these animals were in good health and suitable for use. The rabbits-were individually housed in stainless steel wire-mesh cages (46 cm x 61 cm x 36 cm SL 042571 Report 50-514 Page 4 high) with certified feed (Prolab Certified Rabbit Food, Batch No. 6134 SMI and 6227 SMI, Agway, Inc., St. Marys, OH) and water (Municipal Authority of Westmoreland County, Greensburg, PA) available ad libitum except during exposures. Paperboard (Deotized Animal Cage Board, Shepherd Specialty Papers, Inc., Kalamazoo, MI) was placed beneath the cages and photoperiod (12-hour, 0600 to 1800 hours for light phase), temperature and relative humidity were recorded continuously (Cole-Parmer Hygrothennograph Seven-Day Continuous Recorder, Model .No, 8368-00, Cole Parmer Instrument Co., Chicago, IL). Female rabbits (at: least 2.5 kg and 5.5-6 months old and in good health) were bred, using in-house breeding colony males from the same supplier in animal Room 122. The date of copulation was designated gestational day (gd) 0 (Hafez, 1970). Twenty-four (24) mated females were randomly assigned to each experimental group. The mating period was October 6-9, 1986; therefore gd 0 was also October 6-9, 1986. Mated females were individually housed in Animal Room 122 for the duration of the study (except during exposures). For exposures, mated females were transferred to stainless steel wire-mesh exposure cages (35 cm x 37 cm x 41 cm high) in cage carriers which were moved into the chambers in room 139. Food and water were withheld during exposures. Exposures were for six hours per day, gd 6 through 18, in Room 139, October 12 through October 27, 1986. After each exposure, animals were returned to their original cages and ''home" room. The animals were observed daily for clinical signs throughout the study (gd 0 through 29). Maternal body weights were taken on gd 0, 6, 9, 12, 15, 19 and 29. At scheduled sacrifice on gd 29, rabbits were sacrificed by intravenous injection of T-61 euthanasia solution (Hoechst Corp., Sumerville, NJ) into the marginal ear vein. Maternal sacrifice period was November 4-7, 1986. Thoracolaparotomy. Maternal and Fetal Examinations The maternal body cavities were opened by midline thoracolaparotomy. The gravid uterus, ovaries (including corpora lutea), cervix, vagina, and abdominal and thoracic cavities were examined grossly. Ovarian corpora lutea of pregnancy were counted. Maternal liver and uterine weights were determined. Sections of maternal and nasal turbinates were retained In fixative for possible subsequent histologic examination. The uteri were immediately ligated at their cervical end to prevent the expulsion of conceptuses by myometrial peristalsis, externally examined for signs of hemorrhage, removed from the peritoneal cavity, and dissected longitudinally to expose their contents. All live and dead fetuses and resorption sites (early and late) were noted and recorded. Uteri from females that appeared nongravid were placed in a 10% ammonium sulfide solution for detection of early resorptions (Salewski, 1964). Each fetus was euthanized immediately upon removal from the uterus by intraperitoneal injection of sodium pentobarbital. All live fetuses were weighed and were examined for external malformations including cleft palate, and variations. All live fetuses in each litter were examined for thoracic and abdominal visceral abnormalities by modification of methods described by Report 50-514 Page 5 Staples (1974). The sex of each fetus was determined at the time of visceral examination. One half of the fetuses (even numbered fetuses from litters with an even number of live fetuses, odd numbered fetuses from litters with an odd number of live fetuses) examined viscerally were decapitated and their heads were fixed in Bouin's solution for examination of craniofacial structures by sectioning methods modified from Wilson (1965; 1973) and van Julsingha and Bennett (1977). Intact fetuses (not decapitated) in each litter were eviscerated, fixed in ethanol, and then processed for skeletal staining with alizarin red S (Crary, 1962; Peltzer and Schardein, 1966), and examined, for skeletal malformations and variations. Decapitated fetuses were also eviscerated, fixed and processed for skeletal staining but were not examined. Statistical Analyses The unit of comparison was the pregnant doe or the litter (Weil, 1970). Results of the quantitative continuous variables (e.g., maternal body weights, liver weights, etc.) were intercompared for the three exposure groups and control group by use of Levene's test for equal variances (Levene, 1960), analysis of variance (ANOVA), and t-tests with Bonferroni probabilities. The t-tests were used when the F value from the ANOVA was significant. When Levene's test indicated homogeneous variances, and the ANOVA was significant, the pooled t-test was used for pairwise comparisons. When Levene's test indicated heterogeneous variances, all groups were compared by an ANOVA for unequal variances (Brown and Forsythe, 1974) followed, when necessary, by the separate variance t-test for pairwise comparisons. Nonparametric data obtained following laparohysterectomy were statistically treated using the Kruskal-Wallis test (Sokal and Rohlf, 1969) followed by the Mann-Whitney U test (Sokal and Rohlf, 1969) when appropriate. Incidence data were compared using Fisher's Exact Test (Sokal and Rohlf, 1969). For all statistical tests, the fiducial limit of 0.05 (two-tailed) was used as the criterion for significance. Personnel The evaluation of 1,1,1-trichloroethane for developmental toxicity in New Zealand white rabbits was conducted at Bushy Run Research Center (BRRC), Export, PA for Halogenated Solvents Industry Alliance, Washington, DC 20006. The Sponsor Liaison was Dr. J. A. Barter, PPG Industries, Pittsburgh, FA. The BRRC personnel indicated below contributed to the completion of this study. Dr. R. Mi Tyl served as Study Director. Developmental Toxicity personnel included D. L. Fait, L. C. Fisher, M. F. Kubena (Study Leader), D. J. McNeil (Tarasi), T. A. Rebick, and T. A. Savine. Inhalation personnel included Dr. D. E. Dodd, Dr. I. M. Pritts, W. J. Kintlgh and J. L. Smith. Quality Assurance personnel included L. J. Calisti, J. R. Bernard and W. J. Conroy. Animal care personnel were supervised by M. J. Cardella. The Final Report was prepared by Dr. R. W. Tyl with assistance from L. C. Fisher on data compilation and statistical analyses. The protocol and amendment detailing the design and conduct of this study are presented in Appendix 5. SL 042573 Report 50-514 Page 6 Storage of Records All original data sheets for the present study are stored in the BRRC Archives along with all biological samples collected during the course of the study which remain the responsibility of BRRC. Work sheets and computer prints which were generated in the statistical analysis of data are stored in the BRRC Archives. Copies of this report will be filed with the BRRC Archives. In accordance with EPA (TSGA), all records, data and reports will be maintained in storage for a mininum of ten (10) years or for as long as the quality of the preparation affords evaluation, whichever is less. The study was performed in compliance with Environmental Protection Agency good laboratory practices regulations (EPA, 1983). RESULTS Chamber Analyses The mean analytical concentrations for the 1000, 3000 and 6000 ppm chambers were 1017 17.7 ppm (101.7% of target), 3122 38.6 ppm (104.1% of target) and 5906 45.1 ppm (98.4% of target) respectively. No test chemical was detected in the control chamber (0 ppm) with a minimum detection limit of 10 ppm. The analytical to nominal concentration ratios were 0.90-0.96, indicating no degradation or adsorption of the test chemical to chamber walls. Temperature was maintained at 24.9-25.3C and relative humidity at 41.4-49.7% for all four chambers (Table 1). Details of the procedures and results of the chamber analyses are presented in Appendix 1A and IB. Rabbits The distribution and fate of all female rabbits on study are presented in Table 2. No does died; one doe at 0 ppm delivered early and was removed from study, one additional doe, at 1000 ppm, received a traumatic injury and was also removed from study. Pregnancy rate was high and equivalent across all groups. One litter at 0 ppm, five (5) at 1000 ppm and one (1) at 6000 ppm were completely resorbed at scheduled sacrifice on gd 29. A total of 16-21 litters were examined in each group. Maternal body weights and weight change are presented in Table 3. There were no differences among groups in periodic body weights. However, there was a significant reduction in weight gain for does at 3000 and 6000 ppm for gd 6-9 (first third of the exposure period), for gd 6-19 (exposure period), and for gd 0-29 (gestation period) (Table 3). There were no treatment-related clinical signs of toxicity during gestation except for loose feces observed in two does at 6000 ppm during the exposure period (Table 4). There were also no gross necropsy observations in does which were treatment related (Table 5). SL 042574 Report 50-514 Page 7 There were no statistically significant changes in maternal body weight at sacrifice (absolute or corrected for gravid uterine weight), maternal corrected gestational weight change, or liver weight (absolute or relative to corrected body weight)* There was, however, an apparent concentrationdependent decrease in corrected gestational weight change, with control does losing an average of 82 g and does at 3000 and 6000 ppm losing an average of 191 g and 134 g over the gestational period, respectively. Gravid uterine weight was significantly reduced at 1000 ppm because of five does with totally resorbed litters (Table 6). Individual maternal djata are presented in Appendix 2. Gestational parameters are presented in Table 7. There were no significant effects of treatment on any parameters examined, including number of ovarian corpora lutea, number of total, nonviable (early or late resorptions, or dead fetuses) or viable implantations, percent pre- or postimplantation loss, or sex ratio (X male fetuses). Corpora lutea per doe were sightly reduced (but not statistically significantly) at 6000 ppm, so total implants and viable implants per litter were also reduced (significant at p < 0.05), with no changes in preimplantation loss or non-viable implants per litter relative to controls. At 1000 ppm, the numbers of viable Implants per litter and the percent live fetuses per litter were significantly reduced and the number of non-viable Implants was significantly increased relative to controls because of the five resorbed litters. Fetal body weights per litter (males, females or total fetuses) were statistically equivalent across all groups. Individual fetal body weights (by uterine location) are presented in Appendix 3. The incidence and frequency of rabbit fetal malformations and variations observed in the present study are presented in Tables 8 (malformations) and 9 (variations). There were no significant differences in the incidence of any individual malformation, of malformations by category (external, visceral or skeletal) or of total malformations. One fetus at 6000 ppm exhibited all of the external malformations observed except for dome-shaped head. One fetus at 1000 ppm exhibited both missing fourth ventricle of the brain and scrambled posterior portion of the cerebellum, and one additional fetus at 1000 ppm exhibited missing semilunar valves of the pulmonary artery and aorta, small right ventricle and enlarged left ventricle of heart. In addition, one fetus at 3000 ppm exhibited missing thoracic arch, fused thoracic arch and diagonally displaced thoracic centrum and arch. There were no significant differences in the incidence of individual external or visceral variations, of variations by category or of total variations. Out of 51 skeletal variations observed in the present study, one, extra thirteenth rib, bilateral, exhibited a significantly increased incidence at 6000 ppm, which may indicate slight fetotoxicity. Malformations and variations by individual fetuses and litters are presented in Appendix 4. SL 042575 Report 50-514 Page 8 DISCUSSION The present study has shown that exposure to 1,1,1-trlchloroethane vapor by inhalation during organogenesis in New Zealand white rabbits results in maternal toxicity at 3000 and 6000 ppm. The only indication of possible fetotoxicity was a significant increase in bilateral extra thirteenth ribs at 6000 ppm observed in the presence of maternal toxicity. This is one of the most common rabbit fetal variations observed (Woo and Hoar, 1982). Khera (1985) reviewed teratology data from rabbit studies and indicated that extra ribs (among other findings) was consistently associated with maternal toxicity regardless of the test agent employed and attributed these fetal findings to maternal toxicity. Kenel et al. (1984) also ascribed the observation of extra ribs in rabbit fetuses to maternal and/or fetotoxicity. The slightly (but not statistically significantly) reduced fetal body weights observed in the present study at 3000 and 6000 ppm (associated with maternal toxicity) may also imply fetotoxicity although the values are within the historical controls for this rabbit strain in this laboratory and these findings are usually not considered evidence for fetotoxicity unless the differences are statistically significant relative to controls. Even if these data were interpreted as indicative of fetotoxicity, the A/D ratio (Fabro et al., 1982), the ratio of the lowest doses resulting in adult and developmental toxicity would be equal to one (1.0), indicating no preferential susceptibility to the conceptus. This conclusion is consistent with the other data for this study indicating effects on the conceptus only at exposure concentrations detrimental to the doe. CONCLUSIONS Exposure to 1,1,1-trichloroethane vapor during organogenesis in New Zealand white rabbits resulted in maternal toxicity at 3000 and 6000 ppm and slight fetotoxicity at 6000 ppm. The "no observable effect level" (NOEL) for maternal toxicity was 1000 ppm and the developmental toxicity NOEL was 3000 ppm. No embryotoxicity or teratogenicity was observed at any exposure concentrations employed. Reviewed and Approved by: Study Director PATH/esk/0949P-4 06-23-87 SL 042576 Director Report 50-514 Page 9 REFERENCES Brown, M. B. and A. B. Forsythe (1974). The Small Sample Behavior of Some Statistics which Test the Equality of Several Means. Technometrics 16. 129-132. Carpenter, C. P., E. R. Kinkead, D. L. Geary, Jr., L. J. Sullivan, and J. M. King (1975). Petroleum Hydrocarbon.Toxicity Studies 1. Methodplogy. toxicol. Appl. Pharmacol. 31, 246-262. Crary, D. D. (1962). Modified Benzyl Alcohol Clearing of Alizarin-Stained Specimens Without Loss of Flexibility. Strain Technol. 31, 124-125. Environmental Protection Agency, (1983). Part III EPA Toxic Substances Control; Good Laboratory Practice Standards; Final Rule. Federal Register 48(230): 53922-53944. Fabro, S., G. Schull, and N. A. Brown (1982). The Relative Teratogenic Index and Teratogenic Potency: Proposed Components of Developmental Risk Assessment. Teratog. Carcinog. Mutagen. Z, 01-76. Hafez, E. S. E. (ed.) (1970). Reproduction and Breeding Techniques for Laboratory Animals. Lea and Febiger, Philadelphia, PA. Kenel, M. F., J. L. Schardein, R. D. Terry and L. G. Miller (1984). Developmental Variations as Markers of Maternal and Embryotoxicity. Teratology 29(2). 40-41A. Khera, K. S. (1985). Maternal Toxicity: A Possible Etiological Factor in Embryo-Fetal Deaths and Fetal Malformations of Rodent-Rabbit Species. Teratology 31 129-153. Levene, H. (1960). Robust Tests for Equality of Variance. In: Contributions to Probability and Statistics. I. Olkin et al., eds., Stanford University Press, Stanford, CA, pp. 278-292. Peltzer, M. A. and J. L. Schardein (1966). A convenient Method for Processing Fetuses for Skeletal Staining. Stain Technol. 41, 300-302. Salewski, E. (1964). Farbemethode Zum Makroskopischen Nachweis Von Implantations-Stellen am Uterus Der Ratte. Naunyn-Schmiedebergs, Arch. Exp. Pathol. Pharmakol. 242, 367. Sokal, R. R. and F. J. Rohlf (1969). Biometry. W. H. Freeman and Co., San Francisco, pp. 369-371, 299-340, 370-372, 589-595. Staples, R. E. (1974). Detection of Visceral Alterations in Mammalian Fetuses Teratology 9, A37. (Continued) SL 042577 Report 50-514 Page 10 REFERENCES (Continued) van Julsingha, E. B.f and C. G. Bennett (1977). A Dissecting Procedure for the Detection of Anomalies in the Rabbit Foetal Head. In: Methods in Prenatal Toxicology. D. Neubert, H. J. Merker, and T. E. Kwasigroch, eds. PSG Publishing Company, Inc., Littleton, Massachusetts, pp. 126-144. Weil, C. S. (1970). Selection of the Valid Number of Sampling Units and a Consideration of Their Combination in Toxicological Studies Involving Reproduction, Teratogenesis dr Carcinogenesis. Fd. Cosmet. Toxicol. |!, 177-182. Wilson, J. G. (1965). Embryological Considereations in Teratology. In: Teratology Principles and Techniques. J. G. Wilson and J. Warkany, eds. The University of Chicago Press, pp. 251-277. Wilson, J. G. (1973). Environment and Birth Defects. Academic Press, NY. Woo, D. C. and R. M. Hoar (1982). Reproductive Performance and Spontaneous Malformations in Control New Zealand White Rabbits: A Joint Study by MARTA. Teratology 25(2). 82A. Tables provided by the authors. PATH/esk/0949P-2 03-24-87 SL 042578 Report 50-514 Page 11 SL 042579 Table 1 Exposure Parameters: Definitive Study Target concentration, ppm Analytical concentration, ppma A/T Ratiob Nominal concentration, ppma A/N Ratio3*c Temperature, Ca Relative humidity, %a ______ 0_______ < MDLd - 25.1 1.08 49.7 3.18 ______ 1000 1017 17.7 1.017 1134 14.6 0.90 0.013 25.1 1.25 44.5 4.76 ______ 3000 3122 38.6 1. 041 3234 0.96 0.96 0.014 24.9 1.10 44.6 4.35 ' a Grand mean of 16 daily means standard deviation b Analytical to target concentration ratio c Analytical to nominal concentration ratio d Less than the minimum detection limit of 10 ppm WPC/esk/0764P-2 06-19-87 ______ 6000 5906 45.1 0.984 6534 48.3 0.90 0.0096 25.3 0.90 41.4 3.74 Report 50-514 Page 12 TABLE 2 DEVELOPMENTAL TOXICITY STUDY OF INHALED 1,1,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS DISTRIBUTION AND FATE OF NEW ZEALAND WHITE RABBITS ON STUDY PPM FEMALES ON STUDY 0 NO.. (%) 24 1000 NO. <%) 24 3000 NO.. (%) 24 FEMALES THAT DELIVERED EARLY .1 ( 4.2)a FEMALES REMOVED FROM STUDY FEMALES THAT DIED 0 ( 0.0) 0 ( 0.0) FEMALES EXAMINED AT LAPAROTOMY NON GRAVID GRAVID RESORPTIONS ONLY DAMS WITH VIABLE FETUSES 23 ( 95.8) 1 ( 4.3) 22 ( 95.7) 1 ( 4.5) 21 ( 95.5) FEMALES THAT WERE GRAVID 22 ( 95.7) a Female removed from study, b Apparent cage accident; trauma. 0 ( 0.0) 1 ( 4.2)b 0 ( 0.0) 23 ( 95.8) 2 C 8.7) 21 ( 91.3) 5 ( 23.8) 16 C 76.2) 21 ( 91.3) 0 ( 0.0) 0 ( 0.0) 0 ( 0.0) 24 (100.0) 4 ( 16.7) 20 ( 83.3) 0 ( 0.0) 20 (100.0) 20 ( 83.3) 6000 NO.. (%) 24 0( 0 0( 0 0 ( 0. 24 (100, 3 ( 12, 21 ( 87 1 ( 4.8) 20 ( 95.2) 21 ( 87.5) SL 042580 Report 50-514 Page 13 TABLE 3 DEVELOPMENTAL toxicity STUDY OF inhaled 1.1,1-TRICMLOROETHANE IN NEW ZEALAND WHITE RABBITS mean gestational BODY WEIGHTS AND WEIGHT CHANGES PPM 0 1000 3000 GESTATIONAL DAY 0 body weights MEAN S.D. N (GRAMS) 3452.4 196.11 22 3506.6 285.12 21 3481.1 269.79 20 DAY 6 MEAN 5.0. N 3601.8 194.87 22 3622.8 316.94 21 3626.3 265.82 20 DAY 9 MEAN S.D. N 3574.3 218.42 22 3567.4 312.48 21 3538.4 220.69 20 DAY 12 mean S.D. N 3590.2 213.82 22 3575.0 324.68 21 3537.3 231.66 20 DAY 15 MEAN S.D. N 3641.3 222.95 22 3612.7 335.46 21 3582.1 233.09 20 DAY 19 mean S.D. N 3691.5 225.12 22 3621.0 358.78 21 3569.0 258.69 20 DAY 29 MEAN S.D. N 3897.9 239.19 22 3819.7 395.15 21 3783.0 285.93 20 GESTATIONAL BODY WEIGHT CHANGES (GRAMS) DAY 0 TO 6 (PRE-TREATMENT) MEAN 149.4 S.D. 112.88 N 22 116.2 120,55 21 145.2 124.62 20 DAY 6 TO 9 MEAN S.D. N -27.5 54.64 22 -55.4 49.30 21 -87,9** 64.05 20 DAY 9 TO 12 MEAN S.D. N 15.9 37.38 22 7.6 59.10 21 -1 .1 61.26 20 DAY 12 TO 15 MEAN S.D. H 51.0 58.97 22 37.7 82.10 21 44.8 61.51 20 DAY 15 TO 19 MEAN S.D. N 50.3 43.64 22 8.3 89.64 21 -13.0* 67.00 20 6000 3502.9 286.73 21 3619.0 277.88 21 3543.9 261.93 21 3524.9 253.39 21 3557.7 232.76 21 3563.3 224.80 21 3797.8 266.90 21 116.1 97.53 21 -75.2* 57.95 21 -19.0 50.12 21 32.8 74.32 21 5.6 73.01 21 042581 SL Report 50-514 Page 14 TABLE 3 DEVELOPMENTAL TOXICITY STUDY OF INHALED 1 , 1, 1-TRICHL0R0ETHANE IN NEW ZEALAND WHITE RABBITS MEAN gestational BODY WEIGHTS AND WEIGHT CHANGES PPM 0 1000 3000 DAY 6 TO 19 (TREATMENT) MEAN S.D. N 89.7 91.23 22 -1.8 140.34 21 -57.3 142.55 20 DAY 19 TO 29 (POST-TREATMENT) MEAN S.D. H 206.4 59.22 22 198.7 109.60 21 214.0 134,78 20 DAY 0 TO 29 (GESTATION) MEAN S.D. N 445.5 165.9 22 313.1 178.39 21 301.9 194.12 20 6000 -55.7 139.30 21 234.5 106.38 21 294.8 183.50 21 MEAN DIFFERENCES CALCULATED FROM INDIVIDUAL DIFFERENCES * p < 0.05 compares to control. ** p < 0.01 compared to control. SL 042582 Report 50-514 SL 042583 TABLE 4 DEVELOPMENTAL TOXICITY STUDY OF INHALED t,t,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS MATERNAL CLINICAL OBSERVATIONS BY GESTATION DAY CATEGORY FINDING (LOCATION) GESTATIONAL DAVS DEAD SCHEDULED SACRIFICE 19- 29 EVES/EARS/NOSE LACRIMAT10N (EVE-LEFT) 6- IB 19- 29 OCULAR DISCHARGE, OTHER (EYE-BOTH, EVE-LEFT) 6- 18 19- 29 EXCRETA LOOSE FECES 6- IB PAPERBOARD DRY 6- IB FOOD/WATER FEED HOPPER FULL 6- IB SKIN 19- 29 ALOPECIA (HIP-BOTH, SIDE-BOTH) 6- ia 19- 29 ABSCESS (NECK) 19- 29 OTHER WATER TIP NOT FUNCTIONING - DRV PAPERBOARD 6- 18 BLOOD (BY HEMASTIX) ON PAPERBOARD 19- 29 TISSUE ON PAPERBOARD (UNRECOGNIZABLE) 19- 29 ppm 0 1000 3000 6000 22 21 20 21 00 1 0 00 1 0 00 00 1 1 1 1 000 00 1 2 0 01 10 01 30 00 0 00 0 1 1 00 1 0 00 1 0 02 0 1 02 0 0 *T3 <fJtQ) It SL 042584 TABLE 5 DEVELOPMENTAL T0XIC1TV STUDY OF INHALED 1,t,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS MATERNAL GROSS NECROPSY OBSERVATIONS PPM 0 1000 3000 6000 NUMBER OF ANIMALS IN DOSE GROUP 22 2) 20 2 1 OVARIES -CYST CORPORA LUTEA -ABSENT 8 UTERUS -CYST -CONTAINS LIQUID FLUID -MISSING RIGHT UTERINE HORN VAGINA -DISCHARGE BLOODY AMNIOTIC SACS -ABSENT 8 PLACENTAS -ABSENT 3 -CONTAINS BLOOD (BY HEMASTIX) a These findings are the result of litters containing early resorptions only or does found to be gravid by ammonium sulfide staining of the uterus. *0 fis fpto OfQt *o o 'I rt Cv l/l O Report 50-514 Page 17 TABLE 6 DEVELOPMENTAL TOXICITY STUDY OF INHALED 1,1,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS MATERNAL ORGAN WEIGHTS AT SACRIFICE PPM INITIAL BODY WEIGHT MEAN S.D. N (g) 0 3452.4 196.11 22 1000 3506.6 285.12 21 3000 34B1.1 269.79 20 6000 3502.9 286.73 21 BODY WEIGHT AT SACRIFICE (g) MEAN 3897.9 S.D. N 239.19 22 GRAVID UTERINE WEIGHT MEAN S.D. N (9) 527.98 140.25 22 CORRECTED BODY WEIGHT MEAN S.D. N <9)" 3369.89 280.33 22 CORRECTED BODY WEIGHT MEAN S.D. N CHANGE (9)b -B2.50 227.72 22 LIVER WEIGHT (g) MEAN S.D. N 89.32 11.54 22 RELATIVE LIVER WEIGHT MEAN S.O. N (%)c 2.65 0.29 22 3819,7 395.15 21 371.30* 214.04 21 3448.40 359.84 21 -58.17 192.74 21 92.46 27.97 21 2.68 0.74 21 3783.0 205.93 20 492.46 139.05 20 3290.57 258.27 20 -190.53 194.55 20 91.11 15.54 20 2.76 0.35 20 3797.6 266.90 21 429.09 124.63 21 3368.70 318.29 21 -134.25 169.02 21 95.49 13.22 21 2.83 0.25 21 a Corrected body weight = body weight at sacrifice minus gravid uterine weight, b Corrected body weight change = corrected body weight minus Initial body weight, c Calculated as a percentage of the corrected body weight. * p < 0.05 compared to control. SL 042585) TABLE 7 DEVELOPMENTAL TOXICITY STUDY OF INHALED 1 .1 ,1-TRICHLORQETHANE IN NEW ZEALAND WHITE RABBITS GESTATIONAL PARAMETERS AT SACRIFICE PPM 0 CORPORA LUTEA/DOE MEAN S.D. N 10.3 1,6 22 TOTAL IMPLANTS/LITTER MEAN S.D. N 9.4 2.3 22 PERCENT PREIMPLANTATION MEAN S.D. N LOSS 9.9 16.5 22 VIABLE IMPLANTS MEAN S.D. N 8.4 2.8 22 NON-VIABLE IMPLANTS MEAN S.D. N 1.0 1.7 22 EARLY RESORPTIONS MEAN S.D. N 0.9 1.7 22 LATE RESORPTIONS MEAN S.D. N 0.0 0.0 22 DEAD FETUSES MEAN S.D. N 0.1 0.3 22 PERCENT LIVE FETUSES MEAN S.D. N 89.6 21.1 22 1000 a.a 3.3 21 8.2 2.2 21 8.8 12.2 21 5.2** 3.6 21 3.0 3.1 21 2.4 2.9 21 0.3 1.3 21 0.3 0.6 21 60.3 40.4 21 3000 10.6 1.8 20 9.3 2.7 20 12.B 19.2 20 8.3 3.1 20 1.0 2.1 20 0.4 0.9 20 0.0 0.0 20 0.6 2.0 20 90.2 20.0 20 Report 50-514 Page 18 6000 9.2 2.2 21 8.2* 1.8 21 10.8 12.0 21 7.0* 2.6 21 1.1 1.6 21 0.9 1.3 21 0.0 0,2 21 0.2 0.6 21 83.8 25.1 21 SL 042586 Report 50-514 Page 19 TABLE 7 DEVELOPMENTAL TOXICITY STUDY OF INHALED 1 .1 .1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS GESTATIONAL PARAMETERS AT SACRIFICE PPM 0 1000 3000 SEX RATIO (%MALE MEAN S.D. N FETUSES) 48.9 16.0 21 59.5 22.1 16 45.7 24.1 20 FETAL BODY WEIGHTS ALL FETUSES MEAN S.D. N PER LITTER fGRAMS) 43.16 5.59 21 44.23 5.22 16 39.46 5.74 20 MALE FETUSES MEAN S.D. N 43.13 5.94 21 44.65 5.90 16 39.14 5.42 18 FEMALE FETUSES MEAN S.D. N 43.25 5.98 21 43.14 5.72 15 39.15 5.77 20 a Percent preimplantation loss = corpora corpora lutea multiplied by 100. * p < 0.05 compared to control . ** p < 0.01 compared to control . lutea minus total implants divided by 6000 48.7 23.0 20 40.19 4.42 20 41.17 4.86 19 39.33 4.18 20 0425871 SL SL 042588 TABLE B DEVELOPMENTAL TOXICITY STUDY OF INHALED 1 , 1 . 1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS MALFORMATIONS OBSERVED IN 1FETUSES AND LITTERS3 NUMBER EXAMINED EXTERNALLY13 GROUP (ppm): F ETUS ES 0 1000 3000 6000 IBS 1 10 165 148 L ITT E R S 0 1000 3000 6000 2 1 16 20 20 DOME-SHAPED HEAD 0 0.0 1 0.9 1 0.6 1 0.7 0 0.0 1 6.2 1 5.0 1 5.0 MICROAGNATHIA 0 0.0 0 0.0 0 0.0 1 0.7 0 0.0 0 0.0 0 0.0 1 5.0 SNOUT SHORT 0 0.0 0 0.0 0 0.0 1 0.7 0 0.0 0 0.0 0 0.0 1 5.0 FOREPAWS RIGIDLY FLEXED - BILATERAL 0 0.0 0 0.0 0 0.0 1 0.7 0 0.0 0 0.0 0 0.0 1 5.0 INVERSION - LEFT HINDPAW 0 0.0 0 0.0 0 0.0 1 0.7 0 0.0 0 0.0 0 0.0 1 5.0 None significantly different from control See footnotes on last page of table. SL 042589 TABLE 8 DEVELOPMENTAL TOXICITY STUDY OF INHALED 1.1,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS MALFORMATIONS OBSERVED IN FETUSES AND LITTERS NUMBER EXAMINED VISCERALLYc GROUP (ppm): FETU SES 0 1000 3000 6000 IBS 1 to 1 65 148 LIT TER s 0 1000 3000 6000 2 1 16 20 20 LATERAL VENTRICLE DILATED - TISSUE DEPRESSED 10 5.4 5 4.5 3 1 .8 5 3.4 5 23.8 3 18.7 3 15.0 4 20.0 CAVITATION ORAL CAVITY 1 0.5 0 0.0 0 0.0 0 0.0 1 4.8 0 0.0 0 0.0 0 0.0 CAVITATION IN MEDULLA 0 0.0 1 0.9 0 0.0 1 0.7 0 0.0 1 6.2 0 0.0 1 5.0 THIRD VENTRICLE DILATED - TISSUE DEPRESSED 0 0.0 0 0.0 t 0.6 1 0.7 0 0.0 0 0.0 1 5.0 1 5.0 FOURTH VENTRICLE MISSING 0 0.0 1 0.9 0 0.0 0 0.0 0 0.0 1 6.2 0 0.0 0 0.0 POSTERIOR PORTION OF CEREBELLUM SCRAMBLED ENLARGED AORTA 0 0.0 0 0.0 1 0.9 1 0.9 0 0.0 a 0.0 0 0.0 1 0.7 0 0.0 0 0.0 1 6.2 1 6.2 0 0.0 0 0.0 0 0.0 1 5.0 SMALL BUT PATENT PULMONARY ARTERY 0 0.0 0 0.0 0 0.0 1 0.7 0 0.0 0 0.0 0 0.0 1 5.0 VENTRICULAR SEPTAL DEFECT 0 0.0 1 0.9 1 0.6 2 1 .4 0 0.0 1 6.2 1 5.0 1 5.0 SEMILUNAR VALVES OF THE PULMONARY ARTERY MISSING SEMILUNAR VALVES OF THE AORTA MISSING RIGHT VENTRICLE OF THE HEART SMALL 0 0.0 0 0.0 0 0.0 1 0.9 1 0.9 1 0.9 0 0.0 0 0.0 0 0.0 1 0.7 1 0.7 1 0.7 0 0.0 0 0.0 0 0.0 1 6.2 1 6.2 1 6.2 0 0.0 0 0.0 0 0.0 1 5.0 1 5.0 1 5.0 None significantly different from control See footnotes on last page of table. po to ro onq> on ho rr m o t i<--|s* 042590 V TABLE 8 DEVELOPMENTAL TOXICITY STUDY OF INHALED I, 1 , 1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS MALFORMATIONS OBSERVED IN FETUSES AND LITTERS8 GROUP (ppm): NUMBER EXAMINED VISCERALLY0 LEFT VENTRICLE OF THE HEART ENLARGED HOLE IN EXTERIOR OF LEFT ATRIUM F ETU SES 0 1000 3000 6000 IBS 1 10 165 146 0 0.0 0 0.0 0 0.0 t 0.7 0 0.0 1 0.9 0 0.0 0 0.0 L I T TER S 0 1000 3000 6000 21 16 20 20 0 0.0 0 0.0 0 0.0 1 5.0 0 0.0 1 6.2 0 0.0 0 0.0 PAPILLARY MUSCLES MISSING - RIGHT VENTRICLE 0 0.0 1 0.9 0 0.0 0 0.0 0 0.0 1 6.2 0 0.0 0 0.0 ECTOPIC TESTIS 0 0.0 0 0.0 0 0.0 1 0.7 0 0.0 0 0.0 0 0.0 1 5.0 GALLBLADDER MISSING I 0.5 0 0.0 1 0.6 0 0.0 1 4.8 0 0.0 1 5.0 0 0.0 TEETH PRIMORDIA MISSING 0 0.0 1 0.9 0 0.0 0 0.0 0 0.0 1 6.2 0 0.0 0 0.0 None sfgnificant 1y different from control See footnotes on last page of table. t*oT3 P(t)O (JO CD o NKJ> rr m u01 i -to TABLE 8 DEVELOPMENTAL TOXICITY STUDY OF INHALED 1 . 1 , 1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS MALFORMATIONS OBSERVED IN FETUSES AND LITTERS NUMBER EXAMINED SKELETALLYd GROUP (ppm): CERVICAL RIB - CERVICAL ARCH #7 - UNILATERAL CERVICAL ARCH #3 MISSING - UNILATERAL EXTRA CERVICAL CENTRUM FUSED TO CENTRUM #7 F ETU SES 0 1000 3000 6000 86 51 76 72 0 0.0 0 0.0 0 0.0 1 1.4 0 0.0 0 0.0 0 0.0 1 1.4 0 0.0 0 0.0 0 0.0 1 1 .4 LIT TER S 0 1000 3000 6000 21 16 20 20 0 0.0 0 0.0 0 0.0 1 5.0 0 0.0 0 0.0 0 0.0 1 5.0 0 0.0 0 0.0 0 0.0 1 5.0 EXTRA CERVICAL ARCH - UNILATERAL - TOTAL OF 7 0 0.0 0 0.0 0 0.0 1 1.4 0 0.0 0 0.0 0 0.0 1 5.0 LATERAL CURVATURE OF SPINE - CERVICAL AREA 0 0.0 0 0.0 0 0.0 1 1 .4 0 0.0 0 0.0 0 0.0 1 5.0 THORACIC ARCH MISSING - UNILATERAL 1 1.2 0 0.0 1 1.3 0 0.0 1 4.8 0 0.0 1 5.0 0 0.0 THORACIC CENTRUM AND ARCHES DIAGONALLY DISPLACED 1 1.2 0 0.0 t t.3 0 0.0 1 4.8 0 0.0 1 5.0 0 0.0 THORACIC ARCHES FUSED - UNILATERAL 0 0.0 0 0.0 1 1 .3 0 0.0 0 0.0 0 0.0 1 5.0 0 0.0 LUMBAR CENTRUM ANO ARCHES DIAGONALLY DISPLACED 1.2 0 0.0 0 0.0 0 0.0 1 4.8 0 0.0 0 0.0 0 0.0 RIB #10 - FORKED - UNILATERAL 0 0.0 0 0.0 0 0.0 1 1 .4 0 0.0 0 0.0 0 0.0 1 5.0 RIB #11 - MISSING - UNILATERAL 1 1 .2 0 0.0 0 0.0 0 0.0 1 4.8 0 0.0 0 0.0 0 0.0 RIB #3 MISSING - UNILATERAL 0 0.0 0 0.0 1 1.3 0 0.0 0 0.0 0 0.0 1 5.0 0 0.0 None significantly different from control See footnotes on last page of table. 0li*0bb0 wFJ Report 50-514 50-514 SL 042592 TABLE 8 DEVELOPMENTAL TOXICITV STUDY OF INHALED t,1,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS MALFORMATIONS OBSERVED IN FETUSES AND LITTERSa NUMBER EXAMINED SKELETALLYd GROUP (ppm): F ETU SES 0 1000 3000 6000 B6 51 76 72 LIT TER S 0 1000 3000 6000 21 16 20 20 EXTRA RIB BETWEEN EXISTING RIBS - UNILATERAL 0 0.0 0 0.0 1 1.3 0 0.0 0 0.0 0 0.0 1 5.0 0 0.0 RIBS #2 THROUGH #5 FUSED 0 0.0 0 0.0 1 1.3 0 0.0 0 0.0 0 0.0 1 5.0 0 0.0 None significantly different from control See footnotes on last page of table. *tJ 50 0(P00> "Otf3t) 1 N) rr TABLE 8 DEVELOPMENTAL TOXICITY STUDY OF INHALED 1,1,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS MALFORMATIONS OBSERVED IN FETUSES AND LITTERS8 Ul r* GROUP (ppm); F ETU SE S 0 1000 3000 6000 L I T TER S 0 1000 3000 6000 042593 total malformations NUMBER WITH EXTERNAL MALFORMATIONS PERCENT WITH EXTERNAL MALFORMATIONS 0 0.0 1 0.9 1 0.6 2 1.4 0 0.0 1 6.2 5.0 2 10.0 NUMBER WITH SOFT TISSUE MALFORMATIONS PERCENT WITH SOFT TISSUE MALFORMATIONS 11 5.9 9 8.2 6 3.6 10 6. B 6 666 28.6 37.5 30.0 30.0 NUMBER WITH SKELETAL MALFORMATIONS PERCENT WITH SKELETAL MALFORMATIONS 1 1 .2 0 0.0 1 1 .3 2 2.8 1 4.8 0 0.0 12 5.0 10.0 TOTAL NUMBER WITH MALFORMATIONS TOTAL PERCENT WITH MALFORMATIONS 12 6.5 10 9.1 8 4.8 13 8.8 6 67 8 28.6 37.5 35.0 40.0 None significantly different from control 8 For all findings, the number (of fetuses affected or litters with one or more affected fetuses) Is presented on top and the percentage of the total (fetuses or litters) examined is presented beneath. A single fetus may be represented more than once in listing Individual defects. Only live fetuses were examined. All fetuses were examined externally. c All fetuses were examined viscerally (Staples, 1974), and approximately 50% of each litter were examined for soft tissue craniofacial defects (Wilson, 1965; van Julslngha and Bennett, 1977), d Approximately 50% of each litter were examined for skeletal defects after staining with alizarin red S. hj JO sb n 0m0 ToH) tLofl rt Ln O Ln t-- SL 042594 NUMBER EXAMINED EXTERNALLY^ ECCHVMOSIS - HEAD ECCHYMOSIS - TRUNK EOCHYMOSIS - EXTREMITIES EYE BULGE FLATTENED TABLE 9 DEVELOPMENTAL TOXICITY STUDY OF INHALED 1,1,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS VARIATIONS OBSERVED IN FETUSES AND LITTERS8 GROUP (ppm): F ETUS ES 0 1000 3000 6000 L I TTERS 0 1000 3000 6000 185 I 0.5 6 3.2 1 10 1 0.9 1 0.9 165 6 3.6 1 0.6 148 9 6. 1 1 0.7 21 1 4.8 6 28.6 16 1 6.2 1 6.2 20 4 20.0 1 5.0 20 5 25.0 1 5.0 3 1 .6 0 0.0 3 1 .8 5 3.4 3 14.3 034 0.0 15.0 20.0 0 0.0 1 0.9 0 0.0 1 0.7 0 0.0 1 6.2 0 0.0 1 5.0 None significantly different from control See footnotes on last page of table. 0tto0o *otoo <NT>y rt VOi 1 HLn SL 042595 TABLE 9 DEVELOPMENTAL TOXICITY STUDY OF INHALED 1,1,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS VARIATIONS OBSERVED IN FETUSES AND LITTERS8 NUMBER EXAMINED VISCERALLYc IRREGULAR RUGAE GROUP (ppm}! F ETUSES 0 1000 3000 6000 1B5 1 10 165 148 3 t .6 4 3.6 3 1.8 1 0.7 L I T TER s 0 1000 3000 6000 21 16 20 20 2 9.5 3 18.7 3 15.0 1 5.0 THIRD VENTRICLE DILATED - NO DEPRESSION 7 3.8 z 1 .8 6 3.6 7 4.7 6 25 6 28.6 12.5 25.0 30.0 LATERAL VENTRICLE DILATED - NO DEPRESSION 6 3.Z 6 5.5 12 7.3 6 4. 1 5 676 23.8 37.5 35.0 30.0 DARK BROWN MATERIAL AMONG BRAIN SECTIONS 0 0.0 2 1.8 2 1 .2 0 0.0 0 0.0 2 12.5 2 10.0 0 0.0 DARK BROWN AREA OF BRAIN TISSUE 0 0.0 2 t .8 0 0.0 0 0.0 0 0.0 1 6.2 0 0.0 0 0.0 THYMUS DARK RED 1 0.5 0 0.0 0 0.0 0 0.0 1 4.8 0 0.0 0 0.0 0 0.0 SMALL BUT PATENT DUCTUS ARTERIOSUS 0 0.0 1 0.9 0 0.0 0 0.0 0 0.0 1 6.2 0 0.0 0 0.0 RIGHT SUBCLAVIAN ARTERY SMALL 0 0.0 1 0.9 0 0.0 0 0.0 0 0.0 1 6.2 0 0.0 0 0.0 MEMBRANOUS VENTRICULAR SEPTUM THINNED 0 0.0 0 0.0 0 0.0 1 0.7 0 0.0 0 0.0 0 0.0 1 5.0 LEFT ATRIUM PALE 0 0.0 1 0.9 0 0.0 0 0.0 0 0.0 1 6.2 0 0.0 0 0.0 FETAL ATELECTASIS 2 1.1 2 1.8 6 3.6 0 0.0 2 9.5 2 12.5 5 25.0 0 0.0 PARTIAL FETAL ATELECTASIS 0 0.0 0 0.0 0 0.0 1 0.7 0 0.0 0 0.0 0 0.0 1 5.0 None significantly different from control See footnotes on last page of table. B<t3 0 5(f0t O(Qft *OV s> n VIS-OS SL 042596 TABLE 9 DEVELOPMENTAL TOXICITY STUDY OF INHALED 1 . 1 , 1 -TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS VARIATIONS OBSERVED IN FETUSES AND LITTERS* GROUP (ppm): F ETU SES 0 1000 3000 6000 LIT TER s 0 1000 3000 6000 NUMBER EXAMINED VISCERALLYc 185 1 to 165 148 21 16 . 20 20 AZYGOUS LUNG LOBE MISSING 8 4,3 1 0.9 3 1.8 7 4.7 5 23.8 i 6.2 3 15,0 4 20.0 LIVER PALE ? 0.5 0 0.0 0 0.0 0 0.0 1 4.8 0 0.0 0 0.0 0 0.0 SPLEEN PALE 20 60 t . 1 0.0 3.6 0.0 1 4.8 0 0.0 1 5.0 0 0.0 DARK GREEN FLUID IN STOMACH 0 0.0 0 0.0 0 0.0 1 0.7 0 0.0 0 0.0 0 0.0 1 5.0 DARK GREEN FLUID IN INTESTINES 0 0.0 0 0.0 0 0.0 1 0.7 0 0.0 0 0.0 0 0.0 l 5.0 DILATED RENAL PELVIS - UNILATERAL 0 0.0 0 0.0 1 0.6 ? 0.7 0 0.0 0 0.0 1 5.0 1 5.0 DILATED URETER - UNILATERAL 0 0.0 0 0.0 1 0.6 0 0.0 0 0.0 0 0.0 1 5.0 0 0.0 RIGHT TESTIS THIN WITH NO CHARACTERISTIC SHAPE 0 0.0 0 0.0 0 0.0 1 0.7 0 0.0 0 0.0 0 0.0 1 5.0 REDUCED NUMBER OF TEETH PRIMORDIA 0 0.0 0 0.0 0 0.0 2 1 .4 0 0.0 0 0.0 0 0.0 2 10.0 TEETH PRIMORDIA SMALLER THAN NORMAL 0 0.0 0 0.0 0 0.0 1 0.7 0 0.0 0 0.0 0 0.0 1 5.0 None significantly different from control See footnotes on last page of table. *ft0 0f0t TpfO3to K> rt 00 U"i O ut--1 i -IN Report 50-514 SL 042597 TABLE 9 DEVELOPMENTAL TOXICITY STUDY OF INHALED 1,I,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS VARIATIONS OBSERVED IN FETUSES AND LITTERS GROUP (ppm): F ETUSES 0 1000 3000 6000 L I T TER S 0 1000 3000 6000 NUMBER EXAMINED SKELETALLYd 86 51 76 72 21 16 20 20 CERVICAL CENTRA A1,#2,#3 AND/OR #4 POORLY OSSIFIED 4 4.7 2 3.9 5 6.6 5 6.9 2 9.5 '2 12.5' 3 15.0 3 15.0 CERVICAL CENTRUM #2 0IAGONALLY DISPLACED 0 0.0 0 0.0 0 0.0 1 1 .4 0 0.0 0 0.0 0 0.0 1 5.0 CERVICAL CENTRUM 47 B1LOBED 0 0.0 0 0.0 0 0.0 1 1 .4 0 0.0 0 0.0 0 0^0 1 5.0 EXTRA 413 THORACIC CENTRUM AND ARCHES 22 1 7 29 43 25.6 33.3 38.2 59.7 13 9 15 IB 61.9 56.2 75.0 90.0 THORACIC CENTRUM 46 BILOBED 1 1.2 0 0.0 1 1.3 2 2.8 1 4.8 0 0.0 1 5.0 2 10.0 EXTRA THORACIC ARCH BETWEEN EXISTING ARCHES 0 0.0 0 0.0 0 0.0 1 1 .4 0 0.0 0 0.0 `0 0.0 1 5.0 EXTRA ARCH SMALL 0 0.0 0 0.0 0 0.0 1 1 .4 0 0.0 00 0.0 . 0.0 1 5.0 THORACIC CENTRUM 47 BILOBED 1 1.2 0 0.0 0 0.0 \ 1.4 1 4.8 0 0.0 -0 0.0 1 5.0 THORACIC CENTRUM 411 POORLY OSSIFIED 1 1 .2 0 0.0 0 0.0 0 0.0 1 4.8 0 Q.O 0 0.0 0 0.0 THORACIC CENTRUM 42 SPLIT 0 0.0 0 0.0 1 1.3 0 0.0 0 0.0 O 0.0 .1 5.0 0 0.0 THORACIC CENTRUM 43 SPLIT 0 0.0 0 0.0 1 .3 0 0.0 0 0.0 0 0.0 1 5.0 0 0.0 THORACIC CENTRUM 44 SPLIT 0 0.0 0 0.0 1 1 .3 0 0.0 0 0.0 0 0.0 1 5.0 0 0.0 None significantly different from control See footnotes on last page of table. <CTQD ft) 1NC> SL 042598 TABLE 9 DEVELOPMENTAL TOXtCITV STUDY OF INHALED 1 , 1 ,1-TR1CHLOROETHANE IN NEW ZEALAND WHITE RABBITS VARIATIONS OBSERVED IN FETUSES AND LITTERS NUMBER EXAMINED SKELETALLYd LUMBAR CENTRUM A1 POORLY OSSIFIED GROUP (ppm): F ETU SES 0 1000 3000 6000 86 51 76 72 0 0.0 0 0.0 0 0.0 1 1 .4 L I T TER s 0 1000 3000 6000 21 0 0.0 16 6 .0.0 . 