液相色谱-质谱联用法快速测定人血浆中他达拉非浓度

Rapid determination of tadalafil concentration in human plasma by liquid chromatography-tandem mass spectrometry

目的采用液相色谱-质谱联用法(LC-MS/MS)测定人血浆中他达拉非的药物浓度。方法以地西泮为内标,血浆样品经甲醇沉淀蛋白后,以甲醇∶0.2%甲酸水溶液=33∶67(v/v)为流动相,Agilent公司ZORBAX Eclipse plus®C_(8)(4.6 mm×100 mm,3.5μm)色谱柱分离,流速为700μl/min,每针7.00 min。样品经电喷雾离子源正离子化后,通过三重四极杆串联质谱仪,在多反应监测模式下测定他达拉非(m/z 390.2→268.2)和内标地西泮(m/z 285.5→154.0)的浓度。结果他达拉非的血浆浓度在0.50~900.00 ng/ml范围内线性良好,定量下限为0.50 ng/ml,批内、批间精密度(RSD)均在4.5%~11.1%以内,相对偏差(RE)在±15%范围内。他达拉非血浆样品室温放置24 h,反复冻融(-20℃)3次及冰冻(-20℃)保存90 d的情况下均稳定。结论该分析方法简便、特异性高、灵敏度高,可用于人血浆中他达拉非的浓度检测。

Objective To determine the tadalafil concentration in human plasma by liquid chromatography-tandem mass spectrometry(LC-MS/MS).Methods With diazepam as internal standard,the plasma samples were precipitated with methanol,and the mobile phase was methanol∶0.2%formic acid aqueous solution=33∶67(v/v).The chromatographic column separation was ZORBAX Eclipse Plus®C_(8)(4.6 mm×100 mm,3.5μm)from Agilent Company,the flow rate was 700μL/min,and each needle was 7.00 min.The samples were positively ionized by electrospray ionization source,and then the concentrations of tadalafil(m/z 390.2→268.2)and internal standard diazepam(m/z 285.5→154.0)were determined by triple quadrupole mass spectrometer in multi-reaction monitoring mode.Results Tadalafil plasma concentration demonstrated good linearity in the range of 0.50-900.00 ng/ml,and the lower limit of quantification was 0.50 ng/ml.The intra-batch and inter-batch relative standard deviations(RSD)were all within 4.5%-11.1%,and the relative error(RE)was within±15%.The tadalafil plasma samples were stable after being placed for 24 hours at room temperature,repeated freezing and thawing(-20℃)for 3 times and frozen(-20℃)for 90 days.Conclusion The method is simple,specific and sensitive,and can be used for the determination of tadalaf il in human plasma.