HPLC-MS/MS同时测定人血浆中对乙酰氨基酚和咖啡因含量的方法学研究

目的建立同时测定人血浆中对乙酰氨基酚和咖啡因浓度的HPLC-MS/MS法。方法以茶碱为内标,血浆样品用甲醇沉淀蛋白后直接进样。用Waters symmetry C18（150 mm×4.6 mm,5μm）为分析柱,甲醇-0.2%醋酸=35∶65（v/v）为流动相,流速为0.9 mL·min^-1,采用柱后分流,0.2 mL·min^-1进入质谱,柱温35℃。选择监测的离子为m/z152.1→109.9（对乙酰氨基酚）、m/z195.3→138.1（咖啡因）和m/z181.1→123.9（茶碱）。结果血浆中对乙酰氨基酚和咖啡因的线性范围分别为0.02-4.04μg·mL^-1,5.05~1 010 ng·mL^-1;日内日间精密度RSD均〈7.94%。结论本方法专属性强,灵敏度高,操作简便、快速,符合生物样品分析要求,适用于临床药动学研究。

Determination of paracetamol and caffeine in human plasma by HPLC-MS/MS

Objective To establish an HPLC-MS/MS method for the determination of paracetamol and caffeine in human plasma.Methods Theophylline was used as the internal standard.The plasma sample was injected directly for determination after being precipitated by methanol.A C18 analytical column（150 mm×4.6 mm,5 μm） was used and the mobile phase was a mixture of methanol-2% acetic acid（35∶65）,the flow rate was 0.9 mL·min^-1 and after the separation,the mobile phase was splitted and 0.2 mL·min^-1 was into the MS.The column temperature was 35℃.HPLC-MS/MS mode had target ions at m/z 152.1→109.9 for paracetamol,m/z 195.3→138.1 for caffeine and m/z 181.1→123.9 for theophylline.Results The linear range of paracetamol and caffeine in the plasma were 0.02-4.04 μg·mL^-1 and 5.05-1010 ng·mL^-1,respectively.The RSDs of intraday and interday were no more than 7.94%.Conclusion This specific,sensitive and reliable HPLC-MS/MS method offers an alternative to monitor and evaluate the pharmacokinetics of paracetamol and caffeine.