Simultaneous Determination of Amoxicillin, Clarithromycin and Esomeprazolein Mice Plasma after Oral Administration by Reverse Phase HPLC Method

A simple and precise reverse phase high-performance liquid chromatography (RP-HPLC) method for simultaneous determination of amoxicillin, clarithromycin and esomeprazole in mice plasma after an oral administration was developed and validated. An isocratic elution was achieved on C18 column with a mobile phase containing buffer, potassium di hydrogen phosphate (KH2PO4 ) 0.05 M at pH 5 and methanol (60:40 v/v) at a flow rate of 1 mL/min, injection volume was 10 µL and UV detection was kept at 205 nm. Linearity was in the range of 0.5-100 µg/mL for amoxicillin, 50-1000 µg/mL for clarithromycin and 0.1-100 µg/mL for esomeprazole. Limit of detection (LOD) and Limit of quantification (LOQ) were 0.26 µg/mL and 0.79 µg/mL for amoxicillin, 8.97 µg/mL and 27.20 µg/mL for clarithromycin, 0.13 µg/mL and 0.39 µg/mL for esomeprazole respectively. All samples were stable at room temperature for 72 hours. The pharmacokinetic studies showed that the maximum plasma concentrations (Cmax) were 1042.17 ± 4.0, 218.67 ± 5.3 and 18.97 ± 3.6 µg/mL for amoxicillin, clarithromycin and esomeprazole respectively. Whereas, the times to reach maximum plasma concentration (Tmax) were 2.0, 4.0 and 2.0 hours respectively. Over all, the validated HPLC method may be used for the determination of such drugs in their pharmaceutical formulation and can be applied for routine quality control analysis.