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Several solvent systems are described for the separation of neutral and phenolic steroids. Benzene and methanol in different proportions, with and without methylene chloride or chloroform, were found to be the most suitable systems for the separation of the 3 classic estrogens. Heptane, isooctane, hexane and cyclohexane were the basic solvents used for the chromatography of various neutral steroids. These systems required the addition of small amounts of either benzene, ethyl acetate or chloroform together with methanol. Sephadex LH-20 column chromatography offers several features which make it particularly useful in steroid assay procedures such as competitive protein binding and radioimmunoassay.
Carr et al. (1971) studied this question.