基于LC-MS/MS法的人血清中美托洛尔药物浓度的测定

Determination of Metoprolol Drug Concentration in Human Serum by LC-MS/MS Method*1

目的:建立基于LC-MS/MS法的人血清中美托洛尔药物浓度的测定方法。方法:以Acquity UPLC BEH C18(2.1 mm×50 mm,1.7μm)色谱柱,0.1%甲酸液(A液)为流动相-A,甲醇-水(95∶5)溶液(含0.1%甲酸,2 mmol/L甲酸铵)(B液)为流动相-B,梯度洗脱条件为0~0.2 min用20%B液、0.2~2.5 min用20%~60%B液、2.5~2.8 min用60%~20%B液、2.8~3 min用20%B液,流速为0.3 mL/min,柱温为40℃;采用甲醇沉淀蛋白法,从20μL血清样本中提取分析物及内标物(艾司洛尔);电喷雾电离(electrospray ionization,ESI)在多反应监测(multiple reaction monitoring,MRM)的正离子模式下运行,检测血清中美托洛尔的药物浓度。结果:美托洛尔标准曲线方程为A=0.173064ρ-0.182573,药物浓度在0.45~180 ng/mL范围内呈良好的线性关系(r=0.9996),定量下限为0.45 ng/mL,日内、日间精密度的相对标准偏差小于15%,提取回收率为88.36%~93.57%。结论:以LC-MS/MS法测定血清中美托洛尔的药物浓度具有较强的专属性,血清消耗量少,操作简单、准确、快捷,可用于临床日常美托洛尔的药物浓度监测。

Objective:To establish the LC-MS/MS method for the determination of metoprolol drug concentration in human serum.Methods:The Acquity UPLC BEH C18(2.1 mm×50 mm,1.7μm)column was used,taking0.1%formic acid solution(solution A)as mobile phase-A,methanol-water(95:5)solution(containing 0.1%formic acid,2 mmol/L ammonium formate)(solution B)as mobile phase-B;the gradient elution conditions were 20%solution B at0~0.2 min,20%~60%solution B at 0.2~2.5 min,60%~20%solution B at 2.5~2.8 min,20%solution B at 2.8~3 min,with a flow rate of 0.3 mL/min and column temperature of 40℃.The analyte and internal standard(esmolol)were extracted from 20μL serum samples by the method of protein precipitation with methanol;the electrospray ionization(ESI)was running in the positive ion mode of multiple reaction monitoring(MRM),to detect the drug concentration of metoprolol in serum.Results:The standard curve equation of metoprolol was A=0.173064ρ-0.182573.The drug concentration exhibited a good linear relationship in the range of 0.45~180 ng/mL(r=0.9996),the lower limit of quantification was 0.45 ng/mL,the RSDs of intra-day and inter-day precision were less than 15%and the extraction recovery rate ranged from 88.36%to 93.57%.Conclusion:The LC-MS/MS method for the determination of the drug concentration of metoprolol in serum has strong specificity,with low serum consumption.It is easy to operate,accurate and fast,and can be clinically used for the routine drug concentration monitoring of metoprolol.