Rates and product identification for trenbolone acetate metabolite biotransformation under aerobic conditions.

Trenbolone acetate metabolites are endocrine-active contaminants discharged into the aquatic environment in runoff from agricultural fields, rangelands, and concentrated animal feeding operations. To investigate the environmental fate of these compounds and their biotransformation mechanisms, the authors used inocula from a variety of different water sources and dosed biologically active microcosms with approximately 1400 ng/L of trenbolone acetate metabolites, including 17ß-trenbolone, trendione, and 17α-trenbolone. To investigate aerobic biotransformation rates and interconversions between known trenbolone acetate metabolites, gas chromatography-tandem mass spectrometry was used to measure concentrations and assess product distributions as a function of time. High-resolution liquid chromatography-tandem mass spectrometry (LC-MS/MS) was used to characterize novel transformation products and potential transformation pathways. Kinetic analysis yields observed half-lives of approximately 0.9 d, 1.3 d, and 2.2 d for 17ß-trenbolone, trendione, and 17α-trenbolone, respectively, at 20 °C, although colder conditions increased half-lives to 8.5 d and biphasic transformation was observed. Relative to reported faster attenuation rates in soils, trenbolone acetate metabolites are likely more persistent in aqueous systems. Product distributions indicate an enzymatic preference for biotransformation between trendione and 17ß-trenbolone. The LC-MS/MS characterization indicates dehydrogenation products as the major detectable products and demonstrates that major structural elements responsible for bioactivity in steroids are likely retained during biotransformation.

Product-to-parent reversion of trenbolone: unrecognized risks for endocrine disruption.

Trenbolone acetate (TBA) is a high-value steroidal growth promoter often administered to beef cattle, whose metabolites are potent endocrine-disrupting compounds. We performed laboratory and field phototransformation experiments to assess the fate of TBA metabolites and their photoproducts. Unexpectedly, we observed that the rapid photohydration of TBA metabolites is reversible under conditions representative of those in surface waters (pH 7, 25°C). This product-to-parent reversion mechanism results in diurnal cycling and substantial regeneration of TBA metabolites at rates that are strongly temperature- and pH-dependent. Photoproducts can also react to produce structural analogs of TBA metabolites. These reactions also occur in structurally similar steroids, including human pharmaceuticals, which suggests that predictive fate models and regulatory risk assessment paradigms must account for transformation products of high-risk environmental contaminants such as endocrine-disrupting steroids.