New sensitive assay of vancomycin in human plasma using high-performance liquid chromatography and electrochemical detection

A method using reversed-phase high-performance liquid chromatography with e lectrochemical detection for the analysis of vancomycin in human plasma was developed. Chromatographic conditions included an octadecyl column, a mobi le phase of acetonitrile-sodium phosphate buffer (pH 7) (12:88), a total ru n time of 12 min, and coulometric electrochemical detection at +700 mV. Lin ear detector response was found in the range 5-100 mug ml(-1) after a 1:80 dilution or from 0.5 to 50 mug ml(-1) after a 1:20 dilution of the samples. In both cases the correlation coefficient (r) of the calibration curve sta ndard was better than 0.995. Vancomycin determination was based on a denatu ration of plasma proteins with methanol, then a dilution with mobile phase was performed. Recovery df vancomycin from plasma was 103.1+/-3.9%, and no interference from commonly used drugs or endogenous compounds was observed. A significant correlation was shown with the EMIT assay (r=0.92, P<0.001) using clinical samples from children. This HPLC technique is simple, sensit ive, rapid, precise, selective and requires only 100 <mu>l of plasma for co mpletion. (C) 2001 Elsevier Science B.V. All rights reserved.