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

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,3 μm)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2.0 μL,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200 μg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-tandem 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 precipitation solvent; salbutamol and d₉-salbutamol were separated on a Luna HILIC column (100 mm×3.0 mm,3 μm)using acetonitrile(consisting of 0.1% formic acid)-5 mmol·L^-1 ammonium acetate (pH 8.5 adjusted with ammonia water)(93∶7,v/v)as a mobile phase with isocratic elution.The injection volume was 2.0 μL,and a chromatographic 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 salbutamol 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.