A high performance liquid chromatography method for simultaneous determination of albendazole metabolites in human serum

Abstract

A simple assay for albendazole (ABZ) main metabolites—albendazole sulphoxide (ABZ-SO), albendazole sulphone (ABZ-SO 2) and albendazole amino sulphone (ABZ-SO 2-NH 2)—in serum using high performance liquid chromatography was developed. The method involves liquid–liquid extraction of the serum by ethyl acetate, clean up with n-hexane and re-extraction with ethyl acetate, followed by separation on RP-C 8 column with a mixture of methanol: acetonitrile: acetic acid: water (40:1:10:49) as the eluting solvent. ABZ-SO and mebendazole—used as internal standard—were detected by UV ( λ=286 nm), and ABZ-SO 2 and ABZ-SO 2-NH 2 with fluorescence spectrophotometer at (Excitation=286 nm, Emission=333 nm) and (Excitation=286 nm, Emission=315 nm), respectively. The assay was accurate and reproducible with a detection limit of 10 ng/ml for ABZ-SO, 2 ng/ml for ABZ-SO 2 and 4 ng/ml for ABZ-SO 2-NH 2. Disregarding ABZ determination, which is not of pharmacokinetic importance as it is not found in human plasma after oral administration, the proposed method is appropriate for further pharmacokinetic and metabolism study of this drug.