超高效液相色谱-质谱联用法测定人血浆中沙丁胺醇浓度

Determination of the concentration of salbutamol in human plasma by ultra performance liquid chromatography-tandem mass spectrometry

目的:建立测定人血浆中沙丁胺醇浓度的超高效液相色谱-质谱联用方法。方法:内标选用沙丁胺醇-d9,以甲醇沉淀血浆中蛋白,取上清液吹干复溶;采用Welch Ultimate XS-C18色谱柱(3.0 mm×100 mm,3μm),预柱选用Welch Ultimate XB-C18柱(2.1 mm×5 mm,5μm),以含0.1%甲酸5 mmol·L^-1乙酸铵的5%乙腈和纯乙腈为流动相,梯度洗脱;应用电喷雾离子化,正离子模式下进行多反应监测沙丁胺醇(m/z 240.2→m/z 147.9)和内标沙丁胺醇-d9(m/z 249.1→m/z 149.1)的浓度。结果:沙丁胺醇的线性范围为5~2000 pg·mL^-1,定量下限为5 pg·mL^-1,日内、日间准确度为101.93%~109.03%,精密度RSD为1.45%~8.50%。结论:本方法简便、准确、灵敏、稳健且经济,适用于硫酸沙丁胺醇吸入气雾剂的临床药动学研究和生物等效性评价。

OBJECTIVE To establish an ultrahigh performance liquid chromatography-tandem mass spectrometry method for the determination of salbutamol concentration in human plasma.METHODS The internal standard was albuterol-d9,and the protein in plasma was precipitated by methanol.The supernatant was dried and reconstituted.The Welch Ultimate XS-C18 column(3.0 mm×100 mm,3μm)was applied with the Welch Ultimate XB-C18 guard column(2.1 mm×5 mm,5μm)being as the guard column.Gradient elution was conducted with the mobile phase 5%acetonitrile containing 0.1%formic acid 5 mmol·L^-1 ammonium acetate and pure acetonitrile.Electrospray ionization and multi-reaction monitoring was applied to determine the salbutamol concentration in positive ion mode(m/z 240.2→m/z 147.9)and the concentration of the internal standard salbutamol-d9(m/z 249.1→m/z 149.1).RESULTS The linear concentration range of the calibration curve was from 5 to 2000 pg·mL^-1(r=0.9992),and the lower limit of quantification was 5 pg·mL^-1.The intra-day and inter-day accuracy was from 101.93%to 109.03%.The RSD of precision ranged from 1.45%to 8.50%.CONCLUSION This method is simple,accurate,sensitive,robust and economical which is suitable for clinical pharmacokinetic study and bioequivalence evaluation of salbutamol sulfate inhalation aerosol.