HPLC-MS法测定人血浆中他林洛尔的浓度及其在中国健康志愿者体内的药动学研究

HPLC-MS determination of talinolol in human plasma and application to pharmacokinetic studies on Chinese healthy volunteers

目的:建立一种快速、特异灵敏的测定人血浆中他林洛尔浓度的高效液相色谱-质谱法(HPLC-MS),为研究他林洛尔在中国健康志愿者体内的药物动力学研究提供手段。方法:以普萘洛尔作为内标(I.S.),血浆样品碱化后用叔丁基甲醚萃取,用 Phenomenex C_(18)反相色谱柱(4.6 mm×250 mm,5 μm,USA)进行分离,以乙腈-缓冲液(10 mmol·L^(-1)醋酸铵和0.1%甲酸)(40:60,v/v)为流动相,柱温40℃,流速0.85 mL·min^(-1),HPLC—MS/ESI^+法选择性监测准分子离子峰(他林洛尔:m/z=364.3,I.S.:m/z=260.3)。结果:他林洛尔及内标普萘洛尔在5.5 min 内完全分离;他林洛尔在1.0～400.0 ng·mL^(-1)时线性关系良好,相关系数为0.9996;萃取回收率大于82%;方法回收率大于98%;最低检测浓度为0.3 ng·mL^(-1);日内日间 RSD均小于8%。主要药动学参数如下:C_(max)(147.8±63.8)ng·mL^(-1);t_(max)(2.0±0.7)h;t_(1/2)(12.0±2.6)h。结论:本方法简便快速,灵敏准确,适用于他林洛尔药物动力学的研究。

Objective： A rapid and sensitive LC - MS method for determination and screening in human plasma of talinolol is described to assess the pharmaeokineties of talinolol tablets. Methods： The analytes in plasma were extracted by liquid - liquid extraction using methyl t - butyl ether as the solvent after samples had been alkalinized with sodium hydroxide and spiked with I. S.. After removing and drying the upper organic phase, the extracts were reconstituted with a fixed volume of water （ ammonium acetate： 10 mmol·L^-1, formic acid 0. 1% ） - aeetonitrile （60： 40, v/v）. The extracts were analyzed by a high performance liquid chromatography coupled to eleetrospray ionization mass spectrometry （ HPLC - MS/ESI）, with positive ion SIM detection of talinolol （ [M ＋ H ]^＋ , m/z = 364.3 ） using propranolol（ [M ＋ H ]^＋ , m/z = 260. 3 ） as internal standard. The HPLC separation of the analytes was performed on a Phenomenex Cl8 （250 mm × 4. 6 mm,5 μm, USA） column, with a flow rate of 0. 85 mL min^-1 Results：The complete elution was obtained in less than 5.5 min. The calibration curve was linear in 1.0 -400. 0 ng· mL^-1 for talinolol, with a coefficient of determination above 0. 9996. The average extraction recovery was above 82%. The methodology recovery was above 98%. The limit of detection（ LOD）was 0.3 ng· mL^-1 for talinolol. The intra - day and inter - day relative standard deviations were less than 8%. A single 50 mg dose of talinolol tablet was administered to 12 healthy Chinese volunteers, the main pharmaeokinetie data are as follows ：Cmax （147. 8 ± 63.8） ng· mL^-1 ; tmax （ 2.0 ± 0. 7 ） h ; t1/2 （ 12. 0 ± 2.6 ） h. Conclusion： The method is accurate, sensitive and simple for the pharmaeokinetie study of talinolol.