Abstract
Steroid extracts from the uring of newborn infants can be separated into two fractions according to their mode of conjugation: metabolites excreted in the sulphate and glucuronide fractions appeared to be of different biological significance. Both fractions still represent complex mixtures of structurally related compounds which are not completely resolved by gas-liquid chromatography (GLC). Methods are presented for the fractionation of the extracts by lipophilic gel chromatography on a highly substituted hydroxyalkoxypropyl Sephadex derivative. A straight-phase system eluted with benzene permits the separation of groups of steroids according to the number of hydroxyl groups in the molecule. A reversed-phase system eluted with methanol- n-heptane (9:1) permits the rapid isolation of the polar steroids. Both systems provide additional structural information of the chromatographed steroids and form the basis of useful separation and purification procedures prior to GLC and gas chromatography—mass spectrometry (GC-MS). An example is given of the application of the straight-phase system to the fractionation of a urinary extract for GC-MS and mass fragmentography.
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