High Performance Liquid Chromatographic Separation of Sensitive Fluorescent Derivatives of Bile Acids with Cyclodextrin-Containing Mobile Phase

Abstract

The high-performance liquid chromatographic behavior of new bile acid fluorescent derivatives, the 7-methoxy-1,4-benzoxazin-2-one-3-methy 1 esters, with a cyclodextrin-containing mobile phase is compared to those of the bile acid-pyrenacyl and -anthroyl esters. Compared with conventional methods, inclusion chromatography gives a much more satisfactory separation of the bile acid fluorescent derivatives in a short time, but the chromatographic behavior of these derivatives has not been much influenced by the fluorophore used. The detection limits of the new derivatives are in the range of 10–20 fmol at a signal to noise ratio of 5. The application of this method for the separation of g1ycine-conjugated bile acids in human bile is also described.