3 alpha-, 7 alpha-, and 12 alpha-OH group specific enzymic analysis of biliary bile acids: comparison with gas-liquid chromatograpy

3 alpha-Hydroxysteroid dehydrogenase (3 alpha-HSDH) from P. testosteroni, 7 alpha-HSDH (Escherichia coli ATCC No. 29532) and 12 alpha-HSDH (Clostridium group P, strain C48-50, ATCC No. 29733) were used to directly measure 3 alpha-, 7 alpha-, and 12 alpha-OH groups in extracted human bile-rich duoden...

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Bibliographic Details
Published in:Journal of lipid research 1980-03, Vol.21 (3), p.381-385
Main Authors: Macdonald, I A, Williams, C N, Musial, B C
Format: Article
Language:English
Subjects:
3-alpha-Hydroxysteroid Dehydrogenase (B-Specific)
3-Hydroxysteroid Dehydrogenases
Bile - analysis
Bile Acids and Salts - analysis
Chenodeoxycholic Acid - analysis
Cholic Acids - analysis
Chromatography, Gas
Deoxycholic Acid - analysis
Humans
Hydroxysteroid Dehydrogenases
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Summary:3 alpha-Hydroxysteroid dehydrogenase (3 alpha-HSDH) from P. testosteroni, 7 alpha-HSDH (Escherichia coli ATCC No. 29532) and 12 alpha-HSDH (Clostridium group P, strain C48-50, ATCC No. 29733) were used to directly measure 3 alpha-, 7 alpha-, and 12 alpha-OH groups in extracted human bile-rich duodenal aspirates. Twelve samples chosen from widely differing ratios of cholic/chemodeoxycholic/deoxycholic were computed by solving three simultaneous equations. Comparison of these ratios with those obtained by a) thin-layer chromatography and 3 alpha-, 7 alpha-HSDH assays and b) gas-liquid chromatographic analysis showed no significant difference. Addition of known amounts of pure cholic, chenodeoxycholic, deoxycholic, or lithocholic acid to individual bile extracts gave an appropriate yield of 3 alpha-, 7 alpha-, and 12 alpha-OH groups. The direct (non-chromatographic) enzymic method has the advantages of being rapid, convenient, and inexpensive, and thus suitable for clinical use.
ISSN:0022-2275
1539-7262
DOI:10.1016/S0022-2275(20)39818-7