Enzymic color development of urinary 3 alpha-hydroxysteroids on thin-layer chromatograms.
I describe the use of 3 alpha-hydroxysteroid dehydrogenase (EC 1.1.1.50) for enzymic color development of 3 alpha-hydroxysteroids. Urinary 3 alpha-hydroxysteroids are converted to 3-ketosteroids by the enzyme, and the NADH formed is reacted with 2-p-iodophenyl-3-p-nitrophenyl-5-phenyl-tetrazolium chloride and "diaphorase" to form the colored formazan. The dye so formed on a thin layer chromatographic plate is measured by densitometoric scanning. I illustrate use of this method by showing results for urinary excretion of 3 alpha-hydroxysteroids by some various sorts of patients.
D015096:3-Hydroxysteroid Dehydrogenases; D002855:Chromatography, Thin Layer / Q000379:methods; D006801:Humans; D006914:Hydroxysteroids / Q000652:urine*; D013053:Spectrophotometry / Q000379:methods; D013379:Substrate Specificity
D006914:Hydroxysteroids; D015096:3-Hydroxysteroid Dehydrogenases