用UPLC-MS/MS法测定人血浆中厄他培南的浓度

Determination of ertapenem in human plasma by UPLC-MS/MS

目的 建立一种简单、灵敏、快速的超高效液相色谱-串联质谱法,用于测定人血浆中厄他培南浓度。方法 以厄他培南-D4为内标,用乙腈溶液沉淀血浆中的蛋白;色谱柱:ACQUITY HSS T3 (2.1 mm×50.0 mm,1.8μm);流动相为0.1%甲酸水溶液(含有2 mmol·L^(-1)甲酸铵)-乙腈(0.1%甲酸),梯度洗脱方式;流速:0.4 mL·min^(-1);进样量:1μL;柱温:45℃;分析时间:4.5 min;采用电喷雾离子源(ESI),正离子选择反应监测模式(SRM)模式扫描。考察该方法的专属性、标准曲线和定量下限、精密度与回收率、基质效应、稀释效应、稳定性。结果 厄他培南在0.5~80.0μg·mL^(-1)线性关系良好,其标准曲线方程为y=4.25×10^(-1)x-2.64×10^(-2)(r~2=0.999 0);定量下限为0.5μg·mL^(-1),批内、批间相对标准偏差为1.39%~4.15%。提取回收率为58.36%~64.57%,稀释效应的相对标准偏差为3.30%,基质效应为99.71%~103.23%。室温、反复冻融、4℃、长期稳定性的相对标准偏差均小于10%。结论 该方法灵敏、快速、专属性强,适用于厄他培南的临床药物治疗监测。

Objective A simple,sensitive and rapid ultra high performance liquid chromatography tandem mass spectrometry(UPLC-MS/MS) method was developed and validated for the determination of ertapenem in human plasma.Methods Using ertapenem-D4 as internal standard,the protein in plasma was precipitated with acetonitrile;chromatographic column:ACQUITY HSS T3(2.1 mm×50.0 mm,1.8 μm);the mobile phase was 0.1% formic acid aqueous solution(containing 2 mmol·L^(-1) ammonium formate)-acetonitrile(0.1% formic acid),using a gradient elution;flow rate:0.4 mL·min^(-1),injection volume:1 μ L,column temperature:45 ℃,the analysis time was 4.5 min,the scanning mode is positive ion selective reaction monitoring mode(SRM) with an electric spray ion source(ESI).The specificity,standard curve and lower limit of quantification,precision and recovery,matrix effect,dilution effect and stability were investigated.Results Ertapenem had a good linearity within 0.5-80.0 μg·mL^(-1),and the standard curve was y=4.25×10^(-1)x-2.64×10^(-2)(r^(2)=0.999 0),the lower limit of quantification was 0.5 μg·mL^(-1),the relative standard deviation within and between batches is 1.39%-4.15%.The extraction recovery rate was 58.36%-64.57%,and the relative standard deviation of dilution effect was 3.30%,and the matrix effect was 99.71%-103.23%.The relative standard deviation of room temperature,repeated freeze-thaw,4 ℃,and long-term stability are all less than 10%.Conclusion The method is sensitive,rapid and specific,which is suitable for clinical monitoring of Ertapenem.