Department of Pharmacy, Eastern Hepatobiliary Surgery Hospital, Second Military Medical University, Shanghai 200438, China; Shan dong Institute of Pharmaceutical Industry, Ji'nan 250101, China.
Steroids. 2013 Dec 11;78(12-13):1164-70. doi: 10.1016/j.steroids.2013.08.009. Epub 2013 Sep 5.
A rapid, simple and sensitive high-performance liquid chromatography tandem mass spectrometry method was developed and validated for simultaneous determination of six main steroidal saponins in Paris polyphylla in rat plasma. Ginsenoside Rg3 was selected as the internal standard (IS). Plasma samples were pretreated with protein precipitation, and the separation was achieved on a reverse phase Agilent poroshell120 EC-C18 column using a gradient mobile phase system of acetonitrile-water containing 0.1% formic acid. The triple quadruple mass spectrometer was set in negative electrospray ionization mode and multiple reaction monitoring (MRM) was used for six steroidal saponins quantification. The precursors to produce ion transitions monitored for polyphyllin I, polyphyllin II, polyphyllin VI, polyphyllin VII, dioscin, gracillin and IS were m/z 899.5>853.4, 1059.5>1013.5, 783.4>737.4, 1075.5>1029.5, 913.5>867.4, 929.5>883.4 and 819.5>783.4, respectively. The intra- and inter-day precisions (RSD%) were less than 13% and the average extraction recoveries ranged from 85% to 97.0% for each analyte. Six steroidal saponins were proved to be stable during sample storage, preparation and analytical procedures. The established method was employed for simultaneous quantification and successfully used for the first time for the pharmacokinetics evaluation of the six main compounds after intragastric administration of P. polyphylla extract in Sprague-Dawley rats.
建立并验证了一种快速、简单、灵敏的高效液相色谱-串联质谱法，用于同时测定大鼠血浆中 6 种主要甾体皂苷的含量。以人参皂苷 Rg3 为内标（IS）。血浆样品经蛋白沉淀预处理，采用含 0.1%甲酸的乙腈-水梯度洗脱系统，在 Agilent poroshell120 EC-C18 反相柱上进行分离。三重四极杆质谱仪在负离子电喷雾电离模式下，采用多反应监测（MRM）模式对 6 种甾体皂苷进行定量分析。监测到的 Polyphyllin I、Polyphyllin II、Polyphyllin VI、Polyphyllin VII、Dioscin、Gracillin 和 IS 的前体离子跃迁分别为 m/z 899.5>853.4、1059.5>1013.5、783.4>737.4、1075.5>1029.5、913.5>867.4、929.5>883.4 和 819.5>783.4。日内和日间精密度（RSD%）均小于 13%，平均提取回收率在 85%至 97.0%之间。6 种甾体皂苷在样品储存、制备和分析过程中均稳定。该方法用于同时定量分析，并首次成功应用于大鼠灌胃昆明山海棠提取物后 6 种主要成分的药代动力学评价。