20 0 0.0 20 1 5.0 LUMBAR CENTRUM #2 POORLY OSSIFIED 0 0.0 0 0.0 0 0.0 1 1 .4 0 0.0 -0 0.0 0 0.0 1 5.0 LUMBAR ARCHES #6 POORLY OSSIFIED 0 0.0 0 0.0 1 1 .3 0 0.0 0 0.0 0 0.0 1 5 .0 0 0.0 LUMBAR ARCHES #1 POORLY OSSIFIED 0 0.0 0 0.0 1 1.3 0 0.0 0 0.0 0 0.0 1 5.0 0 0.0 LUMBAR ARCHES #2 POORLY OSSIFIED 0 0.0 0 0.0 1 1 .3 0 0.0 0 0.0 0 0.0 1 5.0 0 0.0 LUMBAR CENTRUM #7 POORLY OSSIFIED 1 1 .2 0 0.0 0 0.0 0 0.0 1 4.8 0 0.0 0 0.0 0 0.0 RUDIMENTARY RIB #13 - FIRST LUMBAR ARCH - UNILATERAL 13 15.1 5 9.8 3 3.9 4 5.6 10 5 3 4 47.6 31 ;2 15.0 20.0 RUDIMENTARY RIB #13 - FIRST LUMBAR ARCH - BILATERAL 3 3.5 2 3.9 A 5.3 1 1.4 3 14.3 2 12.5 3 15.0 1 5.0 EXTRA RIB #13 - FIRST LUMBAR ARCH - UNILATERAL EXTRA RIB #13 - FIRST LUMBAR ARCH - BILATERAL 9 10.5 23 26.7 5 9.8 9 17.6 2 2.6 22 2B .9 6 8.3 18 25.0 6 2B. 6 17 8 1.0 . 5' 2 31.. 2 10.0 . a ' 14 50.0 70... 0 6 30.0 10 50.0 RUDIMENTARY RIB #13 - THORACIC ARCH #13 - UNILATERAL 1 1 .2 0 0.0 ! 1 .3 1 1 .4 1 4.8 01 0.0 . 5.0 1 5.0 RUDIMENTARY RIB #13 - THORACIC ARCH #13 - BILATERAL 0 0.0 1 2.0 0 0.0 0 0.0 0 0.0 1 6.2 -. 0 0.0 0 0.0 None significantly different from control See footnotes on last page of table. T) p0 (fJtQ) Tft3) {ft O OU) rr OI --* Report 50- SL 042599 TABLE 9 DEVELOPMENTAL TOX1CITV STUDY OF INHALED 1,1,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS VARIATIONS OBSERVED IN FETUSES AND LITTERS NUMBER EXAMINED SKELETALLYd GROUP (ppm): F ETU SES 0 1000 3000 6000 66 51 76 72 L I T TER s 0 1000 . 3000 6000 21 16 20 20 EXTRA RIB #13 - THORACIC ARCH #13 - UNILATERAL 0 0.0 0 0.0 3 3.9 1 1.4 0 0.0 0 0.0 3 15.0 1 5.0 EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL 21 15 26 42 24.4 29.4 34.2 58.3 12 - 9 15 18* 57. 1 56.2 75.0 90.0 FRONTAL BONE - POORLY OSSIFIED PARIETAL - POORLY OSSIFIED INTERPARIETAL - POORLY OSSIFIED BODY OF THE HYOID - POORLY OSSIFIED MAXILLARY - POORLY OSSIFIED 0 0.0 1 2.0 2 2.3 0 0.0 0. 0 0.0 0.0 3 3.5 4 7.8 0 0.0 1 2.0 1 1.3 1 1 .3 0 0.0 4 5.3 2 2.6 1 1.4 1 1 .4 1 1 .4 5 6.9 1 1.4 0 0.0 2 9.5 0 0.0 2 9.5 0 0.0 1 6.2 } 5.0 1 5.0 0 0.0. 1 5.0 1 5.0 0 0.0 0 0.0 1 5.0 1 6.2 3 15.0 4 20.0 ,1 6.2 2 10.0 1 5.0 PARIETAL - HOLE IN BONE 1 .2 0 0.0 1 1.3 0 0.0 1 4.8 0 0.0 1 5:0 0 0.0 SOME INTER.PHALANGES (FORELIMB) POORLY OSSIFIED 8 9.3 5 9.8 14 18.4 21 29.2 44 7 10 19.0 25.0 35.0 50.0 MAJORITY INTER.PHALANGES (FORELIMB) POORLY OSSIFIED 0 0.0 0 0.0 0 0.0 1 I .4 0 0.0 0 0.0 , 0 0.0 1 5.0 SOME INTER.PHALANGES (HINDLIMB) POORLY OSSIFIED 0 0.0 0 0.0 0 0.0 1 1 .4 0 0.0 0 0.0 0 0.0 1 5.0 STERNEBRA #2 POORLY OSSIFIED 1 1 .2 0 0.0 1 1 .3 1 1.4 1 4.8 01 0.0 ' 5.0 1 5.0 *= significantly different from control at .05 level using two-tailed Fisher's exact test* See footnotes on last page of table* i 1 V wfDto Report 50-514 SL 042600 TABLE 9 DEVELOPMENTAL TOXICITY STUDY OF INHALED I,1,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS VARIATIONS OBSERVED IN FETUSES AND LITTERS8 NUMBER EXAMINED SKELETALLYd GROUP (ppm): F ETU S E S 0 1000 3000 6000 86 51 76 72 LIT TER s 0 1000 3000 6000 21 16 20 20 STERNE8RA #4 POORLY OSSIFIED 0 0.0 0 0.0 0 0.0 1 1.4 0 0.0 0 0.0 0 0.0 1 5.0 STERNEBRA A5 POORLY OSSIFIED 22 13 26 13 25.6 25.5 34.2 18. 1 9 8 12 9 42.9 50.0 60.0 45.0 STERNEBRA #6 POORLY OSSIFIED 30 34.9 5 9.B 10 13.2 14 19.4 13 4 6 9 61.9 25.0 30,0 45.0 STERNEBRA #5 UNOSSIFIED 2 2.3 t 2.0 3 3.9 5 6.9 2 9.5 1 3* 4 6.2- 15.0 20.0 STERNEBRA *6 UNOSSIFIED 0 0.0 1 2.0 1 1 .3 2 2.8 0 0.0 r 6.2 12 5.0 10.0 STERNEBRAE #1 AND A2 FUSED 00 1 0 0.0 0.0 1.3 0.0 0 0.0 0 0.0 1 5.0 0 0.0 STERNEBRAE #4 AND #5 FUSED 20 1 0 2.3 0.0 1.3 0.0 2 9.5 O' . 1 0.0 . 5.0 0 0.0 EXTRA OSSIF. SITE ABOVE STERNEBRA SEGMENT #1 0 0.0 1 2.0 0 0.0 1 1 .4 0 0.0 1 6.2 0 0.0 1 5.0 STERNEBRA A1 BILOBED 0 0.0 0 0.0 0 0.0 1 1 .4 0 0.0 0 0.0 0 0.0 1 5.0 STERNEBRA #2 BILOBED 0 0.0 0 0.0 0 0.0 1 1.4 0 0.0 0 0.0 0 .0.0 1 5.0 STERNEBRA A3 BILOBED 0 0.0 0 0.0 0 0.0 1 1 .4 0 0.0 0 0.0 0 0.0 1 5.0 STERNEBRA #4 BILOBED 0 0.0 0 0.0 0 0.0 1 1 .4 0 0.0 .0 D.O 0 0.0 1 5.0 None significantly different from control See footnotes on last page of table. O0Q) <t> wNJ 0*2601 f NUMBER EXAMINED SKELETALLYd STERNEBRA #5 BILOBED STERNEBRA #6 BILOBED STERNEBRAE #2 THROUGH #5 FUSED TABLE 9 DEVELOPMENTAL TOXICITY STUDY OF INHALED 1 , 1 ,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS VARIATIONS OBSERVED IN FETUSES AND LITTERS3 GROUP (ppm): F ETU SES 0 1000 3000 6000 L I T' T E R S 0 tooo- 3000 6000 86 51 76 72 21 16 20 20 0 0.0 0 0.0 0 0.0 1 1.4 0 0.0 0 0.0 0 0.0 1 5.0 0 0.0 0 0.0 0 0.0 1 1 *4 0 0.0 00 o: o 0.0 1 5.0 0 0.0 0 0.0 0 0.0 i 1.4 0 0.0 0 0.0 .0 0.0 1 5.0 None significantly different from control See footnotes on last page of table. t^o3 oo mSo (0 on UU>> rt m o Ut--1< SL 042602 TABLE 9 DEVELOPMENTAL TOXICITY STUDY OF INHALED 1.1.1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS VARIATIONS OBSERVED IN FETUSES AND LITTERS GROUP (ppm): F ETU SES 0 1000 3000 6000 L I TfER S 0 1000 3000 6000 TOTAL VARIATIONS NUMBER WITH EXTERNAL VARIATIONS PERCENT WITH EXTERNAL VARIATIONS 9 4.9 3 2.7 to 15 6. 1 10.1 7 33.3 3 IB. 7 78 35.0 40.0 NUMBER WITH SOFT TISSUE VARIATIONS PERCENT WITH SOFT TISSUE VARIATIONS 25 13.5 18 37 16.4 22.4 23 15.5 12 57 . 1 to 62.5 16 80.0 15 75.0 NUMBER WITH SKELETAL VARIATIONS PERCENT WITH SKELETAL VARIATIONS 74 43 68 72 86.0 64.3 89.5 100.0 21 15 19 20 100.0 93.7 95.0 100.0 TOTAL NUMBER WITH VARIATIONS TOTAL PERCENT WITH VARIATIONS 99 60 103 94 53.5 54.5 62.4 63.5 21 15 20 20 100.0 93.7 100.0 100.0 None significantly different from control a For all findings* the number (of fetuses affected or titters with one or more affected fetuses} is,presented on top and the percentage of the total (fetuses or litters) examined is presented beneath. A single fetus may be represented more than once in listing individual defects. Only live fetuses were examined. b AM fetuses were examined externally. c AM fetuses were examined visceral1y (Staples, 1974), and approximately 50% of each litter were examined for soft tissue craniofacial defects (Wilson, 1965; van Julslngha and Bennett, 1977), d Approximately 50% of each litter were examined for skeletal defects after staining ;, with alizarin red S, -' 0*30 pfDa <jq fO on W rr un LOIn Report 50-514 Page 35 BUSHY RUN RESEARCH R. D. 4, Mellon Road, Export, Pennsylvania 15632 CENTER Telephone (4121 733-5200 GOOD LABORATORY PRACTICES COMPLIANCE This study was performed in full compliance with the Good Laboratory Practices (GLP) Standards promulgated by the U. S. Environmental Protection Agency Toxic Substances Control {Federal Register 48(2301. 53922-53944, 1983]. Prepared by: PATH/esk/0949P-1 03-23-87 R. W. Tyl, rti.D., DABT Study Director Date SL 042603^ Bushy Run Research Center A Joint Mellon Institute--Union Carbide Corporation Operation Report 50-514 Page 36 BUSHY RUN RESEARCH R. D. 4, Mellon Road, Export, Pennsylvania 15632 CENTER Telephone (412) 733-5200 Quality Assurance Unit Study Inspection Summary Test Substance: 1,1,1 Trichloroethane Study Dvelprontal Toxicity Study of Inhaled 1,1,1 Trichloroethane in New Zealand White Rabbits Study Director: R. W. Tyl, Ph.D., D.A.B.T. The Quality Assurance Unit of BRRC conducted the inspections listed below and reported the results to the study director and to management on the dates indicated. It is the practice of this Quality Assurance Unit to report the results of each inspection to both the study director and management. Inspection Date Type Date OAU Report Issued To Studv Director To Management 7-14-86 Protocol, Probe Study 7-17-86 7-18-86 7-15-86 Protocol, Full Study 7-17-86 7-18-86 7-31-86 Event-Animal Receipt, Probe Study 8-5-86 8-18-86 8-22-86 Event-Exposure, Probe Study 9-2-86 9-9-86 9-3-86 Event-Sacrifice, Probe Study 9-4-86 9-9-86 9-16-86 Protocol Amendment #1, Probe Study 9-16-86 9-16-86 9-23-86 Event-Animal Receipt, Full Study 9-30-86 10-31-86 10-7-86 Event-Mating, Full Study 10-9-86 10-31-86 10-15-86 Event-Exposure, Full Study 10-22-86 Protocol Amendment #1, Full Study 11-6-86 Event-Necropsy, Full Study 6-2-87 to Raw Data and Reports, Probe 6-8-87 and Full Studies 10-20-86 10-22-86 11-7-86 6-10-87 10-31-86 10-22-86 12-10-86 6-24-87 042604 Bushy Run Research Center A Joint Mellon Institute--Union Carbide Corporation Operation Report 50-514 Page 37 Inspection Date Type Date OAU Report Issued To Studv Director To Management 6-8 to 6-10-87 Analytical Raw Data and Reports, Probe and Full Studies 6-10-87 6-23-87 6-24-87 Archives 6-24-87 6-24-87 Lirfda J. Callst^, Group Leader Date Good Laboratory Practices/Quality Assurance SL 042605 Report 50-514 APPENDIX 1 Inhalation Chamber Operation and Analyses (28 Pages) Paae Appendix 1A Analytical Report - Definitive Study .............................................. 1-9 Appendix IB Methodology and Inhalation Chamber Operation Definitive Study................................................................................. 1-5 Appendix 1C Analytical Report - Exposure Range-Finding Study .................. 1-9 Appenidx ID Methodology and Inhalation Chamber Operation - Exposure Range-Finding Study....................................................... 1-5 SL 042606 Report 50-514 APPENDIX 1A Analytical Report Definitive Study (9 Pages) SL 042607 Report 50-514 APPENDIX 1A Chemical and Chamber Atmosphere Analyses for the Developmental Toxicity Study of Inhaled 1.1.1-Trichloroethane in CD Rats and in New Zealand White Rabbits SUMMARY The concentrations of i,l,l-trichloroethane (TCE) In the 'exposure chambers were monitored throughout the 16 days of exposure by gas chromatography * The concentration in each chamber was determined approximately 13 times during each 6-hour exposureN The overall means (t standard deviations) were 1017 ( 18), 3122 (+ 39), and 5906 ( 45) ppm for target concentrations of 1000, 3000, and 6000 ppm, respectively. No TCE concentration above the estimated minimum detection limit was detected in the control chamber. The test chemical was analyzed before and after the study; no significant compositional changes were detected. CHEMICALS Three 55-gallon containers of 1,1,1-trichloroethane (TCE) test material (CAS Number 71-55-6, Lot Number TA8 606182, BRRC Sample Number 49-228 A,B,C) were received from The Dow Chemical Company (Freeport, TX) on June 25, 1986. Test material from the original container was placed in one-gallon glass bottles and blanketed with nitrogen. The chemical and physical properties of the test material are described in Table 1. The purity and stability were verified by analysis both before and after the study. These analyses, provided by Mr. William D. Tasto, Dow Chemical USA, are presented In Table 2. No significant compositional changes occurred while the study progressed. The test material remained 99.9% pure throughout the study. The gases for the chromatograph were obtained from Linde Corporation, The nitrogen and hydrogen were Ultra High Purity grade and the air was breathing quality. EQUIPMENT A Perkin-Elmer Model 3920B gas chromatograph (GC) equipped with a flame ionization detector was used to monitor the TCE vapor concentrations in the chambers. The GC operating parameters are presented in Table 3. A Spectra-Physics series 4000 central processor, a data interface, and a Perkin-Elmer automatic gas sampling system (station and valve programmer units and a gas sampling valve) were used for the analyses. The chromatogram and chromatographic data were printed on a printer/plotter providing a permanent record of the analyses. The chromatographic data were also transmitted to a Hewlett-Packard Model 2640 B terminal and to a Digital PDP 1170 computer with a RSX 11 M+ operating system. The computer software provided statistical treatment of the data and also triggered an alarm if an exposure chamber concentration deviated more than 10% from the target concentration. -- gl, 0A2608 Report 50-514 Appendix 1A 2- - CALIBRATION Calibration of the gas chromatograph was achieved by injecting gas standards, which were prepared by syringe injection of test material into Tedlar gas bags. These standards were prepared using the mathematical relationship: C x V. x MW x 298 x P d x 24.45 x T X 760 -6 u where: V = required volume of calibration liquid in milliliters (mL) at temperature T in degrees Kelvin (K) C = the desired final calibration concentration in parts per million (ppm) Vjj = volume of container (Liters) MW = molecular weight of the calibration liquid P = barometric pressure in millimeters of mercury (mm Hg) d = density of calibration liquid in grams per milliliter (g/mL) at temperature T 24.45 = molecular volume at 298K and 760 mm Hg, in (Liters) T = temperature in degrees Kelvin (K) A linear calibration curve (Figure 1) was obtained when areas (integration counts) were plotted versus the gas standard concentrations. The calibration factor (KF) was calculated as: _ AREA " CONCENTRATION The mean KF for all valid injections of standards was used in subsequent microprocessor calculations. This calibration factor was checked at least once each week during the exposure period and did not require adjustment. Registered Trademark, DuPont de Nemours, E. I. & Co. SL 042609 Report 50-514 Appendix 1A 3- CHAMBER ATMOSPHERE ANALYSIS Each chamber atmosphere was analyzed for TCE approximately once every 25 minutes during each 6-hour exposure. The daily mean concentrations are listed in Table 4. The means of daily means ( standard deviation) were 1017 ( 18), 3122 ( 39), and 5906 ( 45)'ppm, for target concentrations of.. 1000, 3000, and 6000 ppm, respectively. No concentration of TCE above the estimated minimum detection limit of 10 ppm was found' in the control chamber during the study. The daily analytical/nominal TCE concentration ratios are given in Table 5; the nominal concentration being an estimate calculated from the quantity of test material delivered and the chamber airflow rate. The overall mean analytical/nominal concentration ratios were 0.90, 0.96, and 0.90 for the 1000 ppm, 3000 ppm, and 6000 ppm exposure concentrations, respectively. Acknowledgement: John M. Smith, B.S. Technical Assistance WPC/llf/11892-1 06-23-87 Irvin M. Pritts, Ph.D Research Scientist Date SL 4261o Report 50-514 Appendix 1A -4- Table 1 Chemical and Physical Properties of 1.1.1-Trlchloroethane1 Syn nyms: Bolling Point: Specific Gravity 8 25/25C: Vapor Density: Appearance and Odor: Vapor Pressure 8 20C: Solubility in Water: Flash Point: Methyl Chloroform 74C 1.330-1.335 4.55 Colorless liquid; irritating odor at high concentrations 100 mm Hg 0.07 g/100 g 8 25C None ^-Material Safety Data Sheet, Dow Chemical U.S.A., MSDS:000699, 6/21/86. WPC/llf/1189Z-l 06-23-87 Sl> 042&H Report 50-514 Appendix 1A -5- Table 2 Test Material Analysis^ Pre-Study Post-Study GC Purity Chloroethylene 1,1-dichloroethylene 1,1-dichloroethane 1,2-dichloroethane 1,2-epoxybutane Water Content Acidity (as HC1) Color, APHA Non-Volatile Residue Infrared Scan > 99.9 wt% 0.0009 wt% 0.0355 wt% 0.0029 wt% 0.0050 wt% 0.0284 wt% 35 ppm 4.8 ppm 0 (Pt.-Co.) 4.5 ppm Typical > 99.9 wt% 0.0004 vt% 0.0400 wt% 0.0021 wt% 0.0040 wt% 0.0270 wt% 62 ppm 2.3 ppm 0 (Pt.-Co.) 3.8 ppm Typical (no change) ^Analyses provided by Mr. William D. Tasto, Dow Chemical U.S.A., on September 18, 1986 and on November 12, 1986. WPC/llf/1189Z-l 06-23-87 SL 042612 6- - Report 50-514 Appendix 1A Table 3 Gas Chromatograph Operating Parameters Chromatograph: Perkin-Elmer 3920 Detector: Tlame Ionization Column: 10X SP-2100 on 80/100 mesh Supelcoport (10 ft. x 1/8 in. stainless steel) Column temperature: 125*C Injector temperature: Ambient gas sample valve Detector temperature: 250C Carrier flow rate: 15 mL/minute nitrogen Hydrogen flow rate: 66 mL/minute Air flow rate: 303 mL/minute Sample size: 1 cc Retention time: 90 seconds GC attenuation: 100 x 2 Printer/Plotter attenuation: 50 Chart speed: 1 cm/minute Approximate minimum detection limit: 10 ppm WPC/llf/1189Z-l 06-23-87 SL 042613 Exposure Day 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 Report 50-514 Appendix 1A --7-- Table 4 Summary of Daily Mean Chamber Concentrations ______ 1000 Target Concentration, ppm______ ~ . 3000 6000 0 Daily Mean Concentrations (ppml * SD1 1041 17.7 1035 53.2 1042 16.8 1033 8.2 1031 13.3 1014 23.9 1017 6.1 1031 17.3 990 8.8 986 19.5 1019 HK 21.7 1010 60.9 1019 7.7 1010 30.2 1008 5.1 992 20.8 3132 49.5 3118 30.3 3178 16.8 3167 17.6 3202 11.5 3094 40.6 3093 29.9 3050 25.5 3075 50.3 3119 16.5 3120 17.5 3149 42.3 3116 16.5 3101 10.2 3136 18.1 3107 44.8 5986 83.1 5943 40.4 5964 35.4 5941 41.9 5925 181.3 5880 93.0 5885 98.5 5867 30.6 5886 112.9 5803 266.3 5934 32.6 5927 45.1 5910 88.7 5888 51.8 5892 97.1 5857 183.9 < MDL' < MDL < MDL < MDL < MDL < MDL < MDL < MDL < MDL < MDL < MDL < MDL < MDL < MDL < MDL < MDL Mean of Daily Means Standard Deviation 1017 17.7 3122 38.6 5906 45.1 *SD = Standard deviation 2< MDL = Less than the estimated minimum detection limit. WPC/llf/1189Z-l 06-23-87 < MDL SL 042614 Report 50-514 SL 042615 Analytical to Nominal Concentration Ratios Exposure Day 1000 nom Analytical^ Nominal A/N2 Target Concentration 3000 oom Analytical Nominal A/N Analytical 6000 Dorn Nominal A/N 1 1041 1131 0.92 2 1035 1141 0.91 3 1042 1148 0.91 4 1033 1129 0.91 5 1031 1120 0.92 6 1014 1145 0.89 7 1017 1127 0.90 8 1031 1152 0.89 9 990 1110 0.89 10 986 1109 0.89 11 1019 1141 0.89 12 1010 1150 0.88 13 1019 1145 0.89 14 1010 1138 0.89 15 1008 1146 0.88 16 992 1113 0.89 3132 3118 3178 3167 3202 3094 3093 3050 3075 3119 3120 3149 3116 3101 3136 3107 3179 3180 3252 3261 3224 3224 3235 3228 3168 3275 3252 3270 3258 3270 3252 3224 0.99 0.98 0.98 0.97 0.99 0.96 0.96 0.94 0.97 0.95 0.96 0.96 0.96 0.95 0.96 0.96 5986 5943 5964 5941 5925 5880 5885 5867 5886 5803 5934 5927 5910 5888 5892 5857 6543 6487 6547 6553 6520 6546 6536 6525 6421 6588 6567 6592 6574 6560 6546 6438 0.91 0.92 0.91 0.91 0.91 0.90 0.90 0.90 0.92 0.88 0.90 0.90 0.90 0.90 0.90 0.91 i CtO Mean of Daily Means Standard Deviation 1017 17.7 1134 14.6 0.90 0.013 3122 38.6 3234 33.8 0.96 0.014 5906 , 45.1 6534 48.3 0.90 o oo ^Daily mean concentration (ppm) 2Analytical/nominal concentration ratio WPC/llf/11892-1 06-23-87 >a fDCHXtu* > Report 50-514 -9. Appendix 1A K M Z C O O I m Zl I x SL 042616 Figure 1. Calibration Curve Report 50-514 APPENDIX IB Methodology and Inhalation Chamber Operation 1,1,1-Trichloroethane Vapor Inhalation Definitive Study (5 Pages) SL 042617 Report 50-514 APPENDIX IB Methodology and Inhalation Chamber Operation 1.1.1-Trichloroethane Vapor Inhalation Teratology Full Study METHODS -Inhalation Chamber Description and Operation The inhalation chambers used in this study were located in Room 139* They were constructed from stainless-steel with glass windows for animal observation (Table 1). Chamber volume was approximately 4320 liters and airflow was 1000 L/min (14 air changes per hour). Chamber temperature and relative humidity were recorded using minimum-maximum thermometers (Abbeon Cal, Inc., Santa Barbara, CA and Brooklyn Thermometer Company, Inc., Farmingdale, NY) and an Airguide Humidity Indicator (Airguide Instrument Company, Chicago, IL), respectively. Temperature and relative humidity measurements were recorded eleven times per exposure. 1.1.1-Trichloroethane Vapor Generation Liquid 1,1,1-trichloroethane was metered from a piston pump (Fluid Metering Inc., Oyster Bay, NY) for all target chamber concentrations into a heated glass evaporator similar in design as that described by Carpenter et. al. (Toxicol. AppI. Pharmacol. 12:246-262, 1975). Two evaporators were set-up in series for the 6000 ppm target concentration. This was necessary because of the large volume of test material needed to generate the high level concentration. The temperature in the evaporators were maintained at the lowest level sufficient to vaporize the liquid. The resultant vapor was carried into the chamber by a countercurrent air stream that entered the bottom of the evaporator. Temperature measurements were obtained from the airstream of each evaporator during the exposure regimen. The temperature was measured by using a Doric Trendicator Model 400A probe (Doric Scientific Division, Emerson Electric Co., San Diego, CA). RESULTS Chamber Temperature and Relative Humidity During exposures, the mean of daily mean chamber temperatures for all exposure groups ranged from 24.9 to 25.3C (Table 2), and the relative humidity ranged between 41.4 to 49.7% (Table 3). Evaporator Temperature The temperature for all evaporators ranged from 35 to 64C. 0A2&1S SL 2- - Report 50-514 Appendix IB Acknowledaements David W. Fait, B.S., AALAS Cert. II Master Technologist William J. Kintigh, B.S. AALAS Cert. I Master Technologist Y Date WPC/llf/1116Z-l 03-17-87 SV 0 hi6^ Report 50-514 Appendix IB -3- Table 1 1.1.1-Trichloroethane Vapor Inhalation Teratology Full Study . Inhalation Chamber Design and Operation CHAMBER Location: Room 139 Construction: The chamber Is made of stainless-steel. The door in the front of the chamber and the windows in one of the side walls (through which animals were observed) were made of glass. Shane: Rectangular Dimensions: Height - 207 cm Length 213 cm Width 98 cm Total Volume Approximately 4320 Liters Manufacturer: Wahmann Manufacturing Company Timonium, Maryland Airflow Measurement: An orifice plate was positioned in the exhaust duct of the chamber and was connected to a Dwyer Magnehelic pressure gauge. Airflow: For all exposure groups: 1000 liters/min 99* = 20 min. *199 is the theoretically-derived time required for the chamber to reach 99% of the equilibrium concentration. WPC/llf/1116Z-l 03-17-87 Report 50-514 Appendix IB -4- Table 2 1.1.1-Trichloroethane Vapor Inhalation Teratology Full Study Chamber Temperature (C1 Exposure Dav 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 Mean of Daily Means Standard Deviation 6000 Dt>tn 26.9* 25.5 26.3 26.1 25.5 25.6 26.0 25.2 26.1 24.7 24.7 25.2 25.1 23.8 24.5 23.6 25.3 0.90 Tafeet Chamber Concentrations 3000 ppm 1000 ppm 24.9 24.3 26.5 25.9 26.0 25.7 26.3 24.8 25.8 24.6 24.1 24.5 24.2 23.8 23.7 22.5 24.9 24.5 26.5 26.1 26.1 26.1 26.4 25.5 26.3 25.1 24.5 24.8 25.0 24.1 22.9 22.2 24.9 25.1 1.10 1.25 -0 PPm 25.4 24.2 26.2 25.7 25.8 25.7 26.4 25.6 26.3 24.9 24.4 25.3 25.2 23.7 23.5 22.7 25.1 1.08 * Each value represents the mean of 11 measurements taken during each six-hour exposure. WPC/llf/1116Z-l 03-17-87 Report 50-514 Appendix IB -5- Table 3 1.1.1-Trichloroethane Vapor Inhalation Teratology Full Study Chamber Relative Humidity m Exposure Dav 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 Mean of Daily Means Standard Deviation 6000 DDR] Taraet Chamber Concentrations 3000 Dom 1000 DDm 34.1* 38.7 38.1 37.8 40.8 42.0 37.5 37.6 41.3 45.5 46.3 44.5 44.7 45.5 44.5 44.1 40.5 47.4 37.7 38.6 40.9 45.2 39.8 39.9 45.2 45.9 50.8 49.6 47.5 48.3 48.6 48.1 41.0 42.4 35.5 38.2 40.5 42.7 41.3 40.5 48.8 50.2 51.3 48.2 46.4 46.5 49.4 48.4 41.4 44.6 44.5 3.74 4.35 4.76 0 PPm 55.4 50.3 45.0 46.5 46.8 49.9 44.6 45.0 51.4 50.7 53.2 51.4 51.1 52.0 51.0 51.2 49.7 3.18 * Each value represents the mean of 11 measurements taken during each six-hour exposure. WPC/llf/1116Z-l 03-17-87 SL 042622 Report 50-514 APPENDIX 1C Analytical Report Exposure Range-Finding Study (9 Pages) 042623 SL Report 50-514 APPENDIX 1C Chemical and Chamber Atmosphere Analyses for the Developmental Toxicity Exposure Range-Finding Study of Inhaled 1,1.1-Trichloroethane in CD Rats and New Zealand White Rabbits SUMMARY The concentrations of 1,1,1-trichloroethane (TCE) in the exposure chambers were monitored throughout the 14 days of exposure by gas chromatography. The concentration in each chamber was determined approximately 13 times `during each 6-hour exposureN The overall means ( standard deviations) were 1011 ( 18), 3019 ( 161), and 5907 ( 126) ppm for target concentrations of 1000, 3000, and 6000 ppm, respectively. No TCE above the estimated minimum detection limit of 10 ppm was detected in the control chamber. The test chemical was analyzed before and after the study; no significant compositional changes were detected. CHEMICALS Three 55-gallon containers of 1,1,1-trichloroethane (TCE) test material (CAS Number 71-55-6, Lot Number TA8 606182, BRRC Sample Number 49-228 A,B,C) were received from The Dow Chemical Company (Freeport, TX) on June 25, 1986. Test material from the original container was placed in one-gallon glass bottles and blanketed with nitrogen. The chemical and physical properties of the test material are described in Table 1. The purity and stability were verified by analysis both before and after the study. These analyses, provided by Mr. William D. Tasto, Dow Chemical USA, are presented in Table 2. No significant compositional changes occurred while the study progressed. The test material remained 99.9% pure throughout the study. The gases for the chromatograph were obtained from Linde Corporation. The nitrogen and hydrogen were Ultra High Purity grade and the air was breathing quality. EQUIPMENT A Perkin-Elmer Model 3920B gas chromatograph (GC) equipped with a flame ionization detector was used to monitor the TCE vapor concentrations in the chambers. The GC operating parameters are presented in Table 3. A Spectra-Physics series 4000 central processor, a data interface, and a Perkin-Elmer automatic gas sampling system (station and valve programmer units and a gas sampling valve) were used for the analyses. The chromatogram and chromatographic data were printed on a printer/plotter providing a permanent record of the analyses. The chromatographic data were also transmitted to a Hewlett-Packard Model 2640 B terminal and to a Digital PDP 1170 computer with a RSX 11 M+ operating system. The computer software provided statistical treatment of the data and also triggered an alarm if an exposure chamber concentration deviated more than 10% from the target concentration. 04262** 2- - Report 50-514 Appendix 1C CALIBRATION Calibration of the gas chromatograph was achieved by injecting gas standards, which were prepared by syringe injection of test material into Tedlar gas bags. These standards were prepared using the mathematical relationship: C x V x MW x 298 it P -6 V = --------- ----------------------------- x 10 d x 24.45 X T x 760 where: V = required volume of calibration liquid in milliliters (mL) at temperature T in degrees Kelvin (K) C = the desired final calibration concentration in parts per million (ppm) Vh = volume of container (Liters) MW = molecular weight of the calibration liquid P = barometric pressure in millimeters of mercury (mm Hg) d = density of calibration liquid in grams per milliliter (g/raL) at temperature T 24.45 = molecular volume at 298K and 760 mm Hg, in (Liters) T = temperature in degrees Kelvin (K) A linear calibration curve (Figure 1) was obtained when areas (integration counts) were plotted versus the gas standard concentrations. The calibration factor (KF) was calculated as: v-p _____AREA_____ CONCENTRATION The mean KF for all valid injections of standards was used in subsequent microprocessor calculations. This calibration factor was checked at least once each week during the exposure period and did not require adjustment. Registered Trademark, DuPont de Nemours, E. I. & Co. SL 042625 Report 50-514 Appendix 1C -3- CHAMBER ATMOSPHERE ANALYSIS Each chamber atmosphere was analyzed for TCE approximately once every 25 minutes during each 6-hour exposure. The daily mean concentrations are listed in Table.4. The means of daily means ( standard deviation) were 1011 ( 18), 3019 ( 161), and 5907 ( 126) ppm, for target concentrations of . 1000, 3000, 6000 ppm, respectively. No concentration of TCE above the estimated minimum detection limit of lOppm was found in-the control chamber during the study. The daily analytical/nominal TCE concentration ratios are given in Table 5; the nominal concentration being an estimate calculated from the quantity of test material delivered and the chamber airflow rate. The overall mean analytical/nominal concentration ratios were 0.91, 0.99, and 0.90 for the 1000 ppm, 3000 ppm, and 6000 ppm exposure concentrations, respectively. The low daily mean chamber concentration and large standard deviation (2540 ppm 1113) for the 3000 ppm target concentration for day 13 (Table 4) was caused by a vapor generation problem. As a result, test animals were exposed to less than 60 ppm TCE for nearly one hour of the 6-hour exposure. The other 5 hours of exposure proceeded normally. Acknowledgement: John M. Smith, B.S. Technical Assistance WPC/llf/1189Z-l 06-23-87 Irvin M. Pritts, Ph.D. Research Scientist Date SL 0A2626 Report 50-514 Appendix 1C -4- Table 1 Chemical and Physical Properties of 1.1.1-Trlchloroethanel Synonyms: Boiling Point: Specific Gravity 6 25/25C: Vapor Density: Appearance and Odor: Vapor Pressure 6 20C: Solubility in Water: Flash Point: Methyl Chloroform 74C 1.330-1.335 4.55 Colorless liquid; Irritating odor at high concentrations 100 mm Hg 0.07 g/100 g 0 25C None ^Material Safety Data Sheet, Dow Chemical U.S.A., MSDS:000699, 6/21/86. WPC/llf/1189Z-l 06-23-87 SL 042627 Report 50-514 Appendix 1C -5- Table 2 Test Material Analysis^ ' GC Purity Chioroethylene 1,1-dichloroethylene 1,1-dichloroethane 1,2-dichloroethane 1,2-epoxy butane Water Content Acidity (as HC1) Color, APHA Non-Volatile Residue Infrared Scan Pre-Study > 99.9 wt% 0.0026 wt% 0.0340 vt% 0.0040 wtX 0.0045 wt% 0.0281 wtX 26 ppm 1.8 ppm 0 (Pt.-Co.) 3.9 ppm Typical Post-Study > 99.9 wt% 0.0009 wt% 0.0355 wt% 0.0029 wt* 0.0050 wtS 0.0284 wtX 35 ppm 4.8 ppm 0 (Pt.-Co.) 4.5 ppm Unchanged (typical) ^-Analyses provided by Mr. William D. Tasto, Dow Chemical U.S.A. on July 23, 1986 and on September 18, 1986. WPC/llf/1189Z-l 06-23-87 0A2628 Si* Report 50-514 Appendix 1C 6- - Table 3 Gas Chromatograph Operating Parameters Chromatograph: Detector: Column: Column temperature: Injector temperature: Detector temperature: Carrier flow rate: Hydrogen flow rate: Air flow rate: Sample size: Retention time: GC attenuation: Printer/Plotter attenuation: Chart speed: Approximate minimum detection limit: WPC/llf/1189Z-l 06-23-87 Perkin-Elmer 3920 Flame Ionization 10% SP-2100 on 80/100 mesh Supelcoport (10 ft. x 1/8 in. stainless steel) 125C Ambient gas sample valve 250C 15 mL/minute nitrogen 66 mL/minute 303 mL/minute 1 cc 90 seconds 100 x 2 50 1 cm/minute 10 ppm SL 042629) Report 50-514 Appendix 1C -7- Table 4 Summary of Dally Mean Chamber Concentrations Exposure Day _____________Target Concentration, ppm_____________________ 1000 3000 6000 0 Daily Mean Concentrations (ppm) SD* - 1 993 51.9 3236 38.8 6057 154.5 < MDL 2 998 47.6 3092 34.3 5934 46.2 < MDL 3 1007 6.9 3169 105.0 5945 254.5 < MDL 4 1002 + 30.6 3054 24.3 5994 27 A < MDL 5 1011 19.5 3154 61.1 5893 372.8 < MDL 6 1017 19.0 3051 120.1 5923 162.3 < MDL 7 1011 19.5 3000 78.3 5873 148.5 < MDL 8 999 34.9 3000 * 31.7 6013 154,1 < MDL 9 978 23.5 3045 11.4 5825 379.8 < MDL 10 1006 13.1 3021 13.5 5895 145.0 < MDL 11 1035 27.9 2913 174.9 5779 115.1 < MDL 12 1038 9.0 2986 40.7 5588 508.0 < MDL 13 1039 15.5 2540 1113 5881 86.0 < MDL 14 1024 9.6 3004 12.9 6097 18.5 < MDL Mean of Daily Means Standard Deviation 1011 17. 8 3019 161, 4 5907 126.,1 < MDL *SD = Standard deviation WPC/llf/1189Z-l 06-23-87 C/1 tr 0 > to 01 Table 5 UJ Analytical to Nominal Concentration Ratios Exposure ____________ 1000 ppm______________ Day Analytical1 Nominal A/N2 1 993 1113 0.89 2 998 1106 0.90 3 1007 1124 0.90 4 1002 1117 0.90 5 1011 1101 0.92 6 1017 1113 0.91 7 1011 1120 0.90 8 999 1099 0.91 9 978 1082 0.90 10 1006 1106 0.91 11 1035 1125 0.92 12 1038 1111 0.93 13 1039 1110 0.94 14 1024 1101 0.93 Mean of Daily Means Standard Deviation 1011 17.8 1109 11.4 0.91 0.015 ^Daily mean concentration (ppm) 2Analytical/nominal concentration ratio WPC/llf/1189Z-l : 06-23-87 Target Concentration _____________ 3000 ppm________________ Analytical Nominal A/N 3236 3092 3169 3054 3154 3051 3000 3000 3045 3021 2913 2986 2540 3004 3224 3104 3249 3046 3104 3086 3023 2995 3048 3028 2966 3038 2752 3022 1.00 1.00 0.98 1.00 1.02 0.99 0.99 1.00 1.00 1.00 0.98 0.98 0.92 0.99 3019 161.4 3049 117.0 0.99 0.023 ________________ 6000 ppm Analytical Nominal A/N 6057 5934 5945 5994 5893 5923 5873 6013 5825 5895 5779 5588 5881 6097 6552 6553 6609 6477 6546 6546 6525 6506 6485 6560 6574 6508 6525 6567 0.92 0.91 0.90 0.93 0.90 0.90 0.90 0.92 0.90 0.90 0.88 0.86 0.90 0.93 5907 126.1 6538 36.1 0.90 0.01' TTopC33L o Report 50-514 Report 50-514 -9- Appendix 1C (E flC U ia : U O 3 Z H (/) x w is ts ) SL 042632' Figure 1. Calibration Curve Report 50-514 APPENDIX ID Methodology and Inhalation Chamber Operation 1,1,1-Trichloroethane Vapor Inhalation Exposure Range-Finding Study (5 Pages) SI* 042633 Report 50-514 APPENDIX ID Methodology and Inhalation Chamber Operation 1.1.1-Trichloroethane Vapor Inhalation Teratology Probe Study METHODS Inhalation Chamber Description -and Operation The inhalation chambers used in this study were located in Room 139. They were constructed from stainless-steel with glass windows for animal observation (Table 1). Chamber volume was approximately 4320 liters and airflow was 1000 L/min (14 air changes per hour). Chamber temperature and relative humidity were recorded using minimum-maximum thermometers (Abbeon Cal, Inc., Santa Barbara, CA and Brooklyn Thermometer Company, Inc., Farmingdale, NY) and an Airguide Humidity Indicator (Airguide Instrument Company, Chicago, IL), respectively. Temperature and relative humidity measurements were recorded at least ten (10) times per exposure. 1.1.1 - Trichloroethane Vapor Generation Liquid 1,1,1-trichloroethane was metered from a piston pump (Fluid Metering Inc., Oyster Bay, NY) for the 6000, 3000, and 1000 ppm target chamber concentrations into a heated glass evaporator similar in design as that described by Carpenter et. gl. (Toxicol. ApdI. Pharmacol. 32;246-262. 1975). Two evaporators were set-up in series for the 6000 ppm target concentration. This was necessary because of the large volume of test material needed to generate the high level concentration. The temperature in the evaporators were maintained at the lowest level sufficient to vaporize the liquid. The resultant vapor was carried into the chamber by a countercurrent air stream that entered the bottom of the evaporator. Temperature measurements were obtained from the airstream of each evaporator during the exposure regimen. The temperature was measured by using a Doric Trendicator Model 400A probe (Doric Scientific Division, Emerson Electric Co., San Diego, CA). RESULTS Chamber Temperature and Relative Humidity During exposures, the mean of daily mean chamber temperatures for all exposure groups ranged from 20.3 to 24.5C (Table 2), and the relative humidity ranged between 38.5 to 49.6% (Table 3), Evaporator Temperature The temperature for all evaporators ranged from 28 to 41*C. SL 042634 Report 50-514 Appendix ID 2- - Acknowledgements David W. Fait, B.S., AALAS Cert. II Frank L. Howard, III, AALAS Cert. II Master Technologist Master Technologist William J. Kintigfi, B.S AALAS Cert. I Master Technologist 7 Date WPC/llf/1116Z-l 03-07-87 SL 0A2635 Report 50-514 Appendix ID -3~ Table 1 1.1.1-Trichloroethane Vapor Inhalation Teratology Probe Study Inhalation Chamber Design and Operation CHAMBER Location: Room 139 Construction: The chamber is made of stainless-steel. The door in the front of the chamber and the windows in one of the side walls (through which animals were observed) were made of glass. Shane: Rectangular Dimensions: Height - 207 cm Length - 213 cm Width 98 cm Total Volume - Approximately 4320 Liters Manufacturer: Wahmann Manufacturing Company Timonium, Maryland Airflow Measurement: An orifice plate was positioned in the exhaust duct of the chamber and was connected to a Dwyer Magnehelic pressure gauge. Airflow: For all exposure groups: 1000 liters/min fc99* = 20 min. *^99 is the theoretically-derived time required for the chamber to reach 99% of the equilibrium concentration. #PC/llf/1116Z-l 03-17-87 SL 042636 Report 50-514 Appendix ID -4- Table 2 1.1.1-Trlchloroethane Vapor Inhalation Teratology Probe Study Chamber Temperature ("O Exposure Day 1 2 3 4 5 6 7 8 9 10 11 12 13 14 Mean of Daily Means Standard Deviation 6000 DDm 25.6* 25.5 25.2 25.7 24.7 25.4 25.5 24.3 23.8 23.5 24.6 23.5 23.0 23.3 24.5 0.97 Tareet Chamber Concentrations 3000 DDm 1000 ppm. 22.0 21.9 21.8 21.6 21.0 21.4 21.4 21.6 20.3 20.4 21.6 21.4 ** 20.7 21.2 21.0 21.0 20.8 20.5 20.6 21.0 20.8 20.5 21.0 21.1 20.4 20.0 19.7 21.3 20.7 0.55 0.43 0 DDm 23.0 21.5 21.0 21.0 21.0 19.0 18.8 19.2 19.5 19.5 20.3 20.6 20.0 19.9 20.3 1.14 * Each value represents the mean of at least 10 measurements taken during each six-hour exposure. ** Generator technician unable to see temperature gauge. WPC/llf/1116Z-l 03-17-87 SL 042637 Report 50-514 Appendix ID -5- Table 3 1.1,1-Trlchloroethane Vapor Inhalation Teratology Probe Study Chamber. Relative Humiditv Exposure Day 1 2 3 4 5 6 7 8 9 10 11 12 13 14 Mean of Daily Means Standard Deviation 6000 ppm 36.5* 43.5 33.1 38.4 41.8 41.6 37.1 36.3 36.8 35.5 36.6 41.5 40.3 40.2 38.5 2.49 Tareet Chamber Concentrations . 