LC-MS/MS法测定人血浆中万古霉素的血药浓度

目的: 建立测定人血浆中万古霉素浓度的LC-MS/MS分析方法,用于临床万古霉素血药浓度监测。方法: 以甲哌卡因为内标,将50 μL血浆加入到30 μL的25%(w/v)三氯乙酸溶液中,沉淀蛋白质,经离心处理后,取上清液5 μL进样分析。色谱条件: 采用Shimadzu VP-ODS(2.0 mm×150 mm,5 μm)色谱柱,以乙腈-0.1%甲酸水溶液为流动相,梯度洗脱,流速0.3 mL·min^-1;质谱条件: 电喷雾离子源,多反应监测,离子极性为正离子,用于定量的离子对分别为万古霉素m/z 725.0/144.3,内标甲哌卡因m/z 247.3/98.3。结果: 线性范围为0.5~100 μg·mL^-1,最低定量限为0.5 μg·mL^-1;日内、日间RSD<15%,相对误差为-0.8%~1.9%;提取回收率为74.3%~85.0%。结论: 本方法样品处理方法简单,方法准确,灵敏度高,适用于人血浆中万古霉素浓度的测定。

Objective: To establish an LC-MS/MS method for the content determination of vancomycin in human plasma, and apply to the clinical vancomycin monitoring. Methods: Mepivacaine was used as internal standard. 50 μL of plasma was precipitated with 25% trichloroacetic acid(w/v) and an aliquot of 5 μL of the supernatant was injected into the column for the analysis.The separation was achieved on a Shimadzu VP-ODS column(2.0 mm×150 mm, 5 μm) with a mobile phase containing 0.1% formic acid in water and acetonitrile. The mobile phase was gradient eluted at a flow rate of 0.3 mL·min^-1. ESI source was applied and operated in positive ion mode and multiple reaction monitoring(MRM). The ion combination of m/z 725.0/144.3, m/z 247.3/98.3 was used to qualify vancomycin and mepivacaine(internal standard), respectively. Results: Linear calibration curves were obtained in the concentration range of 0.5-100 μg·mL^-1, with the lower limit of quantification of 0.5 μg·mL^-1. The intra- and inter-day precision(RSD) values were below 15% and relative error was between -0.8% and 1.9% at all QC levels. The extraction recovery was 74.3%-85.0%. Conclusion: The established LC-MS/MS method of vancomycin determination is convenient, accurate and sensitive, which can be applied in analysis of the vancomycin concentration in human plasma.