Document dYBNzK4zYVx4EKn8kMMxzKK0e

CRI NUMBER R & D REPORT^ tRl NIJMHR / wp \ DOW CHEMICAL U.S.A. RESTRICTED; for ust within The Dow Chemical Company only* LABORATORY REPORT CODE TBM-116-1 DATE ISSUED October 15, 1975 department INDUSTRIAL HEALTH AND MEDICINE, BIOMEDICAL AND COMPARATIVE TOXICOLOGY RESEARCH LAB A.laB1. n problem no. 4 1 8 i 5 __|1i111 TITLE EVALUATION OF ETHYLENIMINE BY THE DOMINANT LETHAL TEST 6 pages IN FULL REPORT AUTHOR IS-1 AUDTHO_RJ_tS*1 Kilian*SIGNATURE til M,D. f M. C, Benge, H., N Edwards, and_T. G. Pullln E V HEWER`S sTo N A T URI DESCRIPTIVE SUMMARY WITH CONCLUSIONS: This report n INTERIM and mainly; |y I NEW (Include in this spoce references to doto books, ond to earlier related reports, patents and publications*) The dominant lethal effect of ethylenimine (El) was evaluated by use of a modified screening procedure of Epstein and Shafner. The mutagenic index of El was lower than the corresponding control value in each week for the duration of this study. Statistical analysis indicated this difference was significant (p <0.05). Thus, under the conditions of this test procedure, the results do not indicate that El is mutagenic. DISTRIBUTION DEPARTMENT files R & D ADMINISTRATION central report index (566 Bldg. ~ Midland) 5 COPIES DO 14?606 GONF tdfntt ai Distribution lift is contingod on attached page* EVALUATION OF ETHYLENIMINE BY THE DOMINANT LETHAL TEST D. J, Kilian, M,D,, M, C, Benge, H, N. Edwards and T. G. Pullin INTRODUCTION The dominant lethal test, a method of mutagenicity testing in mammals, has been recommended by the MRAK Commission^ as part of required toxicological investigations for clearance of pesticides. This test may also be incorporated into the group of required toxicological investigations of chemicals in the pro posed Toxic Substances Act. ^ Epstein and Shafner^ and Partington and Bateman^ have already tested several environmental pollutants and chemicals using this procedure. According to Schwetz et al, (5) the most readily detectable mutation in the mouse is the dominant lethal which manifests itself as in utero lethality of embryos developing from gametes in which the muta tion has arisen. This procedure reflects mutagenic activity directly induced in male germ cells and excludes systemic drug effects in females. This data may provide one of the simplest means of relating animal induced mutagenic effects to humans. Although extrapolation of data from mice to man involves risk be cause of the species differences between the two, this technique offers a potential screen for chemical mutagens to which workmen are exposed. This paper presents the results of the screening of one such chemical. 00 ,,14260/ CONFIDENT!AL MATERIALS AND METHODS The screening procedure described by Epstein and Shafner^ with the exception of two modifications was used to test Evaluation of Ethylenimine by the Dominant Lethal Test 2 ethylenimine (El; The Dow Chemical Company, Texas Division) on Dublin ICR mice (Flow Laboratories, Dublin, Virginia). The male mice were given a single intraperitoneal injection of 3.8 mg El/kg body weight. The control group was given the solvent, tricaprylin, intraperitoneally. The modifications to the procedure consisted of the use of proven male breeders rather than breeders of unknown mating history and the extension of the tests for a period of nine consecutive weeks after injection rather than eight weeks. Statistical analyses of the data were conducted under the direction of Dr. E. B. Whorton, Jr., Associate Professor and Director, Medical Education Computer Center, Preventive Medicine and Community Health Biostatistics, University of Texas Medical Branch, Galveston, Texas. RESULTS A tabulation of the results of this study is presented in Table 1. The mutagenic index of El was lower than the corres ponding control value in each week for the duration of this study. Statistical analysis using Analysis of Variance procedures (Table 2) indicated this difference was significant (p <0.05). However, this analysis showed there was no significant trend over time and no significant interaction between time and chemical treat- mGnt' DO 147608 CONFIDENT! Al SUMMARY AND CONCLUSIONS The dominant lethal effect of ethylenimine (El) was evaluated /3\ by use of a modified screening procedure of Epstein and Shafner. The mutagenic index of El was lower than the corresponding control Evaluation of Ethylenimine by the Dominant Lethal Test 3 value in each week for the duration of this study. Statistical analysis indicated this difference was significant (p <0.05). Thus, under the conditions of this test procedure, the results do not indicate that El is mutagenic. DO 142609 CONFIDENTIAL BIBLIOGRAPHY 1. Mrak, Emil M. (Chairman), U.S. Department of Health, Education and Welfare, December 1969. 2. Schulz, Ralph R. (Ed.), (1973) Chemical Week 113(8):14-15. 3. Epstein, Samuel S. and H. Shafner (1968), Nature 219:385-387. 4. Partington, M. and A. J. Bateman (1964), Heredity 19:191-200. 5. Schwetz, B. A. et al. The Dow Chemical Company, Report NBT 1.5- 12- (1). ~ DO 142610 OONFTDFNTTAl. TABLE 1 IMPLANTS, RESORPTIONS AND MUTAGENIC INDEX IN FEMALE MICE BRED WITH EI-TREATED MALES Week After Administration 0 (pre-treatment) 1 2 3 4 5 6 7 8 9 TOTAL (9 weeks) Females Bred Control El* 75 45 57 15 87 24 87 24 72 15 69 15 66 15 57 14 57 15 42 15 594 152 Implants Control El 503 263 385 101 638 224 629 238 590 135 476 86 519 103 333 35 449 146 306 117 4,325 1,185 Resorptions Control El 22 11 23 3 27 5 36 12 35 6 28 2 25 2 14 1 14 3 11 3 213 37 *EI (Ethylenimine; The Dow Chemical Company, Texas Division) **MI = _____ Resorptions_____ x jqq Total Implantations o oo zTf o "oR ^\> -ZH O' 1> Mutagenic Index Control El 4.37 4.18 5.97 2.97 4.23 2.23 5.72 5.04 5.93 4.44 5.88 2.33 4.82 1.94 4.20 2.86 3.12 2.05 3.59 2.56 4.92 3.12 TABLE 2 ANALYSIS OF VARIANCE OF DOMINANT LETHAL DATA OF FEMALE MICE BRED WITH El-TREATED MALES Source Chemical Treatment Time Chemical Treat ment X Time df SS 1 534.896 8 266.149 8 138.394 Error 482 54312.812 MS 534.896 33.268 17.299 112.682 F ratio 4.746 0.295 0.153 P value <0.05 >0.25 >0.25 DO CONFIDENTIAL