An ultra performance liquid chromatographic (UPLC) method with photo-diode array (PDA) detection for the quantification of vancomycin in human serum was developed and validated.Protein precipitation was employed for serum sample preparation,and the extract was then injected into the system.Separation was performed on an ACQUITY UPLC BEH C18 column (50 x 2.1 mm,1.7μm) using a mobile phase composed of acetonitrile and 5 mM KH2PO4 buffer at pH 2.5.Both vancomycin and internal standard were determined using PDA detection at 230 urn.The calibration curve was linear over the range from 1 to 100 mg/L (R2 >0.9990).The specificity,extraction recovery,intra-and inter-day accuracy and precision,and stability were validated in human serum.Both UPLC and FPIA assays were applied to measure the vancomycin concentration in 172 clinical serum samples,which were collected from the hospitalized patients receiving vancomycin in Shanghai,China.Correlation analysis between the measurements of UPLC and FPIA gave the linear regression equation:(Concentration by UPLC) =0.949 * (Concentration by FPIA)-0.517 (R2 =0.923).Owing to the short analysis time and small amount of serum needed,the UPLC method developed in this study can be routinely used for vancomvcin therapeutic drug monitoring (TDM).
收起