UPLC Orbitrap HRMS法分析西洋参蒸参弃液浓缩物中皂苷类成分

采用超高效液相色谱-四极杆静电场轨道阱高分辨质谱(UPLC Orbitrap HRMS)技术测定西洋参蒸参弃液浓缩物中15种人参皂苷单体成分含量。采用Thermo Scientific Syncronis C18色谱柱(100mm×2.1mm×1.7μm),以乙腈-0.1%甲酸水溶液为流动相,使用电喷雾电离源(ESI),四极杆静电场轨道阱质量分析器,高分辨质谱仪采集数据。结果表明,人参皂苷Rg1、Rg2、Rg3、Rb1、Rb2、Rc、Rd、Re、Rh1、Rh2、Rk1、Ro、F1、F2和伪人参皂苷F11在线性范围内均存在良好的线性关系,48h内稳定性实验RSD值均小于5%,平均加样回收率为94.38%~102.10%,RSD值不大于3%。西洋鲜参在不同蒸制温度和蒸制时间下得到9组蒸参弃液,通过分析其浓缩物中15种人参皂苷单体化合物的含量,并比较各成分含量的变化情况,总结了4种类型人参皂苷的化学转化规律。该方法简便、准确、灵敏度高、专属性强、重复性好,适用于西洋参蒸参弃液浓缩物中皂苷成分含量的测定,可为开发蒸参弃液提供有效的检测手段。

Analysis of Ginsenosides in Steamed Panax quinquefoliums L. Liquid Waste Concentrates by UPLC Orbitrap HRMS

A method was applied to determine the contents of 15 ginsenosides in steamed Panax quinquefoliums L. liquid waste concentrates by ultra-performance liquid chromatography-Orbitrap high resolution mass spectrometry (UPLC Orbitrap HRMS). The sample was separated on Thermo Scientific Syncronis C18 column (100 mm×2.1 mm×1.7 μm) with the gradient elution of acetonitril-0.1% formic acid as mobile phases, and the flow rate was 0.2 mL/min. The column temperature was set at 35 ℃. The electrospray ionization (ESI) and the Orbitrap mass analyzer were used with high resolution mass data collection. Ginsenosides Rg₁、Rg₂、Rg₃、Rb₁、Rb₂、Rc、Rd、Re、Rh₁、Rh₂、Rk₁、Ro、F₁、F₂ and pseudo ginsenoside F₁₁ show good linear relationship, respectively. The average RSD value of stability test is less than 5% within 48 h. The average sample recovery was in the range of 94.38%-102.10%, and the RSD value is less than 3%. The contents of 15 ginsenosides in steamed Panax quinquefoliums L. liquid waste concentrates under different temperatures and times were determined and compared. For the protopanaxdiol (PPD) type ginsenosides, with the rising of steaming temperature and time, the contents of Rb₁, Rb₂, Rc, Rd and F₂ decreased gradually. While the contents of Rg₃, Rh₂, Rk₁ and Rd increased firstly and then decreased or decreased firstly and then increased. The ginsenoside transformation could explain the content more or less. The protopanaxtriol (PPT), oleanane and ocotillol types ginsenosides presented the similar conversion pathway. The chemical conversion of four types of ginsenosides were summarized. This method is simple, accurate, sensitive, specific and has good repeatability, which could be used to determine the contents of ginsenosides in steamed Panax quinquefoliums L. liquid waste and proved to be effective for detection of the steamed Panax quinquefoliums L. liquid waste concentrates. The steamed Panax quinquefoliums L. liquid waste concentrates includes a variety of ginsenosides, of which the minor ginsenosides have higher bioactivities. So the steamed Panax quinquefoliums L. liquid waste is of great value to develop. It also provides raw materials for medicine and health products.