3flQH-PPiB 1000 ppm .0 ppm 54.6 53.1 46.9 47.0 49.0 48.4 45.6 46.3 45.5 45.5 47.2 50.4 51.2 51.9 52.2 53.1 47.0 46.5 48.7 48.2 45.9 45.4 45.9 47.5 48.2 49.1 51.0 50.6 55.0 55.5 50.5 50.0 51.5 50.8 48.3 48.8 46.8 46.0 45.9 46.9 50.1 48.2 48.8 48.5 49.6 3.01 2.43 2.99 L * Each value represents the mean of at least 10 measurements taken during each six-hour exposure. WPC/llf/lU6Z-l 03-17-87 SL 042638 Report 50-514 APPENDIX 2 Individual In-Life Maternal Data (9 Pages) 42639 0 SL 042640 DEVELOPMENTAL TOXICITY STUDY OF INHALED 1,1,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS INDIVIDUAL BODY WEIGHTS (GRAMS) DURING GESTATION PREGNANCY STATUS DAY 0 6 9 1 2 15 19 DAMS FROM GROUP 1: G - 0 PPM 15946 15953 15954 15961 15967 15980 15990 15991 15993 15994 15997 16002 16003 16005 16006 16016 16020 16025 16028 16036 16039 16041 16045 16047 G G G G G G G G G G DE NG G G G G G G G G G G G G 3592.5 3584.7 3339.0 3499.4 3756.4 3647.8 3658.5 3352.7 3833.0 3507.0 3669.0 3250.4 3430.3 3159.7 3464.9 3641.4 3279.0 3515.3 3353.9 3399.7 3189.8 3143.8 3149.3 3454.3 3916.6 3801.9 3487.8 3574.7 3608.8 3775.7 3801.8 3465.6 3895.4 3645.0 3808.9 3282.4 3663.1 3295.6 3627.8 3899.6 3599.2 3496.1 3616.1 3624.0 3372.0 3387.0 3209.1 3476.3 3870.6 3828.9 3424.0 3488.6 3636.8 3705.9 3774.7 3456.7 3913.4 3685.2 3816.2 3303.7 3580.7 3190.0 3653.9 3907.8 3639.3 3492.0 3593.4 3556.6 3363.0 3213.1 3179.5 3480.3 3948.2 3793.3 3429.4 3536.0 3581.2 3717.7 3766.8 3481.3 3920.9 3664.4 3767.0 3330.0 3603.1 3268.4 3678.5 3975.6 3635.8 3473.7 3619.9 3572.0 3408.2 3275.1 3195.8 3439.3 3992.6 3857.7 3435.0 3524.4 3530.0 3766.4 3735.4 3614.2 3996.2 3715.9 3830.2 3405.6 3674.1 3357. 1 3732.7 4108.9 3729.2 3528.1 3699.4 3590.5 3519.6 3242.0 3355.0 3403.2 4064.6 3823.6 3456.5 3492.3 3668.6 3756.7 3861.8 3636.4 4061.7 3778.6 3B1 1.3 3506.4 3784.7 3404.6 3776.0 4144.9 3758.8 3595.6 3757.5 3663.5 3568.9 3307.9 3378.7 3471.1 MEAN S.D. N 3461.8 196.9 23 3610.8 195.2 23 3584.8 219.3 23 3597.9 212.1 23 3649.5 221 .4 23 3696.7 221.4 23 G=GRAVID NG=NON GRAVID WEIGHT(S) NOT INCLUDED IN CALCULATION OF MEAN DE=OELIVERED EARLY WEIGHT(S) NOT INCLUDED IN CALCULATION OF MEAN 29 4309.7 4084.4 3723.4 3717.6 3831.3 4017.1 4027.5 3903.8 4321 .4 3986.0 3153.1 3676.1 4011.1 3705.3 3890.5 4353.1 3874.4 3B14.8 3941.7 3876.1 377 1.7 3519.5 3430.0 3640.7 3865.5 280.6 23 Repo r t 50-514 Appendi> 2 Page 1 SL 042641^ DEVELOPMENTAL TOXICITY STUOY OF INHALED 1,1, 1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS INDIVIDUAL BODY WEIGHTS (GRAMS) DURING GESTATION PREGNANCY STATUS DAY 0 6 9 12 15 19 29 DAMS FROM GROUP 2: I - 1000 PPM 15944 15945 15957 15962 15963 15964 15965 15971 15974 15975 15976 15961 15963 15986 15996 16010 16015 16026 16027 16030 16031 16037 16036 16046 G G G G G G G G RS G G G G G G G G G G G NG G G NG 3501.4 3426.4 3262.7 3727.5 3376.6 3914.0 3231.8 3896.9 3445.7 3571.1 3172.1 3530.3 3637.9 3658.6 3433. 1 3153.7 3340.5 4210.8 3552.9 3570.3 3504.3 2974.0 3495.2 3279.6 3713.6 3468.1 3390.4 3861.9 3545.8 3721 . 1 3247.4 3966.4 3476.0 3790.5 3310.8 3744.2 3724.4 3870.9 3403. 1 3271.2 3461.1 4587.6 3487.7 3768.9 3477.8 3161.7 3561.9 3228.7 3692.9 3422.4 3348.5 3737.6 3372.8 3736.6 3189.5 3900.5 3344.1 3699.9 3232 . 1 3669.5 3740. 1 3855.8 3438.1 3198.0 3428.6 4495.2 3480.8 3675.5 3429.6 3112.7 3487.9 3225.5 3681.0 3426.0 338 1 . 1 3809.2 3285.7 3625.6 3230.5 3910.7 3246.3 3667.2 3261.4 3660.0 3716.8 3883.2 3555.2 3203.5 3403.9 4581.7 3477.0 3764.1 3420.4 3153.9 3396.7 3156.5 3772.2 3400.7 3453.9 3583.2 3238.8 3727.0 3266.9 4076.3 3732.8 3302.6 3663.1 3728.6 3971.4 3673.6 3225.1 3417.5 4610.7 3603.2 3798.3 3286.5 3256.2 3363.8 3192.4 3718.5 3300.5 3518.9 3865.4 3184.8 3809.4 3334.8 4068.6 3792.4 3248.3 3622.7 3755.0 3949.3 3650.9 3230.2 3386.2 4639.2 3458.1 3928.6 3391.6 3250.2 3328.0 3220. 1 3797.5 3549.1 3657.7 4029.1 3540.2 4231.3 3510.9 4290.8 3950.9 330 1.5 3920.2 3954.0 4082.2 3793.3 3481.4 3438.4 5008.2 3804.6 4025.5 3742.0 3453.2 3393.7 3422.7 MEAN S.O. N 3503.8 278.6 22 3616.2 310.8 22 3557.2 308.6 22 3560.0 324.5 22 3612.7 335.5 21 3621.0 358.6 21 3819.7 395. 1 21 G=GRAVID NG=NON GRAVID WEIGHT(S) NOT INCLUDED IN CALCULATION OF MEAN RS=REMOVED FROM STUDY WEIGHT(S) NOT INCLUDED IN CALCULATION OF MEAN Report 50"514 Appendix 2 Page 2 SL 042642 DEVELOPMENTAL TOXICITY STUDY OF INHALED 1,I,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS INDIVIDUAL BODY WEIGHTS (GRAMS) DURING GESTATION PREGNANCY STATUS DAY 0 6 9 12 15 19 29 DAMS FROM GROUP 3: M -- 3000 PPM 1594 2 15943 15947 15950 15952 15956 15958 1 5960 1596B 15973 15979 15987 15988 15995 15998 16011 16013 16018 16021 16029 16032 16034 16048 16049 G NG G G G G G G G NG G G G G G G G G G NG G G NG G 3389.9 3501.7 3249. 1 3586.3 31 19.6 3153.4 4310.9 3630.B 3397.9 3755.4 3403. 1 3661.9 3355.2 3598.5 3401.9 3545.2 3284.6 3450.9 3513.2 3811.0 3895.9 3235.0 3480.8 3438.7 3482.7 3781 .7 3405.3 3501 .9 3096.5 3434.4 4293.0 3766.5 3871 .4 3866.5 3570.0 3909.8 3432.7 3713.6 3529.5 3658.1 3317.0 3644.9 3696.8 3712.4 4022.7 3500.2 3625.8 3679.1 3413.5 3720.6 3316.2 3472.6 3039.6 3424.7 4054.8 3695.8 3713.7 377 1 .0 3472.4 3762.6 3351 .5 3580 . 1 3467.1 3523 . 1 3394.9 3545.7 3595.6 3741.4 3918.6 3458.0 3527.6 3567.6 3447.7 3634.8 3294.3 3450.1 2991.3 3493.0 4060.7 3634.4 3631.8 3673.6 3501 .5 3756.9 3361.5 3535.1 3333.4 3634.7 3397.3 3662.0 3636.8 3706.0 3948.2 3453.9 3390.9 3520.9 3471.4 3647.3 3365.8 3503.7 2987.8 3519.5 3985.0 3837.9 3610.2 3695.0 3597.8 3817.9 3436.4 3500.3 3393.2 3737.7 345 1 . 1 3646.4 3630.9 3670.9 4042.3 3535.3 3341.4 357 1.2 3549.1 3710.2 3372.3 3549.3 297 1 .6 342 1 .4 4050.4 3850.7 3530.5 3734.6 3604.0 3875.3 3432.1 3431.2 3190.3 3707.9 3482.5 3566.4 3672.1 3663.5 4042.1 3510.9 3056.2 3570.8 3871.1 3954.0 3661.6 3789.7 3262.2 3705.2 4317.6 4115.3 3508.5 4013.7 3929.7 4037.7 3652.3 3647.2 3405.0 3621.0 3408.8 3997.7 3817.4 3826.8 4318.4 3706.2 3683.5 3888.0 MEAN S. D. N 3481.1 269.8 20 3626.3 265.8 20 3538.4 220.7 20 3537.3 231.7 20 358 2. 1 233. 1 20 3569.0 258.7 20 3783.0 285.9 20 G=GRAV1D NG=N0N GRAVID WEIGHT(S) NOT INCLUDED IN CALCULATION OF MEAN Report 50-514 Appendix 2 Page 3 SL 042643 DEVELOPMENTAL TOXICITY STUDY OF INHALED 1,1.1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS INDIVIDUAL BODY WEIGHTS (GRAMS) DURING GESTATION PREGNANCY STATUS DAY 0 6 9 12 15 19 29 DAMS FROM GROUP 4: R -- 6000 PPM 15948 15951 15955 15959 15972 15978 15982 15984 15989 15992 15999 16001 1600 7 16008 16009 16012 16014 16019 16022 16023 16033 16040 16043 16044 G G G G G G G G G NG G G G NG G NG G G G G G G G G 3346.8 3567.0 3209.6 3049.8 3920.4 3373.7 3635.3 3576.1 3720.4 3527.8 3544.1 3621.8 3487.2 3230.8 3240.2 3657.9 3350.8 3309.0 3407.3 3836.6 3421.3 3112.3 4301.0 3531 . 1 3361.9 3600. 1 3328.2 3273.1 4088.7 3535.4 3802.8 3510.8 3997.1 3590.8 3559.1 3699.3 36B4.9 3143.8 3550.7 3752.9 3425.7 3544.3 3512.6 3907.9 3475.6 3253.9 4353.7 3533.9 3362.0 3531.2 3303.6 3200.5 4013.3 3374.2 3736.7 3516.4 3926.6 3546.2 3399. 1 3637.6 3606.5 3007.1 3411.5 3630.0 3305.6 3547.1 3550.7 3774.3 3408.7 3181.2 4184.2 3450.4 3325.2 3540.4 322 1 . 1 3239.1 3973.3 3358.7 3748.4 3526.9 3934.5 3491 .8 3413.5 3627. 1 3595.2 2920.4 3429.0 3503.4 3345.0 3404.0 3473.9 3764.1 3383.8 3134.0 4078.8 3506.8 3335. 1 3636. 1 3280.3 3370. 1 3951.5 3457.4 3750.5 3538.0 3943.9 3488.3 3561.0 3660.1 3664.6 3026.7 3546.7 3410.4 3403.8 3422.2 3301.3 3741.7 3350.4 3176.8 4012.2 3587.7 3274.3 3750.0 3219.1 3450.0 3916.4 3464.3 3766. 1 3492.9 3952.7 3462.6 3434.1 3668.2 3755.4 3055.9 3538.9 3277.3 3531.5 3436.9 3439.9 3685.7 3366. 1 3159.2 3893.8 3634.0 3510.5 4010.9 3435.4 3642.1 4237.6 3753.0 3865.3 3800.5 4184.9 3841.1 3667.7 3923.8 3970.9 3342.8 3809.2 3768.4 3809.1 3765.1 3520.9 3666.3 3577.3 3354.2 4419.8 3829.0 MEAN S.D. N 3502.9 286.7 21 3619.0 277.9 21 3543.9 261 .9 21 3524.9 253.4 21 3557.7 232.8 21 3563.3 224.8 21 3797.8 266.9 21 G=GRAVID NG=NON GRAVID WEIGHT(S) NOT INCLUDED IN CALCULATION OF MEAN Report 50-514 Appendi* 2 Page 4 SL 042644 DOSAGE GROUP G - 0 PPM ANIMAL DEVELOPMENTAL TOXICITY STUDY OF INHALED 1,1,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS CLINICAL OBSERVATIONS BY GESTATION DAY CATEGORY GESTATIONAL DAYS FINDING ) 6045 15991 16006 15980 15993 15953 15994 15961 16036 16020 16041 16005 15954 16026 1 6047 16016 15967 15990 15946 16025 16003 16039 NORMAL DEAD NORMAL DEAD NORMAL DEAD NORMAL DEAD NORMAL DEAD NORMAL DEAD NORMAL DEAD NORMAL DEAD NORMAL DEAD NORMAL DEAD NORMAL DEAD NORMAL DEAD NORMAL DEAD NORMAL OEAD NORMAL DEAD NORMAL DEAD NORMAL DEAD NORMAL DEAD NORMAL DEAD NORMAL DEAD NORMAL DEAD NORMAL DEAD 029- 029- 029- 029- 029- 029- 029- 029- 029- o29- o29- 029- 029- 029- o29- 029- 029- 029- 029- o29- 029- 029- 29 29 29 29 29 29 29 29 29 29 29 29 29 29 29 29 29 29 29 29 29 29 29 29 29 29 29 29 29 29 29 29 29 29 29 29 29 29 29 29 29 29 29 29 NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE Report 50-5 1 4 Appendi* 2 Page 5 SL 042645 DOSAGE GROUP I - 1000 PPM ANIMAL DEVELOPMENTAL TOXICITV STUDY OF INHALED 1,1,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS CLINICAL OBSERVATIONS BY GESTATION DAY CATEGORY GESTATIONAL DAYS FINDING 16037 15996 15975 159B3 159B6 16026 1590 1 16030 15963 15957 16010 15976 NORMAL DEAD NORMAL DEAD NORMAL DEAD NORMAL DEAD NORMAL DEAD NORMAL DEAD NORMAL DEAD NORMAL DEAD NORMAL DEAD FOOO/WATER NORMAL DEAD NORMAL DEAD NORMAL DEAD OTHER 16015 15945 15944 NORMAL DEAD NORMAL DEAD NORMAL DEAD OTHER 16030 1597 1 15964 15962 16027 NORMAL DEAD NORMAL DEAO NORMAL DEAD NORMAL DEAD NORMAL 029- 029- 029- 029- 029- 029- 029- 029- 02911- 029- 029- 0292221- 029- 029- 0292121- 029- 029- 029- 029- 0- 29 29 29 29 29 29 29 29 29 29 29 29 29 29 29 29 29 29 20 29 29 29 29 29 29 23 23 29 29 29 29 29 29 23 22 29 29 29 29 29 29 29 29 29 NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE FEED HOPPER FULL NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE TISSUE ON PAPERBOARD (UNRECOGNIZABLE) BLOOD (BY HEMASTIX) ON PAPERBOARD NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE BLOOD (BY HEMASTIX) ON PAPERBOARD TISSUE ON PAPERBOARD (UNRECOGNIZABLE) NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE NO SIGNIFICANT CLINICAL OBSERVATIONS Report 50-514 Append i * 2 Page 6 SL 042646 DOSAGE GROUP I - 1000 PPM M - 3000 PPM ' ANIMAL DEVELOPMENTAL TOXICITY STUDV OF INHALED 1,1,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS CLINICAL OBSERVATIONS BV GESTATION DAY CATEGORY GESTATIONAL DAYS FINDING 16027 15965 DEAD NORMAL DEAD 16034 NORMAL DEAO EVES/EARS/NOSE 15947 15979 15995 15960 16032 16049 15968 15988 15956 15952 16013 15998 16018 15950 15958 15987 NORMAL DEAD NORMAL DEAD NORMAL DEAD EXCRETA FOOD/WATER NORMAL DEAD NORMAL DEAD NORMAL DEAD NORMAL DEAD FOOD/WATER NORMAL DEAD NORMAL DEAD NORMAL DEAD OTHER NORMAL DEAO NORMAL DEAD FOOD/WATER SKIN NORMAL DEAD NORMAL DEAD NORMAL DEAD NORMAL 29- 29 0- 29 29- 29 SCHEDULED SACRIFICE NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE 3- 22 29- 29 1 1 - 29 12- 21 0- 29 29- 29 o- 29 29- 29 0- 29 29- 29 1 2- 12 12- 20 o- 29 29- 29 o- 29 29- 29 0- 29 29- 29 2- 28 29- 29 29- 29 0- 29 29- 29 0- 29 29- 29 0- 29 29- 29 1 1- 1 1 o- 29 29- 29 o- 29 29- 29 21- 21 25- 27 0- 29 29- 29 0- 29 29- 29 0- 29 29- 29 o- 29 NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE LACR1MATION {EYL 8) OCULAR DISCHARGE. OTHER (EYL 10) NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE PAPER80ARD DRY FEED HOPPER FULL NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE FEED HOPPER FULL NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE WATER TIP NOT FUNCTIONING - DRY PAPERBOARD NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE FEED HOPPER FULL ABSCESS (NCK 3} NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE NO SIGNIFICANT CLINICAL OBSERVATIONS Report 50-51 A Appendi* 2 Page 7 SL 042647 DOSAGE GROUP M - 3000 PPM R - 6000 PPM 1 ANIMAL DEVELOPMENTAL TOXICITY STUDY OF INHALED 1,l,t-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS CLINICAL OBSERVATIONS BY GESTATION DAY CATEGORY GESTATIONAL DAYS FINDING 15987 16011 16021 15942 DEAD NORMAL DEAD NORMAL DEAD NORMAL DEAD 16009 15978 15999 16001 15972 16044 16007 16022 16019 16040 15959 15948 16014 15951 15982 16023 16043 NORMAL DEAD NORMAL DEAD NORMAL DEAD NORMAL DEAD NORMAL DEAD EYES/EARS/NOSE EXCRETA NORMAL DEAD NORMAL DEAD NORMAL DEAD SKIN NORMAL DEAD OTHER NORMAL DEAO NORMAL DEAD NORMAL DEAD NORMAL DEAD NORMAL DEAD NORMAL DEAD NORMAL DEAD NORMAL 290- 29o- 29- 029- 29 29 29 29 29 29 29 SCHEDULED SACRIFICE NO SIGNIFICANT CLINICAL SCHEDULED SACRIFICE NO SIGNIFICANT CLINICAL SCHEDULED SACRIFICE NO SIGNIFICANT CLINICAL SCHEDULED SACRIFICE OBSERVATIONS OBSERVATIONS OBSERVATIONS 0- 29 29- 29 o- 29 29- 29 0- 29 29- 29 0- 29 29- 29 0- 29 29- 29 19 17- 18 0- 29 29- 29 0- 29 29- 29 2- 15 29- 29 16- 29 2- 24 29- 29 24- 28 0- 29 29- 29 0- 29 29- 29 0- 29 29- 29 0- 29 29- 29 0- 29 29- 29 0- 29 29- 29 0- 29 29- 29 0- 29 NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE OCULAR DISCHARGE, OTHER (EYL 1} LOOSE FECES NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE ALOPECIA (HPB 14, NCK 4, PFL 4, NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE BLOOD (BY HEMASTIX) ON PAPERBOARD NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE NO SIGNIFICANT CLINICAL OBSERVATIONS SCHEDULED SACRIFICE NO SIGNIFICANT CLINICAL OBSERVATIONS Report 50-514 Appendi 2 Page 8 SL 042648 DOSAGE GROUP R - 6000 PPM ANIMAL DEVELOPMENTAL TOXICITY STUDY OF INHALED 1 , 1 , 1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS CLINICAL OBSERVATIONS BY GESTATION DAY CATEGORY GESTATIONAL DAYS FINDING 16043 1 5909 15984 16033 15955 DEAD EYES/EARS/NOSE EXCRETA NORMAL DEAD NORMAL DEAD NORMAL DEAD NORMAL DEAD 298- 029- 029- 029- 029- 29 18 7 29 29 29 29 29 29 29 29 SCHEDULED SACRIFICE OCULAR DISCHARGE, OTHER LOOSE FECES NO SIGNIFICANT CLINICAL SCHEDULED SACRIFICE NO SIGNIFICANT CLINICAL SCHEDULED SACRIFICE NO SIGNIFICANT CLINICAL SCHEDULED SACRIFICE NO SIGNIFICANT CLINICAL SCHEDULED SACRIFICE {EYB 9, EYI OBSERVATIONS OBSERVATIONS OBSERVATIONS OBSERVATIONS Report 50-514 Appendix 2 Page 9 Report 50-514 APPENDIX 3 Necropsy Data Including Individual Fetal Body Weights (20 Pages) SL 042649 Report 50-514 Appendix 3 Page 1 GROUP: ANIMAL DEVELOPMENTAL TOXICITY study OF INHALED 1,1.1-TRICHL0R0ETHANE IN NEW ZEALAND WHITE RABBITS DAM GROSS EXAMINATION AT TIME OF LAPAROTOMY G - 0 PPM FEMALE ALL DEATHS COMBINED 16045 4-N0V-B6 EXAMINED NO SIGNIFICANT LESIONS ANIMAL 16002 4-N0V-86 EXAMINED NO SIGNIFICANT LESIONS ANIMAL 15991 4-N0V-86 EXAMINED NO SIGNIFICANT LESIONS ANIMAL 16006 4-N0V-B6 examined NO SIGNIFICANT LESIONS ANIMAL 15980 4-N0V-86 ANIMAL 15993 4-N0V-86 EXAMINED NO SIGNIFICANT LESIONS EXAMINED NO SIGNIFICANT LESIONS ANIMAL 15953 5-N0V-86 ANIMAL 15994 5-N0V-86 ANIMAL 15961 5-N0V-86 EXAMINED NO SIGNIFICANT LESIONS EXAMINED NO SIGNIFICANT LESIONS EXAMINED NO SIGNIFICANT LESIONS ANIMAL 16036 5-N0V-86 EXAMINED NO SIGNIFICANT LESIONS ANIMAL 16020 5-N0V-86 ANIMAL 16041 5-N0V-86 ANIMAL 16005 6-N0V-86 EXAMINED NO SIGNIFICANT LESIONS OVARIES GROSS: CYST LEFT CLEAR FLUID 2MM IN DIAMETER EXAMINED NO SIGNIFICANT LESIONS ANIMAL 15954 6-N0V-86 ANIMAL 16028 6-N0V-86 EXAMINED NO SIGNIFICANT LESIONS EXAMINED NO SIGNIFICANT LESIONS SL 042650 Report 50-514 Appendix 3 Page 2 GROUP: ANIMAL DEVELOPMENTAL TOXICITY STUDY OF INHALED 1 . 1 . 1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS DAM GROSS EXAMINATION AT TIME OF LAPAROTOMY G - 0 PPM FEMALE ALL DEATHS COMBINED 16047 6-N0V-86 EXAMINED * NO SIGNIFICANT LESIONS ANIMAL . 16016 6-NOV-86 EXAMINED - NO SIGNIFICANT LESIONS ANIMAL 15997 6-N0V-86 GROSS: NOT EXAMINED- REMOVED FROM STUDY ANIMAL 15967 7-N0V-86 EXAMINED - NO SIGNIFICANT LESIONS ANIMAL 15990 7-N0V-86 AMNIOTIC SACS GROSS: PLACENTAS GROSS: ABSENT ABSENT ANIMAL 15946 7-N0V-86 EXAMINED - NO SIGNIFICANT LESIONS ANIMAL 16025 7-N0V-86 EXAMINED - NO SIGNIFICANT LESIONS ANIMAL 16003 7-N0V-86 EXAMINED - NO SIGNIFICANT LESIONS ANIMAL 16039 7-N0V-86 EXAMINED - NO SIGNIFICANT LESIONS SL 042651 Report 50-514 Appendix 3 Page 3 GROUP: ANIMAL DEVELOPMENTAL TOXICITY STUDY of INHALED 1.1,1-TRICHL0R0ETHANE IN NEW ZEALAND WHITE RABBITS DAM GROSS EXAMINATION AT TIME OF LAPAROTOMY I - 1000 PPM FEMALE ALL DEATHS COMBINED 16037 4-N0V-86 EXAMINED - NO SIGNIFICANT LESIONS - ANIMAt, 16046 4-N0V-86 UTERUS GROSS: AMNIQTIC SACS GROSS: PLACENTAS GROSS: 2X2MM CLEAR FLUID FILLED AREA RIGHT HORN' ABSENT ABSENT ANIMAL 15996 4-N0V-86 ANIMAL 15975 4-N0V-86 UTERUS GROSS: CONTAINS LIQUID FLUID BILATERAL +++FOR BLOOD BY HEMASTIX EXAMINED * NO SIGNIFICANT LESIONS ANIMAL 15983 4-N0V-B6 EXAMINED - NO SIGNIFICANT LESIONS ANIMAL 15906 4-N0V-86 EXAMINED NO SIGNIFICANT LESIONS ANIMAL 16026 5-N0V-86 UTERUS GROSS: CYST MULTIPLE BILATERAL ANIMAL 15981 5-N0V-S6 EXAMINED - NO SIGNIFICANT LESIONS ANIMAL 1603B 5-N0V-86 CORPORA LUTEA GROSS: AMNIQTIC SACS GROSS: PLACENTAS GROSS: ABSENT ABSENT ABSENT ANIMAL 15963 5-N0V-B6 EXAMINED - NO SIGNIFICANT LESIONS ANIMAL 15957 5-N0V-86 EXAMINED - NO SIGNIFICANT LESIONS animal 16010 5-N0V-86 EXAMINED - NO SIGNIFICANT LESIONS SL 042652 Report 50-514 Appendix 3 Page 4 GROUP: ANIMAL DEVELOPMENTAL TOXICITY STUDY OF INHALED 1,1,1-TRICHLORQETHANE IN NEW ZEALAND WHITE RABBITS DAM GROSS EXAMINATION AT TIME OF LAPAROTOMY I - 1000 PPM FEMAI ALL DEATHS COMBINED 15976 6-N0V-86 AMNIOTIC SACS GROSS: PLACENTAS GROSS: ABSENT ABSENT ANIMAL 16015 ANIMAL 15945 6-N0V-86 6-N0V-86 examined - no significant lesions EXAMINED - NO SIGNIFICANT LESIONS ANIMAL 15944 6-N0V-88 AMNIOTIC SACS GROSS: PLACENTAS GROSS: ABSENT ABSENT ANIMAL 16030 6-N0V-86 ANIMAL 15971 6-NOV-06 VAGINA GROSS: DISCHARGE BLOODY TAKEN OUT OF ORDER +++BLOOO BY HEMASTIX EXAMINED - NO SIGNIFICANT LESIONS ANIMAL 15964 7-N0V-86 EXAMINED - NO SIGNIFICANT LESIONS ANIMAL 15962 7-N0V-86 AMNIOTIC SACS GROSS: PLACENTAS GROSS: ABSENT ABSENT ANIMAL 16027 7-N0V-86 CORPORA LUTEA GROSS: AMNIOTIC SACS GROSS: PLACENTAS GROSS: ABSENT ABSENT ABSENT ANIMAL 16031 7-N0V-86 CORPORA LUTEA GROSS: AMNIOTIC SACS GROSS: PLACENTAS GROSS: ABSENT ABSENT ABSENT ANIMAL 15974 23-0CT-B6 15965 7-N0V-86 GROSS: NOT EXAMINED- OELIVEREO EARLY EXAMINED - NO SIGNIFICANT LESIONS SL 042653) Report 50-514 Appendix 3 Page 5 GROUP: ANIMAL DEVELOPMENTAL TOXICITY STUDY OF INHALED 1,1,1 -TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS DAM GROSS EXAMINATION AT TIME OF LAPAROTOMY M - 3000 PPM FEMAI ALL DEATHS COMBINED 16034 4-N0V-86 EXAMINED ~ NO SIGNIFICANT LESIONS ANIMAL 15947 4-N0V-86 EXAMINED - NO SIGNIFICANT-LESIONS ANIMAL 15979 4-N0V-86 EXAMINED - NO SIGNIFICANT LESIONS ANIMAL 15995 4-N0V-86 EXAMINED * NO SIGNIFICANT LESIONS ANIMAL 15960 4-N0V-86 EXAMINED - NO SIGNIFICANT LESIONS ANIMAL 16032 4-N0V-86 EXAMINED - NO SIGNIFICANT LESIONS animal 16029 5-N0V-86 CORPORA LUTEA GROSS; AMNI0T1C SACS GROSS: PLACENTAS GROSS; ABSENT ABSENT ABSENT ANIMAL 15943 S-NOV-86 AMNIOTIC SACS GROSS: PLACENTAS GROSS: ABSENT ABSENT ANIMAL 16049 5-N0V-86 EXAMINED - NO SIGNIFICANT LESIONS ANIMAL 15968 5-N0V-86 EXAMINEO - NO SIGNIFICANT LESIONS ANIMAL 15988 5-N0V-86 EXAMINED - NO SIGNIFICANT LESIONS ANIMAL 15956 5-N0V-86 EXAMINEO - NO SIGNIFICANT LESIONS ANIMAL 15952 6-N0V-86 EXAMINED - NO SIGNIFICANT LESIONS AMIUAL 16013 6-N0V-86 EXAMINED - NO SIGNIFICANT LESIONS SL 042654 Report 50-514 Appendix 3 Page 6 GROUP: ANIMAL DEVELOPMENTAL TOXICITY STUDY OF INHALED 1,1,1-TRICHlOROETHANE IN NEW ZEALAND WHITE RABBITS DAM GROSS EXAMINATION AT TIME OF LAPAROTOMY M - 3000 PPM FEMALE ALL DEATHS COMBINED 15998 6-N0V-B6 EXAMINED - NO SIGNIFICANT LESIONS ANIMAL 16018 6-N0V-B6 EXAMINED - NO SIGNIFICANT LESIONS ANIMAL 15950 6-N0V-86 EXAMINED - NO SIGNIFICANT LESIONS ANIMAL 15958 6-N0V-86 EXAMINED - NO SIGNIFICANT LESIONS ANIMAL 15973 7-N0V-86 CORPORA LUTEA GROSS: AMNIOTIC SACS GROSS: PLACENTAS GROSS: ABSENT ABSENT ABSENT ANIMAL 15987 7-N0V-86 EXAMINED - NO SIGNIFICANT LESIONS ANIMAL 16011 7-N0V-86 EXAMINED - NO SIGNIFICANT LESIONS ANIMAL 16021 7-NOV-flfi ANIMAL 16048 7-N0V-86 EXAMINED - NO SIGNIFICANT LESIONS CORPORA LUTEA GROSS: AMNIOTIC SACS GROSS: PLACENTAS GROSS: ABSENT ABSENT ABSENT ANIMAL 15942 7-N0V-B6 EXAMINED - NO SIGNIFICANT LESIONS SL 042655 Report 50-514 Appendix 3 Page 7 GROUP: ANIMAL DEVELOPMENTAL TOXICITY STUDY Of INHALED 1 , 1 , 1 -TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS DAM GROSS EXAMINATION AT TIME OF LAPAROTOMY R - 6000 PPM FEMALE ALL DEATHS COMBINED 16000 4-N0V-86 CORPORA LUTEA GROSS: UTERUS GROSS: AMNIOTIC SACS GROSS: PLACENTAS GROSS; ABSENT CYST - RIGHT HORN 2X2MM CLEAR ABSENT ' ABSENT ANIMAL 16009 4-N0V-B6 EXAMINED - NO SIGNIFICANT LESIONS ANIMAL 15978 4-N0V-86 EXAMINED - NO SIGNIFICANT LESIONS ANIMAL 15999 4-N0V-86 EXAMINED - NO SIGNIFICANT LESIONS ANIMAL 16001 4-N0V-B6 EXAMINED - NO SIGNIFICANT LESIONS ANIMAL 15972 4-N0V-86 EXAMINED - NO SIGNIFICANT LESIONS ANIMAL 16012 5-N0V-B6 UTERUS GROSS: AMNIOTIC SACS GROSS: PLACENTAS GROSS: CYST RIGHT MULTIPLE CLEAR FLUID ABSENT ABSENT ANIMAL 16044 5-N0V-B6 EXAMINED - NO SIGNIFICANT LESIONS ANIMAL 16007 5-N0V-B6 EXAMINED - NO SIGNIFICANT LESIONS ANIMAL 16022 5-N0V-B6 EXAMINED - NO SIGNIFICANT LESIONS ANIMAL 16019 5-N0V-86 ANIMAL 16040 5-N0V-86 PLACENTAS GROSS: CONTAINS BLOOO (BY HEMASTIX) +++FOR BLOOD EXAMINED - NO SIGNIFICANT LESIONS SL 042656 Report 50--51A Appendix 3 Page 8 GROUP: ANIMAL DEVELOPMENTAL TOXICITY STUDY OF INHALED 1.1. 1 -TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS DAM GROSS EXAMINATION AT TIME OF LAPAROTOMY R - 6000 PPM FEMALE ALL DEATHS COMBINED 1S9S9 6-N0V-B6 UTERUS GROSS: MISSING filGHT UTERINE HORN FALLOPIAN TUBE AND OVARY NORMAL' ANIMAL 15948 6-N0V-86 EXAMINED - NO SIGNIFICANT LESIONS ANIMAL 16014 6-N0V-B6 EXAMINED - NO SIGNIFICANT LESIONS ANIMAL 15951 6-NOV-86 EXAMINED - NO SIGNIFICANT LESIONS ANIMAL 15982 6-N0V-86 EXAMINED - NO SIGNIFICANT LESIONS ANIMAL 16023 6-N0V-86 EXAMINED - NO SIGNIFICANT LESIONS ANIMAL 16043 7-N0V-86 AMNIOTIC SACS GROSS: PLACENTAS GROSS: ABSENT ABSENT ANIMAL 15989 ANIMAL 15984 7-N0V-H6 7-N0V-86 EXAMINED - NO SIGNIFICANT LESIONS EXAMINED - NO SIGNIFICANT LESIONS ANIMAL 15992 7-N0V-86 CORPORA LUTEA GROSS: AMNIOTIC SACS GROSS: PLACENTAS GROSS: ABSENT ABSENT ABSENT ANIMAL 16033 7-N0V-86 EXAMINED - NO SIGNIFICANT LESIONS ANIMAL 15955 7-N0V-B6 EXAMINED - NO SIGNIFICANT LESIONS SL 042657 s S9Z$q Co E~* DEVELOPMENTAL TOXICITY STUDY OF INHALED 1,1,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS GRAVID UTERINE WEIGHT AND NET MATERNAL BODY WEIGHT IN GRAMS FEMALE GROUP: 0 PPM PREGNANCY INITIAL STATUS BODY WT. TERMINAL BODY WT. UTERUS NET BODY WEIGHT NET BODY WT. CHANGE LIVER 16045 16002 15991 16006 15960 15993 15953 15994 15961 16036 16020 1604! 16005 15954 16028 16047 16016 15967 15990 15946 16025 16003 16039 G NG G G G G G G G G G G G G G G G G G G G G G 3149.30 3250.40' 3352.70 3464.90 3647.60 3633.00 3584.70 3507.00 3499.40 3399.70 3279.00 3143.60 3159.70 3339.00 3353.90 3454.30 3641.40 3756.40 3656.50 3592.50 3515.30 3430.30 3189.80 3430.00 3676.10 3903.80 3890.50 4017.10 4321.40 4064.40 3968.00 3717.60 3876.10 3874.40 3519.50 3705.30 3723.40 3941.70 3640.70 4353.10 3831.30 4027.50 4309.70 3814.80 4011.10 3771 . 70 553.66 30.63 594.00 473.89 594.88 613.41 583.06 553.77 586.43 623. 16 603. 15 466.52 604.84 580.02 450.62 603.24 630.14 610.32 56.57 555.63 646.22 274.95 356.93 2676.14 3645.27 3309.60 3416.6! 3422.22 3707.99 3501.32 3434.23 3131.17 3252.94 3271.25 3052.98 3100.46 3143.38 3491.08 3037.46 3722.96 3220.96 3970.93 3754.07 3168.58 3736.15 3414.77 -273.16 394.87 -42.90 -46.29 -225.56 -125.01 -83.38 -72.77 -368.23 -146.76 -7.75 -90.82 -59.24 -195.62 137.18 -416.84 8 1.56 -535.42 312.43 161.57 -346.72 305.85 224.97 89.40 102.31 82.47 94.00 85.06 91 .63 61.87 96.53 76.49 87.89 85.80 74.49 92.60 77.24 94.49 70.76 99.70 83.54 91.13 108.47 79.38 100.75 119.30 MEAN s.o. N 3452.38 196.11 22 3897.67 239. 19 22 527.98 140.25 22 3369.89 280.33 22 -82.50 227.72 22 89.32 11.54 22 G=GRAVID NG=NON GRAVID WEIGHT(S) NOT INCLUDED IN CALCULATION OF MEAN Report 50~5 1 A Appendix 3 Page 9 SL 042659 DEVELOPMENTAL TOXICITV STUDV OF INHALED 1 , 1 , 1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS GRAVID UTERINE WEIGHT AND NET MATERNAL BOOV WEIGHT IN GRAMS FEMALE GROUP: 1000 PPM PREGNANCY INITIAL STATUS BODY WT. TERMINAL BODY WT. UTERUS NET BOOV WEIGHT NET BODY WT. CHANGE LIVER 16037 16046 15996 15975 15983 15986 16026 1598 1 16038 15963 15957 16010 15976 16015 15945 15944 16030 15971 15964 15962 16027 16031 15965 G NG G G G G G G G G G G G G G G G G G G G NG G 2974.00 3279.60 3433. 10 3571.10 3637.90 3658.80 4210.80 3530.30 3495.20 3376.60 3262.70 3t53.70 3172.10 3340.50 3426.40 3501.40 3570.30 3896.90 3914.00 3727.50 3552.90 3504.30 3231.80 3453.20 3422.70 3793.30 3950.90 3954.00 4082.20 5008.20 3920.20 3393.70 3540.20 3657.70 3481.40 3301.50 3438.40 3549.10 3797.50 4025.50 4290.80 4231.30 4029.10 3804.60 3742.00 3510.90 436.22 34.76 234.31 586.3 1 417.03 383.46 630.59 373.40 33.84 416.59 422.42 397.74 48.84 331.49 496.89 45.98 602.63 649.83 574.56 62.94 43.63 31.35 608.59 3016.98 3387.94 3558.99 3364.59 3536.97 3698.74 4377.61 3546.80 3359.86 3123.61 3235.28 3083.66 3252.66 3106.91 3052.21 3751.52 3422.87 3640.97 3656.74 3966.16 3760.97 3710.65 2902.31 42.98 108.34 125.89 -206.51 - 100.93 39.94 166.81 16.50 -135.34 -252.99 -27.42 -70.04 80.56 -233.59 -374. 19 250.12 -147.43 -255.93 -257.26 238.66 208.07 206.35 -329.49 99.83 86.36 0.00 96.16 99.32 97.48 116.51 118.68 79.99 97.65 69.99 8 1.20 96.80 87.33 69.88 138.64 93.18 86.85 131.23 112.88 90.98 134.92 77.07 MEAN S.D. N 3506.57 285. 12 21 3819.70 395.15 21 371.30 214.04 21 3448.40 359.84 21 -58 . 1 7 192.74 21 92.46 27.97 21 G=GRAVID NG=NON GRAVID WEIGHT(S) NOT INCLUDED IN CALCULATION OF MEAN Repo rt 50-514 Appendi 3 Page to C/1 r 042660 DEVELOPMENTAL TOXICITY STUDY OF INHALED t,1,I-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS GRAVID UTERINE WEIGHT AND NET MATERNAL BODY WEIGHT IN GRAMS FEMALE GROUP: 3000 PPM PREGNANCY INITIAL STATUS BODY WT. TERMINAL BODY WT. UTERUS NET BODY WEIGHT NET BODY WT. CHANGE LIVER 16034 15947 15979 15995 15960 16032 16029 15943 16049 15968 15988 15956 15952 16013 15998 16018 15950 15958 15973 15987 16011 16021 16048 15942 G G G G G G NG NG G G G G G G G G G G NG G G G NG G 3235.00 3249.10 3403.10 3598.50 3630.80 3895.90 3811.00 .3501.70 3438.70 3397.90 3355.20 3153.40 3119.60 3284.60 3401.90 3450.90 3586.30 4310.90 3755.40 3661.90 3545.20 3513.20 3480.80 3389.90 3706.20 3661.60 3929.70 3647.20 4115.30 4318.40 3826.80 3954.00 3888.00 3508.50 3652.30 3705.20 3262.20 3408.80 3405.00 3997.70 3789.70 4317.60 4013.70 4037.70 3621.00 3817.40 3683.50 3871.10 491.91 483.54 668.38 426.B1 672.00 586.90 47.06 40.98 497.00 484.75 536.18 490.88 413,49 541.01 235.68 638.63 334.37 678.97 54.21 204.92 496.49 324.98 33.72 642.34 3214.29 3178.06 3261.32 3220.39 3443.30 3731.50 3779.74 3913.02 3391.00 3023.75 3116.12 3214.32 2848.71 2867.79 3169.32 3359.07 3455.33 3638.63 3959.49 3832.78 3124.51 3492.42 3649.78 3228.76 -20.71 -71.04 -141.78 -378.11 -187.50 -164.40 -31.26 411.32 -47,70 -374. 15 -239.08 60.92 -270.89 -416.81 -232.58 -91.83 -130.97 -672.27 204.09 170.88 -420.69 -20.78 168.98 -161.14 93.61 88.90 90.19 78.16 8 1.27 106.47 88.65 8 1.71 90.50 78.15 .77.50 97.41 67.50 7 1.39 117.77 9B . 98 91.94 106.24 116.79 117.18 76.21 t)6.13 141.70 76.69 MEAN S.D. N 348 1 . 10 269.79 ' 20 3783.03 285.93 20 492.46 139,05 20 3290.57 258.27 20 - 190.53 194.55 20 91.11 15.54 20 G=GRAVID NG=NON GRAVID WEIGHT(S) NOT INCLUDED IN CALCULATION OF MEAN Report 50~5 1 4 Appendi* 3 Page I 1 SL 042661 DEVELOPMENTAL TOXICITY STUDY OF INHALED 1, 1 , 1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS GRAVID UTERINE WEIGHT AND NET MATERNAL BOOV WEIGHT IN GRAMS FEMALE GROUP: 6000 PPM PREGNANCV INITIAL STATUS BODV WT. TERMINAL BOOV WT. UTERUS NET BODY WEIGHT NET BODV WT. CHANGE LIVER 16008 16009 15978 15999 16001 15972 16012 16044 16007 16022 16019 16040 15959 15948 16014 15951 1598 2 16023 16043 15989 15984 15992 16033 15955 NG G G G G G NG G G G G G G G G G G G G G G NG G G 3230.60 3240.20 3373.70 3544.10 3621.80 3920.40 3657.90 3531 .10 3487.20 3407.30 3309.00 3112.30 3049.80 3346.80 3350.80 3567.00 3635.30 3836.60 4301.00 3720.40 3576.10 3527.80 3421.30 3209.60 3342.80 3809.20 3753.00 3667.70 3923.80 4237.60 3768.40 3829.00 3970.90 3520.90 3765.10 3354.20 3642.10 3510.50 3809.10 4010.90 3865.30 3666.30 4419.80 4184.90 3800.50 3841.10 3577.30 3435,40 25.52 387.76 537.97 506.61 381.40 437.88 47.05 438.70 538.69 339.77 352.94 513.11 472.65 481.82 554.22 531.94 324.3 1 330.91 37.11 595.19 529.25 35.79 318.85 399.79 3317.28 3421.44 3215.03 3161.09 3542.40 3799.72 372!.35 3390.30 3432.21 3181.13 3412.16 2841.09 3169.45 3028.68 3254.88 3478.96 3540.99 3335.39 4382.69 3589.71 3271.25 3805.31 3258.45 3035.61 86.48 181.24 -158.67 -383.01 -79.40 -120.68 63.45 -140.80 -54.99 -226.17 103.16 -271.21 119.65 -318.12 -95.92 -88.04 -94.31 -501 .2 1 81.69 - 130.69 -304.85 277.51 - 162.85 - 173.99 131.46 75.30 101.17 8 1.55 114.36 108.28 131.56 98.96 90.98 92.67 98.58 82.75 88.63 86.86 82.98 85.88 106.87 92.84 131.93 97.79 100.88 127.38 104.23 81.87 MEAN S. 0. N 3502.94 286.73 21 3797.79 266.90 21 429.09 124.63 21 3368.70 318.29 21 -134.25 169.02 21 95.49 13.22 21 G=GRAVID NG=NON GRAVID WEIGHT(S) NOT INCLUOEO IN CALCULATION OF MEAN Repor t bO-5 14 Appendia 3 Page 1 2 M e* 0 N) 01 Oi N) DEVELOPMENTAL TOXICITY STUDY OF INHALED 1,!,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS FETAL DATA AT TIME OF LAPAROTOMY FEMALE GROUP: G - 0 PPM VIABLE FETUSES DEAD FETUSES EARLY RESORPTIONS LATE RESORPTIONS IMPLANTATION SITES CORPORA LUTEA SEX ANIMAL AM F RIGHT LEFT RIGHT LEFT RIGHT LEFT RIGHT LEFT RIGHT LEFT RIGHT LEFT HORN HORN TOTAL HORN HORN TOTAL HORN HORN TOTAL HORN HORN TOTAL HORN HORN.TOTAL OVARY TOTAL 16045 16002 15991 16006 15980 15993 15953 15994 15961 16036 16020 16041 16005 15954 16028 16047 16016 15967 15990 15946 16025 16003 16039 36 5 NON GRAVID 72 1 35 4 54 3 54 3 54 4 36 1 54 4 37 6 55 `7 42 3 65 6 35 3 34 5 73 3 83 6 46 6 00 0 49 8 26 7 21 0 23 3 TOTAL 89 96 MEAN 4.0 4.4 S.D. 1.9!2.2 22 GRAVID 88 4.0 2.3 4 8 4 6 6 5 8 5 4 3 3 5 5 2 7 5 4 0 5 3 3 2 97 4.4 1.9 9 9 8 9 9 9 9 9 10 10 6 11 a 7 10 11 10 0 13 10 3 5 185 8.4 2.8 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 1 0 0 0 0 0 2 0.. 1 0..3 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0..0 0..0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 1 0 0 0 0 0 2 0. 1 0 .3 0 0 1 0 1 0 0 0 0 0 0 0 0 0 0 0 0 3 1 0 0 0 6 0.3 0.7 0 1 1 0 0 1 0 0 0 0 0 1 2 1 0 1 0 5 0 0 0 a 13 0.6 1.1 0 1 2 0 1 1 0 0 0 0 0 I 2 1 0 1 0 6 1 0 0 0 19 0.9 1.7 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0..0 0. 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0,.0 0..0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 .0 0 .0 5 1 5 3 4 4 1 4 6 7 4 6 3 5 3 6 7 3 9 7 0 3 96 4.4 2.2 49 9 10 5 10 69 6 . 10 6 10 8 9 5. 9 4 10 3 ' 10 37 \6 12 7 10 3' 8 7 10 6 12 4 . 11 58 5 14 3 10 3 ' 3. 25 1 to 206 5.0 . 9.4 1.8 '2.3 5 2 5 5 4 4 2 B 6 7 5 6 3 6 3 6 7 3 9 7 6 4 1 13 5.1 1.9 6 11 79 5 10 6 11 6 10 6 10 8 10 5 13 4 10 3 10 38 6 12 8 T1 39 8 )1 6 12 4 11 58 5 14 3 10 4 to 37 1 1 4 227 5 . 2 10.3 1 .. 7 1.6 Report 50-514 Append:* 3 Page l3 r1 F to VIABLE DEVELOPMENTAL TOXICITY STUDY OF INHALED 1,1,1-TRICHL0R0ETHANE IN NEW ZEALAND WHITE RABBITS FETAL DATA AT TIME OF LAPAROTOMY FEMALE GROUP: I - 1000 PPM FETUSES DEAD FETUSES EARLY RESORPTIONS LATE RESORPTIONS IMPLANTATION SITES CORPORA LUTEA SEX ANIMAL # M F RIGHT LEFT RIGHT LEFT RIGHT LEFT RIGHT LEFT RIGHT LEFT RIGHT LEFT HORN HORN TOTAL HORN HORN TOTAL HORN HORN TOTAL HORN HORN TOTAL HORN HORN TOTAL OVARY TOTAL 1603 7 i6 6 1 7 000 0 00 0006 1 76 1 7 16046 NON GRAVID 15996 20 1 1 2 0 0 0 1 2 3 3 3 6 5 6 11 4 8 12 15975 63 639 1 0 1 0 0 0 0 0 0 7 ,3 10 7 4 11 159B3 51 336 000 1 0 1 0004 3 743 7 15986 22 3 1 4000 2 35 0005 4 954 9 16026 55 3 7 10 0 0 0 0 0 0 0 0 0 3 7 10 3 8 1 t 1598 1 22 1 34000 3 25 0 004 59A5 9 16038 00 00 000 0 4 26 0 004 2 600 0 15963 24 42 6 1 1 20 00 0005 3 853 8 15957 16010 25 42 5 2 7 1 0 1 0 0 0 0 0 0 6 2 . 8 6 4 10 5 1 6 000 1 0 1 0006 1 763 9 15976 16015 15945 15944 00 41 54 00 0 0 0 0 0 0 2 5 7 0 0 0 2 5 ' 7 3 5 8 4 1 5 0 0 0 0 0 0 0 0 0 4 1 5 6 A 10 3 6 9 0 0 0 1 0 1 0 0 0 4 6 ' 10 A 6 10 0 0 0 0 0 0 4 5 9 0 0 0 A 5 9 5 6 11 16030 15971 82 63 4 6 10 1 0 \ 0 0 0 0 0 0 5 6 1 1 5 6 1 1 2 7 9 0 0 0 0 0 0 0 0 0 2 7 9 3 8 11 15964 52 3 4 7 0 0 0 3 1 4 0 0 0 6 5 1 1 7 5 12 15962 00 000000 4 37 0004 3 74 3 7 16027 00 000000 2 02 0 00 2 0 200 0 16031 NON GRAVID 15965 54 3 6 9 0 1 I 0 0 0 0 0 0 3 ' 7 10 A 7 11 TOTAL 64 46 MEAN 3 .0 2.. 2 S.O. 2 .4 1 ..9 21 GRAVID 56 54 t 10 4 2 6 28 23 51 3 3 6 91 82. 173 91 93 184 2.7 2.6 5.2 0.. 2 0.. 1 0 .3 1.3 1 . 1 2.4 0 . 1 0 . 1 0 .3 4.3 3.9 8.2 4.3 4 . 4 8.8 2.0 2.5 3.6 0.. 4 0.. 3 0..6 1.5 1.7 2.9 0 .7 0 .7 1 ,.3 1.4 2.2 2. 2 1.9 2 . 4 3.3 Report 50-514 Append i * 3 Page 14 w r o > W DEVELOPMENTAL TOXICITV STUDV OF INHALED 1,1,1-TR1CHLOROETHANE O' IN NEW ZEALAND WHITE RABBITS 2 FETAL DATA AT TIME OF LAPAROTOMV FEMALE GROUP: M - 3000 PPM VIABLE FETUSES DEAD FETUSES EARLV RESORPTIONS LATE RESORPTIONS IMPLANTATION SITES CORPORA LUTEA SEX ANIMAL # M F RIGHT LEFT RIGHT LEFT RIGHT LEFT RIGHT LEFT RIGHT LEFT RIGHT LEFT HORN HORN TOTAL HORN HORN TOTAL HORN HORN TOTAL HORN HORN TOTAL HORN HORN TOTAL OVARV TOTAL 16034 54 2 7 9 0 0 0 0 0 0 0 0 0 2 7 .9 2 7 9 15947 61 2 5700 000 0000 257 268 15979 47 7 4 1 1 0 0 0 0 1 1 0 0 0 7 5 1 2 8 5 13 15995 25 5 2 7 1 0 1 0 0 0 0 0 0 6 '2 8 7 3 10 15960 92 5 6 11 0 0 0 1 0 1 0 0 0 6 6 12 6 6 12 16032 74 4 7 11 0 0 0 0 0 0 0 0 0 4 7 11 4 7 t1 16029 NON GRAVID 15943 NON GRAVID 16049 54 4 5 9 0 0 0 0 0 0 0 0 0 4 5 9 4 6 to 1 5968 53 5 3 8 0 0 0 0 0 0 0 0 0 5 3 .8 4 4 8 1 5966 27 3 6 9 0 0 0 0 0 0 0 0 0 3 6 9 4 6 10 15956 24 4260 1 1 0 0 000 0 437 459 15952 36 2 7 9 0 0 0 0 1 1 0 0 0 2 8 10 2 8 10 16013 55 5 5 10 1 0 1 1 0 1 0 0 0 7 5 1 2 6 6 1 2 15998 02 0 2 2 6 3 9 0 0 0 0 0 0 6 5 11 7 5 1 2 16016 57 6 6 12 0 0 0 0 0 0 0 0 0 6 6 12 6 7 13 15950 23 4 5 0 0 0 3 1 4 0 0 0 7 2 9 7 3 10 15956 47 6 5 1 1 0 0 0 0 0 0 0 0 0 6 5 '11 7 6 13 15973 NON GRAVID 159B7 02 2 0 2 0 0 0 0 0 0 0 0 0 2 0 2 2 10 1 2 16011 35 1 7800 0000000 1 78 1 8 9 16021 41 1 4 5 0 0 0 0 0 0 0 0 0 1 4 .5 4 4 8 16048 NON GRAVID 15942 94 4 9 13 0 0 0 0 0 0 0 0 0 4 . 9 13 ' 4 9 13 TOTAL 82 83 MEAN 4 . 1 4 .2 S.O. 2 .5 2 .0 20 GRAVID 72 3 .6 1 .9 93 4.7 2.4 165 8.3 3. 1 8 0 .4 1 .4 4 0.2 0 .7 12 0.6 2.0 5 0.. 2 0..7 3 0.2 0 .4 8 0,.4 0,.9 0 0..0 0..0 0 0..0 0..0 0 0 .0 0 .0 85 4.3 2. 1 100 5.0 2.2 185 9.3 2.7 91 4.6 2. 1 121 6. 1 1 .8 212 10.6 1 .8 Report 50-514 Appendi* 3 Page 15 SL 042665 VIABLE DEVELOPMENTAL TOXICITY STUDY OF INHALED 1.1,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS FETAL DATA AT TIME OF LAPAROTOMY FEMALE GROUP: R - 6000 PPM FETUSES DEAD FETUSES EARLY RESORPTIONS LATE RESORPTIONS IMPLANTATION SITES CORPORA LUTEA SEX ANIMAL * M F RIGHT LEFT RIGHT LEFT RIGHT LEFT RIGHT LEFT RIGHT LEFT RIGHT LEFT HORN HORN TOTAL HORN HORN TOTAL HORN HORN TOTAL HORN HORN TOTAL HORN HORN TOTAL OVARY TOTAL 16008 NON GRAVID 16009 24 1 5 6 0 0 0 0 0 0 0 0 0 1 5 6 5 5 10 15978 53 7 1 8 000 0000 007 1 8 7 t 8 15999 44 448 000 0000 004 4 8 44 0 16001 42 2 4 6 0 0 0 1 0 1 0 0 0 3 4 7 5 6 1\ 1597Z 53 3 5 e 0 2 2 0 0 0 0 0 0 3 7 10 3 7 1 0 16012 NON GRAVID 16044 17 2 6 8 0 0 0 0 2 2 0 0 0 2 8 10 3 6 1 1 16007 63 4 5 9 0 0 0 0 0 0 0 0 0 4 5 9 4 6 10 16022 32 235 000 0 1 t 0002 4635 8 16019 22 2 2 4 1 1 2 2 1 3 1 0 t 6 4 10 6 4 1 0 16040 64 4 6 10 0 0 0 0 0 0 0 0 0 4 6 10 4 7 \ 1 15959 61 07 7000 0 000 000 7 7 27 9 1594B 44 268 000000 0 002 6 8 268 16014 15951 64 46 5 6 5 10 4 to 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 5 6 5 10 4 10 7 6 5 )2 S 11 1598 2 15 066000 0 1 1 0 000 7 7 07 7 16023 05 1 4 5 000 1 1 20002 5 7 268 16043 00 0000003 t4 0 003 1 r4 303 15989 15984 73 2B 5 6 5 10 4 to 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 5 '5 64 10 10 6 6 5 11 5 \\ 15992 NON GRAVID 16033 15955 31 24 1 3 4 000 2020 003 3 63 3 6 5 1 6 0 0 0 2 t 3 0 0 0 7 ,2 9 7 4 TOTAL 73 75 MEAN 3.5 3. 6 S.D. 2.1 2. 0 62 3.0 2.2 86 4. 1 1 .9 148 7.0 2.6 1 0.0 0.2 3 0. 1 0.5 4 0.. 2 0..6 11 0.5 0.9 B 0..4 0..6 19 0.9 t .3 1 0.. 0 0.. 2 0 0 .0 0 .0 1 0 .0 0.. 2 75 3.6 2. 1 97 4.6 1.9 172 8.2 1.8 8B 4.2 2.0 106 5.0 2.0 1 94 92 2.2 Report 50-514 Append!* 3 Page 1 6 SL 042666 * DEVELOPMENTAL TOXICITV STUDY OF INHALED 1,1.1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS INDIVIDUAL FETAL WEIGHTS (GRAMS) FEMALE GROUP: G - 0 PPM IMPLANT # ANIMAL 1 2 3 4 5 6 7 8 9 10 1 1 12 15946 15953 15954 15961 15967 15980 15990 15991 15993 15994 16003 16005 16006 16016 16020 16025 16026 1 6036 16039 1 6041 16045 16047 28.26 36.92 50.16 47.68 43.13 51.43 E 48.76 52.66 48.56 /58.63 41.44 40.25 41.20 49.62 45.99 43.88 43.89 50.20 52.31 44 . 