Abstract
A selective, sensitive, column-swiching-high-performance liquid chromatography employing direct injection of microserum has been developed for vancomycin quantitation in human serum and applied to clinical situations. The columns consist of a separation column filled with reversed ODS-C18 and a pretreatment column filled with a macroporous hydrophilic copolymer. A mixture of potassium phosphate buffer and acetonitrile is used as the mobile phase, and vancomycin is directly detected at a UV wavelength. The chromatogram showed a high separation factor; with no interference from other endogenous substances or combined drugs in surum. Good linearity was also obtained (0-69μmol/l). The coefficient of variation for within- day reproducibility was 1.24-2.12%, with a recovery rate of 97-106%. The minimum limit of detection was 0.03μmol/l (S/N=3). The correlation coefficient between the fluorescence polarization immunoassay and this method was 0.975. The serum concentrations in patients treated with vancomycin were monitored at regular intervals. This fast, accurate, and in expensive method, can be applied to routine clinical laboratory testing.
