RP-HPLC-DAD同时测定大黄中9种有效成分的含量

RP-HPLC-DAD Method for Simultaneous Determination of Nine Active Constituents in Rhubarb

目的：建立RP-HPLC-DAD同时测定大黄中5种游离蒽醌类成分（芦荟大黄素、大黄酸、大黄素、大黄酚和大黄素甲醚）,2种双蒽酮类成分（番泻苷A、番泻苷B）,以及没食子酸、儿茶素合计9种成分含量的方法。方法：采用Agilent Zorbax SB-C18（4.6 mm×250 mm,5μm）色谱柱,流动相0.05%磷酸水溶液-乙腈,梯度洗脱,流速1 mL·min-1,检测波长280 nm,柱温40℃,进样量10μL。结果：芦荟大黄素、大黄酸、大黄素、大黄酚、大黄素甲醚、番泻苷A、番泻苷B、没食子酸和儿茶素9种成分分别在4.56~45.6,16.3~163,7.38~73.8,7.44~74.4,1.82~18.2,57.6~576,28.2~282,3.27~32.7,118~1.18×103mg·L-1与峰面积呈良好的线性关系（r≥0.999 5）,平均加样回收率为96.4%~101.4%,RSD〈3.15%。不同来源的6批大黄样品中9种有效成分的含量差别较大。结论：该方法简单、重复性良好,准确可靠,可用于大黄药材的快速质量评价。

Objective： To establish the method for simultaneous determination of five free anthraquinones（including aloe-emodin,rhein,emodin,chrysophanol,physcion）,two dianthrones（including sennoside A and sennoside B）,gallic acid and catechin from rhubarb.Method： The analysis was achieved with an Agilent Zorbax SB-C 18 analytical column（4.6 mm × 250 mm,5 μm） by gradient elution of 0.05% phosphoric acid in water and acetonitrile at 40 ℃.The flow rate was 1 mL·min-1,injection volume was 10 μL,and detection wave length was set at 280 nm.Result： All the active constituents＇ calibration curves showed a good linearity（r≥0.999 5） in a relatively wide concentration range.The liner range of the active constituents was 4.56-45.6,16.3-163,7.3873.8,7.44-74.4,1.82-18.2,57.6-576,28.2-282,3.27-32.7,118-1.18 × 103mg·L-1,respectively.And the average recovery was 96.4%-101.4%,RSD 3.15%.Furthermore,contents of the Nine active constituents of rhubarb samples from different sources differ significantly.Conclusion： This method is simple,good reproducibility,accuracy and reliability,it can be used for rapid quality evaluation of rhubarb.