General methodology for the estimation of common neutral urinary steroids by multi-columin liquid chromatography

A method is described for the estimation of common neutral urinary steroids by multi-column liquid chromatography. Hydrolysis is performed in two steps: enzymatically, using beta-glucuronidase, followed by solvolysis. Initial short column chromatography separates the neutral steroids into three fractions according to polarity: 17-oxosteroids, corticosteriods less polar than cortolones, and cortolones and cortols. The cortolone, cortol fraction is oxidized and the different steroid groups are chromatographed on capillary aluminum oxide and silica gel columns. A computerized, spectrophotometric system is used for the quantitation procedure.