Analysis of Tissue Steroids By Liquid-Gel Chromatography and Computerized Gas Chromatography-Mass Spectrometry

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Abstract
A method for analysis of unconjugated neutral steroids of low and medium polarity in tissues is described. Nonpolar lipids in the tissue extract are removed by reversed phase chromatography on hydroxyalkoxyalkyl Sephadex. Further purification is obtained by straight phase chromatography using the same gel. Acidic contaminants are removed by filtration of a reaction mixture with methoxyamine hydrochloride through the anion exchanger Amberlyst A-26. The steroids are then analyzed as MO-TMS derivatives using repetitive scanning gas chromatography-mass spectrometry. Spectra are recorded on magnetic tape and quantification of steroids is based on peak areas in specific fragment ion current chromatograms.