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Abstract
A method of increasing the sensitivity of 3β-hydroxy-5-ene (Δ 5)-steroids in liquid chromatography–atmospheric pressure chemical ionization-mass spectrometry (LC–APCI-MS) based on the structural conversion by cholesterol oxidase (ChO) was demonstrated. The Δ 5-steroids were rapidly converted to their 3-oxo-4-ene (Δ 4)-forms by the treatment with ChO and the obtained Δ 4-forms provided 3–14-fold higher sensitivity compared to intact steroids in the positive-APCI-MS. This enzymic conversion method was also applied to the sulfated conjugates of Δ 5-steroids after solvolysis. The method enabled the detection of trace levels of dehydroepiandrosterone and androstenediol 3-sulfate in human serum, which could not be detected by the usual LC–MS.
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