Simultaneous determination of eighteen steroid hormones in antler velvet by gas chromatography–tandem mass spectrometry

A method for the simultaneous determination of eighteen steroid hormones, including 17α-ethinylestradiol, 17α-estradiol, 19-nortestosterone, estriol, testosterone, androsterone, 17β-estradiol, estrone, progesterone, 17α-hydroxyprogesterone, medroxyprogesterone, norethisterone acetate, testosterone 17-propionate, medroxyprogesterone-17-acetate, corticosterone, megestrol-17-acetate, chlromadinone-17-acetate and 17β-estradiol-benzoate, in antler velvet was developed by solid-phase extraction (SPE) and gas chromatography–tandem mass spectrometry (GC–MS/MS). Prior to the determination by GC–MS/MS, the analytes were derivatized with heptafluorobutyric acid anhydrides (HFBA). The recoveries were in the range of 62.13–104.00% for eighteen compounds at concentration levels (5–500 μg/L) and relative standard deviations (RSDs) were lower than 17.15%. The detection limits were from 0.2 to 22.3 μg/kg for the steroid hormones. The developed method was rapid, sensitive and successfully applied to the determination of steroid hormones in antler velvet samples collected from three countries.