Institute of Forensic Medicine and Toxicology, 1st Faculty of Medicine, Charles University in Prague, 12108 Prague 2, Czech Republic.
J Chromatogr B Analyt Technol Biomed Life Sci. 2010 Nov 15;878(30):3167-73. doi: 10.1016/j.jchromb.2010.09.032. Epub 2010 Oct 23.
A rapid and sensitive method using liquid chromatography-tandem mass spectrometry (LC-MS/MS) for simultaneous determination of doxazosin and verapamil in human serum has been developed. Trimipramine-d₃ as an isotopic labelled internal standard was used for quantification. Serum samples were prepared by simple liquid-liquid extraction with mixture of tert butyl methyl ether and ethyl acetate (1:1, v:v). The analytes and internal standard were separated on C18 column using an isocratic elution with 5 mM ammonium formate with 0.02% formic acid and 0.02% formic acid in acetonitrile (55:45, v:v) at a flow rate of 1.1 mL/min. Positive TurboIonSpray mass spectrometry was used with multiple reaction monitoring of the transitions at: m/z 455.3→165.2 and 150.2 for verapamil, m/z 452.2→344.4 and 247.4 for doxazosin, m/z 298.2→103.1 for trimipramine-d₃. Linearity was achieved between 1 and 500 ng/mL (R² ≥ 0.997) for both analytes. An extensive pre-study method validation was carried out in accordance with FDA guidelines. This assay was successfully applied to determine the serum concentrations of doxazosin and verapamil in suspect non-compliance patients.
一种使用液相色谱-串联质谱（LC-MS/MS）同时测定人血清中多沙唑嗪和维拉帕米的快速灵敏方法已经建立。三咪哒啉-d₃作为同位素内标用于定量。血清样品通过正丁基甲基醚和乙酸乙酯（1:1，v:v）混合物的简单液液萃取进行制备。分析物和内标在 C18 柱上使用 5 mM 甲酸铵与乙腈中的 0.02%甲酸和 0.02%甲酸（55:45，v:v）的等度洗脱进行分离，流速为 1.1 mL/min。采用正 TurboIonSpray 质谱，在以下转换处进行多重反应监测：m/z 455.3→165.2 和 150.2 用于维拉帕米，m/z 452.2→344.4 和 247.4 用于多沙唑嗪，m/z 298.2→103.1 用于三咪哒啉-d₃。对于两种分析物，线性范围为 1 至 500 ng/mL（R²≥0.997）。根据 FDA 指南进行了广泛的预研究方法验证。该测定法成功应用于确定可疑不依从患者的多沙唑嗪和维拉帕米的血清浓度。
