产地加工“发汗”对续断中异绿原酸A、异绿原酸B及异绿原酸C含量的影响

Simultaneous determination of isochlorogenic acid A,sochlorogenic acid B,and sochlorogenic acid C in crude and sweated Radix Dipsaci by HPLC

目的:研究产地加工"发汗"对续断中异绿原酸A、异绿原酸B和异绿原酸C含量的影响。方法:建立同时测定异绿原酸A、异绿原酸B和异绿原酸C含量的HPLC方法,然后比较续断"发汗"前后3个成分含量的变化。采用高效液相色谱法,使用Agilent Eclipse XDB-C_(18)(250 mm×4.6 mm,5.0μm)色谱柱,以乙腈(A)-0.05%磷酸水溶液(B)为流动相,梯度洗脱(0～5 min,2%A→6%A;5～18 min,6%A→10%A;18～40 min,10%A→20%A;40～70 min,20%A→25%A;70～80 min,25%A→35%A;80～90min,35%A→60%A;90～110 min,60%A→70%A;110～120 min,70%A),流速1 m L·min^(-1),检测波长为212 nm,柱温30℃。结果:异绿原酸B、异绿原酸A和异绿原酸C的峰面积与质量浓度的线性关系良好(r≥0.999 8),平均加样回收率(n=6)分别为101.5%、101.7%和102.0%;不同产地的续断未发汗品中异绿原酸B、异绿原酸A和异绿原酸C成分含量分别为0.06%～0.13%、0.45%～0.92%和0.18%～0.73%,发汗品中分别为0.09%～0.76%、0.29%～0.88%和0.24%～0.80%。经产地加工"发汗"后,续断中异绿原酸A的含量略有降低,而异绿原酸B和异绿原酸C的含量有所升高。结论:产地加工"发汗"对续断中的异绿原酸A、异绿原酸B和异绿原酸C 3个酚酸类成分有明显影响,为进一步探讨续断"发汗"机理和制定"发汗"加工工艺提供了依据。

Objective：To study the effects of primary processing＂sweating＂on the contents of isochlorogenic acid A,isochlorogenic acid B,and isochlorogenic acid C in crude and sweated Radix Dipsaci.Methods： The simultaneous determination method of isochlorogenic acid A,isochlorogenic acid B and isochlorogenic acid C in crude and sweated Radix Dipsaci were developed by using high performance liquid chromatography（HPLC）,and then the determination results before and after sweating process were compared and analyzed.The sample solution was determined by HPLC with an Agilent Eclipse XDB- C18 column（250 mm×4.6 mm,5 μm）.The acetonitrile（A）-0.05% phosphoric acid water solution（B） was applied as the mobile phase by gradient elution（0～5 min,2%A → 6%A;5～18 min,6%A → 10%A;18～40 min,10%A → 20%A;40～70 min,20%A → 25%A;70～80 min,25%A → 35%A;80～90 min,35%A → 60%A;90～110 min,60%A → 70%A;110～120 min,70%A）.The analyses were performed at 30℃ with a flow rate of 1 m L·min（-1）,and the UV detection was performed at 212 nm.Results： Isochlorogenic acid B,isochlorogenic acid A and isochlorogenic acid C showed good linearity（r ≥ 0.999 8） in the range of the tested concentration,while the average recoveries（n=6） of isochlorogenic acid B,isochlorogenic acid A and isochlorogenic acid C were 101.5%,101.7% and 102.0%,respectively.The contents of the three compositions in crude Radix Dipsaci from different places of production were 0.06%-0.13%,0.45%-0.92% and 0.18%-0.73%,respectively;and the ones in sweated samples were 0.09%-0.76%,0.29%-0.88% and 0.24%-0.80%,respectively.Through primary processing＂sweating＂,both of the contents of isochlorogenic acid B and isochlorogenic acid C were increased,and isochlorogenic acid A was decreased.Conclusion： The results demonstrated that primary processing＂sweating＂indeed had an effect on the contents of isochlorogenic acid A,isochlorogenic acid B and isochlorogenic acid C,which provided proof for the further study of the＂sweating＂mechanism and process of Radix Dipsaci.