LC-MS/MS法测定人血浆中万古霉素的血药浓度

Determination of vancomycin content in human plasma by LC-MS/MS

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#L血浆加入到30#L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#L进样分析。色谱条件:采用Shimadzu VP-ODS(2.0 mm×150 mm,5μm)色谱柱,以乙腈-0.1%甲酸水溶液为流动相,梯度洗脱,流速0.3mL·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0.5～100μg·mL-1,最低定量限为0.5μg·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 % triehloroacetic 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 × 150mm, 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 rn/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 ob- tained in the concentration range of 0. 5 - 100 pg . 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.