High performance liquid chromatography with immobilized Ru(bpy)32+–KMnO4 chemiluminescence detection and its application in metabolism of repaglinide in pig liver microsomes

A novel high performance liquid chromatography–chemiluminescence (HPLC–CL) method for investigation of in vitro metabolism of repaglinide in pig liver microsomes with microdialysis sampling technique was developed. The chromatographic separation was performed on a Hypersil BDS-C18 column with an isocratic mobile phase consisting of methanol and 0.01 mol/L KH2PO4 (pH 3.0) (volume ratio 75:25) at a flow rate of 1.0 mL/min. The detection was based on the chemiluminescence reaction of repaglinide with acidic potassium permanganate (KMnO4) and tris (2,2′-bipyridyl)ruthenium(III) (Ru(bpy)33+), which was immobilized on the cationic ion-exchange resin for obtaining high sensitivity and reducing consumption of expensive reagent.