Document YMLDVpJ9xnYyXgkVVOgw5y0

r t R S . 6 _ /ms Telomer Research Program Ciariant Report Title: In Vivo Rat Micronucleus Assay Author(s): Gregory L. Erexson, Ph.D Contractor: Covance Laboratories, Inc., Vienna, Virginia Study Dates: Initiation: October 18, 2001 In Life: October 23, 2001 - November 19, 2001 Completion: January 25, 2002 Study Objective: To evaluate 8-2 Alcohol for in vivo clastogenic activity and/or disruption of the mitotic apparatus by examining bone marrow polychromatic erythrocytes in male CRL:CD(SD)IGS BR rats for micronuclei. Materials and Methods: Groups of male rats (5/group) were administered a single oral gavage dose of either 0 (vehicle 0.5% aqueous methyl cellulose), 500, 1000, or 2000 mg 8-2 Alcohol/kg body weight. A positive control group of rats received a single oral dose of 60 mg cyclophosphamide/kg. At both 24 and 48 hours, 5 rats from each group were sacrificed and bone marrow was extracted from the tibia. Slides were prepared and scored for micronuclei and the PCE/NCE (polychromatic erythrocytes) (normochromatic erythrocytes) cell ratio was counted (scored at least the first 500 erythrocytes on each slide). Assay acceptance criteria was met with the criteria for a positive response being the detection of a statistically significant increase in micronucleated PCE's for at least 1 dose level and a statistically significant dose-related response. Findings: No lethality or signs of clinical toxicity were seen in the rats given in either 500, 1000, or 2000 mg/kg. 8-2 Alcohol did not induce a statistically significant increase in micronucleated PCE's at any of the dose levels tested. The positive control material showed the expected response demonstrating the sensitivity of the test system. (Data table follows.) Does not contain TSCA CBI 1 3 TRP 23-Apr-03 T reatm en t D ose H arvest T im e (H R ) Vehicle Control 0.5% MC 24 hr 48 hr Positive Control CP 60.0 mg/kg 24 hr Test Article 500 mg/kg 24 hr 48 hr 1000 mg/kg 24 hr 48 hr 2000 mg/kg 24 hr 48 hr "Significantly greater than the corresponding vehicle control, p<0.01 ''Significantly lower than the corresponding vehicle control, p<0.01 % M icron u cleated P C E 's M ean o f 2000 per A nim al S .E . M ales 0.040.02 0.070.01 3.100.29a 0.040.02 0.090.02 0.150.07 0.090.05 0.100.03 0.090.02 R atio P C E :N C E M ean S .E . M ales 1,260.09 1.11 0.04 0.780.00b 1.130.05 1.060.11 1.170.06 0.950.08 1.100.07 1,200.08 Conclusion: 8-2 Alcohol is considered negative in the rat bone marrow micronucleus assay under the conditions of this assay. Publications / Presentations: A) Covance Study 22900-0-454 OECD, 1/25/02. B) Results cited in: Kennedy, G. L., Jung, R., Iwai, H., Shin-Ya, S., Murphy, S., Drouot, N., Finlay, C., Donner, M., Erexson, G., and Stankowski, L., Jr., "Genetic Toxicology of 8-2 Telomer B Alcohol", Toxicol. Sci. 72(S-1):209:2003. Does not contain TSCA CBI 2 J4 TRP 23-Apr-03