14 46.36 30.59 42.95 43.23 41.95 46.27 45.53 E 7 EP 47.54 /42.63 51.68 41.10 34.09 43.50 42.97 48.03 41.64 40.38 43.13 45.98 43.14 46.96 28.88 39.02 49.56 45.39 41.35 43.80 E 39.33 EP 45.35 58.47 42.46 36.91 44.64 42.18 45.39 41.78 45.80 48.28 0.60D 36.72 38.37 29.97 36.99 7 EP 45.20 36.28 /43.76 7E 34.31 47.45 40.67 41.9 1 EP 44.34 36.00 46.78 38.46 43.19 747.69 48.88 34.40 739.47 31.52 743.85 47.48 741.81 29.84 38.17 E 37.85 749.58 43.33 31.31 43.30 27.84 27.98 47.39 47.39 41.15 49.34 749.50 38.64 37.66 25.28 41.12 40.23 43.07 4.07D 44.70 E 44.55 45.06 35.27 18.53 44.75 49.96 45.06 32.25 47.98 E 47.23 43.07 33.12 36.67 735.90 37.09 34.72 38.71 7 EP 37.47 736.03 34.56 739.51 39.25 43.11 45.69 747.31 52.01 743.77 40.57 59.35 45.7 1 44.5 7 EP 35.40 46.67 40.95 738.54 46.76 E 45.50 41.20 41.27 35.84 35.69 35.0 1 743.40 749.92 44.98 40.25 45.03 40.26 36.20 37.47 52.09 45.83 43.32 48.79 42.56 47.31 45.27 729.55 EP EP 43.18 ' 43.58 54.73 31.83 EP 30.51 44.83 52.00 34.04 42.09 37.76 44.08 52.26 46.41 48.21 47.08 44.07 45.54 24.96 42.73 38.76 EP 31.67 EP 47.33 0= DEAD FETUS, EP= EARLY RESORPTION W/PLACENTA, E = EARLY RESORPTION, LF= LATE RESORPTION W/FETAL TISSUE, LB= LATE RESORPTION W/LIMB BUDS, 7 = CERVIX POSITION 13 33.88 14 31.84 15 Report 50-514 Appendi* 3 Page 1 7 SL 042667 DEVELOPMENTAL TOXICITY STUDY OF INHALED I,1,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS INDIVIDUAL FETAL WEIGHTS (GRAMS) FEMALE GROUP: I - 1000 PPM IMPLANT # ANIMAL 1 2 3 4 5 6 7 8 9 10 1 1 15944 15945 15957 15962 15963 15964 15965 1597 1 15975 15976 1598 1 15983 15986 15996 16010 16015 1 6026 16027 16030 16037 16038 E 44.25 38.74 E 45.5 7 EP 39.06 57.36 8.730 E EP 47.82 59.99 EP 41.11 43.39 46.98 E 45.98 40.56 E 6 38,09 34.87 E 43.21 53.26 45.61 46.79 51.66 E 48.51 41.85 EP 47.24 48.35 41 .88 54.06 E 43.57 44.70 E 27.26 36.78 E 35.74 EP 41.66 /40.10 37.86 /E EP E 55.28 LF 50.46 42.64 45.08 7 36.77 43.84 E E EP 31.94 E 1.24D EP /38,54 41.38 44.19 E EP 40.83 54.47 LF 47.85 38.57 743 . 15 7E 734.54 24.19D 7E 41.81 49.02 33.29 47.23 44.50 E 755.09 739.43 EP LF 42.26 744.00 27.13 E 35.75 35.77 E 739.78 50.81 2 1 .940 50. 19 37.21 E 48.63 45.30 7 EP / EP EP 32.63 E 38.63 736.85 E 5.92D 752.66 36.33 42.28 38.29 E 51.01 48.42 54.68 LF 744.77 44.17 E 37.21 38.28 43.32 51.57 42.51 55.04 745. 18 EP EP LF 46.25 32.47 31.71 E 0.42D 746.96 40.8 1 45.29 7E E 30.14 747.10 39.39 E 41.02 50.97 42.90 56.66 42.94 EP EP LF 49.47 40.93 42.63 EP 51.20 34.18 40.14 44.03 44.55 52.80 EP 52.07 0= DEAD FETUS, EP= EARLY RESORPTION W/PLACENTA, E = EARLY RESORPTION. LF= LATE RESORPTION W/FETAL TISSUE, LB= LATE RESORPTION W/LIMB BUDS, 7 = CERVIX POSITION Report 50-514 Appendi* 3 Page 18 SL 042668 i DEVELOPMENTAL TOXICITV STUDY OF INHALED 1,1,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS INDIVIDUAL FETAL WEIGHTS (GRAMS) FEMALE GROUP; M - 3000 PPM IMPLANT * ANIMAL 1 2 3 4 5 6 7 a 9 10 1 1 12 15942 15947 15950 15952 15956 15958 15960 15968 15979 15987 15988 15995 15998 16011 16013 16016 16021 16032 16034 16049 41.24 50.12 44.10 36.74 55.55 47.77 45.35 45.74 52.96 57.99 47.93 38.86 1 . 07D 42,48 EP 45.13 38.35 43.98 41.59 40.09 40.68 46.68 40.77 28.05 47.11 42.42 42.77 37.19 40.43 54.73 44.03 40.39 1.130 /42.68 34.59 41.66 /42.79 43.65 36.91 37.86 36.70 /43.34 EP /25.15 41.14 35.59 40.34 35.57 41.16 / 42.66 4.710 1.03D 36.49 30.85 37.76 36.05 46.12 /34.69 36.27 24.53 46.37 44.15 20.8 1 42.21 38.92 35.26 37.23 36.63 /41.74 35.07 0.89D 36.50 35. 19 23.12 36.66 41.46 34.29 35.88 /33.57 42.66 EP EP /40. 12 33.23 EP 42.42 35.53 28.54 32.52 1.320 41.43 1 .95D 26.41 40.67 /28.50 39.62 Z42.63 33.65 42.95 26.12 25.21 46.31 40.68 4 1.19 /42.22 36.22 36.07 40.57 1 .41D 39.45 37.13 27.30 24.52 31 .70 30.50 22.72 45.59 EP 22.25 53.570 Z44.68 /40.69 36.79 41.35 32.54 /42.25 / 1.12D 40.49 36.28 /37.56 21.02 29.67 33.50 39.92 /48.44 29.50 41.90 39.11 43.66 /38.42 43.04 43.54 36.49 45.12 /31.00 40.04 28.61 35.08 35.14 28.43 29.52 36.5 1 E 33.81 32.07 41.40 40.36 47.15 43.01 41.03 43.10 37.23 EP 46.77 5 1.43 1 . 17D 0.980 A 30.83 36.06 32.49 26.65 35.66 29.98 31.14 37.66 36.67 41.56 43.16 39.12 47; 61 51.31 34.90 45.19 0- DEAD FETUS, EP= EARLY RESORPTION W/PLACENTA, E = EARLY RESORPTION, LF= LATE RESORPTION W/FETAL TISSUE, LB= LATE RESORPTION W/LIMB BUDS, /= CERVIX POSITION 13 41.93 Report 50-514 Appendix 3 Page 19 * 042669 c/a r< DEVELOPMENTAL TOXICITY STUDY OF INHALED 1,1,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS INDIVIDUAL FETAL WEIGHTS (GRAMS) FEMALE GROUP; R - 6000 PPM IMPLANT * IIMAL 1 2 3 4 5 6 7 8 9 10 15948 15951 15955 15959 15972 15978 1598 2 15984 15989 15999 16001 16007 16009 16014 16019 16022 16023 16033 16040 16043 16044 46. 1 2 44.82 EP /48.22 36.17 48.47 /26.05 35.73 46.22 44.83 40.74 47.42 43.60 35.77 LF 43.48 EP EP 35.41 E 42.48 39.28 38.61 45.30 43.44 36.42 37.49 37.17 37.26 41.47 42.86 EP 43.14 /41.96 31.66 EP 38.40 38.55 EP 4 1.35 E 43.21 / 44.82 36.89 27.28 43.32 34.78 39.32 29.24 33.76 44.29 40.48 40.03 45.67 34.74 32.91 EP /42.5B /32.8B 49.29 35.61 E /24.80 46.15 27.90 45.19 48.25 /38.98 42.11 EP 31.48 35.31 42.67 /44.00 43.98 4 1.63 40.51 38.09 39.22 35.19 /4 7.28 36.03 /E 18.87 46.04 28.11 EP 49.65 27.79 43.4! 29 .92 28.87 36.85 /46.04 43.85 /37.59 46.26 35.67 50.47 EP 41.59 51.99 /3t.95 33.95 33.30 42.01 48. 10 52.70 0.60D 44.15 34.89 31.60 Z36.56 43.94 39.92 31.52 49.65 /32.17 10.15D 46.21 43.93 49.46 35.66 43.71 Z39.52 47.71 43.16 35.88 40.74 43.72 /35.03 38.07 41.84 38.47 35.12 36.77 /33 . 1 1 EP 36.79 EP EP 30.54 37.44 /47.25 21.33 /50.36 39.30 36.07 42.33 39.90 35.83 46.76 35.37 42.00 37.92 EP 41 .71 15.150 36.34 32.40 38.35 36.83 46.59 46.49 39.82 EP 45.10 1 . 7 ID 39.44 33.59 39.67 43.43 0= DEAD FETUS, EP= EARLY RESORPTION W/PLACENTA, E = EARLY RESORPTION. LF= LATE RESORPTION W/FETAL TISSUE, LB= LATE RESORPTION W/LIMB BUDS, /= CERVIX POSITION Report 50~514 Appendix 3 Page 20 Report 50-514 APPENDIX 4 Malformations and Variations by Fetuses and by Litters (71 Pages) Pages 4A Gross (External) Findings ......................................................................... 1-19 4B Visceral (Including Craniofacial) Findings......................................1-29 4C Skeletal Findings............................................................................... 1-23 SL 042670 97^0 C/I FEMALES FROM G - 0 PPM r* 15946 15953 15954 15961 i ! DEVELOPMENTAL TOXICITY STUDY OF INHALED 1,1.1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS GROSS FETAL EXAMINATION AT TIME OF LAPAROTOMY IMPLANT # SEX FETAL GROSS OBSERVATION 1 LF# 1 2 LF# 2 3 LF# 3 4 LF# 4 5 LF# 5 6 LF# 6 7 LF# 7 8 9 LF# 8 10 LF# 9 1 1 LF# 1 0 1 2 LF# 1 1 13 LF# 1 2 14 LF# 13 1 LF# 1 2 LF# 2 3 LF# 3 4 LF# 4 5 LF# 5 6 LF# 6 7 LF# 7 8 LF# a 9 LF# 9 10 1 LF# i 2 LF# 2 3 LF# 3 4 5 LF# 4 6 LF# 5 7 LF# 6 B LF# 7 9 LF# B 10 1 LF# 1 2 LF# 2 3 LF# 3 4 LF# 4 5 LF# 5 6 LF# 6 7 LF# 7 8 LF# 8 9 LF# 9 F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F V ECCHYMOSIS - TRUNK LEFT SIDE EARLY(W/PLACENTAL TISSUE) F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS EARLY(W/PLACENTAL TISSUE) F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS EARLV(W/PLACENTAL TISSUE) F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS EARLY(W/PLACNTAL TISSUE) F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS OBSERVATION CODE: M=MALF0RMATI0N. V=VARIATION Report 50-5 14 Appendix 4A Page 1 SL 042672 1 DEVELOPMENTAL TOXICITY STUDY OF INHALED 1,1,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS GROSS FETAL EXAMINATION AT TIME OF LAPAROTOMY FEMALES FROM G - 0 PPM IMPLANT * SEX FETAL GROSS OBSERVATION 1 5967 1 LF* 1 2 LF* 2 3 LF* 3 4 LF* 4 5 LF* 5 6 7 LF* 6 8 LF* 7 9 LF* 8 10 LF* 9 1 1 LF*10 F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F DEAD DEAD AND MACERATED FETUS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS 15980 1 LF* 1 2 LF* 2 3 LF * 3 4 LF* 4 5 LF A 5 6 LF* 6 7 LF* 7 a LF* a 9 LF* 9 M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS 15991 i LF* 1 2 3 LF* 2 4 LF* 3 5 LF* 4 6 LF* 5 7 LF* 6 8 LF* 7 9 LF* e 10 LF* 9 F NO REMARKABLE OBSERVATIONS EARLY(W/PLACENT AL TISSUE) M V ECCHYMOSIS - TRUNK EXTERNAL GENITALIA F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS 15993 1 LF* 1 2 LF* 2 3 4 LF* 3 5 LF* 4 6 LF* 5 7 LF* 6 a LF* 7 9 LF* e 10 LF* 9 F NO REMARKABLE OBSERVATIONS M V ECCHYMOSIS - EXTREMITIES LEFT HIND FOOT EARLY(W/PLACENTAL TISSUE) M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS 15994 1 LF* 1 2 LF* 2 F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS OBSERVATION CODE: M-MA LFORMATION, V=VARIATION Report 50*5 1 4 Appendi* 4A Page 2 SL 042673 > DEVELOPMENTAL TOXICJTV STUDY OF INHALED 1,1.1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS GROSS FETAL EXAMINATION AT TIME OF LAPAROTOMY FEMALES FROM G - 0 PPM IMPLANT # SEX FETAL GROSS OBSERVATION 15994 3 LF# 3 4 LF# 4 5 LF# 5 6 LF* 6 7 LF# 7 8 LF# a 9 LF* 9 F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F V ECCHYMOSIS - TRUNK RIGHT FLANK ON LOWER BACK M NO REMARKABLE OBSERVATIONS 16003 1 LF# i 2 LF# 2 3 LF# 3 M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS 16005 1 LF# 1 2 LF# 2 3 LF# 3 4 LF* 4 5 LF# 5 6 LF# 6 7 LF# 7 8 LF# e 9 LF# 9 10 LF# 10 1 I LF# 1 t 12 F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS EARLY(W/PLACENTAL TISSUE) 16006 1 LF# 1 2 LF# 2 3 LF# 3 4 5 LF# 4 6 LF# 5 7 LF# 6 8 LF* 7 9 to LF# 8 M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS EARLY(W/PLACENTAL TISSUE) F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS EARLY(W/PLACENTAL TISSUE) M NO REMARKABLE OBSERVATIONS 16016 1 LF# 1 2 LF# 2 3 LF# 3 4 LF# 4 5 LF# 5 6 LF* 6 7 LF# 7 8 LF# 8 9 LF# 9 M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M V ECCHYMOSIS - EXTREMITIES FOR ELIM8 - BILATERAL V ECCHYMOSIS - TRUNK LOWER DORSAL BACK F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M V ECCHYMOSIS - HEAD TOP OF HEAD OBSERVATION CODE: M=MALFORMATION, V=VARIATION Report 50-514 Appendix 4A Page 3 SL 042674 DEVELOPMENTAL TOXICITY STUDY OF INHALED 1,1,t-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS GROSS FETAL EXAMINATION AT TIME OF LAPAROTOMY FEMALES FROM G - 0 PPM IMPLANT # SEX FETAL GROSS OBSERVATION 160)6 10 LF#?0 11 12 LF#1 1 M NO REMARKABLE OBSERVATIONS EARLY(W/PLACENTAL TISSUE) F NO REMARKABLE OBSERVATIONS 16020 1 LF# 1 2 LF# 2 3 LF# 3 4 LF# 4 5 LF# 5 6 LF* 6 7 LF* 7 8 LF* 8 9 LF* 9 10 LF* 1 0 M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS 16025 1 LF* 1 2 LF* 2 3 LF* 3 4 LF* 4 5 LF* 5 6 LF* 6 7 LF* 7 8 LF* 8 9 LF* 9 10 LF * 10 F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS 16028 1 LF* 1 2 LF* 2 3 LF* 3 4 LF* 4 5 LF* 5 6 7 LF* 6 8 LF* 7 F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS EARLY(W/PLACENTAL TISSUE) F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS 16036 1 1 LF* 1 2 LF* 2 3 LF* 3 4 LF* 4 5 LF* 5 6 LF* 6 7 LF* 7 8 LF# 8 9 LF# 9 10 LF# 1 0 M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS 1 6039 1 LF# 1 2 LF# 2 M NO REMARKABLE OBSERVATIONS F V ECCHVMOSIS - TRUNK RIGHT SIDE OBSERVATION CODE: M=MALFORMATION, V=VARIATION Report 50-514 Appendix 4A Page 4 SL 042675 FEMALES FROM G - 0 PPM 16039 16041 1 6045 16047 DEVELOPMENTAL TOXICITY STUDY OF INHALEO 1.1,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS GROSS FETAL EXAMINATION AT TIME OF LAPAROTOMY IMPLANT # SEX FETAL GROSS OBSERVATION 3 LF# 3 4 LF# 4 5 LF# 5 F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS 1 LF# 1 2 LF# 2 3 4 LF# 3 5 L F# 4 6 LF# 5 7 LF# 6 1 LF# 1 2 LF# 2 3 LF# 3 4 LF# 4 5 LF# 5 6 LF# 6 7 LF# 7 B LF# B 9 LF# 9 1 LF# 1 2 LF# 2 3 LF# 3 4 LF# 4 5 LF# 5 6 LF# 6 7 LF# 7 8 LF# 8 9 LF# 9 10 LF# 1 0 F NO REMARKABLE OBSERVATIONS M V ECCHVMOSIS - EXTREMITIES LEFT ARM LEFT LEG U DEAD OEAD AND MACERATED FETUS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M V ECCHYMOSIS - TRUNK LEFT FLANK iAT HIND LEG F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS OBSERVATION CODE: M=MALFORMATION, V=VARIATION Report 50-514 Appendik 4A Page 5 SL 042676 DEVELOPMENTAL TOXICITY STUDY OF INHALED 1,1,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS GROSS FETAL EXAMINATION AT TIME OF LAPAROTOMY FEMALES FROM I - 1000 PPM IMPLANT # SEX FETAL GROSS OBSERVATION 15945 1 LF# 1 2 LF# 2 3 LF# 3 4 5 LF# 4 6 LF# 5 7 LF# 6 B LF# 7 9 LF# 8 10 LF# 9 M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS EARLY(W/PLACENTAL TISSUE) F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS 15957 1 LF# 1 2 LF# 2 3 LF# 3 4 LF# 4 5 6 LF# 5 7 LF# 6 8 LF# 7 F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F DEAD DEAD AND MACERATED M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS 15963 1 LF# 1 2 LF# 2 3 LF# 3 4 5 LF# 4 6 LF# 5 7 B LF# 6 M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS U DEAD DEAD AND MACERATED FETUS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS U DEAD DEAD AND MACERATED FETUS F NO REMARKABLE OBSERVATIONS 15964 t 2 LF# 1 3 4 5 LF# 2 6 LF# 3 7 LF# 4 6 LF# 5 9 LF# 6 10 11 LF# 7 EARLY(W/PLACENTAL TISSUE) M NO REMARKABLE OBSERVATIONS EARLY(W/PLACENTAL TISSUE) EARLV(W/PLACENTAL TISSUE) M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS EARLY(W/PLACENTAL TISSUE) F NO REMARKABLE OBSERVATIONS 15965 ' 1 LF# t 2 LF# 2 3 LF# 3 4 LF# 4 5 LF# 5 6 M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F DEAD OBSERVATION CODE: M=MALFORMATION, V=VARIATION Report 50-514 Appe nd i * 4A Page 6 SL 042677 DEVELOPMENTAt TOXICITY STUDY OF INHALED 1,1,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS GROSS FETAL EXAMINATION AT TIME OF LAPAROTOMY FEMALES FROM I - 1000 PPM IMPLANT # SEX FETAL GROSS OBSERVATION 15965 7 LF# 6 8 LF# 7 9 LF# 8 10 LF# 9 F NO REMARKABLE OBSERVATIONS M EYE BULGE FLATTENED RIGHT F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS 1597 1 t LF# 2 LF# 3 LF# 4 LF# 5 LF# 6 LF# 7 LF# 8 LF# 0 9 LF# 9 M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS 15975 1 LF# 1 LF# 2 LF# 3 LF# 4 LF# 5 LF# 6 8 LF# 7 9 LF# 8 10 LF# 9 DEAO DEAD AND MACERATED FETUS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F ECCHYMOSIS - HEAD DORSAL SKULL 15981 LF# 1 LF# LF# LF# EARLY(W/PLACENTAL TISSUE) NO REMARKABLE OBSERVATIONS EARLY(W/PLACENTAL TISSUE) EARLV(W/PLACENTAL TISSUE) M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS EARLY(W/PLACENTAL TISSUE) EARLY(W/PLACENTAL TISSUE) 15983 1 LF# 1 2 LF# 2 3 4 LF# 3 5 LF# 4 6 LF# 5 7 LF# 6 M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS EARLY M NO REMARKABLE OBSERVATIONS F ECCHYMOSIS - TRUNK EXTERNAL GENITALIA M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS 15986 1 LF# 1 2 NO REMARKABLE OBSERVATIONS EARLY(W/PLACENTAL TISSUE) OBSERVATION CODE: M=MALFORMATION, V=VARIATION Report 50-514 Appendi* 4A Page 7 SL 042678 DEVELOPMENTAL TOXICITY STUDY OF INHALED t,I,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS GROSS FETAL EXAMINATION AT TIME OF LAPAROTOMY FEMALES FROM I - 1000 PPM IMPLANT # SEX FETAL GROSS OBSERVATION 15986 3 LF# 2 4 LF# 3 5 6 7 LF# 4 B 9 F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS EARLY(W/PLACENTAL TISSUE) EARLY(W/PLACENTAL TISSUE) F NO REMARKABLE OBSERVATIONS EARLY(W/PLACENTAL TISSUE) EARLY(W/PLACENTAL TISSUE) 15996 1 2 LF# 1 3 4 5 6 7 a 9 10 LF# 2 11 EARLY(W/PLACENTAL TISSUE) M NO REMARKABLE OBSERVATIONS LATE(W/FETAL TISSUE) LATE(W/FET AL TISSUE) LATE(W/FETAL TISSUE) EARLY(W/PLACENTAL TISSUE) LATE(W/FETAL TISSUE) LATE(W/FET AL TISSUE) LATE(W/FETAL TISSUE) M NO REMARKABLE OBSERVATIONS EARLY(W/PLACENTAL TISSUE) 16010 i LF# 1 2 LF# 2 3 LF# 3 4 LF# 4 5 LF# 5 6 7 LF# 6 M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS EARLY{W/PLACENTAL TISSUE) F NO REMARKABLE OBSERVATIONS 16015 1 LF# 1 2 LF# 2 3 LF# 3 4 LF# 4 5 LF# 5 M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS 160 26 ; 1 LF# t 2 LF# 2 3 LF# 3 4 LF# 4 5 LF# 5 6 LF# 6 7 LF# 7 8 LF# B 9 LF# 9 10 LF# 10 M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M M DOME-SHAPED HEAD SLIGHT F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS 16030 1 LF# 1 2 LF# 2 3 LF# 3 M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS OBSERVATION CODE: M=MALFORMATION, V=VARIATION Report 50-514 Appendi* 4A Page 8 L9 xfl r* vD FEMALES FROM I - 1000 PPM 16030 16037 DEVELOPMENTAL TOXICITY STUDY OF INHALED 1,1,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS GROSS FETAL EXAMINATION AT TIME OF LAPAROTOMY IMPLANT # SEX FETAL GROSS OBSERVATION 4 LF# 4 5 6 LF# 5 7 LF# 6 e LF# 7 9 LF# 6 10 LF# 9 1 1 LF# 1 0 F NO REMARKABLE OBSERVATIONS U DEAD UNABLE TO EXAMINE M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS 1 LF# 1 2 LF# 2 3 LF# 3 4 LF# 4 5 LF# 5 6 LF# 6 7 LF# 7 F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS OBSERVATION CODE: M=MALFORMATION, V=VARIATION Report 50-514 Appendi* 4A Page 9 r* <y8 9 1 '' FEMALES FROM M - 3000 PPM 15942 15947 15950 15952 DEVELOPMENTAL TOXICITY STUDY OF INHALED 1,1,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS GROSS FETAL EXAMINATION AT TIME OF LAPAROTOMY IMPLANT # SEX FETAL GROSS OBSERVATION 1 LF A 1 2 LF# 2 3 LF# 3 4 lf# 4 5 lf# 5 6 LF# 6 7 LF# 7 a LF# B 9 LF* 9 10 LF* 10 1 1 LF* 1 1 12 LF* 1 2 1 3 LF* 1 3 M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M V ECCHYMOSIS - HEAD CENTER OF SNOUT DORSAL SKULL F NO remarkable OBSERVATIONS F NO REMARKABLE OBSERVATIONS M M OOME-SHAPED HEAD SLIGHT M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS 1 LF* 1 2 LF* 2 3 LF* 3 4 LF* 4 5 LF* 5 6 LF* 6 7 LF* 7 M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS 1 LF# 1 2 LF# 2 3 4 LF# 3 5 6 LF# 4 7 8 LF# 5 9 1 LF# 1 2 LF# 2 3 4 LF# LF# 34 5 6 LF# 5 7 LF# 6 8 LF# 7 9 LF# 8 10 LF# 9 F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS EARLY(W/PLACENTAL TISSUE) F NO REMARKABLE OBSERVATIONS EARLV{W/PLACENTAL TISSUE) F NO REMARKABLE OBSERVATIONS EARLY(W/PLACENTAL TISSUE) M NO REMARKABLE OBSERVATIONS EARLY F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS EARLYCW/PLACENTAL TISSUE) M NO REMARKABLE OBSERVATIONS M V ECCHYMOSIS - HEAD TOP OF SNOUT F V ECCHYMOSIS - HEAD TOP OF HEAD F V ECCHYMOSIS - HEAD TOP OF HEAD M NO REMARKABLE OBSERVATIONS OBSERVATION CODE: M=MALFORMATION, V=VARIATION Report 50-514 Appendi* 4A Page 1 0 SL 042681 X . DEVELOPMENTAL TOXICITY STUDY OF INHALED 1,I,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS GROSS FETAL EXAMINATION AT TIME OF LAPAROTOMY FEMALES FROM M - 3000 PPM IMPLANT # SEX FETAL GROSS OBSERVATION 15956 1 LF# 1 2 LF# 2 3 LF# 3 4 LF# 4 5 LF# 5 6 LF# 6 7 M V ECCHYMOSIS - 1EXTREMITIES RIGHT HIND 1LEG M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M DEAD DEAD AND MACERATED FETUS INCISORS NOT ERUPTED 15958 1 LF# 1 2 LF# 2 3 LF# 3 4 LF# 4 5 LF# 5 6 LF# 6 7 LF# 7 8 LF# B 9 LF# 9 10 LF# 1 0 1 1 LF# 1 1 F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS 15960 1 LF# 1 2 LF# 2 3 LF# 3 4 LF# 4 5 6 LF# 5 7 LF# 6 8 LF# 7 9 LF# 8 10 LF# 9 1 1 LF# 10 1 2 LF# 1 1 M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS EARLY(W/PLACENTAL TISSUE) M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS 15968 1 LF# 1 2 LF# 2 3 LF# 3 4 LF# 4 5'' LF# 5 6 LF# 6 7 LF# 7 8 LF# B F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS 15979 : 1 LF# 1 2 LF# 2 3 LF# 3 4 LF# 4 F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS OBSERVATION CODE: M=MALFORMATION, V=VARIATION Report 50-514 Appendix 4A Page 1t SL 042682 1 DEVELOPMENTAL TOXICITV STUDV OF INHALED 1,1,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS GROSS FETAL EXAMINATION AT TIME OF LAPAROTOMY FEMALES FROM M - 3000 PPM IMPLANT A SEX FETAL GROSS OBSERVATION 15979 5 LF A 5 6 LF A 6 7 LF A 7 0 lfa 8 9 lfa 9 10 1 1 lfaio 1 2 lfa 1 1 F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS EARLY(W/PLACENTAL TISSUE) M NO REMARKABLE OBSERVATIONS F V ECCHYMOSIS - 1EXTREMITIES TIP OF TAIL 159B7 1 lfa 1 2 lfa 2 F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS 15980 1 LFA 1 2 LFA 2 3 LFA 3 4 lfa 4 5 LFA 5 6 LFA 6 7 LFA 7 0 LFA 8 9 LFA 9 M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS 15995 1 lfa 1 2 LFA 2 3 4 LFA 3 5 LFA A 6 LFA 5 7 LFA 6 0 LFA 7 F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M DEAD DEAD AND MACERATED FETUS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS 15998 i 2 3 4 5 6 7 8 LF* 1 9 10 U DEAD UNABLE TO EXAMINE U DEAD UNABLE TO EXAMINE U DEAD UNABLE TO EXAMINE U DEAD UNABLE TO EXAMINE U DEAD UNABLE TO EXAMINE U DEAD UNABLE TO EXAMINE U DEAD UNABLE TO EXAMINE F NO REMARKABLE OBSERVATIONS U DEAD UNABLE TO EXAMINE U DEAD UNABLE TO EXAMINE OBSERVATION CODE; M=MA LFORMATION, V=VARIATION Report 5 Appendi* Page l 2 SL 042683 DEVELOPMENTAL TOXICITV STUDY OF INHALED 1,1,t-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS GROSS FETAL EXAMINATION AT TIME OF LAPAROTOMY FEMALES FROM M - 3000 PPM IMPLANT # SEX FETAL GROSS OBSERVATION 15998 11 LF# 2 F V ECCHYMOSIS - HEAD TOP OF HEAD 16011 1 LF# 1 2 LF# 2 3 LF# 3 4 LF# 4 5 LF# 5 6 LF# 6 7 LF# 7 8 LF# 8 F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS 16013 1 2 LF# 1 3 LF# 2 4 LF# 3 5 6 LF# 4 7 LF# 5 8 LF# 6 9 LF# 7 10 LF# 8 1 1 LF# 9 12 LF# 110 EARLY(W/PLACENTAL TISSUE) F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS LI DEAD F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS 1 6018 1 LF# 1 2 LF# 2 3 LF# 3 4 LF# 4 5 LF# 5 6 LF# 6 7 LF# 7 e LF# 8 9 LF# 9 10 LF#110 11 LF# 1 1 12 LF# 11 2 F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F V ECCHYMOSIS - HEAD TOP OF HEAD F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS 16021 1 LF# 1 2 LF# 2 3 LF# 3 4 LF# 4 5 LF# 5 M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS 1 6032 1 LF# 1 2 LF# 2 3 LF# 3 4 LF# 4 M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS OBSERVATION CODE: M = MA LFORMATION, V=VARIATION Report 50-514 Appendix 4A Page 13 SL 042684 lALES FROM M - 3000 PPM 16032 1 6034 16049 DEVELOPMENTAL TOXICITY STUDY OF INHALED 1.1.1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS GROSS FETAL EXAMINATION AT TIME OF LAPAROTOMY IMPLANT # SEX FETAL GROSS OBSERVATION 5 LF# 5 6 LF# 6 7 LF# 7 8 LF# a 9 LF# 9 10 LF# 1 0 11 LF# 11 F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS 1 LF# 1 2 LF# 2 3 LF# 3 4 LF# 4 5 LF# 5 6 LF# 6 7 LF# 7 8 LF# a 9 LF# 9 M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS 1 LF# i 2 LF# 2 3 LF# 3 4 LF# 4 5 LF# 5 6 LF# 6 7 LF# 7 8 LF# 8 9 LF# 9 F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F V ECCHVMOSIS - 1EXTREMITIES RIGHT HIND 1-OOT M V ECCHYMOSIS - TRUNK BASE OF SPINE M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS OBSERVATION CODE: M=MALFORMATION, V=VARIATION Report 50-514 Appendix 4A Page 14 SL 042685 DEVELOPMENTAL TOXICITY 5TUDY OF INHALED 1,1,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS GROSS FETAL EXAMINATION AT TIME OF LAPAROTOMY FEMALES FROM R - 6000 PPM IMPLANT # SEX FETAL GROSS OBSERVATION 1 LF* 1 2 LF* 2 3 LF# 3 4 LF# 4 5 LF* 5 6 LF# 6 7 LF# 7 8 LF# 0 M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F. NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS 1 LF# 1 2 LF# 2 3 LF# 3 4 LF# 4 5 LF# 5 6 LF# 6 7 LF# 7 B LF# 8 9 LF# 9 10 LF# 110 F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F V ECCHYMOSIS - HEAD TOP OF HEAD 15955 1 2 LF A 1 3 LFA 2 4 LF# 3 5 6 LF# 4 7 LF# 5 0 LF* 6 9 EARLY(W/PLACENTAL TISSUE) F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS EARLY tW/PLACENTAL TISSUE) M NO REMARKABLE OBSERVATIONS F V ECCHYMOSIS - EXTREMITIES LEFT HIND LEG AT KNEE M NO REMARKABLE OBSERVATIONS EARLYCW/PLACENTAL TISSUE) 15959 1 LF* 1 2 LF* 2 3 LF* 3 4 LF* 4 5 LF* 5 6 LF* 6 7 LF* 7 M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS 15972 ' 1 LF* 1 2 LF* 2 3 LF* 3 4 LF* 4 5 LF* 5 6 7 LF* 6 0 LF* 7 F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS U DEAD DEAD AND MACERATED FETUS M NO REMARKABLE OBSERVATIONS F V ECCHYMOSIS - HEAD ABOVE LEFT EVE OBSERVATION CODE: M=MALFORMATION, V=VARIATION Report 50~51 A Append 1x 4A Page 15 SL 042686 FEMALES FROM R - 6000 PPM 15972 15978 15982 15984 DEVELOPMENTAL TOXICITV STUDY OF INHALED t,1,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS GROSS FETAL EXAMINATION AT TIME OF LAPAROTOMY IMPLANT # SEX FETAL GROSS OBSERVATION 8 LF# 7 9 io lf# a V ECCHYMOSIS - EXTREMITIES LEFT FRONT PAW M DEAD DEAD AND MACERATED FETUS M V ECCHYMOSIS - TRUNK LOWER BACK 1 LF# t 2 LF# 2 3 LF# 3 4 LF# 4 5 LF# 5 6 LF# 6 7 LF# 7 a LF# a M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M V ECCHYMOSIS - HEAO LEFT SIDE IF SNOUT M NO REMARKABLE OBSERVATIONS 1 LF# 1 2 LF# 2 3 LF# 3 4 5 LF# 4 6 LF# 5 7 LF# 6 F V ECCHYMOSIS - HEAD TOP OF HEAO M V ECCHYMOSIS - HEAD TOP OF HEAO F V ECCHYMOSIS - HEAD TOP OF HEAD EARLY(W/PLACENTAL TISSUE) F M SNOUT SHORT M MICROAGNATHIA V EVE BULGE FLATTENED BILATERAL M FOREPAWS RIGIDLY FLEXED - BILATERAL PALMS DOWNWARD M INVERSION - LEFT HINDPAW F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS 1 LF# 1 2 LF# 2 3 LF# 3 4 LF# 4 5 LF# 5 6 LF# 6 7 LF# 7 8 LF# 8 9 LF# 9 10 LF#10 ECCHYMOSIS - EXTREMITIES RIGHT HIND LEG F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M DOME-SHAPED HEAD SLIGHT ECCHYMOSIS - EXTREMITIES RIGHT HINDPAW DIGITS NO REMARKABLE OBSERVATIONS NO REMARKABLE OBSERVATIONS NO REMARKABLE OBSERVATIONS NO REMARKABLE OBSERVATIONS NO REMARKABLE OBSERVATIONS OBSERVATION CODE: M=MALFORMATION, V=VARIATION Report 50-514 Appendix 4A Page 16 Z .8 ^ 0 c/i DEVELOPMENTAL TOXICITV STUDY OF INHALED 1 , 1,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS GROSS FETAL EXAMINATION AT TIME OF LAPAROTOMY FEMALES FROM R - 6000 PPM IMPLANT # SEX FETAL GROSS OBSERVATION 15989 I LF# 1 2 LF# 2 3 LF# 3 4 LF# 4 5 LF# 5 6 LF# 6 7 LF# 7 B LF# 9 9 LF# 9 10 LF#10 M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS 15999 1 LF# 1 2 LF# 2 3 LF# 3 4 LF# 4 5 LF# 5 6 LF# 6 7 LF# 7 8 LF# B F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS 16001 1 LF# 1 2 3 LF# 2 4 LF# 3 5 LF# 4 6 LF# 5 7 LF# 6 M NO REMARKABLE OBSERVATIONS EARLY(W/PLACENTAL TISSUE) F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M V ECCHYMOSIS - EXTREMITIES RIGHT THIGH M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS 16007 1 LF# 1 2 LF# 2 3 LF# 3 4 LF# 4 5 LF# 5 6 LF# 6 7 LF# 7 B LF# B 9 LF# 9 M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS 16009 1 LF# 1 2 LF# 2 3 LF# 3 4 LF# 4 5 LF# 5 6 LF# 6 F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS 16014 1 LF# 1 2 LF# 2 M V ECCHYMOSIS - HEAD SNOUT F NO REMARKABLE OBSERVATIONS OBSERVATION CODE; M=MALFORMATION, V=VAR!ATION Report 50-514 Append 1 x 4A Page 17 889^0 t t-1 DEVELOPMENTAL TOXICITV STUDY OF INHALED 1,1,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS GROSS FETAL EXAMINATION AT TIME OF LAPAROTOMY FEMALES FROM R - 6000 PPM IMPLANT 0 SEX FETAL GROSS OBSERVATION 16014 3 LF* 3 4 LF# 4 5 LF# 5 6 LF# 6 7 LF# 7 B LF# B 9 LF# 9 10 LF#10 M NO REMARKABLE OBSERVATIONS M V ECCHVMOSIS - HEAD TOP OF HEAD F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F V ECCHVMOSIS - HEAD TOP OF SNOUT 16019 I 2 3 4 LF# 1 5 LF# 2 6 7 LF# 3 8 LF# 4 9 to LATECW/FETAL TISSUE) EARLY(W/PLACENTAL TISSUE) EARLY(W/PLACENTAL TISSUE) F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M DEAD DEAD AND MACERATED FETUS DARK BROWN M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS EARLY(W/PLACENTAL TISSUE) U DEAD DEAD AND MACERATED FETUS 16022 1 LF# 1 2 LF# 2 3 LF# 3 4 LF A 4 5 6 LF# 5 F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS EARLY(W/PLACENT A L TISSUE) M NO REMARKABLE OBSERVATIONS 16023 1 2 3 4 '5 .6 .7 LF# LF# LF# LF# LF# 1 2 3 4 5 EARLY(W/PLACENTAL TISSUE) F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS EARLY(W/PLACENTAL TISSUE) 16033 1 2 3 LF# 1 4 LF# 2 5 LF# 3 6 LF# 4 EARLY(W/PLACENTAL TISSUE) EARLY(W/PLACENTAL TISSUE) M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS 16040 1 LF A 1 2 LF# 2 F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS OBSERVATION CODE: M=MALFORMATION, V = VARI ATION Report 50-514 A ppendi* 4A Page 18 * r* 689ZV0 DEVELOPMENTAL TOXI ClTV STUDY OF INHALED 1.1,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS GROSS FETAL EXAMINATION AT TIME OF LAPAROTOMY ALES FROM R - 6000 PPM IMPLANT # SEX FETAL GROSS OBSERVATION I 6040 3 LF# 3 4 LF A 4 5 LF# 5 6 LF# 6 7 LF# 7 8 LF# 8 9 LF# 9 10 LF# 1 0 M NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS F NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS M NO REMARKABLE OBSERVATIONS 16044 OBSERVATION CODE: 1 LF# 1 2 LF# 2 3 LF# 3 4 LF# 4 5 LF# 5 6 7 8 LF# 6 9 LF# 7 to LF# B M=MALFORMATION, V=VAR1ATI0N F M F F F F F F NO REMARKABLE OBSERVATIONS NO REMARKABLE OBSERVATIONS NO REMARKABLE OBSERVATIONS NO REMARKABLE OBSERVATIONS NO REMARKABLE OBSERVATIONS EARLY(W/PLACENTAL TISSUE) EARLY(W/PLACENTAL TISSUE) NO REMARKABLE OBSERVATIONS NO REMARKABLE OBSERVATIONS NO REMARKABLE OBSERVATIONS Xfi r DEVELOPMENTAL TOXICITY STUDV OF INHALED 1,t,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RA88ITS FETAL VISCERAL FINDINGS 069Z' ' 0 DAMS FROM GROUP 1: G - 0 PPM FETUS# VISCERAL OBSERVATION 15946 1 NO REMARKABLE OBSERVATIONS 2 NO REMARKABLE OBSERVATIONS 3 NO REMARKABLE OBSERVATIONS 4 NO REMARKABLE OBSERVATIONS 5 V SPLEEN PALE V THIRD VENTRICLE DILATED - NO DEPRESSION 6 NO REMARKABLE OBSERVATIONS 7 V ;SPLEEN PALE 9 NO REMARKABLE OBSERVATIONS 10 V 1FETAL ATELECTASIS 1 1 NO REMARKABLE OBSERVATIONS 1 2 NO REMARKABLE OBSERVATIONS 13 NO REMARKABLE OBSERVATIONS 14 NO REMARKABLE OBSERVATIONS 15953 1 NO REMARKABLE OBSERVATIONS 2 NO REMARKABLE OBSERVATIONS 3 NO REMARKABLE OBSERVATIONS 4 NO REMARKABLE OBSERVATIONS 5 NO REMARKABLE OBSERVATIONS 6 NO REMARKABLE OBSERVATIONS 7 NO REMARKABLE OBSERVATIONS 8 NO REMARKABLE OBSERVATIONS 9 NO REMARKABLE OBSERVATIONS 15954 1 NO REMARKABLE OBSERVATIONS 2 NO REMARKABLE OBSERVATIONS 3 NO REMARKABLE OBSERVATIONS 5 NO REMARKABLE OBSERVATIONS 6 NO REMARKABLE OBSERVATIONS GRADE CODE; 1-SLIGHT, 2-MODERATE. 3-MARKED, P-PRESENT M-MALFORMATION V-VARIATION GRADE P 1 P P Report 50-514 Appendi 4B Page 1 yi r* o > {O O' vO DEVELOPMENTAL TOXICITY STUDY OF INHALED 1,1,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS FETAL VISCERAL FINDINGS DAMS FROM GROUP 1 : G - 0 PPM FETUS# VISCERAL OBSERVATION 15954 7 NO REMARKABLE OBSERVATIONS 8 NO REMARKABLE OBSERVATIONS 9 NO REMARKABLE OBSERVATIONS 15961 1 NO REMARKABLE OBSERVATIONS 2 NO REMARKABLE OBSERVATIONS 3 NO REMARKABLE OBSERVATIONS 4 NO REMARKABLE OBSERVATIONS 5 NO REMARKABLE OBSERVATIONS 6 NO REMARKABLE OBSERVATIONS 7 NO REMARKABLE OBSERVATIONS 8 V 1FETAL ATELECTASIS 9 V IRREGULAR RUGAE 15967 1 NO REMARKABLE OBSERVATIONS 2 NO REMARKABLE OBSERVATIONS 3 NO REMARKABLE OBSERVATIONS 4 NO REMARKABLE OBSERVATIONS 5 NO REMARKABLE OBSERVATIONS 7 NO REMARKABLE OBSERVATIONS 8 NO REMARKABLE OBSERVATIONS 9 NO REMARKABLE OBSERVATIONS 10 NO REMARKABLE OBSERVATIONS 1 1 NO REMARKABLE OBSERVATIONS 15980 1 NO REMARKABLE OBSERVATIONS 2 NO REMARKABLE OBSERVATIONS 3 V AZYGOUS LUNG LOBE MISSING 4 NO REMARKABLE OBSERVATIONS 5 NO REMARKABLE OBSERVATIONS GRADE CODE: 1-SLIGHT, 2-MODERATE, 3-MARKED, P-PRESENT, M-MALFORMATION V-VARIATION GRADE P P P Repo rt 50-514 Append:x 48 Page 2 CL Q. Cl. SL 042692 * DEVELOPMENTAL TOXICITV STUDY OF INHALED 1,1,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS FETAL VISCERAL FINDINGS DAMS FROM GROUP 1: G - 0 PPM FETUS# VISCERAL OBSERVATION 15980 6 NO REMARKABLE OBSERVATIONS 7 NO REMARKABLE OBSERVATIONS 8 V AZYGOUS LUNG LOBE MISSING V LIVER PALE 9 V AZYGOUS LUNG LOBE MISSING 15991 1 NO REMARKABLE OBSERVATIONS 3 NO REMARKABLE OBSERVATIONS 4 V IRREGULAR RUGAE 5 V AZYGOUS LUNG LOBE MISSING 6 V IRREGULAR RUGAE 7 NO REMARKABLE OBSERVATIONS 8 NO REMARKABLE OBSERVATIONS 9 NO REMARKABLE OBSERVATIONS 10 NO REMARKABLE OBSERVATIONS 15993 1 V LATERAL VENTRICLE DILATED - NO DEPRESSION BILATERAL 2 NO REMARKABLE OBSERVATIONS 4 M LATERAL VENTRICLE DILATED - TISSUE DEPRESSED BILATERAL 5 NO REMARKABLE OBSERVATIONS 6 M LATERAL VENTRICLE DILATED - TISSUE DEPRESSED BILATERAL 7 NO REMARKABLE OBSERVATIONS 8 NO REMARKABLE OBSERVATIONS 9 NO REMARKABLE OBSERVATIONS 10 NO REMARKABLE OBSERVATIONS 15994 1 NO REMARKABLE OBSERVATIONS GRADE CODE: 1-SLIGHT, 2-MODERATE. 3-MARKED, P-PRESENT, M-MALFORMATION V-VARIATION GRADE P P P 1 1 1 Report 50-514 Appendi* 4B Page 3 SL 042693 DEVELOPMENTAL TOXICITY STUDY OF INHALED 1,1.1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS FETAL VISCERAL FINDINGS DAMS FROM GROUP 1: G - 0 PPM 1 5994 FETUS# 2 3 4 5 6 7 8 9 VISCERAL OBSERVATION GRADE NO REMARKABLE OBSERVATIONS M LATERAL VENTRICLE DILATED - TISSUE DEPRESSED P BILATERAL NO REMARKABLE OBSERVATIONS M LATERAL VENTRICLE DILATED - TISSUE DEPRESSED P BILATERAL V THIRD VENTRICLE DILATED - NO DEPRESSION t M CAVITATION ORAL CAVITY P FLOOR OF ORAL CAVITY WHERE NASOPHARYNX IS LOCATED , EXTENDING ANTERIORLY. (4mm X 2mm x 2mm DEEP) NO REMARKABLE OBSERVATIONS NO REMARKABLE OBSERVATIONS NO REMARKABLE OBSERVATIONS M LATERAL VENTRICLE DILATED - TISSUE DEPRESSED P RIGHT PRESENT, LEFT SLIGHT 1600 3 1 M LATERAL VENTRICLE DILATED - TISSUE DEPRESSED BILATERAL V THIRD VENTRICLE DILATED - NO DEPRESSION 2 NO REMARKABLE OBSERVATIONS 3 M LATERAL VENTRICLE DILATED - TISSUE DEPRESSED BILATERAL V THIRD VENTRICLE DILATED - NO DEPRESSION 1 1 1 16005 1 NO REMARKABLE OBSERVATIONS 2 NO REMARKABLE OBSERVATIONS 3 NO REMARKABLE OBSERVATIONS 4 NO REMARKABLE OBSERVATIONS 5 M GALLBLADDER MISSING 6 V AZYGOUS LUNG LOBE MISSING P P GRADE CODE: 1-SLIGHT, 2-MODERATE, 3-MARKED, P-PRESENT M-MALFORMATION V-VARIATION Repor t 50-514 Appendi* 4B Page 4  o s- to V69 DEVELOPMENTAL TOXICITV STUDV OF INHALED 1,1,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS FETAL VISCERAL FINDINGS DAMS FROM GROUP t: G - 0 PPM FETUS* VISCERAL OBSERVATION 16005 7 V AZVGOUS LUNG LOBE MISSING 8 NO REMARKABLE OBSERVATIONS 9 NO REMARKABLE OBSERVATIONS 10 NO REMARKABLE OBSERVATIONS 1 1 NO REMARKABLE OBSERVATIONS 16006 1 NO REMARKABLE OBSERVATIONS 2 NO REMARKABLE OBSERVATIONS 3 NO REMARKABLE OBSERVATIONS 5 NO REMARKABLE OBSERVATIONS 6 NO REMARKABLE OBSERVATIONS 7 NO REMARKABLE OBSERVATIONS 8 NO REMARKABLE OBSERVATIONS 10 NO REMARKABLE OBSERVATIONS 16016 t M LATERAL VENTRICLE DILATED - TISSUE DEPRESSED BILATERAL 2 NO REMARKABLE OBSERVATIONS 3 NO REMARKABLE OBSERVATIONS 4 NO REMARKABLE OBSERVATIONS 5 V AZVGOUS LUNG LOBE MISSING 6 NO REMARKABLE OBSERVATIONS 7 M LATERAL VENTRICLE DILATED - TISSUE DEPRESSED LEFT V LATERAL VENTRICLE DILATED - NO DEPRESSION RIGHT 8 NO REMARKABLE OBSERVATIONS 9 V THIRD VENTRICLE DILATED - NO DEPRESSION 10 NO REMARKABLE OBSERVATIONS 12 NO REMARKABLE OBSERVATIONS GRADE CODE: 1-SLIGHT, 2-MOOER ATE, 3-MARKED. P-PRESENT, M-MALFORMATION V-VARIATI ON GRADE P P P 1 1 1 Report 50-514 Appendi* 48 $L 042695) DEVELOPMENTAL TOXICITY STUDY OF INHALED 1.1,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS FETAL VISCERAL FINDINGS DAMS FROM GROUP 1: G - 0 PPM FETUS# VISCERAL OBSERVATION 16020 16025 16028 1 NO REMARKABLE OBSERVATIONS 2 NO REMARKABLE OBSERVATIONS 3 NO REMARKABLE OBSERVATIONS 4 NO REMARKABLE OBSERVATIONS 5 NO REMARKABLE OBSERVATIONS 6 NO REMARKABLE OBSERVATIONS 7 NO REMARKABLE OBSERVATIONS 8 V THIRD VENTRICLE DILATED - NO DEPRESSION V LATERAL VENTRICLE DILATED - NO DEPRESSION BILATERAL 9 NO REMARKABLE OBSERVATIONS 10 V THYMUS DARK RED V LATERAL VENTRICLE DILATED - NO DEPRESSION BILATERAL 1 NO REMARKABLE OBSERVATIONS 2 NO REMARKABLE OBSERVATIONS 3 NO REMARKABLE OBSERVATIONS 4 NO REMARKABLE OBSERVATIONS 5 NO REMARKABLE OBSERVATIONS 6 NO REMARKABLE OBSERVATIONS 7 NO REMARKABLE OBSERVATIONS 6 NO REMARKABLE OBSERVATIONS 9 NO REMARKABLE OBSERVATIONS 10 NO REMARKABLE OBSERVATIONS 1 NO REMARKABLE OBSERVATIONS 2 NO REMARKABLE OBSERVATIONS 3 NO REMARKABLE OBSERVATIONS GRADE COOE: 1-SLIGHT, 2-MODERATE, 3-MARKED, P-PRESENT M-MALFORMATION V-VARIATION GRADE 1 1 P 1 Repo rt 50-5 14 Appendix 48 Page 6 69^ tn f* DEVELOPMENTAL TOXICITY STUDY OF INHALED t,1.1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS FETAL VISCERAL FINDINGS DAMS FROM GROUP 1: G - 0 PPM FETUS# VISCERAL OBSERVATION 16028 4 NO REMARKABLE OBSERVATIONS 5 NO REMARKABLE OBSERVATIONS 7 NO REMARKABLE OBSERVATIONS 8 NO REMARKABLE OBSERVATIONS 16036 1 NO REMARKABLE OBSERVATIONS 2 NO REMARKABLE OBSERVATIONS 3 NO REMARKABLE OBSERVATIONS 4 NO REMARKABLE OBSERVATIONS 5 NO REMARKABLE OBSERVATIONS 6 NO REMARKABLE OBSERVATIONS 7 NO REMARKABLE OBSERVATIONS 8 NO REMARKABLE OBSERVATIONS 9 NO REMARKABLE OBSERVATIONS 10 NO REMARKABLE OBSERVATIONS 16039 1 NO REMARKABLE OBSERVATIONS 2 NO REMARKABLE OBSERVATIONS 3 NO REMARKABLE OBSERVATIONS 4 NO REMARKABLE OBSERVATIONS 5 NO REMARKABLE OBSERVATIONS 16041 1 NO REMARKABLE OBSERVATIONS 2 V LATERAL VENTRICLE DILATED - NO DEPRESSION ` LEFT V THIRD VENTRICLE DILATED - NO DEPRESSION 4 NO REMARKABLE OBSERVATIONS 5 NO REMARKABLE OBSERVATIONS 6 NO REMARKABLE OBSERVATIONS 7 NO REMARKABLE OBSERVATIONS GRADE CODE: 1-SLIGHT, 2-MODERATE, 3-MARKED, P-PRESENT. M-MALFORMATION V-VARIATION GRADE 1 1 Report 50-514 Appendi* 4B Page 7 SL 042697 * developmental toxicity study of inhaled 1,1,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS FETAL VISCERAL FINDINGS DAMS FROM GROUP 1: G - 0 PPM FETUS# VISCERAL OBSERVATION 16045 16047 1 NO REMARKABLE OBSERVATIONS 2 NO REMARKABLE OBSERVATIONS 3 NO REMARKABLE OBSERVATIONS 4 NO REMARKABLE OBSERVATIONS 5 NO REMARKABLE OBSERVATIONS 6 NO REMARKABLE OBSERVATIONS 7 NO REMARKABLE OBSERVATIONS 8 NO REMARKABLE OBSERVATIONS 9 NO REMARKABLE OBSERVATIONS 1 NO REMARKABLE OBSERVATIONS 2 V AZYGOUS LUNG LOBE MISSING 3 NO REMARKABLE OBSERVATIONS 4 NO REMARKABLE OBSERVATIONS 5 NO REMARKABLE OBSERVATIONS 6 M LATERAL VENTRICLE DILATED - TISSUE DEPRESSED LEFT 7 NO REMARKABLE OBSERVATIONS 8 V LATERAL VENTRICLE DILATED - NO DEPRESSION RIGHT 9 NO REMARKABLE OBSERVATIONS 10 NO REMARKABLE OBSERVATIONS GRADE CODE: 1-SLIGHT, 2-MODERATE, 3-MARKED, P-PRESENT, M-MALFORMATION V-VARIATION GRADE I 1 Report 50-514 Appendin 4B Page B SL 042698^ t DEVELOPMENTAL TOXICITV STUDY OF INHALEO t,1,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS FETAL VISCERAL FINDINGS DAMS FROM GROUP 2: I - 1000 PPM FETUS# VISCERAL OBSERVATION 15945 15957 15963 15964 1 NO REMARKABLE OBSERVATIONS 2 NO REMARKABLE OBSERVATIONS 3 NO REMARKABLE OBSERVATIONS 5 NO REMARKABLE OBSERVATIONS 6 NO REMARKABLE OBSERVATIONS 7 NO REMARKABLE OBSERVATIONS a NO REMARKABLE OBSERVATIONS 9 NO REMARKABLE OBSERVATIONS 10 NO REMARKABLE OBSERVATIONS 1 V IRREGULAR RUGAE 2 NO REMARKABLE OBSERVATIONS 3 NO REMARKABLE OBSERVATIONS 4 NO REMARKABLE OBSERVATIONS 6 V IRREGULAR RUGAE 7 NO REMARKABLE OBSERVATIONS 0 NO REMARKABLE OBSERVATIONS 1 NO REMARKABLE OBSERVATIONS 2 NO REMARKABLE OBSERVATIONS 3 NO REMARKABLE OBSERVATIONS 5 V LATERAL VENTRICLE DILATED - NO DEPRESSION BILATERAL 6 NO REMARKABLE OBSERVATIONS 8 NO REMARKABLE OBSERVATIONS 2 V DARK BROWN MATERIAL AMONG BRAIN SECTIONS BILATERAL LATERAL VENTRICLES M CAVITATION IN MEDULLA GRADE CODE: 1-SLIGHT, 2-MODERATE, 3-MARKED, P-PRESENT M-MALFORMATION V-VARIATION GRADE P P 1 P P Report 50-514 Appendix 4B Page 9 SL 042699; DEVELOPMENTAL TOXICITY STUDY OF INHALED t,I,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS FETAL VISCERAL FINDINGS DAMS FROM GROUP 2: I - 1000 PPM FETUS* VISCERAL OBSERVATION GRADE 15964 5 NO REMARKABLE OBSERVATIONS 6 NO REMARKABLE OBSERVATIONS 7 NO REMARKABLE OBSERVATIONS a NO REMARKABLE OBSERVATIONS 9 NO REMARKABLE OBSERVATIONS 1 1 NO REMARKABLE OBSERVATIONS 15965 t NO REMARKABLE OBSERVATIONS 2 NO REMARKABLE OBSERVATIONS 3 V LATERAL VENTRICLE DILATED - NO DEPRESSION 1 BILATERAL 4 NO REMARKABLE OBSERVATIONS 5 V FETAL ATELECTASIS P M FOURTH VENTRICLE MISSING P V DARK BROWN AREA OF BRAIN TISSUE .P IN MEDULLA AND EXTENDS INTO THALAMUS 5.0 MM IN,DIAMETER. ENTIRE DARK BROWN AREA HAS A MORE SPONGY AND FRIABLE TEXTURE THAN NORMAL BRAIN TISSUE M POSTERIOR PORTION OF CEREBELLUM SCRAMBLED P 7 NO REMARKABLE OBSERVATIONS 8 V DARK BROWN AREA OF BRAIN TISSUE P TWO AREAS 2.0 MM IN DIAMETER ON EACH SIDE OF THE- THALAMUS AND CEREBELLUM. 