HPLC法同时测定桑枝中6种成分的含量

Simultaneous determination of six bioactive components for quality control of Ramulus Mori by HPLC

[目的]建立高效液相色谱(HPLC)方法同时测定桑枝中桑皮苷A、绿原酸、隐绿原酸、芦丁、异槲皮苷和山奈酚含量。[方法]采用反相高效液相色谱(RP-HPLC)法,色谱柱为Agilent C18柱(4.6 mm×250 mm,5μm),流动相为水(含0.1%甲酸)-乙腈,以柱温25℃,梯度洗脱,检测波长为360 nm,流速1 mL/min,采用外标法计算桑枝中6种成分的含量。[结果]桑皮苷A、绿原酸、隐绿原酸、芦丁、异槲皮苷和山奈酚浓度分别在22.50~900.00、6.25~250.00、3.00~120.00、0.08~3.00、0.10~4.00及0.05~2.00μg/mL范围内与各自峰面积值呈良好的线性关系;稳定性实验RSD为1.04%~3.07%,重复性实验RSD为2.22%~4.70%,精密度实验RSD为1.34%~4.90%;平均加样回收率为97.4%~104.0%(RSD≤1.01%)。[结论]本法简便、准确、灵敏,为桑枝的质量控制提供有力依据。

[Objective] To establish a high pressure liquid chromatography（HPLC） method for simultaneous determination of Mulberroside A, cryptochlorogenic acid, chlorogenic acid, rutin, Isoquercitrin and kaempferol content for quality control of Ramulus Mori. [Methods]The separation was carried out on a Agilent C18 column（4.6 mm×250 mm, 5 μm） column with the mobile phase of water（containing 0.1%formic acid）-acetonitrile. The detection wavelength was 360 nm. The flow rate was set at 1 mL/min. The content of six ingredients of Ramulus Mori were calculated by external standard method. [Results] This method exhibited a good linear relationship. Relative standard deviation（RSD） of stability was 1.04%~3.07%. The RSD of the reproducibility was 2.22%~4.70%. The precision RSD was 1.34%~4.90%.The average recovery was 97.4%~104.0%（RSD=1.01%）. [Conclusion] The simple, accurate, sensitive method established could be used to evaluate the quality of Ramulus Mori.