高效液相色谱/电喷雾质谱法测定血浆中曲美他嗪浓度

Determination of Trimetazidine in Human Plasma by High Performance Liquid Chromatography-Mass Spectrometry

建立了高效液相色谱/电喷雾质谱法（HPLC/ESI-MS）测定人血浆中曲美他嗪浓度。以利多卡因为内标,样品经甲醇沉淀蛋白,取上清液用三氟乙酸酸化后,50℃真空离心,浓缩至干,流动相溶解后进样。色谱柱：Waters Xterra MSC 18（150mm×4.6mm,5μm）,柱温：40℃,流动相：pH2.00的三氟乙酸溶液-甲醇（60∶40,V/V）,流速：0.6mL/min;电喷雾离子源,四级杆质谱检测器,选择离子监控模式,检测离子的质核比分别是235（利多卡因）和267（曲美他嗪）。曲美他嗪的线性范围为2.5-100μg/L,检出限2.5μg/L;日间、日内相对标准差均小于7%;相对回收率96.45%-103.03%,提取回收率72.45%-80.47%。

A sensitive and specific high performance liquid chromatography/electrospray ionization-mass spectrometry （HPLC/ESI-MS） method has been developed and validated for the identification and quantification of trimetazidine （TMZ） in human plasma. The TMZ and internal standard, lidocaine, were isolated from plasma by protein precipitation with methanol. The acidified supematant was dried in a rotational-vacuum-concentrator, and the residue was dissolved with mobile phase prior to analysis. Chromat c separation was performed on a Xterra MS C18 column （ 150 mm ×4.6 mm, with 5 μm particle size） with the mobile phase consisting of methanol and water （pH =2.0 adjusted with trifluoroacetic acid） （40：60, V/V）, and the flow rate was 0.6 mL/min. Detection was performed on a single quadrupole mass spectrometer with selected ion monitoring （SIM） mode via electrospray ionization （ESI） source. The method was linear over the concentration range of 2.5 -100 μg/L for TMZ. The intra- and inter-day precisions were less than 7%, relative recovery ranged from 96.45% to 103. 03% and the exaction recovery was between 72.45% and 80.47%. The presented work offers a relatively simple and sensitive methodology within short turn-around time, which has also been successfully applied to a clinical pharmacoki-netic study.