Abstract
The separation and characterization of unconjugated and conjugated bile acid 3-glucuronides in biological fluids without prior deconjugation by high-performance liquid chromatography (HPLC) are described. A urine sample from a patient with obstructive jaundice was passed through a Sep-Pak C 18 cartridge and was separated into groups by ionexchange chromatography on a lipophilic gel, piperidinohydroxypropyl Sephadex LH-20, providing the glucuronide fraction. Subsequent resolution into individual 3-glucuronides was attained by HPLC on μBondapak C 18 and Shodex ODS Pak F-411 columns. The 3-glucuronides of cholate, deoxycholate, chenodeoxycholate, glycocholate, glycochenodeoxycholate and taurochenodeoxycholate were identified on the basis of their behaviour in HPLC using mobile phases of different pH. The enzymatic hydrolysis of these glucuronides and derivatization of deconjugated bile acids with 1-anthroyl nitrile followed by chromatographic separation on a Cosmosil 5C 18 column with fluorescence detection were carried out for unequivocal characterization. The ratio of unconjugated, glyco- and tauro-conjugated bile acid 3-glucuronides excreted in urine was found to be ca. 2:3:1.
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callMore From: Journal of Chromatography B: Biomedical Sciences and Applications
Disclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
