HPLC-MS/MS法测定人血浆中孟鲁司特浓度及其在药动学中的应用

目的：建立HPLC-MS/MS法测定人血浆中的孟鲁司特浓度，并对健康人群空腹及餐后口服孟鲁司特钠颗粒后的药代动力学进行研究。方法：以孟鲁司特-d₆为内标，经乙腈沉淀蛋白后，采用高效液相色谱分离系统，Phenomenex luna C₁₈（50 mm×2.1 mm，5 μm）色谱柱，流动相为乙腈-水-甲酸（80：20：0.01）。采用质谱检测系统，电喷雾离子源（ESI源），正离子扫描，多反应监测模式（MRM）监测m/z 586.6→422.3（孟鲁司特）及m/z 592.0→427.2（孟鲁司特-d₆）。12名健康受试者单剂量空腹或餐后口服孟鲁司特钠颗粒0.5 g后采集血浆样品，以HPLC-MS/MS法测定血浆中孟鲁司特的浓度，计算药动学参数并进行统计分析。结果：孟鲁司特质量浓度在2.060~309.0 ng·mL^-1范围内与色谱响应呈现良好的线性关系，定量下限为2.060 ng·mL^-1。健康中国受试者单剂量空腹及餐后口服孟鲁司特钠颗粒0.5 g后主要药动学参数分别为C_max （173.8±43.0） ng·mL^-1、（150.5±19.8） ng·mL^-1，T_max （2.56±0.62） h、（5.38±1.49） h，AUC_0-t （1 305±345） ng·h·mL^-1、（1 674±329） ng·h·mL^-1，AUC_0~∞（1 357±368） ng·h·mL^-1、（1 761±356） ng·h·mL^-1。统计结果表明，除T_max外，其余参数在空腹及餐后之间的差异均不具有统计学意义。结论：本方法经完全方法学验证，适用于人体血浆中孟鲁司特的测定，并可用于临床药动学研究。药动学研究表明高脂餐不影响孟鲁司特钠颗粒在体内的吸收程度，但是明显延迟其吸收速度。

Objective: To establish a rapid and simple method for the determination of montelukast in human plasma,and study the pharmacokinetics of montelukast sodium granules in Chinese healthy volunteers.Methods: Montelukast-d₆ acted as the internal standard,and the separation of plasma was performed on a Phenomenex luna C₁₈(50 mm×2.1 mm,5 μm) column after precipitated by acetonitrile with a mobile phase comprising of acetonitrile-water-formic acid(80:20:0.01) at a flow rate of 0.5 mL·min^-1,and then detected on the MS system.Ions monitored in the multiple reaction monitoring(MRM) mode were m/z 586.6→422.3 for montelukast and m/z 592.0→427.2 for montelukast-d₆,respectively.The pharmacokinetic characteristics of montelukast sodium granules were investigated in 12 Chinese healthy volunteers after single oral administration of 0.5 g montelukast sodium granules under the condition of fasting or fed,plasma samples collected were detected by the validated HPLC-MS/MS method.Results: Peak area ratio of the drug to the internal standard was used for the quantification of montelukast in plasma samples,the linearity were found to be in the range of 2.060-309.0 ng·mL^-1 and the limit of quantification was 2.060 ng·mL^-1.The analyte was proved to be stable under all the required conditions.The total runtime of analyte was only 2 minutes.The pharmacokinetic features of montelukast in healthy Chinese people after an oral administration were as follows:C_max(173.8±43.0)ng·mL^-1(fasting),(150.5±19.8) ng·mL^-1(fed);T_max(2.56±0.62)h(fasting),(5.38±1.49)h(fed);AUC_0-t(1 305±345) ng·h·mL^-1(fasting),(1 674±329)ng·h·mL^-1(fed);AUC_0~∞(1 357±368)ng·h·mL^-1(fasting) and(1 761±356) ng·h·mL^-1(fed).The differences in all parameters between fasting and fed were not statistically significant except T_max.Conclusion: This is a rapid and simple method to determine montelukast in human plasma and is applicable to the pharmacokinetic study.The results of pharmacokinetic study indicated that high fat meal did not affect the AUC of montelukast sodium in Chinese healthy volunteers,but it did slow down the absorption rate of montelukast in Chinese people.