Abstract
ABSTRACT A high-performance liquid chromatography method has been developed and validated for the simultaneous determination of amoxicillin, metronidazole, and pantoprazole in human plasma. Solid-phase extraction (SPE) was utilized to extract the analytes along with the internal standard, tinidazole. Baseline resolution was achieved using a 30 : 70 v/v acetonitrile–25 mM potassium phosphate buffer containing 0.25% triethylamine (pH 6.5) mobile phase, at a flow rate of 0.5 mL/min. A 5 μm phenyl column, equipped with a guard column, with detection at 230 nm constituted the HPLC system. The method yields retention times of 3.7, 4.8, 6.9, and 16.2 min for amoxicillin, metronidazole, tinidazole (internal standard), and pantoprazole, respectively. Limits of detection were 200 ng/mL for amoxicillin and metronidazole and 100 ng/mL for pantoprazole. Recoveries from human plasma ranged from 83–92% for amoxicillin, 81–89% for metronidazole, 85–94% for tinidazole, and 93–101% for pantoprazole. Intra-day (n=5) and inter-day (n=15) precision (% RSD) and accuracy (% error) for all analytes ranged from 1.11–5.97% and 0.72–13.5%, respectively.
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