亲水作用色谱-串联质谱法测定气雾剂给药后人血浆中的沙丁胺醇

Determination of salbutamol in human plasma after aerosol inhalation using hydrophilic interaction chromatography (HILIC)-tandem mass spectrometry

目的:建立灵敏、快速的亲水作用色谱-串联质谱法(HILIC-MS/MS)测定给予沙丁胺醇气雾剂后人血浆中的沙丁胺醇。方法:以d9-沙丁胺醇为内标,血浆样品经乙腈蛋白沉淀后,采用Luna HILIC(100 mm×3.0 mm,3μm)色谱柱分离。流动相为乙腈(含0.1%甲酸)-5 mmol.L-1醋酸铵溶液(氨水调节pH为8.5)(93∶7,v/v),等度洗脱,进样体积2.0μL,样品分析时间为3.5 min。采用电喷雾电离(ESI)源正离子模式、多反应监测(MRM),用于定量分析的离子反应分别为m/z 240→(148+166)(沙丁胺醇)和m/z 249→(149+167)(d9-沙丁胺醇)。结果:沙丁胺醇定量方法的线性范围为11.7～2340 pg.mL-1,定量下限达11.7 pg.mL-1,日内、日间精密度(RSD)均小于14.1%,准确度(RE)在-3.8%～-0.8%之间。本法成功应用于健康受试者给予硫酸沙丁胺醇气雾剂200μg后的药动学研究。结论:采用稳定同位素内标的HILIC-MS/MS法快速、灵敏和准确,适用于测定人血浆样品中的沙丁胺醇,可测定给药后24 h的血浆浓度。

Objective： To develop and validate a sensitive and rapid hydrophilic interaction chromatography-tan- dem mass spectrometric （HILIC-MS/MS） method for the determination of salbutamol in human plasma after aerosol inhalation. Methods： Samples were prepared from plasma by protein precipitation using acetonitrile as the precipi-tation solvent; salbutamol and d9-salbutamol were separated on a Luna HILIC column （100 mm×3.0 mm,3μm） using acetonitrile（ consisting of 0. 1% formic acid）-5 mmol·L^-1 ammonium acetate （pH 8.5 adjusted with am-monia water） （93：7 ,v/v）as a mobile phase with isocratic elution. The injection volume was 2. 0 μL,and a chroma-tographic total run time was 3.5 min. Mass spectrometry detection was conducted through electrospray ionization in positive ion multiple reaction monitoring modes, using the transitions of m/z 240 → （148 ＋ 166）and m/z 249 → （ 149 ＋ 167 ） for salhutamol and d9-salbutamol, respectively. Results： The method was linear over the concentration range of 11.7 - 2340 pg · mL^ - 1 and the lower limit of quantification was 11.7 pg · mL^-1. The intra - and inter - day precision in terms of the relative standard deviation were less than 14. 1%. The accuracy was in a range of - 3.8% to -0. 8% in terms of relative error. The method was successfully applied to pharmacokinetic evaluation study in healthy volunteers following inhalation of 200 μg salbutamol aerosol. Conclusion： The established HILIC - MS/ MS method is sensitive, rapid and accuracy, which is suitable for the determination of salbutamol in human plasma and can detect the plasma concentration of 24 h post-dose.