The detection and identification of synthetic steroids in horse urine

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Abstract
A scheme for the detection of synthetic steroids which could be used in the “doping” of race-horses is described. The method involves extraction from urine into ethyl acetate: ether (1:1) followed by initial two-dimensional thin-layer chromatography using (1) ethyl acetate (2) methylene chloride: dioxan: water (100:50:50). Elution of the ultraviolet absorbing spots for extinction measurements is followed by further one way thin-layer chromatography as free alcohols in amyl acetate: acetone (1:1). Further chromatography of the steroid alcohols in chloroform: ether: water (80:20:0·5) on formamide-impregnated plates and chromatography of the acetates in ether helps to identify the unknown steroid. Additional identification is made by colour reactions with (1) a tetrazolium reagent (2) vanillin: perchloric acid.

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