9 M VENTRICULAR SEPTAL DEFECT P LARGE MEMBRANOUS AND MUSCULAR M SEMILUNAR VALVES OF THE PULMONARY ARTERY MISSING P M SEMILUNAR VALVES OF THE AORTA MISSING P M RIGHT VENTRICLE OF THE HEART SMALL P M HOLE IN EXTERIOR OF LEFT ATRIUM P LARGE HOLE, NO BLOOD PRESENT IN PERICARDIUM. APPEARS TO BE GRADE CODE: 1-SLIGHT, 2-MODERATE, 3-MARKED, P-PRESENT M-MALFORMATION V-VARIATION Report 50-514 Append 1 * 4B Page 10 SL 042700) DEVELOPMENTAL TOXICITY STUDY OF INHALED I,t,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS FETAL VISCERAL FINDINGS DAMS FROM GROUP 2: I - 1000 PPM FETUS# VISCERAL OBSERVATION 15965 OPEN INTO THE LEFT VENTRICLE. 9 V LEFT ATRIUM PALE WAS PALE BENEATH THE PERICARDIUM AND REMAINED PALE AFTER PERICARDIUM WAS REMOVED M PAPILLARY MUSCLES MISSING - RIGHT VENTRICLE M ENLARGED AORTA V SMALL BUT PATENT DUCTUS ARTERIOSUS V RIGHT SUBCLAVIAN ARTERY SMALL 10 NO REMARKABLE OBSERVATIONS 15971 1 V LATERAL VENTRICLE DILATED - NO DEPRESSION BILATERAL 2 NO REMARKABLE OBSERVATIONS 3 NO REMARKABLE OBSERVATIONS 4 NO REMARKABLE OBSERVATIONS 5 V IRREGULAR RUGAE 6 NO REMARKABLE OBSERVATIONS 7 NO REMARKABLE OBSERVATIONS B NO REMARKABLE OBSERVATIONS 9 NO REMARKABLE OBSERVATIONS 15975 2 NO REMARKABLE OBSERVATIONS 3 NO REMARKABLE OBSERVATIONS 4 NO REMARKABLE OBSERVATIONS 5 NO REMARKABLE OBSERVATIONS 6 NO REMARKABLE OBSERVATIONS 7 NO REMARKABLE OBSERVATIONS 0 NO REMARKABLE OBSERVATIONS 9 NO REMARKABLE OBSERVATIONS 10 V THIRD VENTR CLE DILATED - NO DEPRESSION GRADE CODE: 1-SLIGHT. 2-MODERATE, 3-MARKED, P-PRESENT M-MALFORMATION V-VARIATION GRADE P P P P P 1 P 1 Report 50-514 Appendi* 40 Page t t SL 042701 DEVELOPMENTAL TOXICITY STUDY OF INHALED 1,1,t-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS FETAL VISCERAL FINDINGS DAMS FROM GROUP 2:1- 1000 PPM FETUS# VISCERAL OBSERVATION 15981 1 5983 1598 6 15996 16010 2 NO REMARKABLE OBSERVATIONS 5 M LATERAL VENTRICLE DILATED - TISSUE DEPRESSED BILATERAL 6 NO REMARKABLE OBSERVATIONS 7 M LATERAL VENTRICLE DILATED - TISSUE DEPRESSED BILATERAL 1 NO REMARKABLE OBSERVATIONS 2 V AZYGOUS LUNG LOBE MISSING 4 NO REMARKABLE OBSERVATIONS 5 V FETAL ATELECTASIS 6 NO REMARKABLE OBSERVATIONS 7 V LATERAL VENTRICLE DILATED - NO DEPRESSION BILATERAL 1 NO REMARKABLE OBSERVATIONS 3 NO REMARKABLE OBSERVATIONS 4 NO REMARKABLE OBSERVATIONS 7 V DARK BROWN MATERIAL AMONG BRAIN SECTIONS BILATERAL LATERAL VENTRICLES V THIRD VENTRICLE DILATED - NO DEPRESSION 2 NO REMARKABLE OBSERVATIONS 10 NO REMARKABLE OBSERVATIONS 1 NO REMARKABLE OBSERVATIONS 2 M LATERAL VENTRICLE DILATED - TISSUE DEPRESSED BILATERAL 3 NO REMARKABLE OBSERVATIONS GRADE CODE: 1-SLIGHT, 2-MODERATE. 3-MARKED, P-PRESENT M-MALFORMATION V-VARIATION- GRADE P P P P 1 P t P Report 50-5 5 4 Appendix 48 Page l2 SL 042702 DEVELOPMENTAL TOXICITY STUDY OF INHALED 1.1,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS FETAL VISCERAL FINDINGS DAMS FROM GROUP 2: I - 1000 PPM FETUS# VISCERAL OBSERVATION 16010 4 V LATERAL VENTRICLE DILATED - NO DEPRESSION BILATERAL 5 NO REMARKABLE OBSERVATIONS 7 NO REMARKABLE OBSERVATIONS 16015 1 NO REMARKABLE OBSERVATIONS 2 NO REMARKABLE OBSERVATIONS 3 M LATERAL VENTRICLE DILATED - TISSUE DEPRESSED BILATERAL 4 NO REMARKABLE OBSERVATIONS 5 M LATERAL VENTRICLE DILATED - TISSUE DEPRESSED BILATERAL 1 6026 1 NO REMARKABLE OBSERVATIONS 2 NO REMARKABLE OBSERVATIONS 3 NO REMARKABLE OBSERVATIONS 4 NO REMARKABLE OBSERVATIONS 5 NO REMARKABLE OBSERVATIONS 6 NO REMARKABLE OBSERVATIONS 7 NO REMARKABLE OBSERVATIONS 8 M TEETH PRIMORDIA MISSING RIGHT g NO REMARKABLE OBSERVATIONS 10 NO REMARKABLE OBSERVATIONS 16030 1 NO REMARKABLE OBSERVATIONS 2 NO REMARKABLE OBSERVATIONS 3 NO REMARKABLE OBSERVATIONS 4 V LATERAL VENTRICLE DILATED - NO DEPRESSION BILATERAL GRADE CODE: 1-SLIGHT, 2-MODERATE, 3-MARKED, P-PRESENT M-MALFORMATION V-VARIATION- GRADE 1 P 1 P 1 Repo r t 50-514 Append i * 4B Page 13 SL 042703 DEVELOPMENTAL TOXICITY STUDY OF INHALED 1,1,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS FETAL VISCERAL FINDINGS DAMS FROM GROUP 2: I - 1000 PPM FETUS* VISCERAL OBSERVATION 16030 6 NO REMARKABLE OBSERVATIONS 7 NO REMARKABLE OBSERVATIONS B NO REMARKABLE OBSERVATIONS 9 V IIRREGULAR RUGAE 10 NO REMARKABLE OBSERVATIONS 1 1 NO REMARKABLE OBSERVATIONS 16037 1 NO REMARKABLE OBSERVATIONS 2 NO REMARKABLE OBSERVATIONS 3 NO REMARKABLE OBSERVATIONS 4 NO REMARKABLE OBSERVATIONS 5 NO REMARKABLE OBSERVATIONS 6 NO REMARKABLE OBSERVATIONS 7 NO REMARKABLE OBSERVATIONS GRADE CODE: 1-SLIGHT, 2-MODERATE, 3-MARKED, P-PRESENT, M-MALFORMATION V-VARIATION GRADE P Report 50-514 Appendi* 4B Page 14 SL 042704 DEVELOPMENTAL TOXICITY STUDY OF INHALED 1,1.1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS FETAL VISCERAL FINDINGS DAMS FROM GROUP 3: M - 3000 PPM FETUS# VISCERAL OBSERVATION 15942 15947 15950 1 NO REMARKABLE OBSERVATIONS 2 NO REMARKABLE OBSERVATIONS 3 NO REMARKABLE OBSERVATIONS 4 NO REMARKABLE OBSERVATIONS 5 NO REMARKABLE OBSERVATIONS 6 NO REMARKABLE OBSERVATIONS 7 NO REMARKABLE OBSERVATIONS S NO REMARKABLE OBSERVATIONS 9 V THIRD VENTRICLE OILATEO - NO DEPRESSION 10 NO REMARKABLE OBSERVATIONS 1 1 V AZYGOUS LUNG LOBE MISSING 1 2 NO REMARKABLE OBSERVATIONS 1 3 NO REMARKABLE OBSERVATIONS 1 M LATERAL VENTRICLE DILATED - TISSUE DEPRESSED BILATERAL 2 NO REMARKABLE OBSERVATIONS 3 NO REMARKABLE OBSERVATIONS 4 NO REMARKABLE OBSERVATIONS 5 NO REMARKABLE OBSERVATIONS 6 NO REMARKABLE OBSERVATIONS 7 V LATERAL VENTRICLE DILATED - NO DEPRESSION BILATERAL 1 V FETAL ATELECTASIS 2 NO REMARKABLE OBSERVATIONS 4 NO REMARKABLE OBSERVATIONS 6 V FETAL ATELECTASIS M GALLBLADDER MISSING GRADE CODE: 1-SLIGHT, 2-MODERATE, 3-MARKED, P-PRESENT M-MALFORMATION V-VARIATION GRADE t 1 P P P Report 50-514 Appendi* 4B Page 15 SL 042705 DEVELOPMENTAL TOXICITV STUDV OF INHALED 1,1.1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS FETAL VISCERAL FINDINGS DAMS FROM GROUP 3; M - 3000 PPM FETUS* VISCERAL OBSERVATION 15950 B V DARK BROWN MATERIAL AMONG BRAIN SECTIONS BILATERAL LATERAL VENTRICLES 15952 1 NO REMARKABLE OBSERVATIONS 2 V SPLEEN PALE 3 V SPLEEN PALE 4 V SPLEEN PALE 6 V SPLEEN PALE 7 V SPLEEN PALE e V SPLEEN PALE 9 NO REMARKABLE OBSERVATIONS 10 V FETAL ATELECTASIS 15956 t NO REMARKABLE OBSERVATIONS 2 NO REMARKABLE OBSERVATIONS 3 NO REMARKABLE OBSERVATIONS 4 NO REMARKABLE OBSERVATIONS 5 NO REMARKABLE OBSERVATIONS 6 NO REMARKABLE OBSERVATIONS 15958 1 V FETAL ATELECTASIS M LATERAL VENTRICLE DILATED - TISSUE DEPRESSED BILATERAL 2 NO REMARKABLE OBSERVATIONS 3 V THIRD VENTRICLE DILATED - NO DEPRESSION 4 NO REMARKABLE OBSERVATIONS 5 NO REMARKABLE OBSERVATIONS 6 NO REMARKABLE OBSERVATIONS 7 NO REMARKABLE OBSERVATIONS 8 NO REMARKABLE OBSERVATIONS GRADE CODE: 1-SLIGHT, 2-MODERATE, 3-MARKED, P-PRESENT M-MALFORMATION V-VARIATION GRADE P P P P P P P P P 1 1 Report 50-514 Append t * 4B Page 16 SL 042706 DEVELOPMENTAL TOXICITY STUDY OF INHALED I,1,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS FETAL VISCERAL FINDINGS DAMS FROM GROUP 3: M - 3000 PPM FETUS# VISCERAL OBSERVATION 15958 9 V LATERAL VENTRICLE DILATED - NO DEPRESSION BILATERAL V THIRD VENTRICLE DILATED - NO DEPRESSION 10 NO REMARKABLE OBSERVATIONS 1 1 V LATERAL VENTRICLE DILATED - NO DEPRESSION LEFT 15960 1 V FETAL ATELECTASIS Z NO REMARKABLE OBSERVATIONS 3 NO REMARKABLE OBSERVATIONS 4 NO REMARKABLE OBSERVATIONS 6 NO REMARKABLE OBSERVATIONS 7 NO REMARKABLE OBSERVATIONS 8 V THIRD VENTRICLE DILATED - NO DEPRESSION 9 NO REMARKABLE OBSERVATIONS 10 NO REMARKABLE OBSERVATIONS 1 1 NO REMARKABLE OBSERVATIONS 12 V DARK BROWN MATERIAL AMONG BRAIN SECTIONS BILATERAL LATERAL VENTRICLES 15968 1 NO REMARKABLE OBSERVATIONS 2 NO REMARKABLE OBSERVATIONS 3 NO REMARKABLE OBSERVATIONS 4 NO REMARKABLE OBSERVATIONS 5 NO REMARKABLE OBSERVATIONS 6 NO REMARKABLE OBSERVATIONS 7 NO REMARKABLE OBSERVATIONS a NO REMARKABLE OBSERVATIONS 15979 NO REMARKABLE OBSERVATIONS GRADE CODE: 1-SLIGHT, 2-MODERATE, 3-MARKED, P-PRESENT, M-MALFORMATION V-VARIATION GRADE 1 1 I P 1 P Report 50-514 Appendix 4B Page 1 7 SL 042707 m DEVELOPMENTAL TOXICITV STUDY OF INHALED 1.1.1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS FETAL VISCERAL FINDINGS DAMS FROM GROUP 3: M - 3000 PPM 15979 15987 15988 FETUS# 2 3 4 5 6 7 8 9 11 12 1 2 1 2 3 4 5 6 7 8 9 VISCERAL OBSERVATION NO REMARKABLE OBSERVATIONS NO REMARKABLE OBSERVATIONS NO REMARKABLE OBSERVATIONS NO REMARKABLE OBSERVATIONS NO REMARKABLE OBSERVATIONS NO REMARKABLE OBSERVATIONS NO REMARKABLE OBSERVATIONS M THIRD VENTRICLE DILATED NO REMARKABLE OBSERVATIONS NO REMARKABLE OBSERVATIONS TISSUE DEPRESSED NO REMARKABLE OBSERVATIONS NO REMARKABLE OBSERVATIONS NO REMARKABLE OBSERVATIONS NO REMARKABLE OBSERVATIONS M LATERAL VENTRICLE DILATED - TISSUE DEPRESSED 81ALTERAL NO REMARKABLE OBSERVATIONS NO REMARKABLE OBSERVATIONS V DILATED URETER - UNILATERAL RIGHT V DILATED RENAL PELVIS - UNILATERAL RIGHT V LATERAL VENTRICLE DILATED - NO DEPRESSION BILATERAL NO REMARKABLE OBSERVATIONS V LATERAL VENTRICLE DILATED - NO DEPRESSION - BILATERAL GRADE CODE: 1-SLIGHT, 2-MODERATE. 3-MARKED P-PRESENT M-MALFORMATION V-VARIATION GRADE I 1 P P t 1 Report 50-514 Appendix 4B Page 18 SL 042708) DEVELOPMENTAL TOXICITY STUDY OF INHALED 1.1.1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS FETAL VISCERAL FINDINGS DAMS FROM GROUP 3: M - 3000 PPM FETUS# VISCERAL OBSERVATION 15995 V IRREGULAR RUGAE NO REMARKABLE OBSERVATIONS NO REMARKABLE OBSERVATIONS NO REMARKABLE OBSERVATIONS NO REMARKABLE OBSERVATIONS NO REMARKABLE OBSERVATIONS V THIRD VENTRICLE DILATED - NO DEPRESSION 15998 8 NO REMARKABLE OBSERVATIONS 1 1 V LATERAL VENTRICLE DILATED - NO DEPRESSION LEFT 16011 NO REMARKABLE OBSERVATIONS NO REMARKABLE OBSERVATIONS NO REMARKABLE OBSERVATIONS V LATERAL VENTRICLE DILATED NO DEPRESSION BILATERAL NO REMARKABLE OBSERVATIONS V LATERAL VENTRICLE DILATED - NO DEPRESSION LEFT NO REMARKABLE OBSERVATIONS V LATERAL VENTRICLE DILATED - NO DEPRESSION BILATERAL 16013 NO REMARKABLE OBSERVATIONS NO REMARKABLE OBSERVATIONS NO REMARKABLE OBSERVATIONS NO REMARKABLE OBSERVATIONS NO REMARKABLE OBSERVATIONS V AZYGOUS LUNG LOBE MISSING GRADE CODE: 1-SLIGHT. 2-MODERATE, 3-MARKED, P-PRESENT. il-MALFORMATION V-VARIATION GRADE P Report 50-514 Appendi* 4B Page 19 (60Zz*o DEVELOPMENTAL TOXICITY STUDY OF INHALED 1.1,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS FETAL VISCERAL FINDINGS DAMS FROM GROUP 3: M - 3000 PPM FETUS# 16013 W 0 9 10 11 12 16018 1 2 3 4 5 6 7 6 9 10 11 12 16021 1 2 3 4 5 VISCERAL OBSERVATION M VENTRICULAR SEPTAL DEFECT MUSCULAR NO REMARKABLE OBSERVATIONS NO REMARKABLE OBSERVATIONS NO REMARKABLE OBSERVATIONS NO REMARKABLE OBSERVATIONS NO REMARKABLE OBSERVATIONS V IRREGULAR RUGAE NO REMARKABLE OBSERVATIONS NO REMARKABLE OBSERVATIONS NO REMARKABLE OBSERVATIONS NO REMARKABLE OBSERVATIONS NO REMARKABLE OBSERVATIONS NO REMARKABLE OBSERVATIONS NO REMARKABLE OBSERVATIONS V LATERAL VENTRICLE DILATED - NO DEPRESSION BILATERAAL NO REMARKABLE OBSERVATIONS NO REMARKABLE OBSERVATIONS V LATERAL VENTRICLE DILATED - NO DEPRESSION LEFT NO REMARKABLE OBSERVATIONS NO REMARKABLE OBSERVATIONS NO REMARKABLE OBSERVATIONS V FETAL ATELECTASIS V LATERAL VENTRICLE DILATED - NO DEPRESSION BILATERAL GRADE CODE: 1-SLIGHT, 2-MODERATE, 3-MARKED, P-PRESENT M-MALFORMATION V-VARIATION GRADE P P 1 1 P 1 Report 5Q-5T4 Appendix 4B Page 20 O T Z -^0 # DEVELOPMENTAL TOX1CITV STUDY OF INHALED 1.1,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS FETAL VISCERAL FINDINGS DAMS FROM GROUP 3: M - 3000 PPM FETUS# VISCERAL OBSERVATION 16032 1 NO REMARKABLE OBSERVATIONS 2 NO REMARKABLE OBSERVATIONS 3 V AZYGOUS LUNG LOBE MISSING 4 NO REMARKABLE OBSERVATIONS 5 NO REMARKABLE OBSERVATIONS 6 NO REMARKABLE OBSERVATIONS 7 NO REMARKABLE OBSERVATIONS e NO REMARKABLE OBSERVATIONS 9 NO REMARKABLE OBSERVATIONS 10 NO REMARKABLE OBSERVATIONS 1 1 NO REMARKABLE OBSERVATIONS 16034 1 NO REMARKABLE OBSERVATIONS 2 NO REMARKABLE OBSERVATIONS 3 NO REMARKABLE OBSERVATIONS 4 NO REMARKABLE OBSERVATIONS 5 NO REMARKABLE OBSERVATIONS 6 NO REMARKABLE OBSERVATIONS 7 V THIRD VENTR]!CLE DILATED - NO DEPRESSION S NO REMARKABLE OBSERVATIONS 9 NO REMARKABLE OBSERVATIONS 16049 1 NO REMARKABLE OBSERVATIONS 2 NO REMARKABLE OBSERVATIONS 3 NO REMARKABLE OBSERVATIONS 4 NO REMARKABLE OBSERVATIONS 5 NO REMARKABLE OBSERVATIONS 6 NO REMARKABLE OBSERVATIONS 7 NO REMARKABLE OBSERVATIONS 8 NO REMARKABLE OBSERVATIONS GRADE CODE: 1-SLIGHT, 2-MODERATE, 3-MARKED, P-PRESENT, M-MALFORMATION V-VARIATION GRADE P 1 Report 50-514 Appendi* 4B Page 2 1 DEVELOPMENTAL TOXICITY STUDY OF INHALED 1,1,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS FETAL VISCERAL FINDINGS DAMS FROM GROUP 3; M - 3000 PPM FETUS# VISCERAL OBSERVATION .. 16049 9 V IRREGULAR RUGAE GRADE CODE: 1-SLIGHT, 2-MODERATE, 3-MARKED. P-PRESENT, M-MALFORMATION V-VARIATION GRADE p SL 042711 Report 50-514 Appendix 4B Page 22 aa SL 042712 DEVELOPMENTAL TOXICITY STUDY OF INHALED 1,1,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS FETAL VISCERAL FINDINGS DAMS FROM GROUP 4: R - 6000 PPM FETUS# VISCERAL OBSERVATION 15948 15951 15955 1 NO REMARKABLE OBSERVATIONS 2 NO REMARKABLE OBSERVATIONS 3 NO REMARKABLE OBSERVATIONS 4 NO REMARKABLE OBSERVATIONS 5 NO REMARKABLE OBSERVATIONS 6 NO REMARKABLE OBSERVATIONS 7 NO REMARKABLE OBSERVATIONS B V REDUCED NUMBER OF TEETH PRiMORDIA TWO LEFT, ZERO RIGHT 1 NO REMARKABLE OBSERVATIONS 2 NO REMARKABLE OBSERVATIONS 3 NO REMARKABLE OBSERVATIONS 4 NO REMARKABLE OBSERVATIONS 5 NO REMARKABLE OBSERVATIONS 6 NO REMARKABLE OBSERVATIONS 7 NO REMARKABLE OBSERVATIONS B NO REMARKABLE OBSERVATIONS 9 NO REMARKABLE OBSERVATIONS 10 NO REMARKABLE OBSERVATIONS 2 NO REMARKABLE OBSERVATIONS 3 V AZYGOUS LUNG LOBE MISSING 4 V DILATED RENAL PELVIS - UNILATERAL RIGHT V AZYGOUS LUNG LOBE MISSING 6 NO REMARKABLE OBSERVATIONS 7 NO REMARKABLE OBSERVATIONS B NO REMARKABLE OBSERVATIONS GRADE CODE: 1-SLIGHT, 2-MODERATE, 3-MARKED, P-PRESENT M-MALFORMATION V-VARIATION GRADE P P Report 50-5 1 4 Appendix 4B Page 23 SL 042713 DEVELOPMENTAL TOXICITY STUDY OF INHALED 1.1,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS FETAL VISCERAL FINDINGS DAMS FROM GROUP 4: R - 6000 PPM FETUS# VISCERAL OBSERVATION GRADE 15959 15972 15978 1 NO REMARKABLE OBSERVATIONS 2 NO REMARKABLE OBSERVATIONS 3 NO REMARKABLE OBSERVATIONS 4 NO REMARKABLE OBSERVATIONS 5 NO REMARKABLE OBSERVATIONS 6 NO REMARKABLE OBSERVATIONS 7 NO REMARKABLE OBSERVATIONS 1 NO REMARKABLE OBSERVATIONS 2 V LATERAL VENTRICLE DILATED - NO DEPRESSION RIGHT 3 NO REMARKABLE OBSERVATIONS 4 NO REMARKABLE OBSERVATIONS 5 NO REMARKABLE OBSERVATIONS 7 NO REMARKABLE OBSERVATIONS 8 NO REMARKABLE OBSERVATIONS 10 NO REMARKABLE OBSERVATIONS 1 NO REMARKABLE OBSERVATIONS 2 V THIRD VENTRICLE DILATED - NO DEPRESSION V IRREGULAR RUGAE 3 NO REMARKABLE OBSERVATIONS 4 NO REMARKABLE OBSERVATIONS 5 M ENLARGEO AORTA M SMALL BUT PATENT PULMONARY ARTERY LEFT ANO RIGHT PULMONARY ARTERIES ANO DUCTUS ARERIOSUS ARE NORMAL V MEMBRANOUS VENTRICULAR SEPTUM THINNED M SEMILUNAR VALVES OF THE PULMONARY ARTERY MISSING GRADE CODE: 1-SLIGHT. 2-MODERATE. 3-MARKED, P-PRESENT, M-MALFORMATION V-VARIATION t t P P P P P Report 50-514 Appendix 4B Page 24 SL 042714 DEVELOPMENTAL TOXICITV STUDY OF INHALED 1 , 1 , 1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS FETAL VISCERAL FINDINGS DAMS FROM GROUP 4: R - 6000 PPM FETUS# VISCERAL OBSERVATION 15978 5 M SEMILUNAR VALVES OF THE AORTA MISSING M RIGHT VENTRICLE OF THE HEART SMALL M LEFT VENTRICLE OF THE HEART ENLARGED 6 NO REMARKABLE OBSERVATIONS 7 NO REMARKABLE OBSERVATIONS 8 NO REMARKABLE OBSERVATIONS 15982 1 V AZYGOUS LUNG LOBE MISSING 2 NO REMARKABLE OBSERVATIONS 3 NO REMARKABLE OBSERVATIONS 5 V AZYGOUS LUNG LOBE MISSING M LATERAL VENTRICLE DILATED - TISSUE DEPRESSED BILATERAL 6 NO REMARKABLE OBSERVATIONS 7 V AZYGOUS LUNG LOBE MISSING 15984 1 NO REMARKABLE OBSERVATIONS 2 NO REMARKABLE OBSERVATIONS 3 NO REMARKABLE OBSERVATIONS 4 NO REMARKABLE OBSERVATIONS 5 NO REMARKABLE OBSERVATIONS 6 NO REMARKABLE OBSERVATIONS 7 NO REMARKABLE OBSERVATIONS a V AZYGOUS LUNG LOBE MISSING 9 NO REMARKABLE OBSERVATIONS 10 V THIRD VENTRICLE DILATED - NO DEPRESSION 15989 1 NO REMARKABLE OBSERVATIONS 2 NO REMARKABLE OBSERVATIONS 3 NO REMARKABLE OBSERVATIONS GRADE CODE: t-SLIGHT. 2-MODERATE, 3-MARKED, P-PRESENT, M-MALFORMATION V-VARIATION GRADE P P P P 1 P P 1 Report 50-514 Appendi* 4B Page 25 SL 042715 DEVELOPMENTAL TOXICITY STUDY OF INHALED 1 , 1 . 1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS FETAL VISCERAL FINDINGS DAMS FROM GROUP 4: R - 6000 PPM FETUS# VISCERAL OBSERVATION 15989 4 NO REMARKABLE OBSERVATIONS 5 NO REMARKABLE OBSERVATIONS 6 NO REMARKABLE OBSERVATIONS 7 NO REMARKABLE OBSERVATIONS 8 NO REMARKABLE OBSERVATIONS 9 NO REMARKABLE OBSERVATIONS 10 NO REMARKABLE OBSERVATIONS 15999 NO REMARKABLE OBSERVATIONS 2 NO REMARKABLE OBSERVATIONS 3 NO REMARKABLE OBSERVATIONS 4 NO REMARKABLE OBSERVATIONS 5 NO REMARKABLE OBSERVATIONS 6 NO REMARKABLE OBSERVATIONS 7 NO REMARKABLE OBSERVATIONS 8 NO REMARKABLE OBSERVATIONS 16001 1 NO REMARKABLE OBSERVATIONS 3 NO REMARKABLE OBSERVATIONS 4 NO REMARKABLE OBSERVATIONS 5 V LATERAL VENTRICLE DILATED - NO DEPRESSION BILATERAL 6 V PARTIAL FETAL ATELECTASIS 7 NO REMARKABLE OBSERVATIONS 16007 1 M VENTRICULAR SEPTAL DEFECT MEMBRANOUS 2 NO REMARKABLE OBSERVATIONS 3 NO REMARKABLE OBSERVATIONS 4 M VENTRICULAR SEPTAL DEFECT GRADE CODE: 1-SLIGHT, 2-MODERATE, 3-MARKEO, P-PRESENT, M-MALFORMATION V-VARIATION GRADE 1 P P P Repo rt 50-51 A Append i k 4B Page 26 r 042716 DEVELOPMENTAL TOXICITY STUDY OP INHALED 1,t,I-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS FETAL VISCERAL FINDINGS DAMS FROM GROUP 4: R - 6000 PPM 16007 FETUS# 5 6 7 16009 16014 B 9 1 2 3 4 5 6 1 2 3 4 5 6 7 B 9 10 VISCERAL OBSERVATION MEMBRANOUS NO REMARKABLE OBSERVATIONS NO REMARKABLE OBSERVATIONS M ECTOPIC TESTIS LEFT - TOWARD KIDNEY RIGHT - CAUOALLY, LYING NEXT TO URINARY V RIGHT TESTIS THIN WITH NO CHARACTERISTIC NO REMARKABLE OBSERVATIONS NO REMARKABLE OBSERVATIONS BLADDER SHAPE NO REMARKABLE OBSERVATIONS NO REMARKABLE OBSERVATIONS NO REMARKABLE OBSERVATIONS V LATERAL VENTRICLE DILATED - NO DEPRESSION BILATERAL V THIRD VENTRICLE DILATED - NO DEPRESSION NO REMARKABLE OBSERVATIONS NO REMARKABLE OBSERVATIONS NO REMARKABLE OBSERVATIONS NO REMARKABLE OBSERVATIONS NO REMARKABLE OBSERVATIONS NO REMARKABLE OBSERVATIONS NO REMARKABLE OBSERVATIONS NO REMARKABLE OBSERVATIONS NO REMARKABLE OBSERVATIONS NO REMARKABLE OBSERVATIONS NO REMARKABLE OBSERVATIONS V AZYGOUS LUNG LOBE MISSING GRADE CODE: 1-SLIGHT. 2-MODERATE, 3-MARKED, P-PRESENT M-MALFORMATION V-VARIATION' GRADE P P 1 1 P Report 50-514 Appendix 4B Page 27 a. a SL 042717 DEVELOPMENTAL TOXICITV STUDY OF INHALED t.1,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS FETAL VISCERAL FINDINGS DAMS FROM GROUP 4: R - 6000 PPM FETUSA VISCERAL OBSERVATION 16019 4 NO REMARKABLE OBSERVATIONS 5 NO REMARKABLE OBSERVATIONS 7 V DARK GREEN FLUIO IN STOMACH V DARK GREEN FLUIO IN INTESTINES 8 V LATERAL VENTRICLE DILATED - NO DEPRESSION LEFT 16022 1 V LATERAL VENTRICLE DILATED - NO DEPRESSION' BILATERAL V THIRD VENTRICLE DILATED - NO DEPRESSION 2 NO REMARKABLE OBSERVATIONS 3 M LATERAL VENTRICLE DILATED - TISSUE DEPRESSED ' BILATERAL V THIRD VENTRICLE DILATED - NO DEPRESSION M CAVITATION IN MEDULLA 4 NO REMARKABLE OBSERVATIONS 6 NO REMARKABLE OBSERVATIONS 16023 2 NO REMARKABLE OBSERVATIONS 3 NO REMARKABLE OBSERVATIONS 4 NO REMARKABLE OBSERVATIONS 5 NO REMARKABLE OBSERVATIONS 6 NO REMARKABLE OBSERVATIONS 16033 NO REMARKABLE OBSERVATIONS M LATERAL VENTRICLE DILATED RIGHT V LATERAL VENTRICLE DILATED LEFT V THIRD VENTRICLE DILATED - - TISSUE DEPRESSED - NO DEPRESSION NO DEPRESSION GRADE CODE: 1-SLIGHT, 2-MODERATE, 3-MARKED, P-PRESENT M-MALFORMATION V-VARIATION GRADE 1 1 1 P t P 1 1 1 Repor t 5 0-514 Appendi* 4B Page 28 SL 042718 DEVELOPMENTAL TOXICITV STUDY OF INHALED 1 , 1 . 1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS FETAL VISCERAL FINDINGS DAMS FROM GROUP 4: R - 6000 PPM FETUS# VISCERAL OBSERVATION 16033 5 NO REMARKABLE OBSERVATIONS 6 M LATERAL VENTRICLE DILATED - TISSUE DEPRESSED BILATERAL 16040 1 NO REMARKABLE OBSERVATIONS 2 NO REMARKABLE OBSERVATIONS 3 NO REMARKABLE OBSERVATIONS 4 NO REMARKABLE OBSERVATIONS 5 NO REMARKABLE OBSERVATIONS 6 V THIRD VENTRICLE DILATED - NO DEPRESSION 7 NO REMARKABLE OBSERVATIONS S NO REMARKABLE OBSERVATIONS 9 NO REMARKABLE OBSERVATIONS 10 NO REMARKABLE OBSERVATIONS 16044 1 NO REMARKABLE OBSERVATIONS 2 M LATERAL VENTRICLE DILATED - TISSUE DEPRESSED BILATERAL 3 NO REMARKABLE OBSERVATIONS 4 V REDUCED NUMBER OF TEETH PRIMORDIA ONE RIGHT V TEETH PRIMORDIA SMALLER THAN NORMAL M THIRD VENTRICLE DILATED - TISSUE DEPRESSED 5 NO REMARKABLE OBSERVATIONS a NO REMARKABLE OBSERVATIONS 9 NO REMARKABLE OBSERVATIONS to NO REMARKABLE OBSERVATIONS GRADE CODEt 1-SLIGHT, 2-MODERATE. 3-MARKED, P-PRESENT M-MALFORMATION V-VARIAT10N GRADE P P P P 1 Report 50-514 Appendix 4B Page 29 a a a acl a a a acl SL 042719 DEVELOPMENTAL TOXICITY STUDY OF INHALED 1,1,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS FETAL SKELETAL FINDINGS FEMALES FROM G - 0 PPM IMPLANT # SKELETAL OBSERVATION 15946 V STERNEBRA #6 POORLY OSSIFIED V SOME INTER.PHALANGES (FORELIMB) POORLY OSSIFIED ONE BILATERAL V EXTRA RIB 413 - FIRST LUMBAR ARCH - UNILATERAL RIGHT V RUDIMENTARY RIB #13 - FIRST LUMBAR ARCH - UNILATERAL LEFT V SOME INTER.PHALANGES (FORELIMB) POORLY OSSIFIED ONE BILATERAL V STERNEBRA #5 POORLY OSSIFIED V STERNEBRA #6 POORLY OSSIFIED V CERVICAL CENTRA #1,#2,#3 AND/OR #4 POORLY OSSIFIED #1 V EXTRA RIB #13 - FIRST LUMBAR ARCH - BILATERAL V PARIETAL - POORLY OSSIFIED BILATERAL V BODY OF THE HYOID - POORLY OSSIFIED V STERNEBRA #5 POORLY OSSIFIED V STERNEBRA #6 POORLY OSSIFIED V CERVICAL CENTRA #1,#2,#3 AND/OR #4 POORLY OSSIFIED V EXTRA RIB #13*- FIRST LUMBAR ARCH - UNILATERAL LEFT V STERNEBRA #6 POORLY OSSIFIED V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL V BODY OF THE HYOID - POORLY OSSIFIEO V SOME INTER.PHALANGES (FORELIMB) POORLY OSSIFIED. ONE BILATERAL V STERNEBRA #5 POORLY OSSIFIED V STERNEBRA #6 POORLY OSSIFIED 13 V SOME INTER.PHALANGES (FORELIMB) POORLY OSSIFI ED ONE BILATERAL V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL V STERNEBRA #5 POORLY OSSIFIEO V STERNEBRA #6 POORLY OSSIFIED 15953 NO REMARKABLE OBSERVATIONS V STERNEBRA #6 POORLY OSSIFIED V RUDIMENTARY RIB #13 - FIRST LUMBAR ARCH V EXTRA #13 THORACIC CENTRUM AND ARCHES .BILATERAL GRADE CODE: 1-SLIGHT, 2-MOOERATE. 3-MARKED, P-PRESENT, M-MALFORMATION V-VARIATION. GRADE (P) P P P P P P P P P P P Repor t 50-5 14 Appendi 4C Page 1 aa 042720 co DEVELOPMENTAL TOXICITY STUDY OF INHALED 1 , 1 . 1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS FETAL SKELETAL FINDINGS FEMALES FROM G - 0 PPM IMPLANT # SKELETAL OBSERVATION 15953 6 V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL 8 V EXTRA RIB #13 - FIRST LUMBAR ARCH - BILATERAL 15954 1 V STERNEBRA #5 UNOSSIFIED V STERNEBRA #6 POORLY OSSIFIED V RUDIMENTARY RIB #13 - FIRST LUMBAR ARCH - UNILATERAL RIGHT 3 V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL 6 V STERNEBRA #5 POORLY OSSIFIED 8 V STERNEBRA #5 POORLY OSSIFIED V RUDIMENTARY RIB #13 - FIRST LUMBAR ARCH - UNILATERAL LEFT 15961 2 V STERNEBRAE #4 AND #5 FUSED V RUDIMENTARY RIB #13 - FIRST LUMBAR ARCH -'BI LATERAL 4 V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL 6 V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL 8 V STERNEBRA #5 POORLY OSSIFIED V EXTRA RIB #13 - FIRST LUMBAR ARCH - BILATERAL 15967 1 V EXTRA RIB #13 - FIRST LUMBAR ARCH - BILATERAL 3 V STERNEBRA #6 POORLY OSSIFIED V EXTRA RIB #13 - FIRST LUMBAR ARCH - BILATERAL 5 VCERVICAL CENTRA #t,#2,#3 AND/OR #4 POORLY OSSIFIED #1 V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL 8 NO REMARKABLE OBSERVATIONS 10 V STERNEBRA #6 POORLY OSSIFIED V EXTRA RIB #13 - FIRST LUMBAR ARCH - BILATERAL V CERVICAL CENTRA #1,#2,#3 AND/OR #4 POORLY OSSIFIED #1 15980 2 V EXTRA RIB #13 - FIRST LUMBAR ARCH - BILATERAL 4 V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL 6 V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL 8 V STERNEBRA #6 POORLY OSSIFIED GRADE CODE: 1-SLIGHT, 2-MODERATE, 3-MARKED, P-PRESENT M-MALFORMATION V-VARIATION GRADE P P P P P P P P P P P P P P P P P P P P P P P P P P P P P P P Repor t 50-5 1 4 Appendix 4C Page 2 SL 042721 FEMALES FROM G - 0 PPM DEVELOPMENTAL TOXICITY STUDY OF INHALED 1,1,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS FETAL SKELETAL FINDINGS IMPLANT # SKELETAL OBSERVATION 15991 15993 15994 16003 16005 3 NO REMARKABLE OBSERVATIONS 5 NO REMARKABLE OBSERVATIONS 7 V EXTRA RIB #13 - FIRST LUMBAR ARCH - BILATERAL 9 V EXTRA RIB #13 - FIRST LUMBAR ARCH - UNILATERAL RIGHT V RUDIMENTARY RIB #13 - FIRST LUMBAR ARCH - UNILATERAL LEFT 2 NO REMARKABLE OBSERVATIONS 5 V EXTRA RIB #13 - FIRST LUMBAR ARCH - BILATERAL M RIB #11 - MISSING - UNILATERAL LEFT M THORACIC ARCH MISSING - UNILATERAL #11 LEFT M THORACIC CENTRUM AND ARCHES DIAGONALLY DISPLACED #10 AND #12 M LUMBAR CENTRUM AND ARCHES DIAGONALLY DISPLACED. #1 V THORACIC CENTRUM #11 POORLY OSSIFIED ONLY ONE OSSIFICATION CENTER 7 V STERNEBRA #6 POORLY OSSIFIEO V RUDIMENTARY RIB #13 - FIRST LUMBAR ARCH - UNILATERAL LEFT 9 NO REMARKABLE OBSERVATIONS 2 V STERNEBRA #5 POORLY OSSIFIED V STERNEBRA #6 POORLY OSSIFIEO V EXTRA RIB #13 - FIRST LUMBAR ARCH - BILATERAL 4 V STERNEBRA #5 POORLY OSSIFIED V STERNEBRA #6 POORLY OSSIFIED V EXTRA RIB #13 - FIRST LUMBAR ARCH - BILATERAL 6 V EXTRA RIB #13- FIRST LUMBAR ARCH - BILATERAL 8 V STERNEBRA #5 POORLY OSSIFIEO V STERNEBRA #6 POORLY OSSIFIED V RUDIMENTARY RIB #13 - FIRST LUMBARARCH - BILATERAL V EXTRA RIB #13 - FIRST LUMBAR ARCH - BILATERAL V STERNEBRA #5 POORLY OSSIFIED V STERNEBRA #6 POORLY OSSIFIEO V EXTRA #13 THORACIC CENTRUM AND ARCHES GRADE CODE: 1-SLIGHT. 2-MODERATE, 3-MARKED, P-PRESENT M-MALFORMATION V-VARIATION GRADE P P P P P P P P 3 P P P P P P P P P P P P P P P P Report 50-514 Append i a 4C Page 3 SL 042722 DEVELOPMENTAL TOXICITY STUDY OF INHALED 1,1,1-TRICHLOROETHANE IN NEW ZEALANO WHITE RABBITS FETAL SKELETAL FINDINGS FEMALES FROM G - 0 PPM IMPLANT # SKELETAL OBSERVATION 16005 2 V RUDIMENTARY RIB #13 - THORACIC ARCH #13 - UNILATERAL LEFT 4 V STERNEBRA #5 POORLY OSSIFIEO 6 V STERNEBRA #5 UNOSSIFIED V STERNEBRA #6 POORLY OSSIFIED 8 V PARIETAL - POORLY OSSIFIED BILATERAL V PARIETAL - HOLE IN BONE BILATERAL V STERNEBRA #5 POORLY OSSIFIED 10 V STERNEBRA #6 POORLY OSSIFIED V EXTRA RIB #13 - FIRST LUMBAR ARCH - BILATERAL 16006 1 V STERNEBRA #5 POORLY OSSIFIED V STERNEBRA #6 POORLY OSSIFIED V EXTRA RIB #13 - FIRST LUMBAR ARCH - BILATERAL 3 V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL 6 V STERNEBRA #5 POORLY OSSIFIED V STERNEBRA #6 POORLY OSSIFIED V EXTRA RIB #13 - FIRST LUMBAR ARCH - BILATERAL B V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL V LUMBAR CENTRUM #7 POORLY OSSIFIED 16016 2 V STERNEBRAE #4 AND #5 FUSED V EXTRA RIB #13 - FIRST LUMBAR ARCH - UNILATERAL LEFT 4 NO REMARKABLE OBSERVATIONS 6 V SOME INTER.PHALANGES (FORELIMB) POORLY OSSIFIEO ONE BILATERAL V EXTRA RIB #13 - FIRST LUMBAR ARCH - BILATERAL 8 V RUDIMENTARY RIB #13 - FIRST LUMBAR ARCH - UNILATERAL RIGHT V EXTRA RIB #13 - FIRST LUMBAR ARCH - UNILATERAL LEFT 10 NO REMARKABLE OBSERVATIONS 16020 1 V STERNEBRA #6 POORLY OSSIFIED V EXTRA RIB #13 - FIRST LUMBAR ARCH - BILATERAL 3 V RUDIMENTARY RIB #13 - FIRST LUMBAR ARCH - UNILATERAL RIGHT GRADE CODE: 1-SLIGHT. 2-MODERATE, 3-MARKEO, P-PRESENT M-MALFORMATION V-VARIATION a aa a a a a. cl a a GRADE P P P P P P P P P P P P P P P P P P P Report 50-514 Appendi x 4C Page 4 SL 042723 DEVELOPMENTAL TOXICITY STUDY OF INHALED 1,1.1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS FETAL SKELETAL FINDINGS FEMALES FROM G - 0 PPM IMPLANT # SKELETAL OBSERVATION 1 6070 3 V EXTRA RIB #13 - FIRST LUMBAR ARCH - UNILATERAL LEFT 5 V STERNEBRA #6 POORLY OSSIFIED V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL 7 NO REMARKABLE OBSERVATIONS 9 V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL 16025 1 NO REMARKABLE OBSERVATIONS 3 V STERNEBRA #5 POORLY OSSIFIED 5 NO REMARKABLE OBSERVATIONS 7 NO REMARKABLE OBSERVATIONS 9 V STERNEBRA #6 POORLY OSSIFIED 16028 2 V SOME INTER.PHALANGES (FORELIMB) POORLY OSSIFIED ONE BILATERAL V EXTRA RIB #13 - FIRST LUMBAR ARCH - BILATERAL 4 V SOME INTER.PHALANGES (FORELIMB) POORLY OSSIFIED ONE BILATERAL V STERNEBRA #6 POORLY OSSIFIED V RUDIMENTARY RIB #13 - FIRST LUMBAR ARCH - UNILATERAL RIGHT V EXTRA RIB #13 - FIRST LUMBAR ARCH - UNILATERAL LEFT 7 V STERNEBRA #6 POORLY OSSIFIED V RUDIMENTARY RIB #13 - FIRST LUMBAR ARCH - UNILATERAL RIGHT V EXTRA RIB #13 - FIRST LUMBAR ARCH - UNILATERAL LEFT 16036 1 V STERNEBRA #6 POORLY OSSIFIED 3 V STERNEBRA #5 POORLY OSSIFIED V STERNEBRA #6 POORLY OSSIFIED V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL 5 V BODY OF THE HYOID - POORLY OSSIFIED V STERNEBRA #6 POORLY OSSIFIED 7 V STERNEBRA #5 POORLY OSSIFIED V STERNEBRA #6 POORLY OSSIFIED V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL GRADE CODE: 1-SLIGHT, 2-MODERATE. 3-MARKED, P-PRESENT M-MALFORMATION V-VARIATION V V T3 TJ T ? T 7 ~V V JJ * 0 T 3 TJ TJ 'O "U GRADE P P P P P P P P P P P P Repor t 50-51 A Append 1 * AC Page 5 0 - q. O .Q .Q . a a a a. SL 042724 DEVELOPMENTAL TOXICITY STUOY OF INHALED 1,1.1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS FETAL SKELETAL FINDINGS FEMALES FROM G - 0 PPM IMPLANT # SKELETAL OBSERVATION 16036 9 V STERNEBRA 05 POORLY OSSIFIED V STERNEBRA *6 POORLY OSSIFIED V RUDIMENTARY RIB #13 - FIRST LUMBAR ARCH - UNILATERAL LEFT 16039 16041 2 v EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL 4 V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL 1 v EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL 4 V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL 6 V EXTRA RIB #13 - FIRST LUMBAR ARCH - BILATERAL 16045 2 V STERNEBRA #5 POORLY OSSIFIED V RUDIMENTARY RIB #13 - FIRST LUMBAR ARCH - UNILATERAL LEFT 4 V STERNEBRA #2 POORLY OSSIFIED V STERNEBRA #5 POORLY OSSIFIED V RUDIMENTARY RIB #13 - FIRST LUMBAR ARCH - UNILATERAL RIGHT 6 V STERNEBRA #5 POORLY OSSIFIED B V EXTRA RIB #13 - FIRST LUMBAR ARCH - BILATERAL 16047 1 V EXTRA RIB #13 - FIRST LUMBAR ARCH - BILATERAL 3 V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL 5 V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL 7 V EXTRA RIB #13 - FIRST LUMBAR ARCH - BILATERAL 9 V SOME INTER.PHALANGES (FORELIMB) POORLY OSSIFIED ONE BILATERAL V THORACIC CENTRUM #6 BILOBED V THORACIC CENTRUM #7 BILOBED V RUDIMENTARY RIB *13 - FIRST LUMBAR ARCH - UNILATERAL RIGHT V EXTRA RIB #13 - FIRST LUMBAR ARCH - UNILATERAL LEFT GRADE CODE: 1-SLIGHT. 2-MODERATE. 3-MARKED, P-PRESENT M-MALFORMATION V-VARIATION GRADE P P P P P P P P P P P P P P P P P P P P P Report 50-514 A ppendi x 4C Page 6 SL 042725 FEMALES FROM I - 1000 PPM DEVELOPMENTAL TOXICITY STUDY OP INHALED 1,1.1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS FETAL SKELETAL FINDINGS IMPLANT 0 SKELETAL OBSERVATION 15945 15957 15963 15964 15965 Z V STERNEBRA 05 POORLY OSSIFIED 5 V STERNEBRA #5 POORLY OSSIFIED V EXTRA RIB 013 - FIRST LUMBAR ARCH - BILATERAL 7 V SOME INTER.PHALANGES (FORELIMB) POORLY OSSIFIED ONE BILATERAL V STERNEBRA #5 POORLY OSSIFIED 9 V SOME INTER.PHALANGES (FORELIMB) POORLY OSSIFIED ONE BILATERAL V EXTRA 013 THORACIC CENTRUM AND ARCHES V EXTRA RIB 013 - THORACIC ARCH 013 - BILATERAL 2 V MAXILLARY - POORLY OSSIFIED RIGHT V SOME INTER.PHALANGES (FORELIMB) POORLY OSSIFIEO ONE BILATERAL V STERNEBRA 05 POORLY OSSIFIED 4 V EXTRA 013 THORACIC CENTRUM AND ARCHES V EXTRA RIB 013 - THORACIC ARCH 013 - BILATERAL 7 V RUDIMENTARY RIB 013 - FIRST LUMBAR ARCH - UNILATERAL LEFT V EXTRA RIB 013 - FIRST LUMBAR ARCH - UNILATERAL RIGHT 1 NO REMARKABLE OBSERVATIONS 3 V EXTRA 013 THORACIC CENTRUM AND ARCHES V EXTRA RIB 013 - THORACIC ARCH 013 - BILATERAL 6 V EXTRA 013 THORACIC CENTRUM AND ARCHES V EXTRA RIB 013 - THORACIC ARCH 013 -BILATERAL 5 NO REMARKABLE OBSERVATIONS 7 V EXTRA RIB 013 - FIRST LUMBAR ARCH - BILATERAL 9 NO REMARKABLE OBSERVATIONS 2 V EXTRA OSSIF. SITE ABOVE STERNEBRA SEGMENT 01 4 V STERNEBRA 05 POORLY OSSIFIED V EXTRA RIB 013 - FIRST LUMBAR ARCH - BILATERAL 7 V EXTRA RIB 013 - FIRST LUMBAR ARCH - UNILATERAL LEFT 9 V EXTRA 013 THORACIC CENTRUM AND ARCHES V EXTRA RIB 013 - THORACIC ARCH 013 - BILATERAL GRADE CODE; 1-SLIGHT. 2-MODERATE. 3-MARK ED, P-PRESENT M-MALFORMATION V-VARIATION GRADE P P P P P P P P P P P P P P P P P P P P P P P P P P Repor t 50-5 1 4 -Append 1x 4C Page 7 SL 042726) DEVELOPMENTAL TOXICITY STUDY OF INHALED 1,1,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS FETAL SKELETAL FINDINGS FEMALES FROM I - 1000 PPM IMPLANT # SKELETAL OBSERVATION 15971 2 V EXTRA *13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL V STERNEBRA #5 POORLY OSSIFIED V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA *13 THORACIC CENTRUM AND ARCHES V RUDIMENTARY RIB *13 - THORACIC ARCH #13 - BILATERAL V STERNEBRA #5 POORLY OSSIFIED V STERNEBRA #6 POORLY OSSIFIED 15975 V STERNEBRA #5 UNOSSIFIED V RUDIMENTARY RIB *13 - FIRST LUMBAR ARCH - BILATERAL V STERNEBRA #5 POORLY OSSIFIED V EXTRA RIB #13 - FIRST LUMBAR ARCH - BILATERAL V STERNEBRA #5 POORLY OSSIFIED V RUOIMENTARV RIB #13 - FIRST LUMBAR ARCH - UNILATERAL RIGHT 15981 NO REMARKABLE OBSERVATIONS V EXTRA RIB #13 - FIRST LUMBAR ARCH - BILATERAL 1598 3 V CERVICAL CENTRA *t,#2,#3 AND/OR #4 POORLY OSSIFIED #1 V EXTRA RIB #13 - FIRST LUMBAR ARCH - UNILATERAL RIGHT V RUOIMENTARV RIB #13 - FIRST LUMBAR ARCH - UNILATERAL LEFT V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB *13 - THORACIC ARCH #13 - BILATERAL 15986 NO REMARKABLE OBSERVATIONS V STERNEBRA #5 POORLY OSSIFIED 15996 NO REMARKABLE OBSERVATIONS 16010 NO REMARKABLE OBSERVATIONS V EXTRA RIB #13 - FIRST LUMBAR ARCH - UNILATERAL RIGHT V EXTRA RIB #13 - FIRST LUMBAR ARCH - BILATERAL 16015 V RUDIMENTARY RIB #13 - FIRST LUMBAR ARCH - UNILATERAL GRADE CODE: 1-SLIGHT, 2-MODERATE, 3-MARKED, P-PRESENT, M-MALFORMATION V-VARIATION GRADE P P P P P P P P P P P P P P Report 50-514 Append 1 * 4C Page 8 SL 042727 # DEVELOPMENTAL TOXICtTV STUDY OF INHALED 1.1,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS FETAL SKELETAL FINDINGS FEMALES FROM I - 1000 PPM IMPLANT # SKELETAL OBSERVATION 16015 RIGHT 4 V FRONTAL BONE - POORLY OSSIFIED LEFT V SOME INTER.PHALANGES (FORELIMB) POORLY OSSIFIED ONE BILATERAL V RUDIMENTARY RIB #13 - FIRST LUMBAR ARCH - BILATERAL 16026 1 V SOME INTER.PHALANGES (FORELIMB) POORLY OSSIFIED ONE BILATERAL V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL V STERNEBRA #6 POORLY OSSIFIED 3 V STERNEBRA #5 POORLY OSSIFIED V EXTRA RIB #13 - FIRST LUMBAR ARCH - BILATERAL 5 V STERNEBRA #5 POORLY OSSIFIED V STERNEBRA #6 POORLY OSSIFIED V EXTRA RIB #13 - FIRST LUMBAR ARCH - BILATERAL 7 V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL 9 V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL 16030 1 V BODY OF THE HYOIO - POORLY OSSIFIED V STERNEBRA #6 UNOSSIFIED V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL V EXTRA #13 THORACIC CENTRUM AND ARCHES V CERVICAL CENTRA #1,#2,#3 AND/OR #4 POORLY OSSIFIED #1 3 V BODY OF THE HYOID - POORLY OSSIFIED V STERNEBRA #6 POORLY OSSIFIED V EXTRA RIB #13 - FIRST LUMBAR ARCH - UNILATERAL RIGHT V RUDIMENTARY RIB #13 - FIRST LUMBAR ARCH - UNILATERAL LEFT 6 NO REMARKABLE OBSERVATIONS B V BOOV OF THE HYOID - POORLY OSSIFIED 10 V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL V BOOV OF THE HYOID - POORLY OSSIFIED 16037 2 V STERNEBRA #5 POORLY OSSIFIED V STERNEBRA #6 POORLY OSSIFIED GRADE CODE: 1-SLIGHT. 