Journal of Lipid Research, Vol 27, 208-214, Copyright © 1986 by Lipid Research, Inc.
Capillary gas-liquid chromatography of glycine-conjugated bile acids without prior hydrolysis
JM Street, DJ Trafford and HL Makin
A method is described for the gas-liquid chromatographic (GLC) analysis of
intact glycine conjugates of the major bile acids present in human plasma.
It is, therefore, now possible to analyze glycine-conjugated and
unconjugated bile acids together on a single GLC column without the
necessity for a hydrolytic step. A large number of derivatives of bile acid
glycine conjugates were examined, but only acetate- and silyl
ether-derivatives of carboxylic acid methyl esters were found initially to
be suitable. It was not possible to make acetates consistently, and
trimethylsilyl ethers did not allow resolution of the glycine conjugates of
cholic and chenodeoxycholic acids. Dimethylethylsilyl ether methyl ester
derivatives were subsequently found to give the best results.
Chromatographic conditions for successful analysis of these derivatives
were examined and it was found to be necessary to use wall- coated
capillary columns of thin film thickness (0.12 micron) and very high
carrier gas flow rates (ca. 20 ml/min hydrogen). Using acetonitrile and
Bond Elut extraction, fractionation on Sep-Pak SIL cartridges, and
derivatization as dimethylethylsilyl ether methyl esters, the capillary
gas-liquid chromatography of intact glycine- conjugated bile acids from
human plasma was demonstrated for the first time.
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