School of Pharmacy, Sungkyunkwan University, Suwon 440-746, Republic of Korea.
J Chromatogr B Analyt Technol Biomed Life Sci. 2012 Dec 12;911:59-63. doi: 10.1016/j.jchromb.2012.10.027. Epub 2012 Oct 29.
We have developed and validated a simple, rapid, and sensitive liquid chromatography analytical method employing tandem mass spectrometry (LC-MS/MS) for the determination of tolperisone, a centrally acting muscle relaxant, in human plasma. After liquid-liquid extraction with methyl t-butyl ether, chromatographic separation of tolperisone was performed using a reversed-phase Luna C(18) column (2.0mm×50mm, 5μm particles) with a mobile phase of 10mM ammonium formate buffer (pH 3.5) - methanol (12:88, v/v) and quantified by tandem mass detection in ESI positive ion mode. The flow rate of the mobile phase was 250μL/min and the retention times of tolperisone and the internal standard (IS, dibucaine) were both 0.6min. The calibration curves were linear over a range of 0.5-300ng/mL (r>0.999). The lower limit of quantification, using 200μL human plasma, was 0.5ng/mL. The mean accuracy and precision for intra- and inter-day validation of tolperisone were within acceptable limits. The LC-MS/MS method reported here showed improved sensitivity for quantification of tolperisone in human plasma compared with previously described analytical methods. Lastly, the validated method was successfully applied to a pharmacokinetic study in humans.
我们开发并验证了一种简单、快速和灵敏的液相色谱分析方法，采用串联质谱法（LC-MS/MS）测定人血浆中的托哌松，一种中枢作用的肌肉松弛剂。经过甲基叔丁基醚液-液萃取后，采用 Luna C(18)反相柱（2.0mm×50mm，5μm 颗粒）进行色谱分离，流动相为 10mM 甲酸铵缓冲液（pH 3.5）-甲醇（12:88，v/v），并在 ESI 正离子模式下进行串联质谱检测定量。流动相的流速为 250μL/min，托哌松和内标（IS，丁卡因）的保留时间均为 0.6min。校准曲线在 0.5-300ng/mL 范围内呈线性（r>0.999）。使用 200μL 人血浆，定量下限为 0.5ng/mL。托哌松的日内和日间验证的平均准确度和精密度均在可接受范围内。与以前描述的分析方法相比，本报告的 LC-MS/MS 方法显示出提高的检测灵敏度。最后，该验证方法成功应用于人体药代动力学研究。
