Analytical Chemistry related to Biofunctional Research. Development of an analytical method for intermediates in bile acid biosynthesis by GC/MS.

A method for the analysis of bile alcohols and hydroxycholesterols as intermediates in monohydroxyl bile acid biosynthesis has been developed by gas chromatography/mass spectrometry (GC/MS) with selected ion monitoring in a high-resolution mode. The linearity of the calibration curves for the bile alcohols, hydroxycholesterol and bile acids ranged from 10 similar to 250 pmol (r > 0.999), and the detection limits were 50 similar to 100 fmol (S/N= 2 similar to 6). The recoveries of the bile alcohols, hydroxycholesterol and bile acids from the rat liver mitochondrial fraction ranged over 97.9 similar to 111%, respectively, of the added amounts of their standard samples. The present method was made available for the determination of bile alcohols, hydroxycholesterol and related monohydroxy bile acids in incubation mixtures after only a simple extraction procedure using a solid-phase extraction cartridge.