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

关键词:  桑枝  液相色谱法  异槲皮苷  山奈酚

Key words:  Ramulus Mori  liquid chromatography  Isoquercitrin  kaempferol

摘要:

[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，5 μm），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0.05~2.00 μg/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。

Abstract:

[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.