Steroid metabolism in teleost gonads: purification and identification of metabolites by high-performance liquid...

A simple, efficient, and comprehensive technique for the purification, identification, and quantitation of the common steroid metabolites synthesized by the gonads of teleosts involving five systems of high-performance liquid chromatography (HPLC) was developed. Steroid standards were identified in HPLC by UV absorption at 254 nm or 280 nm, by differential refractive index, or by using radioactive standards. Metabolites that do not absorb UV light and are not resolved in the isocratic HPLC systems were identified in thin-layer chromatography following purification by HPLC. By using this technique, most of the steroid metabolites, including some polar metabolites, synthesized by the gonadal tissues of the teleosts can be purified within three steps of chromatography. The HPLC systems reported here are also useful in identifying the chromium trioxide oxidized products of metabolites, such as triols and tetrols, which considerably narrows down the number of probable metabolites.