Wang Xin-Shuai, Sun Jia-Chun, Yang Rui-Na, Ren Jing, Liang Shuo, Sun Ming, Wang Ying-Fei, Gao She-Gan
Department of Oncology, Cancer Institute, The First Affiliated Hospital of Henan University of Science and Technology, Luoyang, Henan 471003, PR China.
Medical College of Henan University of Science and Technology, Luoyang, Henan 471003, PR China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2014 Jul 15;963:70-4. doi: 10.1016/j.jchromb.2014.05.045. Epub 2014 Jun 2.
A rapid, sensitive and selective ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) was developed and validated for the determination and pharmacokinetic investigation of cephalomannine in rat plasma. Sample preparation was accomplished through a simple one-step deproteinization procedure with 0.2 mL of perchloric acid-methanol (1:9, v/v) to a 0.1 mL plasma sample. Plasma samples were separated by UPLC on an Acquity UPLC BEH C18 column using a mobile phase consisting of acetonitrile-0.1% formic acid in water with gradient elution. The total run time was 2.0 min and the elution of cephalomannine was at 1.60 min. The detection was performed on a triple quadrupole tandem mass spectrometer in the multiple reaction-monitoring (MRM) mode using the respective transitions m/z 832.8→264.1 for cephalomannine and m/z 812.6→286.0 for 10-DAT (internal standard), respectively. The calibration curve was linear over the range of 10-2,000 ng/mL with a lower limit of quantitation (LLOQ) of 10 ng/mL. Mean recovery of cephalomannine in plasma was in the range of 80.9-85.3%. Intra-day and inter-day precision were both <11.2%. This method was successfully applied in pharmacokinetic study after intravenous administration of 5.0mg/kg cephalomannine in rats.
建立了一种快速、灵敏且选择性高的超高效液相色谱串联质谱法（UPLC-MS/MS），用于测定大鼠血浆中头孢曼宁并进行药代动力学研究。通过向0.1 mL血浆样品中加入0.2 mL高氯酸-甲醇（1:9，v/v）的简单一步去蛋白程序来完成样品制备。血浆样品在Acquity UPLC BEH C18柱上通过UPLC进行分离，流动相由乙腈-0.1%甲酸水溶液组成并进行梯度洗脱。总运行时间为2.0分钟，头孢曼宁在1.60分钟洗脱。在三重四极杆串联质谱仪上采用多反应监测（MRM）模式进行检测，头孢曼宁的相应跃迁为m/z 832.8→264.1，10-DAT（内标）的相应跃迁为m/z 812.6→286.0。校准曲线在10-2000 ng/mL范围内呈线性，定量下限（LLOQ）为10 ng/mL。头孢曼宁在血浆中的平均回收率在80.9-85.3%范围内。日内和日间精密度均<11.2%。该方法成功应用于大鼠静脉注射5.0mg/kg头孢曼宁后的药代动力学研究。
