Rapid enzymatic determination of 0-3-oxo-bile acids separated by thin-layer chromatography

A method for the rapid quantification of 3-oxo-5-β-cholan-24-oic acids has been developed. The acids are separated on silica gel G and located using a water spray or iodine vapor. Each oxo acid is eluted from the gel and reduced with sodium borohydride. The resulting α- and β-hydroxy acids are then...

Full description

Saved in:
Bibliographic Details
Published in:Journal of Chromatography A 1978-08, Vol.155 (2), p.421-426
Main Authors: Beher, W.T., Sanfield, J., Stradnieks, S., Lin, G.J.
Format: Article
Language:English
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:A method for the rapid quantification of 3-oxo-5-β-cholan-24-oic acids has been developed. The acids are separated on silica gel G and located using a water spray or iodine vapor. Each oxo acid is eluted from the gel and reduced with sodium borohydride. The resulting α- and β-hydroxy acids are then oxidized in a reaction catalyzed by 3-hydroxysteroid dehydrgenase during which NAD is reduced to NADH. The absorbance of the reaction mixture is determined at 340 nm and is directly proportional to the amount of 3-oxo acid originally present on the thin-layer plate.
ISSN:0021-9673
DOI:10.1016/S0021-9673(00)88004-8