2-MOOERATE, 3-MARKED, P-PRESENT, M-MALFORMATION V-VARIATION O. Cl Cl Cl Q- q. &Cl. Cl GRADE P P P P P P P P P P P P P P P P P P P P P Report 50-514 Append t * 4C Page 9 SL 042728 DEVELOPMENTAL TOXICITY STUDY Of INHALED 1,1,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS FETAL SKELETAL FINDINGS FEMALES FROM I - 1000 PPM IMPLANT # SKELETAL OBSERVATION 16037 2 V EXTRA RIB #13 - FIRST LUMBAR ARCH - BILATERAL 4 V EXTRA #t3 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL 6 V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL GRADE CODE: 1-SLIGHT, 2-MODERATE, 3-MARKED, P-PRESENT, M-MALFORMATION V-VARIAT10N GRAOE P P P P P Report 50-514 Append!* 4C Page 10 q. a SL 042729 FEMALES FROM M - 3000 PPM DEVELOPMENTAL TOXICITV STUDY OF INHALED 1,1,1-TR1CHL0R0ETHANE IN NEW ZEALAND WHITE RABBITS FETAL SKELETAL FINDINGS IMPLANT # SKELETAL OBSERVATION 15942 15947 2 V STERNEBRA #5 POORLY OSSIFIED V RUDIMENTARY RIB #13 - FIRST LUMBAR ARCH - UNILATERAL RIGHT .* V EXTRA RIB #13 - FIRST LUMBAR ARCH - UNILATERAL LEFT 4 V SOME INTER.PHALANGES (FORELIMB) POORLY OSSIFIEO BILATERAL V STERNEBRA #5 UNOSSIFIEO V EXTRA RIB #13 - FIRST LUMBAR ARCH - BILATERAL V PARIETAL - POORLY OSSIFIED BILATERAL 6 V SOME INTER.PHALANGES (FORELIMB) POORLY OSSIFIED BILATERAL V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL B V EXTRA RIB #13 - FIRST LUMBAR ARCH - BILATERAL 10 V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL 12 V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL 2 V EXTRA RIB #13 - FIRST LUMBAR ARCH - BILATERAL 4 V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL 6 V THORACIC CENTRUM #6 BILOBED V THORACIC CENTRUM #2 SPLIT DIAGONALLY DISPLACED V THORACIC CENTRUM #3 SPLIT DIAGONALLY DISPLACED V THORACIC CENTRUM #4 SPLIT DIAGONALLY DISPLACED M THORACIC ARCH MISSING - UNILATERAL #3 LEFT M THORACIC CENTRUM AND ARCHES DIAGONALLY DISPLACED #2,#3,#4 AND #5 M THORACIC ARCHES FUSEO - UNILATERAL #4 TO #5 LEFT M RIB #3 MISSING - UNILATERAL LEFT M EXTRA RIB BETWEEN EXISTING RIBS - UNILATERAL BETWEEN #4 AND #5 RIGHT GRAOE CODE: 1-SLIGHT, 2-MODERATE, 3-MARKED. P-PRESENT. M-MALFORMATION V-VARIATI ON GRADE P P P P P P P P P P P P P P P P P p P P P P P P P Report 50-514 Append lx 4 C Page 1t Q. a SL 042730 DEVELOPMENTAL TOXICITY STUDY OF INHALED 1,t,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS FETAL SKELETAL FINDINGS FEMALES FROM M - 3000 PPM IMPLANT # SKELETAL OBSERVATION 15947 6 M RIBS *2 THROUGH #5 FUSED #2 TO #3 RIGHT #4 TO #5 BILATERAL 15950 2 V BODY OF THE HYOID - POORLY OSSIFIED V SOME INTER.PHALANGES (FORELIMB) POORLY OSSIFIED BILATERAL V STERNEBRA #5 POORLYOSSIFIED V STERNEBRA #6 POORLYOSSIFIED V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL' V LUMBAR ARCHES #6 POORLY OSSIFIED BILATERAL 6 V BODY OF THE HYOID - POORLY OSSIFIED V SOME INTER.PHALANGES (FORELIMB) POORLY OSSIFIED BILATERAL V STERNEBRA #2 POORLY OSSIFIED V STERNEBRA #5 POORLYOSSIFIED V STERNEBRA #6 POORLYOSSIFIED V STERNEBRAE #1 AND #2 FUSED V STERNEBRAE #4 AND #5 FUSED 15952 2 V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - UNILATERAL LEFT 4 V BODY OF THE HYOID - POORLY OSSIFIED V SOME INTER.PHALANGES (FORELIMB) POORLY OSSIFIED TWO BILATERAL V STERNEBRA #6 UNOSSIFIEO V EXTRA RIB #13 - FIRST LUMBAR ARCH - BILATERAL V CERVICAL CENTRA #1,#2,#3 AND/OR #4 POORLY. OSSIFI ED #1 7 V SOME INTER.PHALANGES (FORELIMB) POORLY OSSIFIED TWO BILATERAL V CERVICAL CENTRA #1,#2,#3 AND/OR #4 POORLY OSSIFIEO #1 V RUDIMENTARY RIB #13 - FIRST LUMBAR ARCH - UNILATERAL LEFT 9 V SOME INTER.PHALANGES (FORELIMB) POORLY OSSIFIEO ONE BILATERAL V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL GRADE CODE: 1-SLIGHT. 2-MODERATE, 3-MARKED, P-PRESENT . M-MALFORMATION V-VARIAtION GRADE P P P P P P P P P P P P P P P P P P P P P P P P P P Repon t 50-5 1 4 Appendix 4C o. a SL 042731 FEMALES FROM M - 3000 PPM DEVELOPMENTAL TOXICITY STUDY OF INHALED 1,1.1-TR1CHLOROETHANE IN NEW ZEALAND WHITE RABBITS FETAL SKELETAL FINDINGS IMPLANT # SKELETAL OBSERVATION 15956 15958 15960 15966 1 V EXTRA #13 THORACIC CENTRUM AND ARCHES V RUDIMENTARY RIB #13 - THORACIC ARCH #13 -UNILATERAL RIGHT V EXTRA RIB #13 - THORACIC ARCH #13 - UNILATERAL LEFT 3 V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL 5 V SOME INTER.PHALANGES (FORELIMB) POORLY OSSIFIED ONE BILATERAL V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB#13 - THORACIC ARCH #13 - BILATERAL 2 NO REMARKABLE OBSERVATIONS 4 NO REMARKABLE OBSERVATIONS 6 NO REMARKABLE OBSERVATIONS 8 V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL' 10 V RUDIMENTARY RIB #13 - FIRST LUMBAR ARCH - BILATERAL 2 V RUDIMENTARY RIB #13 - FIRST LUMBAR ARCH - UNILATERAL RIGHT V EXTRA RIB #13 - FIRST LUMBAR ARCH - BILATERAL LEFT 4 V STERNEBRA #5 POORLY OSSIFIED V EXTRA RIB #13 - FIRST LUMBAR ARCH - BILATERAL 7 V STERNEBRA #5 POORLY OSSIFIED V EXTRA RIB #13 - FIRST LUMBAR ARCH - BILATERAL V CERVICAL CENTRA #1,#2,#3 ANO/OR #4 POORLY OSSIFIED #1 9 V STERNEBRA #5 POORLY OSSIFIED V RUDIMENTARY RIB #13 - FIRST LUMBAR ARCH - BILATERAL 11 V STERNEBRA #5 POORLY OSSIFIED V CERVICAL CENTRA #1.#2.#3 ANO/OR #4 POORLY OSSIFIED #1 V RUDIMENTARY RIB #13 - FIRST LUMBAR ARCH - BILATERAL 1 V BODY OF THE HYOID - POORLY OSSIFIED V EXTRA RIB #13 - FIRST LUMBAR ARCH - BILATERAL 3 V EXTRA RIB #13 - FIRST LUMBAR ARCH - BILATERAL 5 V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL GRADE CODE: 1-SLIGHT, 2-MODERATE, 3-MARKED, P-PRESENT M-MALFORMATION V-VARIATION GRADE P P P P P P P P P P 3 P P P P P P P P P P P P P P P Report 50-514 Appendt * 4C Page 1 3 SL 042732 DEVELOPMENTAL TOXICITY STUDY OF INHALED 1,1.1 -TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS FETAL SKELETAL FINDINGS FEMALES FROM M - 3000 PPM IMPLANT 4 SKELETAL OBSERVATION 1596B 7 NO REMARKABLE OBSERVATIONS 15979 2 STERNEBRA 45 POORLY OSSIFIED EXTRA 413 THORACIC CENTRUM AND ARCHES EXTRA RIB 413 - THORACIC ARCH 413 - BILATERAL ' 4 EXTRA 413 THORACIC CENTRUM ANO ARCHES EXTRA RIB 413 - THORACIC ARCH 413 - BILATERAL 6 PARIETAL - HOLE IN BONE LEFT SOME INTER .PHALANGES (FORELIMB) POORLY OSSIFIED ONE LEFT EXTRA RIB 413 - FIRST LUMBAR ARCH - BILATERAL 8 EXTRA 413 THORACIC CENTRUM AND ARCHES EXTRA RIB 413 - THORACIC ARCH 413 - BILATERAL 1 1 STERNEBRA 45 POORLY OSSIFIED EXTRA 413 THORACIC CENTRUM AND ARCHES EXTRA RIB 413 - THORACIC ARCH 413 - BILATERAL 15987 1 V EXTRA RIB 413 - FIRST LUMBAR ARCH - BILATERAL 15988 2 V EXTRA 413 THORACIC CENTRUM AND ARCHES V EXTRA RIB 413 - THORACIC ARCH 413 - BILATERAL 4 V EXTRA RIB 413 - FIRST LUMBAR ARCH - BILATERAL 6 V EXTRA 413 THORACIC CENTRUM AND ARCHES V EXTRA RIB 413 - THORACIC ARCH 413 - BILATERAL V EXTRA 413 THORACIC CENTRUM AND ARCHES V EXTRA RIB 413 - THORACIC ARCH 413 - BILATERAL 15995 V EXTRA RIB 413 - FIRST LUMBAR ARCH - BILATERAL V STERNEBRA 45 POORLY OSSIFIED V STERNEBRA 46 POORLY OSSIFIED V EXTRA RIB 413 - FIRST LUMBAR ARCH - BILATERAL V EXTRA 413 THORACIC CENTRUM AND ARCHES V EXTRA RIB 413 - THORACIC ARCH 413 - BILATERAL 15998 NO REMARKABLE OBSERVATIONS 16011 V EXTRA 413 THORACIC CENTRUM AND ARCHES V EXTRA RIB 413 - THORACIC ARCH 413 - BILATERAL V SOME INTER.PHALANGES (FORELIMB) POORLY OSSIFIED ONE BILATERAL V EXTRA RIB 413 - FIRST LUMBAR ARCH - BILATERAL GRADE CODE: 1-SLIGHT, 2-MODERATE, 3-MARKED, P-PRESENT, M-MALFORMATION V-VARIATION GRADE fieport 50"514 Appendix 4C Page 14 Q-Q. a a a a i a a a a . a a a SL 042733 DEVELOPMENTAL TOXICITY STUDY OF INHALED 1,1,I-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS FETAL SKELETAL FINOINGS FEMALES FROM M - 3000 PPM IMPLANT # SKELETAL OBSERVATION 16011 3 V SOME INTER.PHALANGES (FORELIMB) POORLY OSSIFIED ONE BILATERAL 5 V STERNEBRA #5 POORLY OSSIFIED V STERNEBRA #6 POORLY OSSIFIED V EXTRA *13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL V SOME INTER.PHALANGES (FORELIMB) POORLY OSSIFIED ONE BILATERAL 7 V SOME INTER.PHALANGES (FORELIMB) POORLY OSSIFIED ONE BILATERAL V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL 16013 2 V STERNEBRA #5 UNOSSIFIEO 4 V FRONTAL BONE - POORLY OSSIFIED BILATERAL V STERNEBRA #5 POORLY OSSIFIED 7 V STERNEBRA #5 POORLY OSSIFIED 9 V STERNEBRA #5 POORLY OSSIFIED V EXTRA RIB #13 - FIRST LUMBAR ARCH - BILATERAL It V STERNEBRA #5 POORLY OSSIFIED 16018 1 V STERNEBRA #5 POORLY OSSIFIED V RUDIMENTARY RIB #13 - FIRST LUMBAR ARCH - BILATERAL 3 V STERNEBRA #5 POORLY OSSIFIEO V EXTRA RIB #13 - FIRST LUMBAR ARCH - BILATERAL V LUMBAR ARCHES #1 POORLY OSSIFIED BILATERAL V LUMBAR ARCHES #2 POORLY OSSIFIED BILATERAL 5 V EXTRA #13THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL 7 V EXTRA RIB #13 - FIRST LUMBAR ARCH - BILATERAL 9 V STERNEBRA #5 POORLY OSSIFIED V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB#13 - THORACIC ARCH #13 - BILATERAL 1 1 NO REMARKABLE OBSERVATIONS 16021 2 V MAXILLARY - POORLY OSSIFIED BILATERAL V STERNEBRA #5 POORLY OSSIFIED V EXTRA RIB #13 - FIRST LUMBAR ARCH - BILATERAL GRADE CODE: 1-SLIGHT. 2-MODERATE, 3-MARKED, P-PRESENT M-MALFORMATION V-VARIATION GRADE P P P P P P P P P P P P P P P P P Report 50-514 Appendi* 4C Page l 5 SL 042734 DEVELOPMENTAL TOXICITY STUDY OF INHALEO 1,1,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS FETAL SKELETAL FINDINGS FEMALES FROM M - 3000 PPM IMPLANT # SKELETAL OBSERVATION 1602? 4 V SOME INTER.PHALANGES (FORELIMB) POORLY OSSIFIED ONE BILATERAL V STERNEBRA #5 POORLY OSSIFIED V STERNEBRA #6 POORLY OSSIFIED V EXTRA RIB #13 - THORACIC ARCH#13 -BILATERAL V EXTRA #13 THORACIC CENTRUM ANDARCHES . 16032 2 V STERNEBRA #5 POORLY OSSIFIED 4 NO REMARKABLE OBSERVATIONS 6 V STERNEBRA #5 UNOSSIFIED 8 NO REMARKABLE OBSERVATIONS 10 V STERNEBRA #6 POORLY OSSIFIED 16034 2 V EXTRA RIB #13 - FIRST LUMBAR ARCH - BILATERAL 4 V EXTRA RIB #13 - FIRST LUMBAR ARCH - BILATERAL 6 V STERNEBRA #5 POORLY OSSIFIED V EXTRA RIB #13 - FIRST LUMBAR ARCH - BILATERAL 8 V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL 16049 2 V STERNEBRA #5 POORLY OSSIFIED V STERNEBRA #6 POORLY OSSIFIED V CERVICAL CENTRA #1.#2,#3 AND/OR #4 POORLY OSSIFIED #1 V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - UNILATERAL RIGHT 4 V STERNEBRA #5 POORLY OSSIFIED V STERNEBRA #6 POORLY OSSIFIED V EXTRA RIB #13 - FIRST LUMBAR ARCH - UNILATERAL LEFT 6 V MAXILLARY - POORLY OSSIFIED BILATERAL V STERNEBRA #5 POORLY OSSIFIED V STERNEBRA #6 POORLY OSSIFIED V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL 8 V STERNEBRA #5 POORLY OSSIFIED V STERNEBRA #6 POORLY OSSIFIED GRAOE CODE: 1-SLIGHT, 2-MODERATE, 3-MARKED, P-PRESENT, M-MALFORMATION V-VARIAT10N GRADE P P P P P P P P P P P P P P P P P P P P P P P P P P P P P Report 50-514 Appendix 4C Page 16 CO f o N3 u> Ln FEMALES FROM R - 6000 PPM DEVELOPMENTAL TOXICITY STUDY OF INHALED 1,1,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS FETAL SKELETAL FINDINGS IMPLANT # SKELETAL OBSERVATION 15948 15951 15955 15959 1 V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL 3 V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL 5 V SOME INTER.PHALANGES (FORELIMB) POORLY OSSIFIED ONE BILATERAL V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL 7 V SOME INTER.PHALANGES (FORELIMB) POORLY OSSIFIED ONE BILATERAL V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL 1 V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL 3 V SOME INTER.PHALANGES (FORELIMB) POORLY OSSIFIEDONE BILATERAL V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL 5 V SOME INTER.PHALANGES (FORELIMB) POORLY OSSIFIED ONE BILATERAL V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL 7 V STERNEBRA #6 POORLY OSSIFIEO V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL 9 V STERNEBRA #6 POORLY OSSIFIED V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL 2 V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL 4 V EXTRA RIB #13 - FIRST LUMBAR ARCH - BILATERAL 7 V THORACIC CENTRUM #6 B1L0BED V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL 2 V STERNEBRA #5 POORLY OSSIFIED V RUDIMENTARY RIB #13 - FIRST LUMBAR ARCH - UNILATERAL RIGHT 4 V EXTRA RIB #13 - FIRST LUMBAR ARCH - BILATERAL GRADE CODE: t-SLIGHT, 2-MOOERATE, 3-MARKED, P-PRESENT, M-MALFORMATION V-VARIATION GRADE P P P P P P P P P P P P P P P P P P P P P P P P P P P P P P P P P Report 50-5 t4 Appendi* 4C Page l7 $ DEVELOPMENTAL TOXICITY STUDY OF INHALED I,1,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS FETAL SKELETAL FINDINGS FEMALES FROM R - 6000 PPM IMPLANT # SKELETAL OBSERVATION 15959 6 V STERNEBRA #5 POORLY OSSIFIED V RUDIMENTARY RIB fl3 - FIRST LUMBAR ARCH - BILATERAL 15972 1 V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL 3 V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL 5 V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL 8 V FRONTAL BONE - POORLY 0SSIF1E0 BILATERAL V PARIETAL - POORLY OSSIFIED BILATERAL V INTERPARIETAL - POORLY OSSIFIED V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL 15978 1 V EXTRA RIB #13 - FIRST LUMBAR ARCH - UNILATERAL LEFT 3 V EXTRA #13 THORACIC CENTRUM ANO ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL 5 V STERNEBRA #5 POORLY OSSIFIED V STERNEBRA #6 POORLY OSSIFIED V CERVICAL CENTRA #1,#2,#3 AND/OR #4 POORLY OSSIFIED #1 V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL 7 V STERNEBRA #6 POORLY OSSIFIED V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL 15982 1 V SOME INTER.PHALANGES {FORELIMB) POORLY OSSIFIED ONE BILATERAL V STERNEBRA #5 POORLY OSSIFIED V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL 3 V SOME INTER.PHALANGES (FORELIMB) POORLY OSSIFIED ONE BILATERAL V STERNEBRA #5 UNOSSIFIED V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL 6 V SOME INTER.PHALANGES (FORELIMB) POORLY OSSIFIED GRADE CODE: 1-SLIGHT, 2-MODERATE, 3-MARKED, P-PRESENT, M-MALFORMATION V-VARIATION aaa GRADE P P P P P P P P P P P P P P P P P P P P P P P P P P P P P P Repo rt 50-514 Append l * 4C Page 18 a a a cl cl q. a cl a q. a a a. a a a a a SL 042737 DEVELOPMENTAL TOXICITY STUDY OF INHALED 1 ,1 .1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS FETAL SKELETAL FINDINGS FEMALES FROM R - 6000 PPM IMPLANT # SKELETAL OBSERVATION 15982 ONE BILATERAL 6 V EXTRA A13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL 15984 1 V STERNEBRA #5 POORLY OSSIFIED V STERNEBRA #6 POORLY OSSIFIED V RUDIMENTARY RIB #13 - FIRST LUMBAR ARCH - UNILATERAL RIGHT 3 V STERNEBRA #5 UNOSSIFIED V STERNEBRA #6 POORLY OSSIFIEO V EXTRA RIB #13 - FIRST LUMBAR ARCH - BILATERAL 5 V BODY OF THE HYOID - POORLY OSSIFIED V SOME INTER.PHALANGES (FORELIMB) POORLY OSSIFIEO ONE BILATERAL V STERNEBRA #5 POORLY OSSIFIED V STERNEBRA #6 UNOSSIFIED V EXTRA RIB #13 - FIRST LUMBAR ARCH - BILATERAL 7 V SOME INTER.PHALANGES (FORELIMB) POORLY OSSIFIED ONE BILATERAL V STERNEBRA #6 POORLY OSSIFIED V EXTRA RIB #13 - FIRST LUMBAR ARCH - UNILATERAL LEFT 1 . 9 V STERNEBRA #5 POORLY OSSIFIED V THORACIC CENTRUM #6 BILOBED 15989 1 V SOME INTER.PHALANGES (FORELIMB) POORLY OSSIFIED ONE BILATERAL V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL. 3 V SOME INTER.PHALANGES (FORELIMB) POORLY OSSIFIEO ONE BILATERAL V EXTRA #13 THORACIC CENTRUM AND ARCHES .. V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL 5 V SOME INTER.PHALANGES (FORELIMB) POORLY OSSIFIED ONE BILATERAL V STERNEBRA #5 POORLY OSSIFIED V EXTRA RIB #13 - FIRST LUMBAR ARCH - BILATERAL 7 V SOME INTER.PHALANGES (FORELIMB) POORLY OSSIFIED ONE BILATERAL V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - UNILATERAL RIGHT GRADE CODE: 1-SLIGHT, 2-MODERATE, 3-MARKED, P-PRESENT M-MALFORMATI ON V-VARIATION GRADE P P P P P P P P P P P P Report 50-514 Appendix 4C Page 19 SL 042738 DEVELOPMENTAL TOXICITV STUDY OF INHALED 1,1,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS FETAL SKELETAL FINDINGS FEMALES FROM R - 6000 PPM IMPLANT # SKELETAL OBSERVATION 15989 7 V RUDIMENTARY RIB #13 - THORACIC ARCH #13 - UNILATERAL LEFT 9 V SOME INTER.PHALANGES (FORELIMB) POORLY OSSIFIED ONE BILATERAL V STERNEBRA #5 POORLY OSSIFIED V EXTRA RIB #13 - FIRST LUMBAR ARCH - BILATERAL 15999 1 V EXTRA RIB #13 - FIRST LUMBAR ARCH - BILATERAL 3 V EXTRA RIB #13 - FIRST LUMBAR ARCH - BILATERAL 5 V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL 7 V RUDIMENTARY RIB #13 - FIRST LUMBAR ARCH - UNILATERAL LEFT V EXTRA RIB #13 - FIRST LUMBAR ARCH - UNILATERAL RIGHT 16001 1 V SOME INTER.PHALANGES (FORELIMB) POORLY OSSIFIED ONE BILATERAL V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL 4 V STERNEBRA #6 POORLY OSSIFIED V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL 6 V STERNEBRA #5 POORLY OSSIFIED V STERNEBRA #6 POORLY OSSIFIED V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL 16007 2 V EXTRA RIB #13 - FIRST LUMBAR ARCH - BILATERAL 4 V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL 6 V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL 8 V EXTRA RIB #13 - FIRST LUMBAR ARCH - BILATERAL 16009 1 V STERNEBRA #6 POORLY OSSIFIED V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL 3 V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL V MAXILLARY - POORLY OSSIFIED BILATERAL GRADE CODE: 1-SLIGHT. 2-MODERATE, 3-MARKEO, P-PRESENT, M-MALFORMATION V-VAR I A T1ON GRADE P P P P P P P P P P P P P P P P P P P P P P P P P P P P P P P P Report 50-5 t 4 Appendli 4C Page 20 a a aCl a. a. a. a o_ a a a a SL 042739 DEVELOPMENTAL TOXICITY STUDY OF INHALED 1,1,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS FETAL SKELETAL FINDINGS FEMALES FROM R - 6000 PPM IMPLANT # SKELETAL OBSERVATION 16009 5 V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL 16014 1 V BODY OF THE HYOID - POORLY OSSIFIED V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL 3 V CERVICAL CENTRA #1,#2,#3 AND/OR #4 POORLY OSSIFIED #1 V EXTRA THORACIC ARCH BETWEEN EXISTING ARCHES ' BETWEEN THORACIC ARCHES #8 AND #9 - RIGHT V EXTRA ARCH SMALL EXTRA ARCH SMALLER THAN NORMAL M RIB #10 - FORKED - UNILATERAL RIGHT - IN ASSOCIATION WITH THE EXTRA THORACIC ARCH 5 V CERVICAL CENTRA #1,#2,#3 AND/OR #4 POORLY OSSIFIED #1 V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL 7 V CERVICAL CENTRA #1,#2,#3 AND/OR #4 POORLY OSSIFIED #1 V RUDIMENTARY RIB #13 - FIRST LUMBAR ARCH - UNILATERAL RIGHT V EXTRA RIB #13 - FIRST LUMBAR ARCH - UNILATERAL LEFT V THORACIC CENTRUM #7 BILOBED 9 V STERNEBRA #5 POORLY OSSIFIED V EXTRA RIB #13 - FIRST LUMBAR ARCH - BILATERAL 16019 4 V SOME INTER.PHALANGES (FORELIMB) POORLY OSSIFIED ONE BILATERAL V STERNEBRA #5 UNOSSIFIED V STERNEBRA #6 POORLY OSSIFIED V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL 7 V STERNEBRA #5 UNOSSIFIED V EXTRA RIB #13 - FIRST LUMBAR ARCH - UNILATERAL RIGHT 16022 2 V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL 4 V SOME INTER.PHALANGES (FORELIMB) POORLY OSSIFIED ONE BILATERAL GRADE CODE: 1-SLIGHT, 2-M00ERATE, 3-MARKED, P-PRESENT M-MALFORMATION V-VARIATION GRADE P P P P P P P P P P P P P P P Report 50-514 Append!> 4C Page 2l L t*i0 CD c* PO DEVELOPMENTAL TOXICITY STUDY OF INHALED 1,I,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS FETAL SKELETAL FINOINGS FEMALES FROM R - 6000 PPM IMPLANT # SKELETAL OBSERVATION 16022 4 V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL V LUMBAR CENTRUM #1 POORLY OSSIFIED V LUMBAR CENTRUM #2 POORLY OSSIFIED 16023 3 V STERNEBRA #2 POORLY OSSIFIED V STERNEBRA #4 POORLY OSSIFIED V STERNEBRA #5 UNOSSIFIED V STERNEBRA #6 UNOSSIFIED V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL 5 V BODY OF THE HYOID - POORLY OSSIFIED V STERNEBRA #5 POORLY OSSIFIED V STERNEBRA #6 POORLY OSSIFIED V EXTRA RIB #13 - FIRST LUMBAR ARCH - BILATERAL 16033 3 V CERVICAL CENTRA #1,#2,#3 AND/OR #4 POORLY OSSIFIED ONLY ONE OSSIFICATION CENTER - #3 V CERVICAL CENTRUM #2 DIAGONALLY DISPLACED V CERVICAL CENTRUM #7 BILOBED M CERVICAL RIB - CERVICAL ARCH #7 - UNILATERAL LEFT M CERVICAL ARCH #3 MISSING - UNILATERAL RIGHT M EXTRA CERVICAL CENTRUM FUSED TO CENTRUM #7 EXTRA CENTRUM BENEATH #7 AND FUSED DIAGONALLY M EXTRA CERVICAL ARCH - UNILATERAL - TOTAL OF 7 RIGHT SIDE - ASSOCIATED WITH EXTRA CERVICAL CENTRUM M LATERAL CURVATURE OF SPINE - CERVICAL AREA CERVICAL VERTEBRAE #1 THROUGH #7 V EXTRA RIB #13 - FIRST LUMBAR ARCH - UNILATERAL RIGHT V EXTRA OSSIF. SITE ABOVE STERNEBRA SEGMENT #1 V STERNEBRA #1 BILOBED V STERNEBRA #2 BILOBED V STERNEBRA #3 BILOBED V STERNEBRA #4 BILOBED V STERNEBRA #5 BILOBED V STERNEBRA #6 BILOBED V STERNEBRAE #2 THROUGH #5 FUSED 5 V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL GRADE CODE: 1-SLIGHT, 2-MODERATE, 3- ED, P-PRESENT, M-MALFORMATION V-VARIATION a a. a. Q. cl a a a a. cl cl GRADE P P P P P P P P P P P P P P 3 P P p P P P P Report 50-5 1 4 Appendix 4C Page 22 a cl SL 042741 FEMALES FROM R - 6000 PPM DEVELOPMENTAL TOXICITY STUDY OF INHALED t,t,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS FETAL SKELETAL FINDINGS IMPLANT # SKELETAL OBSERVATION 16040 16044 1 V BODY OF THE HYOID - POORLY OSSIFIED V EXTRA RIB #13 - FIRST LUMBAR ARCH - BILATERAL 3 V BODY OF THE HYOID - POORLY OSSIFIED V SOME INTER.PHALANGES (FORELIMB) POORLY OSSIFIED ONE BILATERAL V EXTRA RIB #13 - FIRST LUMBAR ARCH - BILATERAL 5 V SOME INTER.PHALANGES (FORELIMB) POORLY OSSIFIED' ONE BILATERAL V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL 7 V EXTRA RIB #13 - FIRST LUMBAR ARCH - BILATERAL 9 V EXTRA RIB #13 - FIRST LUMBAR ARCH - BILATERAL V STERNEBRA #5 POORLY OSSIFIED V STERNEBRA #6 POORLY OSSIFIED 1 V SOME INTER.PHALANGES (FORELIMB) POORLY OSSIFIED TWO BILATERAL V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL 3 V MAJORITY INTER.PHALANGES (FORELIMB) POORLY OSSIFIED THREE BILATERAL V SOME INTER.PHALANGES (HINDLIMB) POORLY OSSIFIED ONE BILATERAL V EXTRA #13 THORACIC CENTRUM AND ARCHES V EXTRA RIB #13 - THORACIC ARCH #13 - BILATERAL 5 V EXTRA RIB #13 - FIRST LUMBAR ARCH - BILATERAL 9 V SOME INTER.PHALANGES (FORELIMB) POORLY OSSIFIED ONE BILATERAL V STERNEBRA #6 POORLY OSSIFIED V EXTRA RIB #13 - FIRST LUMBAR ARCH - BILATERAL GRADE CODE: 1-SLIGHT, 2-MODERATE, 3-MARKED, P-PRESENT M-MALFORMATION V-VAR1ATI0N GRADE P P P P P P P P P P P P P P P P P P P P P Report 50-5M Appendi* 4C Page 23 Report 50-514 APPENDIX 5 Protocols and Amendments (39 Pages) SL 042742 Report 50-514 Appendix 5 Page 1 BUSHY RUN RESEARCH R. D. 4, Mellon Road, Export, Pennsylvania 15632 CENTER Telephone (412) 733-5200 PROTOCOL TITLE: Developmental Toxicity Study of Inhaled 1,1,1-Trichloroethane in New Zealand White Rabbits BRRC PROJECT NUMBER: 86-95-80604 SPONSOR: Halogenated Solvents Industry Alliance 1612 K Street, NW, Suite 300 Washington, DC 20006 TESTING FACILITY: Bushy Run Research Center (BRRC) R.D. #4, Mellon Road Export, PA 15632 Attention: R. W. Tyl (412) 733-5277 Reviewed and Approved by: Bushy Run Research Center: YViM.Q i-J 1` ,ll//liSl Rochelle W. Tyl, Fh.D., DABT Study Director/Manager Reproductive and Developmental Toxicology Section Date c. Linda J. CalisTlT B.S., Group Leader, Good Laboratory Practices/Quality Assurance -- / Date 0619P-2 Sponsor: Fred R. Frank, Ph.D. Director Date 042743 Bushy Run Research Center A Joint Mellon Institute --Union Carpide Corporation Operation Report 50-514 Appendix 5 Page 2 Developmental Toxicity Study of Inhaled 1.1.1-Trlchloroethane In New Zealand White Rabbits PURPOSE The purpose of this study is to evaluate the potential for developmental, toxicity of the test article administered by inhalation of the vapor during organogenesis in pregnant New Zealand white rabbits as required by the U. 5. Environmental Protection Agency under Section 4(a) of the Toxic Substances Control Act and promulgated in the final test rule for 1,1,1-trichloroethane under 40 CFR Part 999.4400. MATERIALS AND METHODS Test Article Liquid 1,1,1-trichloroethane (methylchloroform, TCEA; CAS No. 71-55-6) stabilized with less than 0.1% butylene oxide (1,2-epoxybutane; CAS No. 106-88-7) will be used. The test chemical, supplied by Dow Chemical USA, Texas Operations, Freeport, TX, arrived at BRRC on June 25, 1986 in three 55-gallon sealed drums bearing the Lot. No. TA8606182. It received the BRRC No. 49-228 A, B, and C. Pertinent chemical and physical properties of the test chemical are presented in Attachment 1. Purltv and Stability A compositional analysis of the liquid 1,1,1-trichloroethane to be used for this study will be performed by the Sponsor prior to the exposure period. A sample will be sent to the Sponsor at the end of the exposure period for compositional analysis to affirm stability. Handling Precautions Precautions will be undertaken to avoid contact of the test chemical to the clothing, skin or eyes. Rubber or plastic gloves and safety glasses will always be used in handling this compound. Cleaning Contaminated Equipment: Equipment contaminated with the test chemical will be cleaned with acetone. Storage Conditions: The test material will be kept in a chemical storage area. Samples used for the study will be stored in one-gallon glass bottles blanketed with nitrogen and kept in an exhaust hood. A reserve sample will be taken and kept at BRRC. Animals One hundred eight (108) virgin female New Zealand white rabbits (2.5 - 3.5 kg, approximately 5 1/2 months old) will be ordered from Hazelton Dutchland Laboratories, Inc. (Denver, PA). Male New Zealand white rabbits from the BRRC breeding colony will be used for breeding. The New Zealand white rabbit (a 042744 Sl> Report 50-514 Appendix 5 Page 3 lagajnorph) has been selected due to its consistent high fecundity in various laboratories and known incidence of spontaneous malformations (Woo and Hoar, 1982; Stadler et al., 1983). In addition, the gestational stages and morphogenic process of this species, as well as its sensitivity to known teratogens have been described extensively in the literature (Nishimura and Shiota, 1977; Hartman, 1974; Desesso and Jordan, 1977; McClain and Langhoff, 1980; Palmer, 1978). A data base for historical controls is available for this rabbit-strain in this laboratory with regard to reproductive indices and incidence of fetal malformations and variations. Quarantine Period All animals will be housed in BRRC animal rooms 105 (breeding colony males) and 102 (for study females). The animals will be received at least two weeks before the initiation of the mating period. During the quarantine period the animals will be observed regularly for general health status and ability to adapt to the watering system. Body weights will be ascertained at least once prior to the mating period. Female rabbits in good health at least 2.5 kg in weight will be used for this study. Quality Control Quality control will be performed upon the receipt of the animals. Rabbits will be examined by a veterinarian and rabbit fecal samples will be analyzed for intestinal parasites. The quality control evaluation of these animals will be used to determine their health status. Husbandry During the quarantine period, rabbits will be housed individually in stainless steel wire-mesh cages mounted in steel racks. After mating, females will be housed one per cage in stainless steel wire-mesh cages mounted in steel racks. Deotized paperboard (Shepherd Specialty Papers, Inc., Kalamazoo, MI) will be placed under each row of cages to collect solid and liquid excreta. An automatic watering system with demand control valves mounted on each rack will provide water for the rabbits. Water and food will be available ad libitum at all times except during exposures. The light cycle In these rooms will be 12 hours light/12 hours dark. Specific information on the cages (dimensions, etc.) will be included in the study data. The study females will be housed in Room 102 for acclimation, breeding, prior to, between and after the exposures until scheduled sacrifice on gd 29. Animal Identification Each rabbit will be assigned a unique number and will be identified with a Monel metal ear tag (Gey Band and Tag Co,, Morristown, PA). Temperature and Relative Humidity Temperature and relative humidity will be recorded continuously for "each animal holding room. Room temperature will be maintained at 19 3"C, and relative humidity will be maintained between 40% and 60%. SL 042745 Report 50-514 Appendix 5 Page 4 Food and Water The water available to the animals will be tap water (Municipal Authority of Westmoreland County, Greensburg, PA). The incoming water was analyzed for contaminants by the NUS Corporation (Pittsburgh, PA). The water quality analyses performed per EPA specifications included the 129 "priority'' pollutants, which were identified in the Federal. Register, Vol, 45, No.. 98 Appendix D, Part 122. The food available to the rabbits will.be Prolab Certified Babbit Food (Agway Inc., St. Marys, OH). The analyses of the1 food for possible contaminants (including nitrosamine) will be performed by the supplier. Results of water and food analyses will be reviewed by the Study Director. INHALATION CHAMBER OPERATION The animals will be exposed by inhalation since that is the probable route of human exposure and this was the route employed in the exposure range finding study. Chambers Four, approximately 4300-liter stainless steel chambers with glass doors and windows for animal observation will be employed, located in Room 139. Air sampling probes will be positioned in the areas of animal placement. Airflow Chambers will be provided with filtered air at a flow rate of approximately 750-1000 liters/min (approximately 12-14 air changes per hour). All chambers will be maintained at a slight negative pressure to prevent any vapor from entering the room. Chamber Temperature and Relative Humidity Temperature and relative humidity will be recorded at least every 30 minutes during each exposure using a maximum-minimum thermometer (Abbeon Cal., Inc., Sant Barbara, CA) and an Airguide Humidity Indicator (Airguide Instrument Co., Chicago, IL), respectively. Cage Position and Rotation During Exposure In case of undetected differences in environment or 1,1,1-trichloroethane concentration within the chamber, all exposure cage positions will be rotated daily. A description of the rotation will be provided in the raw data. Vapor Generation The liquid test article will be metered from a piston or pump (Fluid Metering Inc., Oyster Bay, NY) into a heated glass evaporator similar in design as described by Carpenter et al. (1975). Temperatures in the evaporator will be maintained at the lowest level sufficient to vaporize the liquid. The resultant vapor will be carried into the chambers by a counter current air stream which enters the bottom of the evaporator. 042746 Sb Report 50-514 Appendix 5 Page 5 Analysis of Chamber Atmosphere Chamber concentrations will be determined approximately once each hour by a gas chromatographic technique. The chamber sample probes will be placed in the breathing zone of the animals. The daily nominal (estimated) chamber concentrations will also be reported. Mating New Zealand white rabbit females (at least 2.5 kg and 5 1/2-6 months old) will be mated to "proven", fertile males from the BRRC breeding colony. Following copulation, the animals will be returned to their respective cages and the date of the copulation will be designated gd 0 (Hafez, 1970). Records will be maintained on the identification numbers of each male used and any relevant comments regarding the mating of each female. Mated female rabbits will be housed individually. Successfully mated females will be randomly assigned to one of three 1,1,1-trichloroethane exposure groups or a control group by a stratified randomization procedure by body weight on gd 0. Each treatment group will consist of twenty-four (24) mated female rabbits. Exposure Groups The target concentrations for the three 1,1,1-trichloroethane-exposed groups will be added to the protocol by amendment based on the results of the exposure range-finding study. The negative control group (animals handled the same as the 1,1,1-trichloroethane-exposed animals) will be exposed to ambient air only (0 ppm). Length of Exposures Rabbits will be exposed on gd 6 through 18 (13 consecutive days) for 6 hours/day. Mptemal Weights Maternal weights will be recorded on gd 0, 6, 9, 12, 15, 19 and 29. All weights taken during the treatment period will be obtained prior to daily exposure. Maternal Observations All study females will be observed daily from gd 0 to scheduled sacrifice for any clinical signs of toxicity. Maternal Sacrifice and Laparotomy Rabbits will be sacrificed on gd 29. Rabbits will be euthanized by intravenous injection of T-61 euthanasia solution (Hoechst Corp., Somerville, NJ) into the marginal ear vein. The maternal body cavities will be opened by a midline thoracolaparotomy. The gravid uteri, ovaries (including corpora lutea), cervices, vagina, and peritoneal and thoracic cavities of each pregnant SL 042747 Report 50-514 Appendix 5 Page 6 animal will be examined grossly. Ovarian corpora lutea of pregnancy will be counted, and maternal liver weight will be determined. Sections of maternal liver and nasal turbinates will be preserved in neutral bufferd 10% formalin for possible subsequent histologic examination. The gravid uteri will be immediately ligated at their cervical end to prevent the expulsion of concept! by myometrial peristalsis. The ligated uteri with ovaries and oviducts attached will be externally examined for signs of hemorrhage and,then removed from the peritoneal cavity, weighed, and dissected longitudinally to expose the contents. All live.and dead fetuses and resorption sites will be recorded. Uteri from females that appear nongravid will be placed in a 10% ammonium sulfide solution for confirmation of pregnancy status (Salewski, 1964), All live rabbit fetuses will be immediately euthanized by intraperitoneal injection of sodium pentobarbital and then will be weighed and examined for external malformations and cleft palate. All of the live fetuses in each litter will be examined for thoracic and abdominal visceral abnormalities by modification of methods described by Staples (1974) and sex determined. Approximately one-half of the fetuses in each litter examined viscerally (odd numbered fetuses from litters with an odd number of live fetuses and even numbered fetuses from litters with an even number of fetuses) will be decapitated and their heads will be fixed in Bouin's solution for examination of craniofacial structures by the free-hand sectioning method modified from Wilson (1965, 1973) and van Julsingha and Bennett (1977). The remaining intact fetuses in each litter will be eviscerated, air-dried and processed for skeletal staining with alizarin red S (Crary, 1962; Peltzer and Schardein, 1966). Decapitated bodies of fetuses will also be processed for skeletal staining but will not be examined unless results warrant, with concurrence of the Sponsor and at additional cost. Dead or Moribund Animals A gross necropsy will be performed on all animals not surviving to the scheduled sacrifice in an attempt to determine the cause of death. Pregnancy status will be determined, if possible, by examination of uterine contents or by staining apparent nongravid uteri with 10% ammonium sulfide. Maternal tissues will be preserved for microscopic examination in neutral buffered 10% formalin only as deemed necessary by the gross findings. Culled Animals Animals not used in the study will be euthanized or used for methods development and training of the BRRC staff. Statistical Analysis The unit of comparison will be the pregnant doe or the litter (Weil, 1970). Results of the quantitative continuous variables (e.g. maternal body weights, liver weights, etc.) will be intercompared for the three exposure groups and one air-control group by use of Levene's test for equal variances (Levene, 1960), analysis of variance (ANOVA), and t-tests with Bonferroni probabilities. The t-tests will be used when the F value from the AH0VA is significant. When Levene's test indicates homogeneous variances and the ANOVA SL 042748 Report 50-514 Appendix 5 Page 7 is significant, the pooled t-test will be used for pairwise comparisons. When Levene's test indicates heterogeneous variances, all groups will be compared by aui ANOVA for unequal variances (Brown and Forsythe, 1974) followed, when necessary, by the separate variance t-test. Nonparametric data obtained following laparohysterectomy will be Statistically evaluated using the Kruskal-Wallis test (Sokal and Rohlf, 1969) followed by the Maim-Whitney U test for pairwise comparisons (Sokal and Rohlf, 1969) when appropriate. Frequency data will be compared using the Fisher's Exact Test (Sokal and Rohlf, 1969). For all statistical tests, the fiducial limit of 0.05 (two-tailed) will be used as the critical level of significance. Personnel Health and Safety Normal safety precautions will be employed in the handling of the test article. The Sponsor will advise BRRC of any additional precautions that should be taken. Protocol Alterations Alterations of this protocol may be made as the study progresses. No major changes in the protocol will be made without the specific written request or consent of the sponsor. In the event that the Sponsor authorizes a protocol change verbally, such change will be made by BRRC. However, it then becomes the responsibility of the Sponsor to follow such verbal change with a written verification. All protocol modifications will be issued as amendments and will be signed by the Study Director and Sponsor. Scheduling Animals arrive at BRRC: September 23, 1986 Acclimatization period begins: September 23, 1986 Mating period begins: October 6, 1986 (first gd 0 also October 6, 1986) Treatment period begins: October 12, 1986 Sacrifice period begins: November 4, 1986 Draft of final report: Approximately three months after the completion of the evaluation of fetal heads and skeletons, on or about April 15, 1987. Records and Data Retention All records pertaining to this study will be permanently retained and placed in the BRRC Archives upon completion of the report. All data, including reports from this study, will be retained for at least ten (10) years after completion of the study in the BRRC Archives and will be made available for inspection upon request by authorized personnel of the sponsor. An appropriate sample of the test article will be retained for ten (10) years following completion of the study. 