高效液相色谱-质谱联用法检测患者血浆中替考拉宁的浓度

Determination of teicoplanin in the patients′plasma by high performance liquid chromatography-mass spectrometry

目的:建立高效液相色谱-质谱联用法(HPLC-MS)定量测定人血浆中替考拉宁的浓度。方法:采用乙腈蛋白沉淀法处理50μL血浆,以达托霉素为内标,色谱柱:Welch Ultimate XB C18(2.1 mm×50 mm,3μm);柱温:40℃;流动相:乙腈-水(0.5%甲酸),梯度洗脱5.5 min。采用ESI离子源,正离子模式多反应监测方式(MRM)检测血浆中替考拉宁浓度,替考拉宁定量离子对分别为m/z 940.5→316.2,内标达托霉素离子对为m/z 811.0→313.0,碰撞电压分别为20 eV和42 eV。考察方法的选择性、残留、基质效应、线性范围、准确度与精密度和样本稳定性等。结果:血浆替考拉宁在1.0~100.0 mg·L^-1线性关系良好,定量下限为1.0 mg·L^-1。批内、批间精密度的标准偏差(RSD)均≤10.9%,替考拉宁的内标归一化基质效应在92.7%~109.3%且RSD≤12.0%。血浆样本可在-20℃条件下长期冻存至少41 d。临床实际样本检测中未发现干扰,方法稳定性良好。结论:该方法简便灵敏、准确可靠,适用于人体血浆中替考拉宁浓度的测定。

OBJECTIVE To establish a high performance liquid chromatography-mass spectrometry(HPLC-MS)method for the quantitative determination of teicoplanin in human plasma.METHODS Plasma of 50μL was prepared by protein precipitation with acetonitrile,used daltomycin as internal standard(IS).Chromatographic separation was performed on a Welch Ultimate XB C18 column(2.1 mm×50 mm,3μm);column temperature:40℃;mobile phase:acetonitrile-water(0.5%formic acid),gradient elution within 5.5 min.The concentration of teicoplanin in plasma was determined by ESI ion source and positive ion mode multi-reaction monitoring method(MRM).The ion transitions were performed at m/z 940.5→316.2 for teicoplanin and m/z 811.0→313.0 for daltomycin,with the collision voltage of 20 eV and 42 eV,respectively.The specificity,carryover,matrix effect,linearity,accuracy and precision of the method,and the stability of plasma samples were investigated.RESULTS The linear relationship of plasma teicoplanin was good in the range of 1.0~100.0 mg·L^-1,and the lower limit of quantification was 1.0 mg·L^-1.The standard deviation(RSD)of intra-batch and inter-batch precision was≤10.9%,and the normalized matrix effect of teicoplanin was 92.7%~109.3%and the RSD was≤12.0%.Plasma samples may be stored frozen at-20℃for at least 41 days.No interference was found in clinical samples with good method stability.CONCLUSION The method is simple,sensitive,accurate and reliable,and is suitable for the determination of teicoplanin in human plasma.