SIMPLE ANALYSIS OF AMOXICILLIN IN PLASMA BY HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY WITH INTERNAL STANDARDIZATION AND ULTRAVIOLET DETECTION

A simple high performance liquid chromatographic method with ultraviolet detection at 229 nm is described for quantitation of amoxycillin in plasma. After deproteination of plasma samples with perchloric acid and adjustment of the pH to 4.9, the supernatant was injected onto a reversed phase C18 column, using acetonitrile: phosphate buffer (0.01 M, pH 7.4) (1:25 v/v) as the mobile phase. Amoxycillin and the internal standard, cefadroxil, were eluted at 23 min and 12 min, respectively, without interference from endogenous substances. Processed samples were stable for at least 24 h at room temperature which permitted automated batch processing overnight. Calibration plots of the amoxycillin to cefadroxil peak-height ratio vs. amoxycillin concentration were linear (P<0.0001; r greater-than-or-equal-to 0.995) from 0.25 mg/L to at least 16.0 mg/L. Between-day and within-day imprecision (CV) ranged between 3.7% and 17.7%. Absolute recovery for amoxycillin and cefadroxil exceeded 82%. The application was demonstrated by the analysis of amoxycillin in human plasma after a single oral dose of amoxycillin (250 mg) suspension.