Ji Pei-ying, Li Zhi-wen, Yang Qing, Wu Rong
Department of Pharmacy, Yangpu Hospital, Tongji University School of Medicine, Shanghai 200090, China.
Department of Pharmacy, Changzheng Hospital, Second Military Medical University, Shanghai 200003, China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2015 Mar 15;985:71-4. doi: 10.1016/j.jchromb.2015.01.022. Epub 2015 Jan 23.
A sensitive and rapid ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method was developed to determine ruscogenin in rat plasma using midazolam as the internal standard (IS). Sample preparation was accomplished through a liquid-liquid extraction procedure with ethyl acetate to 0.2mL plasma sample. The analyte and IS were separated on an Acquity UPLC BEH C18 column (2.1mm×50mm, 1.7μm) with the mobile phase of acetonitrile and 1% formic acid in water with gradient elution at a flow rate of 0.40mL/min. Ruscogenin and IS were eluted at 1.74 and 1.11min, respectively. The detection was performed on a triple quadrupole tandem mass spectrometer equipped with positive-ion electrospray ionization (ESI) by multiple reactions monitoring (MRM) of the transitions at m/z 431.2→287.0 for ruscogenin and m/z 326.2→291.1 for IS. The linearity of this method was found to be within the concentration range of 2-1000ng/mL with a lower limit of quantification of 2ng/mL. Only 2.0min was needed for an analytical run. The matrix effect was 92.4-107.3% for ruscogenin. The intra- and inter-day precision (RSD%) were less than 11.2% and accuracy (RE%) was within ±9.8%. The recovery ranged from 75.4% to 86.3%. Ruscogenin was sufficiently stable under all relevant analytical conditions. The method was also successfully applied to the pharmacokinetic study of ruscogenin in rats.
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，以咪达唑仑为内标（IS）测定大鼠血浆中的鲁斯可皂苷元。通过用乙酸乙酯对0.2mL血浆样品进行液-液萃取程序来完成样品制备。分析物和内标在Acquity UPLC BEH C18柱（2.1mm×50mm，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0.40mL/min。鲁斯可皂苷元和内标分别在1.74和1.11分钟洗脱。检测在配备正离子电喷雾电离（ESI）的三重四极杆串联质谱仪上进行，通过多反应监测（MRM）对鲁斯可皂苷元的m/z 431.2→287.0和内标的m/z 326.2→291.1的跃迁进行监测。该方法的线性范围为2-1000ng/mL，定量下限为2ng/mL。一次分析运行仅需2.0分钟。鲁斯可皂苷元的基质效应为92.4-107.3%。日内和日间精密度（RSD%）小于11.2%，准确度（RE%）在±9.8%以内。回收率在75.4%至86.3%之间。鲁斯可皂苷元在所有相关分析条件下均足够稳定。该方法还成功应用于鲁斯可皂苷元在大鼠体内的药代动力学研究。
