Configuration at C-25 in 3 alpha, 7 alpha, 12 alpha-trihydroxy-5 beta-cholestan-26-oic acid by X-ray crystallography

The two diastereoisomers at carbon-25 of 3 alpha, 7 alpha, 12 alpha-trihydroxy-5 beta-cholestan-26-oic acid, a key intermediate in the biosynthetic pathway of cholic acid, were obtained in pure form by a combination of fractional crystallization and thin-layer chromatography. The configuration at C-25 of these two isomers was established by X-ray crystallography as 25S for one diastereoisomer (mp 199-201 degrees C) and 25R for the other (mp 180-182 degrees C). These findings permit us to determine, unequivocally, the configuration of this naturally occurring C27-bile acid in man and other animals and to establish the stereospecificity of the microsomal and mitochondrial omega-hydroxylation pathway for the side-chain oxidation of cholesterol to bile acids.