0427^9 Sh Report 50-514 Appendix 5 Page 8 Good Laboratory Practice Compliance The Bushy Run Research Center through the administration of a quality assurance program by the Good Laboratory Practices Committee and Quality Assurance Unit, assures compliance of all phases of toxicological studies with existing regulations and generally accepted good laboratory practices according to EPA (TSCA), 1983. Study Personnel Study Director/Manager, Teratology Section: Attending Veterinary Pathologist: Pathologist: Teratology Personnel: Inhalation Personnel: Quality Assurance Personnel: Animal Care Personnel: R. W. Tyl, Ph.D., DABT P. E. Losco, VMD, Diplomate ACVP E. H. Fowler, DVM, Ph.D., Diplomate ACVP D. L. Fait, AALAS Cert. II K. A. France, B.A. L. C. Fisher, B.S. M. F. Kubena, B.S. T. A. Rebick, A.H.T., A.S., AALAS Cert. I D. J. McNeil, A.H.T., A.S., AALAS Cert. I. T. A. Savlne, HT(ASCP) D. E. Dodd, Ph.D., DABT, Manager Inhalation I. M. Pritts, Ph.D. D. R. Klonne, Ph.D. W. J. Kintigh, B.S., AALAS Cert. I L. J. Calisti, B.S,, AALAS Cert. Ill, Group Leader J. R. Bernard, B.S. W. J. Conroy, B.S. M. J. Cardella, AALAS Cert. Ill, Supervisor In addition, other BRRC personnel may participate in this study. Report A report will be prepared upon completion of the study. The report (containing a summary of results from the exposure range-finding study on the test chemical conducted at BRRC in 1986) will include mean maternal body weights of pregnant animals, mean number of corpora lutea per dam, total, viable and nonviable implantations, embryonic resorptions (early, later), preimplantation losses, fetal sex ratios, mean fetal body weights, and malformations and variations observed in this study. All means will be accompanied by standard deviations. The report will Include the following tables and appendices: A. Tables Summary of Chamber Analyses Animal Distribution and Fate Summary of Maternal Body Weight and Weight Changes Summary of Maternal Clinical Signs SL 042750 Report 50-514 Appendix 5 Page 9 Tables (Continued-) Summary of Laparohysterotomy Findings Summary of Fetal External, Visceral (including results from craniofacial examinations), and Skeletal Examinations Dates of Quality Assurance inspections and report to management of significant deviations from protocol B. Appendices Protocol and Amendments(s) Chamber Operation and Analyses Exposure Range-Finding Study Report Individual Animal Data for Fetal Body Weights, Fetal Malformations and Fetal Variations WPG/esk/0619P-2 07-01-86 SL 042751 Report 50-514 Appendix 5 Page 10 REFERENCES Brown, M. B. and A. B. Forsythe (1974). The small sample behavior of some statistics which test the equality of several means. Technometrics 129-132. Carpenter, C. P,, E. R. Kinkead, D. L. Geary, Jr., L. J. Sullivan and J. M. King (1975). Petroleum Hydrocarbon Toxicity Studies. I. Methodology. Toxicol. Appl. Pharmacol. 22, 246-262. Crary, D.D, (1962). Modified benzyl alcohol clearing of Alizarin-stained specimens without loss of flexibility. Stain Technology XL, 124-125. DeSesso, J. M. and R. L. Jordan (1977). Drug-induced limb dysplasias in fetal rabbits. Teratology 22, 199-212, Environmental Protection Agency, (1983). Part III EPA Toxic Substances Control; Good Laboratory Practice Standards; Final Rule. Federal Register 48(230)t 53922-53944. Hafez, E. S. E. (ed.) (1970). Reproduction and Breeding Techniques for Laboratory Animals. Lea and Febiger, Philadelphia, PA. Hartman, H. A. (1974). The fetus in experimental teratology. In: The Biology of the Laboratory Rabbit, S. H. Weisbroth, R. E. Flatt, and A. L. Kraus, eds., Academic Press, NY, pp. 92-153. Levene, H. (1960). Robust tests for equality of variance. In: Contributions to Probability and Statistics. I. Olkin et al., eds., Stanford University Press, Stanford, CA, pp. 278-292. McClain, R. M. and L. Langhoff (1980). Teratogenicity of diphenylhydantoin in the New Zealand white rabbit. Teratology 21. 371-379. Nishimura, H. and K. Shiota (1977). Summary of comparative embryology and teratology. In: Handbook of Teratology, J. G. Wilson and F. C. Fraser, eds., Plenum Press, NY, Vol. 3, pp. 119-154. Palmer, A. K. (1978). The design of subprimate animal studies. In: Handbook of Teratology. J. G. Wilson and F. C. Fraser, eds.. Plenum Press, NY, Vol. 4, PP 215-253. Peltzer, M.A. and J. L. Schardein (1966). A convenient method for processing fetuses for skeletal staining. Stain Technology 41, 300-302. o<>*752 Report 50-514 Appendix 5 Page 11 REFERENCES (Continued) Salevski, E. (1964). Farbemethode Zum Makroskopischen Nachweis Von Implantations-Stellen am Uterus Der Ratte. Naunyn-Schmiedebergs, Arch. Exp. Pathol. Pharmakol. 241, 367. Sokal, R. R. and F. J. Rohlf (1969). Biometry. W. H. Freeman and Co., San Francisco, pp. 369-371, 299-340, 370-372, 589-595. Stadler, J., M.-J. Kessedjian and J. Perraud (1983). Use of the New Zealand white rabbit in teratology: Incidence of spontaneous and drug-induced malformations. Fd. Cosmet. Toxicol. 21(5)f 631-636. Staples, R. E. (1974) Detection of visceral alterations in mammalian fetuses. Teratology A37. van Julslngha, E. B. and C. G. Bennett (1977). A dissecting procedure for the detection of anomalies in the rabbit foetal head. In: Methods in Prenatal Toxicology. D. Neubert, H. J. Merker, and T. E. Kwasigroch, Eds. PSG Publishing Company, Inc., Massachusetts, pp. 126-144. Weil, C. S. (1970). Selection of the valid number of sampling units and a consideration of their combination in toxicological studies involving reproduction, teratogenesis or carcinogenesis. Fd. Cosmet. Toxicol. 177-182. Wilson, J. G. (1965). Embryological Considerations in Teratology. In: Teratology Principles and Techniques. J. G. Wilson and J. Warkany, eds., The University of Chicago Press, pp. 251-277. Wilson, J. G. (1973). Environment and Birth Defects. Academic Press, NY. Woo, D. C. and R. M. Hoar (1982). Reproductive performance and spontaneous malformations in control New Zealand White rabbits: A Joint study by MARTA. Teratology 25(2). 82A. (Tables provided by the authors). WPC/esk/0619P-l 07-01-86 SL 042753) ATTACHMENT 1 Page 1 of 5 Report 50-514 Appendix 5 Page 12 MATERIAL SAFETY DATA SHEET Dow Chemical U.S.A. Midland, MI 48674 Emergency Phone: 517-636-4400 Product Code: 55352 Page: 1 PRODUCT NAME: METHYL CHLOROFORM, LOW STABILIZED Effective Date: 11/20/85 Date Printed: 06/21/86 M$DS:000699 1. INGREDIENTS: 1,1,1-Trichloroethane CAS# 000071-55-6 99.9% 2. PHYSICAL DATA: BOILING POINT: I65F (74C) VAP PRESS: 100 mmHg g 20C VAP DENSITY: 4.55 SOL. IN WATER: 0.07 g/100 g 25C SP. GRAVITY: 1.330-1.335 25/25C APPEARANCE: Colorless liquid. ODOR: Irritating odor at high concentrations. 3. FIRE AND EXPLOSION HAZARD DATA: FLASH POINT: None METHOD USED: TOC, TCC, COC FLAMMABLE LIMITS LFL: 7.5* 6 25C UFL: 153; 8 25C EXTINGUISHING MEDIA: Water fog. FIRE EXPLOSION HAZARDS: Not available. FIRE-FIGHTING EQUIPMENT: Positive-pressure, self-contained respiratory equipment. (Continued on Page 2) (R) Indicates a trademark of The Dow Chemical Company SL 042754 ATTACHMENT 1 Page 2 of 5 Report 50-514 MATERIAL SAFETY DATA SHEET ----------------------------------------------------------------------------------------------------------------------- Appendix 5 Page 13 Dow Chemical U.S.A. Midland, MI 48674 Emergency Phone: 5X7-636-4400 Product Code: 55352 Page: 2 PRODUCT NAME: METHYL CHLOROFORM, LOW STABILIZED Effective Date: 11/20/85 Date Printed: 06/21/86 MSDS:000699 4. REACTIVITY DATA: STABILITY: (CONDITIONS TO AVOID) Autoignition temperature 998F (537C). Avoid open flames, welding arcs, or other high temperature sources which induce thermal decomposition. INCOMPATIBILITY: (SPECIFIC MATERIALS TO AVOID) Water - slow hydrolysis produces corrosive acid. Avoid contact with or storage in aluminum and its alloys. Definitely avoid contact with metallic aluminum and zinc powders. HAZARDOUS DECOMPOSITION PRODUCTS: Hydrogen chloride and very small amounts of phosgene and chlorine. HAZARDOUS POLYMERIZATION: Will not occur. 5. ENVIRONMENTAL AND DISPOSAL INFORMATION: ACTION TO TAKE FOR SPILLS/LEAKS: Small leaks: Mop up, wipe up, or soak up immediately. Remove to out-of-doors. Large spills: Evacuate area. Contain liquid; transfer to closed metal containers. Keep out of water supply. DISPOSAL METHOD: When disposing of the unused contents, the preferred options are to send to licensed reclaimer, permitted incinerators, or to evaporate very small quantities in compliance with local, state, and federal regulations including Subtitle C of the Resource Conservation and Recovery Act. Dumping into sewers, on the ground, or into any body of water is strongly discouraged, and may be illegal. Consult The Dow Chemical Company for further information. 6. HEALTH HAZARD DATA: EYE: May cause pain. May cause slight transient (temporary) irritation with slight transient corneal injury. Vapors may irritate eyes. SKIN CONTACT: Prolonged or repeated exposure may cause skin irritation. Repeated contact may cause drying or flaking of (Continued on Page 3) (R) Indicates a trademark of The Dow Chemical Company SI 42755) MATERIAL SAFETY DATA SHEET ATTACHMENT 1 Page 3 of 5 Report 50-514 Appendix 5 Page 14 Dow Chemical U.S.A. Midland, MI 48674 Emergency Phone: 517-636-4400 Product Code: 55352 Page: 3 PRODUCT NAME: METHYL CHLOROFORM, LOW STABILIZED Effective Date: 11/20/85 Date.Printed:. 06/21/86 MSD$:000699 6. HEALTH HAZARD DATA: (CONTINUED) skin. SKIN ABSORPTION: A single prolonged skin exposure is not likely to result in absorption of harmful amounts. The LD50 for rabbits is about 15*000 mg/kg. INGESTION: Single dose oral toxicity is low. The LD50 for rats is >10,000 mg/kg. If aspirated (liquid enters the lung), may be rapidly absorbed through the lungs and result in injury to other body systems. INHALATION: Minimal anesthetic or narcotic effects may be seen in the range of 500-1000 ppm trichloroethane. Progressively higher levels over 1000 ppm may cause dizziness, drunkenness; concentrations as low as 10,000 ppm can cause unconsciousness and death. In confined or poorly ventilated areas, vapors which readily accumulate can cause unconsciousness and death. These high levels may also cause cardiac arrhythmias (irregular heartbeats). SYSTEMIC & OTHER EFFECTS: Based on available data, repeated exposures are not anticipated to cause any significant adverse effects. 1,1,1-Trichloroethane and similar mixtures did not cause cancer in long-term animal studies. Birth defects are unlikely. Exposures having no adverse effects on the mother should have no effect on the fetus. In animal studies, has been shown not to interfere with reproduction. Results of in vitro ('test tube') mutagenicity tests on 1,1,1trichloroethane have been negative. Results of mutagenicity tests in animals have been negative. 7. FIRST AID: EYES: Irrigate immediately with water for at least 5 minutes. SKIN: Wash off in flowing water or shower. Remove contaminated clothing and wash before reuse. INGESTION: Do not induce vomiting. Call a physician and/or (Continued on Page 4) (R) Indicates a trad mark of The Dow Chemical Company SL 042756 MATERIAL SAFETY DATA SHEET ATTACHMENT 1 Page A of 5 Report 50-514 Appendix 5 Page 15 Dow Chemical U.S.A. Midland. MI 48674 Emergency Phone: 517-636-4400 Product Code: 55352 Page: 4, PRODUCT NAME: METHYL CHLOROFORM, LOW STABILIZED Effective Date: .11/20/85 Date Printed: .06/21/86 MSDS:000699 7. FIRST AID: (CONTINUED) transport to emergency facility immediately. INHALATION: Remove to fresh air. If not breathing, give mouth-to-mouth resuscitation. If breathing is difficult, give oxygen. Call a physician. NOTE TO PHYSICIAN: Because rapid absorption may occur through lungs if aspirated and cause systemic effects, the decision of whether to induce vomiting or not should be made by an attending physician. If lavage is performed, suggest endotracheal and/or esophageal control. Danger from lung aspiration must be weighed against toxicity when considering emptying the stomach. Exposure may increase "myocardial irritability". Do not administer sympathomimetic drugs unless v absolutely necessary. No specif it antidote. Supportive care. Treatment based on judgment of the physician in response to reactions of the patient. 8. HANDLING PRECAUTIONS: EXPOSURE GUIDELINE (S): 1,1,1-Trichloroethane - 0SHA standard is 350 ppm and current ACGIH TLV is 350 ppm (450 ppm STEL). VENTILATION: Control airborne concentrations below the exposure guideline. Use only with adequate ventilation. Local exhaust ventilation may be necessary for some operations. Lethal concentrations may exist in areas with poor ventilation. RESPIRATORY PROTECTION: Atmospheric levels should be maintained below the exposure guideline. When respiratory protection is required for certain operations, use an approved air-purifying respirator. For emergency and other conditions where the exposure guideline may be greatly exceeded, use an approved positive pressure self-contained breathing apparatus. In confined or poorly ventilated areas, use an approved positive pressure self-contained breathing apparatus. SKIN PROTECTION: For brief contact, no precautions other than clean body-covering clothing should be needed. When prolonged (Continued on Page 5) (R) Indicat s a trademark of The Dow Chemical Company SL 042757 MATERIAL . Page 16 SAFETY DATA SHEET ATTACHMENT 1 Page 5 of 5 Report 50-514 . Appendix 5 Dow Chemical U.S.A. Midland, MI 48674 Emergency Phone: 517-636-4400 , Product Code: 55352 #Page: 5 PRODUCT NAME: METHYL CHLOROFORM, LOW STABILIZED Effective Date: 11/20/85 Date Printed: 06/21/86 MSDS:000699 . 8. HANDLING PRECAUTIONS: (CONTINUED) or frequently repeated contact could occur, use protective clothing impervious to this material. Selection of specific items such as gloves, boots, apron, or full body suit will depend on operation. EYE PROTECTION: Use safety glasses. Where contact with liquid is likely, chemical goggles are recommended because eye contact with this material may cause pain, even though it is unlikely to cause injury. 9. ADDITIONAL INFORMATION: SPECIAL PRECAUTIONS TO BE TAKEN IN HANDLING AND STORAGE: Handle with reasonable care. Avoid breathing vapors. Store in a cool dry place. Concentrated vapors of methyl chloroform, low stabilized - are heavier than air and will collect in tow areas such as pits, degreasers, storage tanks, and other confined areas. Do not enter these areas where vapors of this product are suspected unless special breathing apparatus is used and an observer is present for assistance. 1.1.1-Trichloroethane products should not be packaged in aluminum aerosol cans or with finely divided aluminum or its alloys in an aerosol can. Aluminum is not an acceptable material of construction for pumps, mixers, fittings, storage tanks for 1.1.1-trichloroethane products or formulations. Metallic aluminum and zinc powders should be avoided. For additional information on toxicity, handling precautions, and first aid, refer to chlorinated solvents literature form no. 100-5792. MSDS STATUS: Revised 1, 5, 6, 7. and 9- (R) Indicates a trademark of The Dow Chemical Company The Information Herein Is Given In Good Faith, But No Warranty, Express Or Implied, Is Made. Consult The Oow Chemical Company For Further Information. BUSHY RUN RESEARCH R D, 4, Mellon Road, Export. Pennsylvania 15632 Report 50-514 Appendix 5 Page 17 CENTER Telephone (412) 733-5200 PROTOCOL AMENDMENT #1 TITLE: Developmental Toxicity Study of Inhaled 1,1,1-Trichloroethane in New Zealand White Rabbits BRRC PROJECT NUMBER: 86-95-80604 SPONSOR: Halogenated Solvents Industry Alliance 1612 K Street, NW, Suite 300 Washington, DC 20006 TESTING FACILITY: Bushy Run Research Center (BRRC) R.D. #4, Mellon Road Export, PA 15632 Attention: R. W. Tyl (412) 733-5277 Reviewed and Approved by: Bushy Run Research Center: iJ 111 Rochelle W. Tyl, Ph.D., TABT Study Director/Manager Reproductive and Developmental Toxicology Section / Date Auq CjL Linda J. Callst^t B.S., Group Leader, Good Laboratory Practices/Quality Assurance Tate 0816P-1 042759 Sponsor: Fred R. Frank, Ph.D. Director Date nes A. Barter, Ph.D., DABT Bushy Run Research Center A Joint Mellon Institute--Union Carbide Corporation Operation Report 50-514 Appendix 5 Page 18 Protocol Amendment //I Page 2 The protocol is amended as follows: 1. Location of Protocol Change: MATERIALS AND METHODS, Quarantine Period (Pace 31 Description of Protocol Change: From: "All animals will be housed in BRRC animal rooms 105 (breeding colony males) and 102 (for study females). The animals will be received at least two weeks before the initiation of the mating period. During the quarantine period the animals will be observed regularly for general health status and ability to adapt to the watering system. Body weights will be ascertained at least once prior to the mating period. Female rabbits in good health at least 2.5 kg in weight will be used for this study." To: "All animals will be housed in BRRC animal rooms 105 (breeding colony males) and 122 (for study females). The animals will be received at least two weeks before the initiation of the mating period. During the quarantine period the animals will be observed regularly for general health status and ability to adapt to the watering system. Body weights will be ascertained at least once prior to the mating period. Female rabbits in good health at least 2.5 kg in weight will be used for this study," Rationale; The animal room for the study females was changed prior to their arrival. The rooms have the same environmental controls and the change will not affect the conduct of the study. 2. Location of Protocol Change: MATERIALS AMD METHODS. Husbandry (Page 3) Description of Protocol Change: From: "During the quarantine period, rabbits will be housed individually in stainless steel wire-mesh cages mounted in steel racks. After mating, females will be housed one per cage in stainless steel wire-mesh cages mounted in steel racks. Deotized paperboard (Shepherd Specialty Papers, Inc., Kalamazoo, MI) will be placed under each row of cages to collect solid and liquid excreta. An automatic watering system with demand control valves mounted on each rack will provide water for the rabbits. Water and food will be available libitum at all times except SL 042760 Report 50-514 Appendix 5 Page 19 Protocol Amendment #1 Page 3 during exposures. The light cycle in these rooms will be 12 hours light/12 hours dark. Specific information on the cages (dimensions, etc.) will be included in the study data. The study females will be housed in Room 102 for acclimation, breeding, prior t.o, between and after the exppsures until scheduled sacrifice on gd 29." To: "During the quarantine period, rabbits will be housed individually in stainless steel wire-mesh cages mounted in steel racks. After mating, females will be housed one per cage in stainless steel wire-mesh cages mounted in steel racks. DeotizedQ paperboard (Shepherd Specialty Papers, Inc., Kalamazoo, MI) will be placed under each row of cages to collect solid and liquid excreta. An automatic watering system with demand control valves mounted on each rack will provide water for the rabbits. Water and food will be available d libitum at all times except during exposures. For exposures, mated females will be transferred, one per cage, into stainless steel wire-mesh cages and moved into the appropriate chambers in room 139. Stainless steel shelf pans will be placed under each row of cages to prevent urinary and fecal contamination of animals at lower levels. After exposure, the animals will be returned to their original cages and "home" room. The light cycle in these rooms will be 12 hours light/12 hours dark. Specific information on the cages (dimensions, etc.) will be included in the study data. The study females will be housed in Room 102 for acclimation, breeding, prior to, between and after the exposures until scheduled sacrifice on gd 29." Rationale: The procedure for transferring females to exposure cages for the exposure period was added for completeness. 3. Location of Protocol Change: INHALATION CHAMBER OPERATION. Exposure Groups (Page 5) Description of Protocol Change: From: "The target concentrations for the three 1,1,1-trichloroethane-exposed groups will be added to the protocol by amendment based on the results of the exposure range-finding study. The negative control group (animals handled the same as the 1,1,1-trichloroethane-exposed animals) will be exposed to ambient air only (0 ppm)." To: "The target concentrations for the three 1,1,1-trichloroethane-exposed groups will be 1000, 3000 and 6000 ppm. The negative control group (animals handled the same as the 1,1,1-trichloroethane-exposed animals) will be exposed to ambient air only (0 ppm)." SL 042761 Report 50-514 Appendix 5 Page 20 Protocol Amendment #1 Page 4 Rationale: The three target exposure concentrations were selected by the Sponsor (with concurrence of the Study Director) after examination of the draft results from the exposure range-finding study on September 17, 1986 via conference call with Dr. J. A. Barter, Dr. K. S. Rao, .Dr. J. P. Lyon, and Dr. R. W. Tyl. The Study Director was notified by the Sponsor on September 29, 1986 that the U.S. EPA had given verbal agreement to these selected target exposure concentrations. PATH/esk/0816P-l 10-22-86 SL 042762 Report 50-514 Appendix 5 Page 21 BUSHY RUN RESEARCH R. 0. 4, Mellon Road, Export, Pennsylvania 15632 CENTER Telephone (4121 733-5200 PROTOCOL TITLE: Developmental Toxicity Exposure Range-Finding Study of Inhaled 1,1,1-Trichloroethane in New Zealand White Rabbits BRRC PROJECT NUMBER: 86-95-80603 SPONSOR: Halogenated Solvents Industry Alliance 1612 K Street, NW, Suite 300 Washington, DC 20006 TESTING FACILITY: Bushy Run Research Center (BRRC) R.D. #4, Mellon Road Export, PA 15632 Attention: R. W. Tyl (412) 733-5277 Reviewed and Approved by: Bushy Run Research Center: J/vJaiit U. KJ Rochelle W. Tyl, Ph.D'., DABT Study Director/Manager Reproductive and Developmental Toxicology Section ljnl'51 Date L, Linda J. Calibti, B.S., Group Leader, Good Laboratory Practices/Quality Assurance Date 0618P-3 Sponsor: Fred R. Frank, Ph.D, Director Date k.u-,0 ___fa-tibs -11& Jainrfs A. Barter, Ph.D., DABT te 042763 SL Bushy Run Research Center A Joint Mellon Institute--Union Carbide Corporation Operation Report 50-514 Appendix 5 Page 22 Page 2 Developmental Toxicity Exposure Range-Finding Study of Inhaled 1.11-Trlchloroethane In Hew Zealand White Rabbits PURPOSE The purpose of this study is to establish the exposure-response range of the test article for maternal and/or embryonic toxicity in pregnant New Zealand white rabbits in order to select appropriate exposure concentrations for use in the definitive developmental toxicity study. Test Article MATERIALS AND METHODS Liquid 1,1,1-trichloroethane (methylchloroform; TCEA; CAS No. 71-55-6) stabilized with less than 0.1% butylene oxide (1,2-epoxybutane; CAS No. 106-88-7) will be used. The test chemical, supplied by Dow Chemical USA, Texas Operations, Freeport, TX, arrived at BRRC on June 25, 1986 in three 55-gallon sealed drums bearing the Lot No. TA8606182. It received the BRRC No. 49-228 A, B, and C. Pertinent chemical and physical properties of the test chemical are presented in Attachment 1. Purity and Stability A compositional analysis of the liquid 1,1,1-trichloroethane to be used for this study will be performed by the Sponsor prior to the exposure period. A sample will be sent to the Sponsor at the end of the exposure period for compositional analysis to affirm stability. Handling Precautions Precautions will be undertaken to avoid contact of the test material to the clothing, skin or eyes. Plastic or rubber gloves and safety glasses will always be used in handling this compound. Cleaning Contaminated Equipment: Equipment contaminated with the test chemical will be cleaned with acetone. Storage Conditions: The test material will be kept in a chemical storage area. Samples used for the study will be stored in one-gallon glass bottles blanketed with nitrogen and kept in an exhaust hood. A reserve sample will be taken and kept at BRRC. Animals Forty (40) virgin female New Zealand white rabbits (2.5 - 3.5 kg, approximately 5 1/2 months old) will be ordered from Hazelton Dutchland Laboratories, Inc. (Denver, PA). Male New Zealand white rabbits from the BRRC breeding colony will be used for breeding. The New Zealand white rabbit (a lagamorph) has been selected due to its consistent high fecundity in various SL 042764 Report 50-514 Appendix 5 Page 23 Page 3 laboratories and known incidence of spontaneous malformations (Woo and Hoar, 1982; Stadler, e al., 1983). In addition, the gestational stages and morphogenic process of this species, as well as its sensitivity to known teratogens have been described extensively in the literature (Nishimura and Shiota, 1977; Hartman, 1974; Desesso and Jordan, 1977; McClain and Langhoff, 1980; Palmer, 1978). A data base for historical controls is available for this rabbit strain in this laboratory with regard to reproductive Indices and incidence of fetal malformations and variations. Quarantine Period All animals will be housed in BRRC animal rooms 102 (females) and 105 (for male rabbit colony). The animals will be received at least two weeks before the initiation of the mating period. During the quarantine period the animals will be observed regularly for general health status and ability to adapt to the watering system. Body weights will be ascertained at least once prior to the mating period. Female rabbits in good health at least 2.5 kg in weight will be used for this study. Quality Control Quality control will be performed upon the receipt of the animals. Rabbits will be examined by a veterinarian and rabbit fecal samples will be analyzed for intestinal parasites. The quality control evaluation of these animals will be used to determine their health status. Husbandry During the quarantine period, rabbits will be housed individually in stainless steel wire-mesh cages mounted in steel racks. After mating, females will be housed one per cage in stainless steel wire-mesh cages mounted in steel racks. Deotized paperboard (Shepherd Specialty Papers, Inc., Kalamazoo, MI) will be placed under each row of cages to collect solid and liquid excreta. An automatic watering system with demand control valves mounted on each rack will provide water for the rabbits. Water and food will be available ad libitum at all times except during exposures. All study females will remain in Room 102 during quarantine, mating, prior to, between, and after exposures until scheduled sacrifice on gd 19. The light cycle in this room will be 12 hours light/12 hours dark. Specific information on the cages (dimensions, etc.) will be included in the study data. Animal Identification Each rabbit will be assigned a unique number and will be identified with a Monel metal ear tag (Gey Band and Tag Co., Morristown, PA). Temperature and Relative Humidity Temperature and relative humidity will be recorded continuously for each animal holding room. Room temperature will be maintained at 19 3#C, and relative humidity will be maintained between 40% and 60%. SL 042765 Report 50-514 Appendix 5 Page 24 Page 4 Food and Water The water available to the animals will be tap water (Municipal Authority of Westmoreland County, Greensburg, PA). The incoming water was analyzed for contaminants by the NUS Corporation (Pittsburgh, PA). The water quality analyses performed per EPA specifications included the 129 "priority" pollutants, which were identified in the Federal Register, Vol, 45, No. 98 Appendix D, Part 122. The food available to the rabbits will be Prolab Certified Rabbit Food (Agway Inc., Sf. Marys, OH). The analyses of the food for possible contaminants (including nitrosamine) will be performed by the supplier. Results of water and food analyses will be reviewed by the Study Director. INHALATION CHAMBER OPERATION Chambers Four, approximately 4300-liter stainless steel chambers with glass doors and windows for animal observation will be employed, located in Room 139. Air sampling probes will be positioned in the areas of animal placement. Airflow Chambers will be provided with filtered air at a flow rate of approximately 750-1000 liters/min (approximately 12-14 air changes per hour). All chambers will be maintained at a slight negative pressure to prevent any vapor from entering the room. Chamber Temperature and Relative Humidity Temperature and relative humidity will be recorded at least once every 30 minutes during each exposure using a maximum-minimum thermometer (Abbeon Cal., Inc., Sant Barbara, CA) and an Airguide Humidity Indicator (Airguide Instrument Co., Chicago, IL), respectively. Cage Position and Rotation Purine Exposure In case of undetected differences in environment or 1,1,1-trichloroethane concentration within the chamber, all exposure cage positions will be rotated daily. A description of the rotation will be provided in the raw data. Vapor Generation The liquid test article will be metered from a piston or pump (Fluid Metering Inc., Oyster Bay, NY) into a heated glass evaporator similar in design as described by Carpenter et al. (1975). Temperatures in the evaporator will be maintained at the lowest level sufficient to vaporize the liquid. The resultant vapor will be carried into the chambers by a counter current air stream which enters the bottom of the evaporator. SL 0^2766 Report 50-514 Appendix 5 Page 25 Page 5 Analysis of Chamber Atmosphere Chamber concentrations will be determined approximately once each hour by a gas chromatographic technique. The chamber sample probes will be placed in the breathing zone of the animals. The daily nominal (estimated) chamber concentrations will also be reported. Metjpg New Zealand white rabbit females (at least 2.5 kg and 5 1/2-6 months old) will be mated to "proven", fertile males from the BRRC breeding colony. Following copulation, the animals will be returned to their respective cages and the date of the copulation will be designated gd 0 (Hafez, 1970). Records will be maintained on the identification numbers of each male used and any relevant comments regarding the mating of each female. Mated female rabbits will be housed individually. Successfully mated females will be randomly assigned to one of three 1,1,1-trichloroethane exposure groups or a control group by a stratified randomization procedure by body weight on gd 0. Each treatment group will consist of eight mated female rabbits. Exposure Groups The target concentrations for the three 1,1,1-trichloroethane-exposed groups will be 1000, 3000 and 6000 ppm. The negative control group (animals handled the same as the 1,1,1-trichloroethane-exposed animals) will be exposed to ambient air only (0 ppm). Length of Exposures Rabbits will be exposed on gd 6 through 18 (13 consecutive days) for 6 hours/day. Maternal Weights Maternal weights will be recorded on gd 0, 6, 9, 12, 15 and 19. All weights taken during the treatment period will be obtained prior to daily exposure. Maternal Observations All animals will be observed daily throughout the study (mating to necropsy) for clinical signs of toxicity. Maternal Sacrifice and Laparotomy Rabbits will be sacrificed on the day following the last exposure on gd 19. Rabbits will be euthanized by intravenous injection of T-61 euthanasia solution (Hoechst Corp., Somerville, NJ) into the marginal ear vein._ Immediately after euthanization, the eyes will be examined In situ by gently SL 042767 Report 50-514 Appendix 5 Page 26 Page 6 pressing a glass slide against the cornea and examining the eye under fluorescent light illumination. The complete necropsy examination, external and internal, will be performed by a veterinary pathologist. The maternal body cavities will be opened by a midline thoracolaparotomy. The gravid uteri, overies (including corpora lutea), cervices, vagina, and peritoneal and thoracic cavities of each pregnant animal will be examined grossly, j Ovarian corpora lutea of pregnancy will he counted, and maternal liver weight will be determined. Sections of maternal liver, along with other tissues deemed appropriate by the study pathologist at the time of necropsy, will be preserved in buffered neutral 10% formalin but will not be examined histologically unless deemed necessary to interpret other observations from this study. The gravid uteri will be immediately ligated at their cervical end to prevent the expulsion of concepti by myometrial peristalsis. The ligated uteri with ovaries and oviducts attached will be externally examined for signs of hemorrhage and then removed from the peritoneal cavity, weighed, and dissected longitudinally to expose the contents. All live and dead fetuses and resorption sites will be recorded. Uteri from females that appear nongravid will be placed in a 10% ammonium sulfide solution for confirmation of pregnancy status (Salewski, 1964). Dead or Moribund Animals A gross necropsy will be performed on all animals not surviving to the scheduled sacrifice in an attempt to determine the cause of death. Pregnancy status will be determined, if possible, by examination of uterine contents or by staining apparent nongravid uteri with 10% ammonium sulfide. Maternal tissues will be preserved for microscopic examination in neutral buffered 10% formalin only as deemed necessary by the gross findings. Culled Animals Animals not used in the study will be euthanized or used for methods development and training of the BRRC staff. Guidelines for Exposure Concentrations Based on the results of this study, three exposure levels of the test substance will be determined for the definitive developmental toxicity study. The "high" concentration should cause "minimal" adult animal toxicity (e.g. loss of body weight, change in relative organ weight, etc.) without exceeding a mortality rate of 10%. The low exposure level will be determined which will cause no observable adult animal toxicity. The intermediate exposure level will provide information on any embryo/fetal toxicity (including teratogenicity). The Study Director will make recommendations to the Sponsor as to exposure levels for the full teratogenicity study, but the final decision as to exposure levels will rest with the Sponsor. Statistical Analysis The unit of comparison will be the pregnant doe or the litter (Well, 1970). Results of the quantitative continuous variables (e.g. maternal body weights, liver weights, etc.) will be intercompared for the three treatment SL 042768 Report 50-514 Appendix 5 Page 27 Page 7 groups and one control group by use of Levene's test for equal variances (Levene, 1960), analysis of variance (ANOVA), and t-tests with Bonferroni probabilities. The t-tests will be used when the F value from the ANOVA is significant. When Levene's test indicates homogeneous variances and the ANOVA is significant, the pooled t-test will be used for pairwise comparisons. When Levene's test indicates heterogeneous variances, all groups will be compared by an ANOVA for unequal variances (Brown and Forsythe, 1974) followed, when necessary, by the separate variance t-test. Nonparametric data obtained following laparohysterectomy will be statistically evaluated using the Kruskal-Wallis test (Sokal and Rohlf, 1969) followed by the Maim-Whitney TJ test for pairwise comparisons (Sokal and Rohlf, 1969) when appropriate. Frequency data will be compared using the Fisher's Exact Test (Sokal and Rohlf, 1969). For all statistical tests, the fiducial limit of 0.05 (two-tailed) will be used as the critical level of significance. Personnel Health and Safety Normal safety precautions will be employed in the handling of the test article. The Sponsor will advise BRRC of any additional precautions that should be taken. Protocol Alterations Alterations of this protocol may be made as the study progresses. No major changes in the protocol will be made without the specific written request or consent of the sponsor. In the event that the Sponsor authorizes a protocol change verbally, such change will be made by BRRC. However, it then becomes the responsibility of the Sponsor to follow such verbal change with a written verification. All protocol modifications will be issued as amendments and will be signed by the Study Director and Sponsor. Scheduling Animals arrive at BRRC: July 29, 1986 Acclimatization period begins: July 29, 1986 Mating period begins: August 16, 1986 (first gd 0 also August 16, 1986) Treatment period begins: August 22, 1986 Sacrifice period begins (gd 19): September 4, 1986 Submission of draft data to sponsor: The data in draft form will be submitted to the Sponsor within two (2) weeks after the last sacrifice day for selection of the appropriate exposure concentrations for the definitive development toxicity study. A complete report f this study will be included as an appendix in the Final Report. Records and Data Retention All records pertaining to this study will be permanently retained and placed in the BRRC Archives upon completion of the report. All data. Including reports from this study, will be retained for at least ten (10) years after completion of the study in the BRRC Archives and will be made available for inspection upon request by authorized personnel of the sponsor. An appropriate sample of the test article will be retained for ten (10) years following completion of the study. Report 50-514 Appendix 5 Page 28 Page 8 Good Laboratory Practice Compliance The Bushy Bun Research Center through the administration of a quality assurance program by the Good Laboratory Practices Committee and Quality Assurance Unit, assures compliance of all phases of toxicological studies with existing regulations and generally accepted good laboratory practices according to EPA (TSCA), 1983. Study Personnel. Study Director/Manager, Teratology Section: Attending Veterinary Pathologist: Pathologist: Teratology Personnel: Inhalation Personnel: Quality Assurance Personnel: Animal Care Personnel: R. W. Tyl, Ph.D., DABT P. E. Losco, VMD, Diplomate ACVP E. H, Fowler, DVM, Ph.D., Diplomate ACVP D. L. Fait, AALAS Cert, II K. A. France, B.A. L. C. Fisher, B.S. M. F. Kubena, B.S. T. A. Rebick, A.H.T., A.S., AALAS Cert. I D. J. McNeil, A.H.T., A.S., AALAS Cert. I. T. A. Savine, HT(ASCP) D. E. Dodd, Ph.D., DABT, Manager I. M. Pritts, Ph.D. D. R. Klonne, Ph.D. W. J. Kintlgh, B.S., AALAS Cert. I L. J. Calisti, B.S., AALAS Cert. Ill, Group Leader J. R. Bernard, B.S. W. J. Conroy, B.S. M. J. Cardella, AALAS Cert. Ill, Supervisor In addition, other BRRC personnel may participate in this study, ggport A draft summary of the results will be prepared upon the completion of the study and provided to the sponsor. Summary tables will be prepared including: chamber analytical data distribution and fate of all animals on study maternal clinical signs maternal body weights and weight changes maternal liver weight (absolute and relative) maternal gravid uterine weight ovarian corpora lutea of pregnancy status of uterine implantation sites (early, late resorptions, dead fetuses, live fetuses) These data will be used to determine the exposure concentrations for use in the definitive developmental toxicity evaluation of 1,1,1-trlchloroethane. A complete description of materials and methods, results (including tables) and conclusions of this exposure range-finding study will be included as. an appendix in the Final Report of the definitive developmental toxicity study. WPC/esk/0618P-2 07-01-86 St A2'?o Report 50-514 Appendix 5 Page 29 Page 9 REFERENCES Brown, M. B, and A. B. Forsythe (1974). The small sample behavior of some statistics which test the equality of several means. Technometrics 16, 129-132. Carpenter, C. P., E, R. Kinkeadj D. L. Geary, Jr., L. j. Sullivan and J. M. King (1975). Petroleum Hydrocarbon Toxicity Studies. I. Methodology. Toxicol. Appl. Pharmacol. 