超高效液相色谱-四极杆静电场轨道阱高分辨质谱法测定米炒人参炮制前后皂苷成分及其裂解规律的研究

Determination of saponin components and their cleavage laws in rice-fried ginseng before and after concoction by ultra performance liquid chromatography-quadrupole-orbitrap-mass spectrometry

中文关键词:  米炒人参  超高效液相色谱-四极杆静电场轨道阱高分辨质谱  人参皂苷  模拟炮制；裂解规律

英文关键词:rice-fried ginseng  ultra performance liquid chromatography-quadrupole-orbitrap-mass spectrometry  ginsenoside  simulated processing  law of cleavage

中文摘要:

      目的 建立超高效液相色谱-四极杆静电场轨道阱高分辨质谱法(ultra performance liquid chromatography-quadrupole-orbitrap-mass spectrometry, UPLC-Q-Orbitrap-MS)检测米炒人参炮制前后皂苷成分的方法, 并研究其裂解规律。方法 采用Supelco C18色谱柱, 以乙腈-0.1%甲酸水溶液梯度洗脱, 应用电喷雾离子源, 负离子全扫描模式采集一、二级质谱数据, 扫描范围为150~2000 m/z。结合质谱数据库及相关文献信息, 运用X Calibur2.2软件对米炒人参中皂苷类成分进行鉴定。以6种人参皂苷Re、Rg1、Rb1、Rc、Rb2、Rb3进行模拟炮制, 确定皂苷类成分裂解产物, 明确皂苷成分的裂解规律。结果 从人参中检测出14个成分, 鉴定出13种人参皂苷成分; 米炒人参中检测出23个成分, 鉴定出20种人参皂苷成分。通过比较人参米炒前后的皂苷类成分, 发现米炒人参中存在人参中未检测到的8种稀有人参皂苷20(S)-Rg2、20(S)-Rh1、20(R)-Rh1、F2、20(S)-Rg3、20(R)-Rg3、20(S)-Rs3、20(R)-Rs3。模拟炮制结果表明, 人参皂苷Re脱去C-20糖基, 转化为稀有人参皂苷20(S)-Rg2; 人参皂苷Rg1脱去C-20位糖基, 转化为稀有人参皂苷20(S)-Rh1、20(R)-Rh1; 人参皂苷Rb1、Rb2、Rb3、Rc脱去C-20或C-3位糖基, 转化为稀有人参皂苷20(S)-Rg3、20(R)-Rg3或F2。结论 人参经米炒后, 稀有人参皂苷成分增加, 产生的稀有皂苷为原型皂苷发生苷键裂解而获得, 模拟炮制可作为其裂解规律研究的有效方法。

英文摘要:

      Objective To establish a method for determination of saponin components in rice-fried ginseng before and after concoction by ultra performance liquid chromatography-quadrupole-orbitrap-mass spectrometry, and study their cleavage laws. Methods A Supelco C18 column was used with gradient elution of acetonitrile-0.1% formic acid aqueous solution, and an electrospray ionization was applied to collect the primary and secondary mass spectrometry data in negative-ion full-scan mode, with a scanning range of 150–2000 m/z. Combined with the information from the mass spectrometry database and the related literatures, the identification of the saponin constituents in rice fried ginseng was carried out by using the X Calibur 2.2 software was used to identify the saponin components in ginseng fried in rice. Six ginsenosides Re, Rg1, Rb1, Rc, Rb2 and Rb3 were used to simulate processing, to determine the cleavage products of the saponin-like components and to clarify the cleavage laws of the saponin components. Results Fourteen components were detected and 13 kinds of ginsenoside components were identified from ginseng; 23 kinds of components were detected and 20 kinds of ginsenoside components were identified from rice-fried ginseng. By comparing the ginsenoside components of ginseng before and after rice-frying, it was found that 8 kinds of rare ginsenosides 20(S)-Rg2, 20(S)-Rh1, 20(R)-Rh1, F2, 20(S)-Rg3, 20(R)-Rg3, 20(S)-Rs3 and 20(R)-Rs3, which were not detected in ginseng, were found in the rice-fried ginseng. The results of simulated processing indicated that the ginsenosides Re was converted to the rare ginsenoside 20(S)-Rg2 by removing the C-20 sugar group; the ginsenoside Rg1 was converted to the rare ginsenosides 20(S)-Rh1, 20(R)-Rh1 by removing the sugar group at the C-20 position; and the ginsenosides Rb1, Rb2, Rb3, Rc were converted to the rare ginsenosides 20(S)-Rg3, 20(R) -Rg3 or F2 by removing the sugar group at the C-20 or C-3 position. Conclusion Rare ginsenoside components increased after ginseng is fried in rice, and the resulting rare saponins are obtained by glycosidic bond cleavage of the prototypical saponins, and simulated processing can be an effective method for the study of its cleavage pattern.