Analysis of bile acids by high performance liquid chromatography after pre-column derivatization with 9-(2-hydroxyethyl)-acridone

9-(2-hydroethyl) -acridone (hea), a novel ultraviolet and fluorescence labelling reagent, has been utilized as pre-column derivatization reagent for the analysis of 10 bile acids by high performance liquid chromatography ( hplc) with fluorescence detection. hea reacts with bile acids in the presence of the coupling agent 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide hydrochloride ( edc . hcl) and the base catalyst 4-dimethylaminopyridine ( dmap) to give the corresponding sensitively fluorescent derivatives with an excitation maximum at lambda(ex) 404 nm and an emission maximum at lambda(em) 440 nm. the labeled derivatives exhibit high stability under reversed-phase conditions. the method, in conjunction with a gradient elution, offers a baseline resolution of the 10 bile acids derivatives on a reversed-phase hypersil bds c-18 column. the average detection limit ( at a signal-to-noise ratio of 3) for per injection is 8.3 fmol.