Development and Validation of a Rapid High-Performance Liquid Chromatography Method with UV Detection for the Determination of Vancomycin in Mouse Plasma

Abstract

A rapid high-performance liquid chromatography (HPLC) method for the quantification of vancomycin in mouse plasma samples was developed and validated. Norvancomycin was used as the internal standard. Chromatographic separation was achieved on a Vydac C18 column (4.6×50 mm, 3 µm particle size) and the detection was made at 214 nm. A gradient elution was programmed with the mobile phases of 0.1% v/v trifluoroacetic acid (A) and 95:5 v/v acetonitrile: 0.1% TFA (B) and a flow rate of 1 ml/min. The total run time was 15 min. The calibration curve was linear over the range of 0.1-20 µg/ml, with a correlation coefficient (r) higher than 0.997 and the lower limit of quantitation (LLOQ) of 0.1 µg /ml. The intra-day accuracy values were between 90 and 112% and the inter-day ones ranged from 96 to 104%. Precision values ranged from 1.7 to 9.5% for intra-day and 6.3 to 9.4% for inter-day. Stability studies indicated that the mouse plasma samples containing vancomycin could be stored in freezer at -80°C and handled under normal laboratory conditions without significant loss of the drug. The assay was successfully applied to the pharmacokinetics and bio-distribution study of novel formulations of vancomycin in mice.