Analysis of Fecal Bile Acids and Neutral Steroids Using Gas-Liquid Chromatography

Abstract

In the present pilot study, for investigating the physiological effects of different types of nondigestible oligosaccharides, we have validated the application of methodologies for the analysis of bile acids and neutral steroids in feces of human subjects. The accuracy of the extraction and chromatographic procedures for the analyses of bile acids and neutral steroids was determined by recovery of added compounds to fecal homogenate. The precision of the above procedures was checked by analyzing these compounds in samples (n = 5) of the same fecal homogenate. Recoveries of added bile acids ranged from 86 and 96%, and those of neutral steroids varied from 81 to 97%. The precision expressed as coefficients of variation of bile acids and neutral steroids ranged from 2.3 to 8.3% and from 6.3 to 11.8%, respectively. The intra- and interindividual variabilities expressed as coefficients of variation of bile acids varied from 1 to 58 and from 0 to 74%, respectively. The same variabilities for neutral steroids ranged from 0.5 to 107% and from 1 to 168%, respectively. The methods validated in the present pilot study were adequate for applying to our forthcoming European Union coordinated major study on the physiological effects of different types of nondigestible oligosaccharides and involving large numbers of samples.

Fructooligosaccharide and Soy Isoflavone Suppress Colonic Aberrant Crypt Foci and Cyclooxygenase-2 Expression in Dimethylhydrazine-Treated Rats

“…One microliter was injected into a gas chromatograph (Star 3600X, Varian Inc., Palo Alto, CA), which was equipped with a DB-5 capillary column (30 m ϫ 0.32 mm ϫ 1 m film thickness) and a flame ionization detector. 27 The injector and detector temperature were set at 300°C. The oven temperature was programmed with the following steps: (1) increase temperature from 50°C to 230°C at 30°C/minute; (2) increase temperature from 230°C to 285°C at 1°C/minute and hold for 10 min-…”

Determination of underivatised sterols and bile acid trimethyl silyl ether methyl esters by gas chromatography–mass spectrometry–single ion monitoring in faeces

“…1). A variety of different methods of sample preparation for gas chromatographical analysis of faecal sterols have been published [3][4][5][6][7][8]. Most procedures include derivatisation, often silylation, of the sterols.…”