超高压液相色谱-串联质谱法检测万古霉素血药浓度及与化学发光法的比较
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摘要
目的建立超高压液相色谱-串联质谱(UPLC-MS/MS)检测人血清万古霉素浓度的新方法。方法采用Waters ACQUITY UPLC/ABSCIEX QTRAP4000超高效液相色谱串联质谱仪和ACQUITY UPLC BEH Amide色谱柱,以去甲万古霉素为内标,电喷雾离子源、正离子多反应监测模式检测万古霉素血药浓度。方法学评价实验包括:专属性、线性、精密度、准确度和与微粒子化学发光分析法,并进行方法学比较。结果 UPLC-MS/MS法专属性好,反应时间为2 min,线性范围为1～100 mg/L (R~2=0. 997),定量下限为1 mg/L,日内、日间精密度小于15%,准确度范围在96. 2%～108%。UPLC-MS/MS法与常规开展的微粒子化学发光免疫分析方法比较,结果相关性良好,相关方程y=0. 576x+2. 367(R~2=0. 911),但存在21%的负偏倚。结论 UPLC-MS/MS可简便、特异地定量人血清中万古霉素浓度,与化学发光方法具有较好的可比性,可用于临床检测。
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 QTRAP4000 ultra high-performance liquid chromatography tandem mass spectrometry,with ACQUITY UPLC BEH Amide chromatography column and norvancomycin as the 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 (R~2= 0. 997) with the quantitative limit of 1 mg/L. The inter-day and inner-day precision were less than 15%,and the accuracy was in therange 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. 576 x + 2. 367 (R~2=0. 911). There was 21% negative bias when compared UPLC-MS/MS 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,which has good correlation with chemiluminescence method and is appropriate for clinical application.
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 QTRAP4000 ultra high-performance liquid chromatography tandem mass spectrometry,with ACQUITY UPLC BEH Amide chromatography column and norvancomycin as the 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 (R~2= 0. 997) with the quantitative limit of 1 mg/L. The inter-day and inner-day precision were less than 15%,and the accuracy was in therange 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. 576 x + 2. 367 (R~2=0. 911). There was 21% negative bias when compared UPLC-MS/MS 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,which has good correlation with chemiluminescence method and is appropriate for clinical application.
引文
[1]Rybak MJ,Lomaestro BM,Rotschafer JC,et al.Therapeutic monitoring of vancomycin in adults summary of consensus recommendations from the American Society of Health-System Pharmacists,the Infectious Diseases Society of America,and the Society of Infectious Diseases Pharmacists[J].Pharmacotherapy,2009,29:1275-1279.
[2]陈耀龙,陈恳,叶志康,等.中国万古霉素治疗药物浓度监测指南的制定[J].中国询证医学杂志,2015,15:236-239.
[3]Zhao W,Kaguelidou F,Biran V,et al.External evaluation of population pharmacokinetic models of vancomycin in neonates:the transferability of published models to different clinical settings[J].Br J Clin Pharmacol,2013,75:1068-1080.
[4]Oyaert M,Peersman N,Kieffer D,et al.Novel LC-MS/MS method for plasma vancomycin:comparison with immunoassays and clinical impact.[J].Clin Chim Acta,2015,441:63-70.
[5]Cao Y,Yu J,Chen Y,et al.Development and validation of a new ultra-performance liquid chromatographic method for vancomycin assay in serum and its application to therapeutic drug monitoring[J].Ther Drug Monit,2014,36:175-181.
[6]范捷,郭志磊,于洋,等.高效液相色谱法测定万古霉素血药浓度[J].中国医院药学杂志,2012,32:1590-1592.
[7]裔照国,岳峰,季宏建,等.HPLC法测定人血清中去甲万古霉素浓度及临床应用[J].药学与临床研究,2009,17:522-524.
[8]Shou D,Dong Y,Shen L,et al.Rapid quantification of tobramycin and vancomycin by UPLC-TQD and application to osteomyelitis patient samples[J].J Chromatogr Sci,2014,52:501-507.
[2]陈耀龙,陈恳,叶志康,等.中国万古霉素治疗药物浓度监测指南的制定[J].中国询证医学杂志,2015,15:236-239.
[3]Zhao W,Kaguelidou F,Biran V,et al.External evaluation of population pharmacokinetic models of vancomycin in neonates:the transferability of published models to different clinical settings[J].Br J Clin Pharmacol,2013,75:1068-1080.
[4]Oyaert M,Peersman N,Kieffer D,et al.Novel LC-MS/MS method for plasma vancomycin:comparison with immunoassays and clinical impact.[J].Clin Chim Acta,2015,441:63-70.
[5]Cao Y,Yu J,Chen Y,et al.Development and validation of a new ultra-performance liquid chromatographic method for vancomycin assay in serum and its application to therapeutic drug monitoring[J].Ther Drug Monit,2014,36:175-181.
[6]范捷,郭志磊,于洋,等.高效液相色谱法测定万古霉素血药浓度[J].中国医院药学杂志,2012,32:1590-1592.
[7]裔照国,岳峰,季宏建,等.HPLC法测定人血清中去甲万古霉素浓度及临床应用[J].药学与临床研究,2009,17:522-524.
[8]Shou D,Dong Y,Shen L,et al.Rapid quantification of tobramycin and vancomycin by UPLC-TQD and application to osteomyelitis patient samples[J].J Chromatogr Sci,2014,52:501-507.