Enzymatic Determination and Thin-layer Chromatography of Bile Acids in Blood

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Abstract
It has been found that NAD-linked β-steroid dehydrogenase prepared from Pseudomonas testosteroni is useful for the determination of bile acids, the method being similar, in principle, to that described by Talalay et al. The bile acid, free, esterified or conjugated, having C-3-hydroxyl group (either 3α or 3β is determined by this method, regardless of either the number or the positions of the other hydroxyl group or groups in the molecules. This method is applicable for the determination of bile acids in blood and the total concentrations on the average of human blood were found to be 220±54 μ./100 ml. those of rabbits 727±202 μg./100 ml., and those of rats 230±175 μg./100ml. The bile acids in the ether-soluble acidic material of blood extract, hydrolyzed or non-hydrolyzed, were identified by thin-layer chromatography.