Simultaneous quantification of amoxycillin and metronidazole in plasma using high-performance liquid chromatography with photodiode array detection

Abstract

A simple, robust HPLC method was developed to measure simultaneously the plasma concentrations of amoxycillin and metronidazole in order to assess their disposition in the eradication of Helicobacter pylori. Plasma samples were protein precipitated, pH adjusted and the supernatant injected onto the HPLC system which used a C18 column, paired-ion aqueous mobile phase and photodiode array detection of amoxycillin at 230 nm and metronidazole at 313 nm. Intra- and inter-day precision and inaccuracy were less than 10% for concentrations between 5 and 20 mg/l. The limit of quantification was 1 mg/l. Samples were stable on the HPLC injector for 48 h at room temperature and multiple freeze-thaw cycles led to no decomposition.