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  • Title: Derivatization in liquid chromatography/mass spectrometric analysis of neurosteroids.
    Author: Mitamura K, Shimada K.
    Journal: Se Pu; 2001 Nov; 19(6):508-12. PubMed ID: 12545460.
    Abstract:
    Liquid chromatography/mass spectrometry (LC/MS) is now considered to be the most promising analytical method for the determination of biological substances, especially nonvolatile or highly polar substances. However, some compounds do not show enough sensitivity in LC/MS and soft-ionization methods commonly used in LC/MS, such as electrospray ionization (ESI) and atmospheric pressure chemical ionization (APCI), sometimes do not give satisfactory structural information. This report presents an overview of the derivatization methods in the LC/MS analysis of neurosteroids or neuroactive neurosteroids, which are synthesized and accumulated in the nervous system. The derivatization of pregnenolone 3-sulfate, one of these steroids, with 4-(N,N-dimethylaminosulfonyl)-7-hydrazino-2,1,3-benzoxadiazole gave a satisfactory sensitivity during the quantitative analysis using LC/ESI-MS. The obtained results are much lower than those previously obtained using gas chromatography/mass spectrometry or radioimmunoassay. On the other hand, the derivatization to acetate was useful for the treatment of labile catechol estrogens in rat brains and gave enough structural information in LC/APCI-MS, which confirmed the existence of catechol estrogens in mammalian brains.
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