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  • Title: Enzymic conversion of 3beta-hydroxy-5-ene-steroids and their sulfates to 3-oxo-4-ene-steroids for increasing sensitivity in LC-APCI-MS.
    Author: Higashi T, Takayama N, Shimada K.
    Journal: J Pharm Biomed Anal; 2005 Sep 15; 39(3-4):718-23. PubMed ID: 15905063.
    Abstract:
    A method of increasing the sensitivity of 3beta-hydroxy-5-ene (Delta5)-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 Delta5-steroids were rapidly converted to their 3-oxo-4-ene (Delta4)-forms by the treatment with ChO and the obtained Delta4-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 Delta5-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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