THE DETERMINATION OF DELTA-5-ANDROSTENEDIOL AND ITS SULFATE IN SERUM AND URINE BY GAS-CHROMATOGRAPHY MASS-SPECTROMETRY

OBJECTIVES We developed an assay for delta-5-androstenediol (adiol) an d delta-5-androstenediol-3-sulphate (adiol-3S) in serum and adiol and total adiol sulphate (adiol-S) in urine. DESIGN An analytical procedur e using HPLC and gas chromatography-mass spectrometry was devised and tested for its reliability. MEASUREMENTS After addition of deuterated androstenediol as internal standard, serum and urine samples were extr acted. Steroid sulphates were hydrolysed. The extracts and hydrolysate s were purified on HPLC, adiol was derivatized using heptafluorobutyri c anhydride and finally quantified by gas chromatography-mass spectrom etry. RESULTS The assay is accurate and reproducible. The coefficient of variation (CV) for the determination of adiol in serum samples is 4 % (intra-assay) and 9% (interassay) and for urine samples 3 and 8% res pectively. The intraassay CV for the adiol-3S analyses is 5% for serum and 2% for urine samples while the interassay CV values for adiol-3S are 10% for serum and 7% for urine samples. The recovery of adiol and adiol-3S from serum and urine samples is 97%. CONCLUSIONS The develope d assay meets the analytical demands needed for clinical applications.