HPLC-ESI/MS法测定续断"发汗"前后绿原酸和川续断皂苷Ⅵ的含量

目的: 建立续断"发汗"前后绿原酸和川续断皂苷Ⅵ的高效液相色谱-电喷雾-三重四极杆质谱联用测定方法。方法: 采用Agilent Zorbax SB C₁₈(3.5 mm×100 mm,3.5 μm)色谱柱,以水(A)-甲醇(B)为流动相,梯度洗脱(0～3 min,60%B→100%B;3～7 min,100%B;7～10 min,100%B→60%B)。绿原酸以m/z 353.0为母离子,m/z 191.0、127.0为子离子,川续断皂苷Ⅵ以m/z 927.5为母离子,m/z 603.3、323.3为子离子,采用多反应监测(MRM)负离子模式对续断"发汗"前后进行检测。结果: 5个不同批次的续断经"发汗"后,绿原酸含量均有所降低,分别降低了41.18%、20.54%、20.51%、54.27%、5.02%;川续断皂苷Ⅵ的含量均有所增加,分别增加了15.45%、1.18%、12.08%、9.22%、119.72%。结论: 产地加工—"发汗"影响续断中绿原酸和川续断皂苷Ⅵ的含量,实验结果为进一步研究"发汗"机理提供依据。

Objective: To establish an HPLC-electrospray ionisation tandem QQQ mass spectrometry method for simultaneous determination of chlorogenic acid and asperosaponin Ⅵ in crude and sweated Radix Dipsaci. Methods: The two components were analyzed simultaneously with a Zorbax C₁₈ column by gradient elution using water-methanol as the mobile phase (0-3 min, 60%B→100%B;3-7 min, 100%B;7-10 min, 100%B→60%B).The mass spectrometer was operated in the negative-ion mode using multiple reaction monitoring (MRM), and the precursor ion of chlorogenic acid at m/z 353.0 yielded two main daughter ions of m/z 191.0, 127.0, and the precursor ion of asperosaponin Ⅵ at m/z 927.5 yielded two main daughter ions of m/z 603.3, 323.3. Results: After being sweated, the content of chlorogenic acid in five batches of Radix Dipsaci decreased by 41.18%, 20.54%, 20.51%, 54.27%, 5.02%, respectively, and the content of asperosaponin Ⅵ increased by 15.45%, 1.18%, 12.08%, 9.22%, 119.72%, respectively. Conclusion: The method of origin processing "sweating" could change the contents of chlorogenic acid and asperosaponin Ⅵ in Radix Dipsaci.The experimental results provided the basis for further study on sweating mechanism.

1 ChP(中国药典).2010.Vol Ⅰ(一部):309
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