A quantitative HPLC–MS method for the simultaneous determination of testosterone, 11-ketotestosterone and 11- hydroxyandrostenedione in fish serum

BLASCO, M.; CARRIQUIRIBORDE, P.; MARINO, D.; RONCO, A.E.; SOMOZA, G.M.

JOURNAL OF CHROMATOGRAPHY B

ELSEVIER

Año: 2009 vol. 877 p. 1509 - 1515

0378-4347

A simple and novel HPLC–MS method for the simultaneous quantification of testosterone, 11-ketotestosterone, and 11[1]-hydroxyandrostenedione in fish serum was developed and validated. Separation was achieved on a C-18 column using a water–acetonitrile mobile-phase with a cycle time of 12 min. Ion detectionwas performed using ESI positive SIM at [M+H] (m/z 303, 303, 289). The linear ranges (0.2–50 ng/ml), limits of detection (0.1–0.2 ng/ml) and quantification (0.2–0.5 ng/ml) were established. The method was validated by measuring the three androgens in goldfish sera, displaying comparable values to those reported by other analytical techniques (RIA, EIA).