Abstract
A system of paper chromatography has been presented in which the stationary phase is a high-boiling hydrocarbon mixture (odorless kerosene) and the mobile phase is 60% aq. propanol. This system is not only useful for separation and identification of the cholesterol class of compounds but may be conveniently used to free tissue extracts of so-called fat. The use of the system in preparative chromatography has been discussed. Special application of two color reagents, phosphomolybdic acid and phosphotungstic acid, has been described. Proper use of the former can make it fairly specific for some steroids with secondary alcohol functions. Phosphotungstic acid was shown to give a characteristic pink color with many steroids having Δ 5, 3β-ol functions if warmed gently lavender a vivid lavender if heated more strongly. Certain steroids with allylic alcohol functions were also very chromogenic but with varying results. Precautions and limitations in the use of the reagents were discussed.
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