21, 246-262. DeSesso, J. M. and R. L. Jordan (1977). Drug-induced limb dysplasias in fetal rabbits. Teratology 15, 199-212. Environmental Protection Agency (EPA) (1983). Part III EPA Toxic Substances Control; Good Laboratory Practice Standards; Final Rule. Federal Register 48(230). 53922-53944. Hafez, E. S. E. (ed.) (1970). Reproduction and Breeding Techniques for Laboratory Animals. Lea and Febiger, Philadelphia, PA. Hartman, H, A. (1974). The fetus in experimental teratology. In: The Biology of the Laboratory Rabbit, S. H, Weisbroth, R. E. Flatt, and A. L. Kraus, eds,. Academic Press, NY, pp. 92-153. Levene, H. (1960). Robust tests for equality of variance. In: Contributions to Probability and Statistics, I. Olkin et al., eds., Stanford University Press, Stanford, CA, pp. 278-292. McClain, R. M. and L. Langhoff (1980). Teratogenicity of dlphenylhydantoin in the New Zealand white rabbit. Teratology 21, 371-379. / Nishimura, H. and K. Shiota (1977). Summary of comparative embryology and teratology. In: Handbook of Teratology, J. G. Wilson and F. C. Fraser, eds., Plenum Press, NY, Vol. 3, pp. 119-154. Palmer, A. K. (1978). The design of subprimate animal studies. In: Handbook of Teratology. J. G. Wilson and F. C. Fraser, eds., Plenum Press, NY, Vol. 4, pp 215-253. Salewski, E. (1964). Farbemethode Zura Makroskoplschen Nachweis Von Implantations-Stellen am Uterus Der Ratte. Naunyn-Schmiedebergs, Arch. Exp. Pathol. Pharmakol. 247. 367. Sokal, R. R. and F. J. Rohlf (1969). Biometry. W. H. Freeman and Co., San Francisco, pp. 369-371, 299-340, 370-372, 589-595. 042771 SL Report 50-514 Appendix 5 Page 30 Page 10 REFERENCES (Continued'! Stadler, J,f M.-J. Kessedjian and J. Perraud (1983). Use of the New Zealand white rabbit in teratology: Incidence of spontaneous and drug-induced malformations. Fd. Cosmet. Toxicol. 21(5). 631-636.. Weil, C. S. (1970). Selection of the valid number of sampling units and a consideration of their combination in toxicological studies involving reproduction, teratogenesis or carcinogenesis. Fd. Cosmet. Toxicol. &, 177-182. Woo, D. C. and R. M. Hoar (1982). Reproductive performance and spontaneous malformations in control New Zealand White rabbits: A joint study by MARTA. Teratology 25(2). 82A. (Data provided by the authors). WPC/esk/0618P-l 07-01-86 SL 042772 ATTACHMENT 1 Page 1 of 5 Report 50-514 Appendix 5 Page 31 MATERIAL SAFETY DATA SHEET Dow Chemical U.S.A. Midland, MI 48674 Emergency Phone: 517-636-4400 Product Code: 55352 Page: 1 PRODUCT NAME: METHYL CHLOROFORM, LOW STABILIZED Effective Date: 11/20/85 Date Printed: 06/21/86 MSDS:000&99 1, INGREDIENTS: 1,1,1-Trichloroethane CAS# 000071-55-6 99-9% 2. PHYSICAL DATA: BOILING POINT: l65F (74C) VAP PRESS: 100 irnnHg g 20C VAP DENSITY: 4.55 SOL. IN WATER: 0.07 g/100 g g 25C SP. GRAVITY: 1.330-1-335 @ 25/25C APPEARANCE: Colorless liquid. ODOR: Irritating odor at high concentrations. 3, FIRE AND EXPLOSION HAZARD DATA: FLASH POINT: None METHOD USED: TOC, TCC. COC FLAMMABLE LIMITS LFL: 7.5* 25C UFL: 15% @ 25C EXTINGUISHING MEOIA: Water fog. FIRE & EXPLOSION HAZARDS: Not available. FIRE-FIGHTING EQUIPMENT: Positive-pressure, self-contained respiratory equipment. (Continued on Page 2) (R) Indicates a trademark of The Dow Chemical Company 042773 SL MATERIAL ATTACHMENT 1 Page 2 of 5 Report 50-514 SAFETY DATA SHEET Appendix 5 ______________________________________ Page 32 Dow Chemical U.S.A. Midland, MI 48674 Emergency Phone: 517-636-4400 Product Code: 55352 Page: 2 PRODUCT NAME: METHYL CHLOROFORM, LOW STABILIZED Effective Date: 11/20/85 Date Prihted: 06/21/86, MSDS:000699 4. REACTIVITY DATA: STABILITY: (CONDITIONS TO AVOID) Autoignition temperature 998F (537C). Avoid open flames, welding arcs, or other high temperature sources which induce thermal decomposition. INCOMPATIBILITY: (SPECIFIC MATERIALS TO AVOID) Water - slow hydrolysis produces corrosive acid. Avoid contact with or storage in aluminum and its alloys. Definitely avoid contact with metallic aluminum and zinc powders. HAZARDOUS DECOMPOSITION PRODUCTS: Hydrogen chloride and very small amounts of phosgene and chlorine. HAZARDOUS POLYMERIZATION: Will not occur. 5. ENVIRONMENTAL AND DISPOSAL INFORMATION: ACTION TO TAKE FOR SPILLS/LEAKS: Small leaks: Mop up, wipe up, or soak up immediately. Remove to out-of-doors. Large spills: Evacuate area. Contain liquid; transfer to closed metal containers. Keep out of water supply. DISPOSAL METHOD: When disposing of the unused contents, the preferred options are to send to licensed reclaimer, permitted incinerators, or to evaporate very small quantities in compliance with local, state, and federal regulations including Subtitle C of the Resource Conservation and Recovery Act. Oumping into sewers, on the ground, or into any body of water is strongly discouraged, and may be illegal. Consult The Dow Chemical Company for further information. 6. HEALTH HAZARD DATA: EYE: May cause pain. May cause slight transient (temporary) irritation with slight transient corneal injury. Vapors may irritate eyes. SKIN CONTACT: Prolonged or repeated exposure may causd skin irritation. Repeated contact may cause drying or flaking of (Continued on Page 3) (R) Indicates a trademark of The Dow Chemical Company SL 042774 ATTACHMENT 1 Page 3 of 5 Report 50-514 MATERIAL SAFETY DATA SHEET Appendix 5 --------------------------------------------- -------------------------------------------------------------------------- Page 33 Dow Chemical U.S.A. Midland, MI 48674 Emergency Phone: 517-636-4400 Product Code: 55352 Page: 3 PRODUCT NAME: METHYL CHLOROFORM, LOW STABILIZED Effective Date: 11/20/85 Date Printed: 06/21/86 MSDS:000699 6. HEALTH HAZARD DATA: (CONTINUED) skin. SKIN ABSORPTION: A single prolonged skin exposure is not likely to result in absorption of harmful amounts. The LD50 for rabbits is about 15*000 mg/kg. INGESTION: Single dose oral toxicity is low. The LD50 for rats is >10,000 mg/kg. If aspirated (liquid enters the lung), may be rapidly absorbed through the lungs and result in injury to other body systems. INHALATION: Minimal anesthetic or narcotic effects may be seen in the range of 500-1000 ppm trichloroethane. Progressively higher levels over 1000 ppm may cause dizziness, drunkenness; concentrations as low as 10,000 ppm can cause unconsciousness and death. In confined or poorly ventilated areas, vapors which readily accumulate can cause unconsciousness and death. These high levels may also cause cardiac arrhythmias (irregular heartbeats). SYSTEMIC 6 OTHER EFFECTS: Based on available data, repeated exposures are not anticipated to cause any significant adverse effects. 1,1,1-Trichloroethane and similar mixtures did not cause cancer in long-term animal studies. Birth defects are unlikely. Exposures having no adverse effects on the mother should have no effect on the fetus. In animal studies, has been shown not to interfere with reproduction. Results of in vitro ('test tube') mutagenicity tests on 1,1,1-- trichloroethane have been negative. Results of mutagenicity tests in animals have been negative. 7. FIRST AID: EYES: Irrigate immediately with water for at least 5 minutes. SKIN: Wash off in flowing water or shower. Remove contaminated clothing and wash before reuse. INGESTION: Do not induce vomiting. Call a physician and/or (Continued on Page 4) (R) Indicates a trademark of The Dow Chemical Company 042775 SL ATTACHMENT 1 Page 4 of 5 MATERIAL SAFETY DATA SHEET Report 50-514 Appendix 5 Page 34 Dow Chemical U.S.A. Midland, MI 48674 Emergency Phone: 517-636-4400 Product Code: 55352 Page: 4 PRODUCT NAME: METHYL CHLOROFORM, LOW STABILIZED Effective Date: 11/20/85 Oate Printed: 06/21/86 MSDS:000699 7. FIRST AID: (CONTINUED) transport to emergency facility immediately. INHALATION: Remove to fresh air. If not breathing, give mouth-to-mouth resuscitation. If breathing is difficult, give oxygen. Call a physician. NOTE TO PHYSICIAN: Because rapid absorption may occur through lungs if aspirated and cause systemic effects, the decision of whether to induce vomiting or not should be made by an attending physician. If lavage is performed, suggest endotracheal and/or esophageal control. Danger from lung aspiration must be weighed against toxicity when considering emptying the stomach. Exposure may increase "myocardial irritability". Do not administer sympathomimetic drugs unless absolutely necessary. No specific antidote. Supportive care. Treatment based on judgment of the physician in response to reactions of the patient. 8. HANDLING PRECAUTIONS: EXPOSURE GUI DELINE (S): 1,1,1-Trichloroethane - OSHA standard is 350 Ppm and current ACGIH TLV is 350 ppm (450 ppm STEL). VENTILATION: Control airborne concentrations below the exposure guideline. Use only with adequate ventilation. Local exhaust ventilation may be necessary for some operations. Lethal concentrations may exist in areas with poor ventilation. RESPIRATORY PROTECTION: Atmospheric levels should be maintained below the exposure guideline. When respiratory protection is required for certain operations, use an approved air-purifying respirator. For emergency and other conditions where the exposure guideline may be greatly exceeded, use an approved positive pressure self-contained breathing apparatus. In confined or poorly ventilated areas, use an approved positive pressure self-contained breathing apparatus. SKIN PROTECTION: For brief contact, no precautions other than clean body-covering clothing should be needed. When prolonged (Continued on Page 5) (R) Indicates a trademark of The Dow Chemical Company MATERIAL SAFETY DATA SHEET ATTACHMENT 1 Page 5 of 5 Report 50-514 . Appendix 5 Page 35 Dow Chemical U.S.A. Midland, MI 48674 Emergency Phone: 517-636-4400 , Product Code: 55352 'Page: 5 PRODUCT NAME: METHYL CHLOROFORM, LOW STABILIZED Effective Date: 11/20/85 Date Printed; 06/21/86 MSDS:000699 8. HANDLING PRECAUTIONS: (CONTINUED) or frequently repeated contact could occur, use protective clothing impervious to this material. Selection of specific items such as gloves, boots, apron, or full body suit will depend on operation. EYE PROTECTION: Use safety glasses. Where contact with liquid is likely, chemical goggles are recommended because eye contact with this material may cause pain, even though it is unlikely to cause injury. 9. ADDITIONAL INFORMATION: SPECIAL PRECAUTIONS TO BE TAKEN IN HANDLING AND STORAGE: Handle with reasonable care. Avoid breathing vapors. Store in a cool dry place. Concentrated vapors of methyl chloroform, low stabilized - are heavier than air and will collect in low areas such as pits, degreasers, storage tanks, and other confined areas. Do not enter these areas where vapors of this product are suspected unless special breathing apparatus is used and an observer is present for assistance. 1.1.1-Trichloroethane products should not be packaged in aluminum aerosol cans or with finely divided aluminum or its alloys in an aerosol can. Aluminum is not an acceptable material of construction for pumps, mixers, fittings, storage tanks for 1.1.1-trichloroethane products or formulations. Metallic aluminum and zinc powders should be avoided. For additional information on toxicity, handling precautions, and first.aid, refer to chlorinated solvents literature form no. 100-5792. MSDS STATUS: Revised 1, 5, 6, 7. and 9. (R) Indicates a trademark of The Dow Chemical Company The Information Herein Is Given In Good Faith, But No Warranty, Express Or Implied, Is Made. Consult The Dow Chemical Company For Further Information. Report 50-514 Appendix 5 Page 36 BUSHY RUN RESEARCH R. 0. 4, Mellon Road, Export, Pennsylvania 15632 CENTER Telephone (412) 733-5200 PROTOCOL AMENDMENT tfl TITLE: Developmental Toxicity Exposure Range-Finding Study of Inhaled 1,1,1-Trichloroethane in New Zealand White Rabbits BRRC PROJECT NUMBER: 86-95-80603 SPONSOR: Halogenated Solvents Industry Alliance 1612 K Street, NW, Suite 300 Washington, DC 20006 TESTING FACILITY: Bushy Run Research Center (BRRC) R.D. #4, Mellon Road Export, PA 15632 Attention: R. W. Tyl (412) 733-5277 Reviewed and Approved by: Bushy Run Research Center: )(\,(u i j <-)!?//I*( fa Rochelle W. Tyl, Ph.D.' DABT Study Director/Manager Reproductive and Developmental Toxicology Section Date /-1 '-A GJ--.r.j Linda J. Calisti, B.S., Group Leader, Good Laboratory Practices/Quality Assurance Date */;ck. Fred R. Frank, Ph.D. Director Date 0759P-1 Sponsor: SL 042778 Bushy Run Research Center A Joint Melton Institute--Union Carbide Corporation Operation Report 50-514 Appendix 5 Page 37 Protocol Amendment it 1 Page 2 The protocol is amended as follows: 1. Location of Protocol Change: MATERIALS AND METHODS. Quarantine Period (Page 3) Description of Protocol Change: From: All animals will be housed in BRRC animal rooms 102 (females) and 105 (for male rabbit colony). The animals will be received at least two weeks before the initiation of the mating period. During the quarantine period the animals will be observed regularly for general health status and ability to adapt to the watering system. Body weights will be ascertained at least once prior to the mating period. Female rabbits in good health at least 2.5 kg in weight will be used for this study." To: All animals will be housed in BRRC animal rooms 107 (females) and 105 (for male rabbit colony). The animals will be received at least two weeks before the initiation of the mating period. During the quarantine period the animals will be observed regularly for general health status and ability to adapt to the watering system. Body weights will be ascertained at least once prior to the mating period. . Female rabbits in good health at least 2.5 kg in weight will be used for this study." Rationale for Change: The animal room was changed from 102 to 107 for the females prior to their arrival. The rooms are essentially indentical with all of the same environmental conditions and controls. This change will not impact on the conduct of the study. 2. Location of Protocol Change: MATERIALS AMD METHODS. Husbandry (Page 33 Description of Protocol Change: From: "During the quarantine period, rabbits will be housed individually in stainless steel wire-mesh cages mounted in steel racks. After mating, females will be housed one per cage in stainless steel wire-mesh cages mounted in steel racks. Deotlzed paperboard (Shepherd Specialty Papers, Inc., Kalamazoo, MI) will be placed under each row of cages to collect solid and liquid excreta. An automatic watering system with demand control valves mounted on each rack will provide water for the rabbits. Water and food will be available ad libitum at all times except during exposures. All study females will remain in Room 102 during quarantine, mating, prior to, between, and after exposures until-scheduled sacrifice on gd 19. The light cycle in this room will be 12 hours light/12 hours dark. Specific information on the cages (dimensions, etc.) will be included in the study data." SL 042779 Report 50-514 Appendix 5 Page 38 Protocol Amendment //l Page 3 To: "During the quarantine period, rabbits will be housed individually in stainless steel wire-mesh cages mounted in steel racks. After mating, females will be housed one per cage in stainless steel wire-mesh cages mounted in steel racks. Deotized paperboard (Shepherd Specialty Papers, Inc., Kalamazoo, MI) will be placed under each row of cages to collect solid and liquid excreta. An automatic watering system with demand control valves mounted on each rack will provide water for the rabbits. Water and food will be available ad libitum at all times except during exposures. All study females will remain in Room 107 during quarantine, mating, prior to, between, and after exposures until scheduled sacrifice on gd 19. The light cycle in this room will be 12 hours light/12 hours dark. Specific Information on the cages (dimensions, etc.) will be included in the study data." Rationale for Change: The animal room was changed from 102 to 107 for the females prior to their arrival. The rooms are essentially lndentical with all of the same environmental conditions and controls. This change will not impact on the conduct of the study. 3. Location of Protocol Change: INHALATION CHAMBER OPERATION. Scheduling fPage 7) Description of Protocol Change: From: "Animals arrive at BRRC: July 29, 1986 Acclimatization period begins: July 29, 1986 Mating period begins; August 16, 1986 (first gd 0 also August 16, 1986) Treatment period begins: August 22, 1986 Sacrifice period begins (gd 19): September 4, 1986 Submission of draft data to sponsor: The data in draft form will be submitted to the Sponsor within two (2) weeks after the last-sacrifice day for selection of the appropriate exposure concentrations for the definitive development toxicity study. A complete report of this study will be included as an appendix in the Final Report." To: "Animals arrive at BRRC: July 30, 1986 Acclimatization period begins: July 30, 1986 Mating period begins: August 16, 1986 (first gd 0 also August 16, 1986) Treatment period begins: August 22, 1986 Sacrifice period begins (gd 19): September 4, 1986 Submission of draft data to sponsor: The data in draft form will be submitted to the Sponsor within two (2) weeks after the last sacrifice day for selection of the appropriate exposure concentrations for the definitive development toxicity study. A complete report of this study will be included as an appendix in the Final Report." SL 042780 Report 50-514 Appendix 5 Page 39 Protocol Amendment til Page A Rationale for Change: The arrival date of the animals was delayed one day from July 29 to July 30, 1986, at the request of the supplier. This change still allows for at least two weeks of acclimatization and will not impact on the conduct of the study. WFC/esk/0759P-l 09-16-86 Sl> O*271 Report 50-514 APPENDIX fi Developmental Toxicity Exposure Range-Finding Study of Inhaled 1,1,1-Trichloroethane in New Zealand White Rabbits (11 Pages) Report 50-514 APPENDIX 6 Developmental Toxicity Exposure Range-Finding Study of 1,1,1-Trichloroethane in New Zealand White Rabbits OBJECTIVE The objective of this study was to establish the exposure-response range of the test article for maternal and/or embryonic toxicity in pregnant New Zealand white rabbits in order to select appropriate exposure concentrations . for use in the definitive developmental toxicity study. MATERIALS AND METHODS The test chemical, lot number and BRRC Sample Number were the same as in the definitive study. Forty (40) virgin female New Zealand white (NZW) rabbits (birthdate February, 1986) were received from the same supplier as that used in the definitive study on July 30, 1986, and were housed in animal room 102 upon arrival and moved to room 107 on the following day for the remainder of the study. Certified feed from the same supplier (Batch No. 6134 SMI) and water were available gd libitum except during exposures. Females were singly housed throughout the study. Quarantine, mating, exposures, and observational procedures were the same as those employed in the definitive study. Natural mating occurred on August 16 and 17, 1986; gestational day (gd) 0 was also August 16 and 17, 1987. Eight (8) mated females were assigned to each treatment group by stratified randomization procedures by body weight on gd 0. Exposures were for six hours per day, for 13 consecutive days, gd 6 through 18, August 22 through September 4, 1986. Exposure concentrations were 0, 1000, 3000 and 6000 ppm. Inhalation procedures including exposure of animals and generation and analysis of test atmospheres were the same as those employed in the definitive study. Exposures took place in the chambers in room 139. Animals were observed daily for clinical signs of toxicity and weighed on gd 0, 6, 9, 12, 15 and 19. On gd 19 (the day following the last exposure), all study females were sacrificed by intravenous injection of T-61 euthanasia solution, on September 4 and 5, 1986. Maternal eyes were immediately examined in situ by gently pressing a glass slide against the cornea and examining the eye under fluorescent light illumination. A complete necropsy examination was performed on each animal by Dr. E. H. Fowler, DVM, Ph.D., Veterinary Pathologist. Ovarian corpora lutea of pregnancy were counted, the gravid uterus and maternal liver were weighed and sections of the liver fixed for possible subsequent histopathology. The gravid uterus was examined for implantation sites, resorptions (early and late) and live and dead fetuses. Apparent nonpregnant uteri were stained with 10% ammonium sulfide for confirmation of pregnancy status. The protocol and amendment describing the design and conduct of this study are presented in Appendix 5. SL 042783 Report 50-514 Appendix 6 Page 2 RESULTS Analytical exposure concentrations were 98.4-101.1% of target concentrations; analytical to nominal concentration ratios were 0.90-0.99 indicating no degeneration or adsorption to chamber walls. Temperature and relative humidity in the chambers was maintained at 20.3-24.5C and 38.5-49.6%, respectively (Table 1). Details of chamber and exposure parameters are presented in Appendix 1C and ID. No females died, aborted, were removed from study or delivered early. Pregnancy rate was high and equivalent across all groups. One doe at 6000 ppm resorbed her entire litter (Table 2). Maternal body weights (Table 3) were depressed in a concentration-related pattern (predominantly at 6000 ppm) on gd 9, 12, 15 and 19, although no pairwise comparisons were statistically significantly different. Maternal body weight gain was depressed with increasing concentrations for gd 6-19 (treatment period), 9-12, 12-15, 15-19 and 0-19 (gestation period), notably at 3000 and 6000 ppm, although no pairwise comparisons were statistically significantly different (Table 3). Clinical signs of toxicity, including ocular discharge, loose feces and red material on paper board, were observed in three different does at 6000 ppm (Table 4). There were no maternal eye lesions observed at necropsy. No maternal observations at necropsy (Table 5) appeared treatment related except possibly the fully resorbed litter at 6000 ppm (Table 5). Maternal body and organ weight parameters at necropsy (Table 6) exhibited clear concentration-related changes: absolute body weight, body weight corrected for gravid uterine weight, and gestational body weight change (corrected for initial body weight and weight of the gravid uterus), all exhibited concentration-related decrements, although no pairwise comparisons were statistically signficantly different. Absolute and relative liver weight appeared reduced for all chemical-exposed groups. The value at 1000 ppm (but not at 3000 or 6000 ppm) was significantly reduced relative to that of controls. Gestational parameters are presented In Table 7. Ovarian corpora lutea and uterine implantation sites were slightly (but not statistically significantly) reduced at 3000 and 6000 ppm. There were no effects of treatment on preimplantation loss or on non-viable implants per litter; there was a significant reduction in non-viable implants at 1000 and 3000 ppm and a significant Increase in percent live fetuses per litter at 1000 ppm. There were no dead fetuses in this study. At 6000 ppm, the percent live fetuses per litter was reduced (but not significantly), due, at least in large part, to one doe with a fully resorbed litter. SL 042784 Report 50-514 Appendix 6 Page 3 CONCLUSIONS Based on the results of this exposure range-finding study the Sponsor selected target exposure concentrations for the definitive study, with concurrence of the Study Director on September 17, 1986. The exposure concentrations selected were 0, 1000, 3000 and 6000 ppm (the same as in the range-finding study) with the following rationale. At 6000 ppm in the range-finding study, there was evidence of-maternal toxicity: reduced body weights and weight gain, clinical signs of toxicity, and reduced corrected body weight at sacrifice (separate and distinct'from contributions of the gravid uterus). There was also possible developmental toxicity (reduced percent live fetuses per litter). These changes were not statistically significantly different from controls, but this may be due to the small size of each group (seven-eight does). For the definitive study with 24 females per group, these effects will probably attain statistical significance. At 3000 ppm, the same evidence of maternal toxicity was observed, with the likelihood that this exposure concentration will also produce effects in the definitive study. The 1000 ppm exposure concentration should be a "no observable effect level" (NOEL) for both maternal and developmental toxicity. PATH/esk/0949P-2 03-24-87 0A2785 Si' Report 50-514 Appendix 6 Page 4 SL 042786 Table 1 Exposure Parameters: Exposure Range-Finding Study Target concentration, ppm Analytical concentration, ppma A/T Ratiob Nominal concentration, ppma A/N Ratioac Temperature, Ca Relative humidity, %a 0 < MDLd - - 20.3 1.14 49.6 2.99 1000 1011 17.8 1.011 1109 11.4 0.91 0.015 20.7 * 0.43 48.5 2.43 3000 3019 161.4. 1.006 3049 117.0 0.99 0.023 21.3 0.55 48.8 * 3.01 a Grand mean of 14 daily means standard deviation b Analytical to target concentration ratio c Analytical to nominal concentration ratio d Less than the minimum detection limit of 10 ppm WPC/esk/0764P-2 03-17-87 6000 5907 126.1 0.984 6538 36.1 0.90 0.019 24.5 0.97 38.5 2.49 TABLE 2 DEVELOPMENTAL TOXICITY EXPOSURE RANGE-FINDING STUDY OF INHALED 1.1.1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS DISTRIBUTION AND FATE Report 50-514 Appendix 6 Page 5 PPM FEMALES ON STUDY 0 NO . (*) a 1000 NO . (X) 8 3000 NO . (X) 8 6000 NO . (X) 8 FEMALES THAT DIED 0 0 0 0 FEMALES THAT ABORTED 0 0 0 d FEMALES THAT DELIVERED EARLY 0 0 0 0 FEMALES REMOVED FROM STUDY 0 0 0 0 females examineo at laparotomy nonpregnant PREGNANT DOES WITH RESORPTIONS ONLY a DOES WITH VIABLE FETUSES 8 (100.0) 0 ( 0.0) a (100.0) 0 ( 0.0) 8 (100.0) 8 (100.0) 0 ( 0.0) 8 (100.0) 0 ( 0.0) B (100.0) 8 (100.0) 0 ( 0.0) 8 (100.0) 0 ( 0.0) B (100.0) 8 (100.0) 0 ( 0.0) 8 (100.0) ( 12.5) 7 ( 87.5) a The totally resorted litter was detected by ammonium sulfide staining and examination of the uterus at scheduled sacrifice. SL 042787 TABLE 3 DEVELOPMENTAL TOXICITY EXPOSURE RANGE-FINOING STUDY OF INHALED 1,1.1-TRICHLOROETHANE in NEW ZEALAND WHITE RABBITS BODY WEIGHTS AND WEIGHT CHANGE DURING GESTATION PPM 0 1000 3000 PPPY WEIGHT? (IJRAM?) gestational day MEAN 0 S.O. N 3767.0 257.76 8 3727.8 260.19 8 3764.2 240.95 8 gestational day MEAN S.D. N 6 3958.5 267.53 8 3936.4 257.62 8 3915.1 257.66 8 GESTATIONAL day MEAN S.D. N 9 3830.9 285.52 8 3856.8 281.07 8 3801.0 242.31 B GESTATIONAL DAY MEAN S.D. N 12 3828.6 252.62 8 3858.0 277.78 8 3779.9 253.15 8 GESTATIONAL DAY MEAN S.D. N 15 3866.7 271.46 a 3895.8 293.36 8 3805.0 277,69 e GESTATIONAL DAY MEAN S.D. N 19 3844.8 256.52 8 3846.4 256.80 8 3745.2 301.83 8 BODY WEIGHT CHANGE fGRAMS) GESTATIONAL DAYS 0 TO 6 (PRE-TREATMENT) MEAN 191.5 S.D. 91.76 N8 208.6 66.97 8 150.9 67.38 8 GESTATIONAL DAYS (TREATMENT) MEAN S.D. N 6 TO 19 -113.8 211.22 8 -90.0 55.32 8 -170.0 144.36 8 GESTATIONAL DAYS MEAN S.O. N 6 TO 9 -127.6 73.48 8 -79.6 73.35 8 -114.1 78.59 8 GESTATIONAL DAYS MEAN S.O. N 9 TO 12 -2.4 78.13 8 1.2 90.48 8 -21.1 56.42 8 GESTATIONAL DAYS MEAN S.D. N 12 TO 15 38.1 127.93 8 37.9 82.18 8 25.1 77.59 8 Report 50-514 Appendix 6 Page 6 6000 3720.5 231.47 8 3065.6 271.21 8 3758.6 219.46 8 3711.5 180.87 8 3722.6 147.87 8 3631.5 176.91 8 145.1 66.73 8 -234.1 185.66 8 -107.1 86.65 8 -47.1 61.29 8 11.1 42.88 8 SL 042788 TABLE 3 (CONTINUED) DEVELOPMENTAL TOXICITY EXPOSURE RANGE-FINDING STUDY of inhaled 1,1,1-TRICHLQROETHANE IN NEW ZEALAND WHITE RABBITS BOOY WEIGHTS AND WEIGHT CHANGE DURING GESTATION PPM 0 1000 3000 GESTATIONAL oays mean S.D. N 15 TO 19 -21.9 141.32 e -49.4 .94.69 a -59.8 39.69 8 GESTATIONAL DAYS (GESTATION) MEAN S.D. N 0 TO 19 77.7 213.02 8 118.7 68.61 a -19.0 167.05 8 MEAN DIFFERENCES CALCULATED FROM INDIVIDUAL DIFFERENCES Report 50-514 Appendix 6 Page 7 6000 -91.0 52. 14 8 -89.0 138.71 8 042789 Sh SL 042790 TABLE 4 DEVELOPMENTAL TOXICITV EXPOSURE RANGE-FINDING STUDY OF INHALED 1,1,1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS INCIDENCE OF CLINICAL OBSERVATIONS BY GESTATION DAY CATEGORY FINDING (LOCATION) GESTATIONAL DAYS 1.1.1 -TRICHLOROETHANE (PPM, 1nhaia11 on) 0 1000 3000 6000 NORMA). NO SIGNIFICANT CLINICAL OBSERVATIONS OEAO SCHEDULED SACRIFICE EVES/EARS/NOSE OCULAR DISCHARGE, OTHER (EYE-RIGHT) EXCRETA LOOSE FECES DRV RED MATERIAL ON PAPERBOARD** UNABLE TO HEMASTIX 0- 5 6- 19 6- 19 6- 19 6- 19 6- 19 8a 8 B 8 Be8B 88 8 8 000 1 000 1 000 1 BLOOD BY HEMASTIX ON TRAY BENEATH CAGE 6- 19 0 1 00 " Number of does exhibiting the observation. Tuts observation mss noted In the mornlna. prior to exposure, on gestation day 6 for that animal X wi O O' I u TABLE 5 DEVELOPMENTAL TOXICTTV EXPOSURE RANGE-FINOING STUDV OF INHALED 1 , 1 ,1 -TR1 CHLOROETHANE IN NEW ZEALAND WHITE RABBITS MATERNAL GROSS OBSERVATIONS AT NECROPSY - INCIDENCE SUMMARY NUMBER OF PREGNANT DOES PPM 0 1000 3000 6000 88B8 CORPORA LUTEA -CORPORA ALBICANS Oa 0 UTERUS -NO BLOCKAGE .LEFT HORN 10 AMNIOTIC SACS -ABSENT 00 PLACENTAS -ABSENT 00 LUNGS -COLOR CHANGE -EDEMA -TISSUE SAVED 00 00 00 * Number of does with gross observatI on. b This doe mss found to be pregnant by amonlum sulfide staining of tne uterus. c Same doe. 0 lb \0 a lb 0 1b ic 0 ic 0 ic 0 t 6 ^ V *0 > 7) TJ H re Tre) Do an \fl B< n X O' 50-514 TABLE 6 DEVELOPMENTAL TOXICITY EXPOSURE RANGE-FINDING STUDY OF INHALED 1.1,1-TRICHLQROETHANE in NEW ZEALAND WHITE RABBITS MATERNAL ORGAN WEIGHTS AT SACRIFICE Report 50-514 Appendix 6 Page 10 PPM 0 1000 3000 6000 INITIAL BODY WEIGHT MEAN S.D. N (GRAMS) 3767,0 257.76 '8 3727.8 260.19 8 3764.2 240.95 8 3720.5 231.47 8 BODY WEIGHT AT SACRIFICE MEAN S.D. N (GRAMS) 3844.8 2S6.52 8 3846:4 256.80 8 3745.2 301.83 B 3631.5 176.91 a GRAVID UTERINE WEIGHT MEAN S.D. N (GRAMS) T63.27 31.65 8 177.91 19.04 8 158.46 28.58 8 144.65 41.69 8 CORRECTED BODY WEIGHT MEAN S.D. N (GRAMS) 3681.51 243.55 8 CORRECTED BODY WEIGHT CHANGE (GRAMS) b MEAN *85.53 S.D. N 220.95 a 3666.53 243.12 B -59.25 73.74 8 3586.73 296.33 8 -177.47 176.11 8 3466.90 187.29 8 -233.63 169.70 8 LIVER WEIGHT (GRAMS) MEAN S.D. N 108.72 17.92 8 84.48 12. B8 8 93.49 26.55 a 88.74 11,74 6 RELATIVE LIVER WEIGHT MEAN S.D. N <*)c 2.96 0,47 a 2.30** 0.29 8 2.58 0.50 a 2.54 0.23 8 a Corrected body weight * body weight at sacrifice minus gravid uterine weight. Corrected body weight change * body weight at sacrifice minus gravid uterine weight minus initial body weight. Calculated as a percentage of the corrected body weight. *4` p < 0.01 compared to control. 042792 SL TABLE 7 DEVELOPMENTAL TOXICITY exposure range-finding STUDY OF INHALED 1,1.1-TRICHLOROETHANE IN NEW ZEALAND WHITE RABBITS GESTATIONAL PARAMETERS AT SACRIFICE Report 50-514 Appendix 6 Page 11 PPM 0 1000 3000 NO. PREGNANT DOES CORPORA LUTEA/DQE MEAN S.D. N B 10.1 1.1 8 8 10.9 1.8 8 a 9.5 1.8 8 TOTAL IMPLANTS/LITTER MEAN S.D. N 8.9 2.7 8 PERCENT PREIMPLANTATION MEAN S.D. N L0SSb 15.0 23.2 8 10.1 1.5 8 6.2 8.1 8 8.3 1 ,4 8 11.5 15.1 8 VIABLE IMPLANTS MEAN S.D. N 8.1 10.1 2.3 1.5 B8 8.1 1.6 8 NON-VIABLE IMPLANTS MEAN S.D. N 0.8 0.7 a 0.0 0.0 8 0.1 0.4 8 EARLY RESORPTIONS MEAN S.D. N 0.5 0.8 8 0.0 0.0 8 0.1 0.4 8 LATE RESORPTIONS MEAN S.D. N 0.2 0.5 8 0.0 0.0 8 0.0 0.0 8 DEAD FETUSES MEAN S.D. N 0.0 0.0 8 0.0 0.0 8 0.0 0.0 8 PERCENT LIVE FETUSES MEAN S.D. N 92.8 6.6 8 100.0 0.0 8 98.2 5.1 8 " The N is reduced because the data from one doe with no corpora lutea (no viable implants) were removed from the calculations. Percent preimplantation loss = corpora lutea minus total Implants divided by corpora lutea multiplied by 100. * p < 0.05 compared to control. ** p < 0.01 compared to control. 6000 8 9.1 2.0 7 8.0 2.6 8 4.3 5.5 7a 7.4 3.2 8 0.6 1.2 8 0.4 1.1 8 0.2 0.7 8 0.0 0.0 8 85.2 35.0 8 O'.n93