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

Determination of chlorogenic acid and asperosaponin Ⅵ in crude and sweated Radix Dipsaci by HPLC-ESI/MS

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,3.5μm)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. Meth- ods： 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 rain, 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 pre- cursor 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 con- tents of chlorogenic acid and asperosaponin VI in Radix Dipsaci. The experimental results provided the basis for fur- ther study on sweating mechanism.