High performance liquid chromatography with immobilized Ru(bpy)32+-KMn04 chemiluminescence detection

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.