Abstract: Objective To establish the liquid chromatography-tandem mass spectrometry method for vancomycin in human serum. Methods The analyses were carried out using the Waters ACQUITY UPLC/AB SCIEX QTRAP 4000 ultra high-performance liquid chromatography tandem mass spectrometry, with ACQUITY UPLC BEH Amide chromatography column and norvancomycin as internal standard. The electrospray ionization and positive ion multiple reaction monitoring mode was used for vancomycin mass spectrometry measurement. The method evaluation tests included specificity, linearity, precision, accuracy and method comparison with micro-particle chemiluminescence method. Results The method of UPLC-MS/MS for vancomycin had good specificity. The retention time of vancomycin was 2.0 minutes. The linear range was 1-100 mg/L (R2 = 0.997) with the quantitative limit of 1mg/L. The inter-day and inner-day precision were less than 15%, and the accuracy was in the range of 96.2%～108%. The correlation analysis showed a good linear between micro-particle chemiluminescence immunoassay method and UPLC-MS/MS, with the correlation equation y=0.576x+2.367 (R2=0.911). There was 21% negative bias when UPLC-MS/MS compared with chemiluminescence method for higher specificity. Conclusions A simple and accurate method of UPLC-MS/MS/MS for determination of vancomycin in human serum is developed in this study, which has good correlation with chemiluminescence method, and is appropriate for clinical application.

Key words: liquid chromatography-tandem mass spectrometry, vancomycin