Clinical Pharmacology Research Center, Peking Union Medical College Hospital and Chinese Academy of Medical Sciences, Beijing 100730, China.
J Pharm Biomed Anal. 2013 Mar 25;76:59-64. doi: 10.1016/j.jpba.2012.11.033. Epub 2012 Dec 24.
A robust and validated ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method has been developed for the simultaneous determination of buagafuran and one metabolite (M1) in human plasma. The two analytes were extracted from plasma samples using tert-butyl methyl ether after addition of the internal standard and chromatographed on an Acquity UPLC BEH C18 column (2.1mm×50mm, 1.7μm) thermostatted at 35°C with methanol-water (75:25, v/v) as the mobile phase at an isocratic flow rate of 0.4mL/min. The detection was performed on an API 5500 mass spectrometer coupled with electrospray ionization (ESI) source in positive mode. The multiple reactions monitoring (MRM) transitions of m/z 245.2→109.1 and m/z 279.1→243.1 were used to quantify buagafuran and M1, respectively. The assay was validated over the concentration range of 0.5-200ng/mL for the two analytes. Precision and accuracy are in accordance with the generally accepted criteria for bioanalytical methods. The extraction recovery and the matrix effect were investigated. This method was successfully applied to support a clinical study where multiple oral doses were administrated to healthy Chinese volunteers to investigate the pharmacokinetics, safety and tolerability of buagafuran.
已开发出一种稳健且经过验证的超高效液相色谱-串联质谱（UPLC-MS/MS）方法，用于同时测定人血浆中的 buagafuran 和一种代谢物（M1）。两种分析物均通过加入内标后，用叔丁基甲基醚从血浆样品中提取，并在 Acquity UPLC BEH C18 柱（2.1mm×50mm，1.7μm）上进行色谱分离，在 35°C 下使用甲醇-水（75:25，v/v）作为流动相以等度流速 0.4mL/min 进行洗脱。检测采用 API 5500 质谱仪，与电喷雾电离（ESI）源在正模式下联用。用于定量分析 buagafuran 和 M1 的多重反应监测（MRM）转换分别为 m/z 245.2→109.1 和 m/z 279.1→243.1。该测定法在两个分析物的浓度范围为 0.5-200ng/mL 内进行了验证。精密度和准确度符合生物分析方法的公认标准。考察了提取回收率和基质效应。该方法成功应用于支持一项临床研究，该研究对健康的中国志愿者进行了多次口服给药，以研究 buagafuran 的药代动力学、安全性和耐受性。
