HPLC Determination of Carboxyl Group Using 2-Bromoacetyltriphenylene as a Pre-labeling Reagent - Separative determination of bile acids by HPLC

2-Bromoacetyltriphenylene 유도체화제를 이용한 카르복실기 함유 성분의 분석법 (II) - 담즙산 혼합물의 HPLC에 의한 분리정량

A high performance liquid chromatographic method was developed for the simultaneous determination of free and glycine conjugated bile acids. Free and glycine conjugated bile acids were extracted from bear gall bladder by methanol and from serum using a Sep-pak $C_{18}$ catridge. The extracted bile acids were labeled with 2-bromoacetyltriphenylene in acetonitrite using 18-crown-6-ether as a catalyst. Derivatized bile acids were separated from the individual bile acids on a reversedphase column (Chemcosorb 5-ODS-H) using acetonitrile-methanol-water(10:50:30) as a mobile phase and monitored by an UV-detector at 254nm. Linearities of calibration curve were obtained between 4 ng and 24 ng, and recoveries from bear gall bladder sample were higher than 94%.