Development of HPLC Methods for the Determination of Vancomycin in Human Plasma, Mouse Serum and Bronchoalveolar Lavage Fluid

Abstract

Two high-performance liquid chromatography methods utilizing a protein precipitation technique were developed to analyze vancomycin in human plasma, mouse serum and bronchoalveolar lavage (BAL) fluid. The mobile phase consisted of ammonium phosphate buffer with acetonitrile. A cross-matrix validation was performed to ensure that the mouse serum was comparable to the original biological matrix of human plasma. Murine BAL samples were run on a saline standard curve. For saline samples, the mobile phase from the human plasma study was used with the addition of 1M sodium hydroxide (0.2%) to avoid interfering peaks. A reversed-phase column was used with an ultraviolet detector set at 240 nm for human plasma and 198 nm for saline to increase peak size. The standard curves were liner over the ranges of 1 to 80 µg/mL for human plasma and 0.1 to 10 µg/mL for saline. These assays are simple, reproducible and accurate. These analytical techniques were successfully applied to analyze vancomycin concentrations in mouse serum and BAL samples.