Wang Zhe, Yang Hui, Sun Wei, Huang Cheng-ke, Cui Xiao, Qiu Xiang-jun, Lian Qing-quan, Wang Zeng-shou
Department of Pharmacy, The Second Affiliated Hospital & Yuying Children's Hospital of Wenzhou Medical University, Wenzhou 325027, China.
Medical College of Henan University of Science and Technology, Luoyang 471003, China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2014 Sep 15;967:235-9. doi: 10.1016/j.jchromb.2014.07.033. Epub 2014 Aug 4.
A sensitive and rapid ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was developed to determine thiamphenicol (TAP) in human plasma using chlorzoxazone as the internal standard (IS). Sample preparation was accomplished through a liquid-liquid extraction procedure with ethyl acetate to precipitation of plasma protein, and to a 0.1 mL plasma sample. The analyte and IS were separated on an Acquity UPLC BEH C18 column (2.1 mm × 50 mm, 1.7 μm) with the mobile phase of acetonitrile and 1% formic acid in water with gradient elution at a flow rate of 0.40 mL/min. The detection was performed on a triple quadrupole tandem mass spectrometer equipped with electrospray ionization (ESI) by multiple reactions monitoring (MRM) of the transitions at m/z 354.3→185.1 for TAP and m/z 168.1→132.1 for IS. The linearity of this method was found to be within the concentration range of 10-8000 ng/mL with a lower limit of quantification of 10 ng/mL. Only 1.5 min was needed for an analytical run. The method herein described was superior to previous methods and was successfully applied to the pharmacokinetic study of TAP in healthy Chinese volunteers after oral administration.
建立了一种灵敏、快速的超高效液相色谱 - 串联质谱（UPLC-MS/MS）法，以氯唑沙宗为内标（IS）测定人血浆中的甲砜霉素（TAP）。样品制备通过液 - 液萃取程序完成，用乙酸乙酯沉淀血浆蛋白，针对0.1 mL血浆样品进行。分析物和内标在Acquity UPLC BEH C18柱（2.1 mm×50 mm，1.7μm）上分离，流动相为乙腈和含1%甲酸的水溶液，采用梯度洗脱，流速为0.40 mL/min。在配备电喷雾电离（ESI）的三重四极杆串联质谱仪上进行检测，通过多反应监测（MRM）模式监测TAP的m/z 354.3→185.1和内标的m/z 168.1→132.1的跃迁。该方法的线性范围为10 - 8000 ng/mL，定量下限为10 ng/mL。一次分析运行仅需1.5分钟。本文所述方法优于以往方法，并成功应用于健康中国志愿者口服给药后TAP的药代动力学研究。
