Abstract
Capillary gas-liquid chromatographic separation was studied for the complete set of the 26 theoretically possible isomers of mono-, di-, and trihydroxylated 5 beta-cholanic acids, which differ from one another in the number, position, and configuration of hydroxyl groups at C-3, C-7, and/or C-12 in the nucleus, as well as for some of their related acids. The bile acid samples were chromatographed as their methyl ester-trimethylsilyl (TMSi) ether derivatives and analyzed on three capillary columns coated with nonpolar OV-1, slightly polar OV-17, and polar SP-2340 as liquid phases. The retention times on capillary gas-liquid chromatography (GLC) responded dramatically to the minor structural differences, and almost complete separation of the positional and stereochemical isomers was achieved by the combined use of SP-2340 and OV-17 (or OV-1) capillary columns.
Highlights
We report here capillary gas-liquid chromatography (GLC) analyses of the 26 acids, as well as of some related acids, as their methyl ester-TMSi ether derivatives, on three columns of differing liquid phase polarities
T h e source of bile acid samples used in this study was as follows: lithocholic, chenodeoxycholic, ursodeoxycholic, deoxycholic, cholic, hyocholic, and hyodesoxycholic acids were available from commercial sources; A combination of gas-liquid chromatography (GLC) and mass spectrometry is well recognized as one of the most powerful tools for separating and identifying the individual components in a mixture of bile acids obtained in metabolic studies
Concerning the GLC of bile acids, a large number of studies have been concerned with the relationship between the structures and the retention times on various liquid phases as well as with the use of suitable derivatives such as acetates, trimethylsilyl (TMSi) ethers, or trifluoroacetates [1,2,3,4]
Summary
T h e source of bile acid samples used in this study was as follows: lithocholic, chenodeoxycholic, ursodeoxycholic, deoxycholic, cholic, hyocholic, and hyodesoxycholic acids were available from commercial sources;. Concerning the GLC of bile acids, a large number of studies have been concerned with the relationship between the structures and the retention times on various liquid phases as well as with the use of suitable derivatives such as acetates, trimethylsilyl (TMSi) ethers, or trifluoroacetates [1,2,3,4]. Those data are of special value in structural elucidations, the studies were almost exclusively carried out by using conventional packed columns which often render in-. The chromatograph was fitted with a support-coated open tubular type of glass capillary column coated with three different polarities of liquid phases: OV-1 (1 00% dimethyl silicone),OV-17 (50%phenyl methyl silicone),
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