Determination of Urinary Pregnanetriol using Thin-layer Chromatography and 3α-hydroxysteroid-dehydrogenase
1976 Volume 5 Issue 1 Pages 140-143
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1976 Volume 5 Issue 1 Pages 140-143
Determination of urinary pregnanetriol using 3α-hydroxysteroid-dehydrogenase was described. It is based on the specific enzymatic convertion of 3α-hydroxy group on the steroid molecule into 3-keto group. Quantitation is obtained by measurment of the absorption at340nm from NADH, following a hydrolysis by β-glucuronidase, extraction with methylenechloride and purification by thin-layer chromatography. The main advantage of the method rests in its specificity of 3α-hydroxysteroid-dehydrogenase in comparison with the reaction of sulfuric acid.
