Capillary gas-liquid chromatography of acetate-methyl esters of bile acids

Gas-liquid chromatographic separations of acetate-methyl esters of several common bile acids with and without a hydroxyl group at C-6 are compared with those of the corresponding trimethylsilyl ether-methyl esters on a CP-Sil-5 CB capillary column. Unlike the trimethylsilyl ether derivatives, the retention indices of the corresponding acetates were greatly influenced by the number of hydroxyl groups in the ring system. Epimeric hydroxyl groups at carbons 6, 7 as well as 12 increased retention index of the acetate-methyl esters of the bile acids, the effect of the 7 beta-hydroxyl group being most prominent. The 6 beta-acetoxyl group increased the retention index more than the 6 alpha-acetoxy group and contrary to the trimethylsilyl ether derivatives, a 6 beta, 7 beta-diacetoxy group showed larger increase in the retention index than the corresponding 6 alpha, 7 beta-diacetoxy group. The acetate derivatives of bile acid-methyl esters show larger retention times and reduced sensitivity than the corresponding trimethylsilyl ether derivatives. However, gas chromatography of bile acid acetate-methyl esters can be very useful for the characterization of bile acids and for bile acid analysis in the rat where muricholic acids and hyodeoxycholic acid are in abundance, since these bile acids are difficult to resolve from each other and from other common bile acids as the trimethylsilyl ether derivatives.