高灵敏度液相色谱-串联质谱法测定人血浆中普拉克索

目的: 建立灵敏、易操作的液相色谱(串联质谱法测定人血浆中普拉克索,并应用于临床药动学研究。 方法: 血浆样品经乙醚-二氯甲烷(3∶ 2,v/v)液-液萃取后,以乙腈-10 mmol·L^-1醋酸铵-甲酸(60∶ 40∶ 0.1,v/v/v)为流动相,Venusil ASB-C₁₈柱(150 mm×4.6 mm,5 μm)进行分离,采用电喷雾电离源,以选择反应监测(SRM)方式进行正离子检测,用于定量分析的离子反应分别为m/z 212→m/z(111 + 126 +153)(普拉克索)和m/z 243→m/z 210(石杉碱甲,内标)。 结果: 测定血浆中普拉克索的线性范围为5.92~740 pg·mL^-1,定量下限为5.92 pg·mL^-1,日内、日间精密度(RSD)均小于9.7%,准确度(RE)在-2.5%~0.6%之间。本法被成功应用于健康受试者单剂量口服0.125 mg和0.25 mg盐酸普拉克索片的药动学研究。 结论: 本方法同时选择3个主要碎片离子作为普拉克索定量产物离子,明显改进了分析方法的灵敏度,适用于人血浆样品中普拉克索的测定。

Objective: To develop and validate a sensitive and convenient liquid chromatographic-tandem mass spectrometric (LC-MS/MS) method for the determination of pramipexole in human plasma and to study the pharmacokinetics of pramipexole. Methods: The plasma sample was extracted with ethyl ether dichlormethane (3∶ 2,v/v),then separated on a Venusil ASB-C₁₈(150 mm×4.6 mm,5 μm)column using acetonitrile-10 mmol·L^-1 ammonium acetate-formic acid (60∶ 40∶ 0.1,v/v/v) as the mobile phase.A tandem mass spectrometer equipped with electrospray ionization source was used as the detector and operated in the positive ion mode.Quantitation was performed using selective reaction monitoring (SRM) of the transitions of m/z 212→m/z(111 + 126 +153)and m/z 243→m/z 210 for pramipexole and the internal standard huperzine A,respectively. Results: The linear calibration curves were obtained in the concentration range of 5.92-740 pg·mL^-1.The lower limit of quantification was 5.92 pg·mL^-1.The intra-day and inter-day relative standard deviation over the entire concentration range was less than 9.7%.The accuracy was in the range of -2.5% to 0.6% in terms of relative error.The method was successfully applied to a pharmacokinetic study of pramipexole dihydrochloride monohydrate tablets containing 0.125 mg or 0.25 mg in healthy volunteers. Conclusion: Three SRM transitions are selected in the quantification of pramipexole,which improve the sensitivity of the method significantly,and the method is proved to be suitable for the determination of pramipexole in human plasma.