Separation and characterization of ursodeoxycholate 7-n-acetylglu-cosaminides in human urine by high-performance liquid chromatography with fluorescence detection

The separation and characterization of unconjugated, glycine- and taurine-conjugated ursodeoxycholate 7-N-acetyl-glucosaminides in human urine without prior deconjugation have been carried out by high-performance liquid chromatography (HPLC) on a reversed phase column. The bile acid fraction was obtained from a urine specimen by the combined use of a Sep-Pakc18 cartridge for solid phase extraction and lipophilic gel, piperidinohydroxypropyl Sephadex LH-20 (PHP-LH-20), for ion-exchange chromatography. Bile acid N-acetyl-glucosaminides were derivatized quantitatively into the corresponding fluorescent esters through the inherent primary hydroxyl group by treatment with 9-anthroyl cyanide. The derivatives were separated into unconjugated, glycine- and taurine-conjugated fractions on PHP-LH-20. Subsequent resolution into individual N-acetylglucosaminides was attained.