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Biology of Reproduction 59, 1023-1028 (1998)
©Copyright 1998 Society for the Study of Reproduction, Inc.

Development and Validation of a Fecal Testosterone Biomarker in Mus musculus and Peromyscus maniculatus1

Joseph E. Billittia, Bill L. Lasleyb, , and Barry W. Wilson2,a

a Departments of Environmental Toxicology and Animal Science b and Department of Population Health and Reproduction, University of California at Davis, Davis, California 95616

This is a report on the development and validation of an ELISA method to determine fecal testosterone levels, and on their evaluation as a biomarker for adverse effects of endocrine-disrupting chemicals on reproductive health using male rodents of the Peromyscus maniculatus and Mus musculus species as an animal model. The ELISA antibody had the highest specificity for testosterone (100%), followed by dihydrotestosterone (57.4%) and androstenediol (0.27%). Radiolabeled testosterone was injected i.p. into three mice. Fecal samples were collected, extracted, and analyzed by liquid scintillation counting. The ELISA was performed to characterize the excretion kinetics and metabolic fate of circulating testosterone. Solubilization of feces with 10% methanol overnight provided an extraction efficiency of 87% for all metabolites; an ethyl ether extraction was more selective for testosterone. The fecal excretion of the testosterone was a biphasic process with a majority of the radioactivity recovered in the first 24 hours. HPLC analysis revealed at least five testosterone metabolites in feces, with most metabolites being less polar than testosterone. This study forms the initial evaluation of what will become a field monitoring tool.

1 This work was supported by the Ecotoxicology Program of the U.C. Toxic Substances Research and Teaching Program, U.S. Air Force (AFOSR-91-0226), NIEHS Center for Environmental Health Sciences (P30-ES-05707), and the U.S. EPA (R819658) Center for Ecological Health Research at U.C. Davis. Although the information in this document has been wholly or in part funded by the United States Environmental Protection Agency, it may not necessarily reflect the views of the Agency and no official endorsement should be inferred.

2 Correspondence: Barry W. Wilson, Environmental Toxicology Dept., 1 Shields Ave., Davis, CA 95616. FAX: 530 752 3394; bwwilson{at}ucdavis.edu






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Copyright © 1998 by the Society for the Study of Reproduction.