[关键词]
[摘要]
目的 建立同时测定黄芪中5种黄酮类成分含量的高效液相色谱(HPLC)分析方法并进行黄芪的指纹图谱研究。方法 采用HPLC-DAD,Waters SymmetryShieldTM RP18色谱柱(250 mm×4.6 mm,5 μm),乙腈(A)-水(0.05%磷酸)(B)为流动相梯度洗脱,体积流量1.0 mL/min,检测波长246、206 nm,柱温25℃,进样量20 μL,对黄芪中毛蕊异黄酮葡萄糖苷、芒柄花苷、(6aR,11aR)-9,10-二甲氧基紫檀烷-3-O-β-D-葡萄糖苷、毛蕊异黄酮、7-羟基-4'-甲氧基异黄酮进行含量测定。结果 方法学考察结果显示,黄芪中5种黄酮类成分线性范围,相关系数良好(r=0.999 8~0.999 9,n=9),各色谱峰间分离度均达到定量要求。精密度(RSD<1.86%),稳定性(RSD<2.01%),重复性(RSD<1.97%),加样回收率(RSD<2.90%,n=9)等参数均达到定量分析要求。建立了黄芪的HPLC指纹图谱,标定了27个共有峰,样品相似度在0.808~0.971,存在一定差异。结论 本实验建立的方法准确、简便、重复性好,可为黄芪药材的质量控制提供参考。
[Key word]
[Abstract]
Objective To establish a method to determine the components of five flavonoids in Radix Astragali and built fingerprint of Radix Astragali by HPLC. Methods HPLC-DAD method was used to determine five flavonoids simultaneously, including calycosin-7-glucoside, ononin, (6aR, 11aR)-9, 10-dimethoxypterocarpan-3-O-β-D-glucoside, calycosin and 7-hydroxyl-4'-methoxyisoflavone. The separation was carried on a Waters Symmetry ShieldTM RP18 column (250 mm×4.6 mm, 5 μm) with the mobile phase consisting of acetonitrile (A)-water (0.05% phosphoric acid) (B) by using gradient elution. The flow rate was 1.0 mL. min-1 and the detection wavelengths were at 246 and 206 nm. The column temperature was set at 25℃ and the injection volume was 20 μL.Results Methodology examination showed that the five flavonoids showed a good linear relationship (r=0.999 8-0.999 9, n=9), precision (RSD < 1.86%), stability (RSD < 2.01%) and repeatability (RSD < 1.97%) conformed to requirements. The fingerprint of Radix Astragali was built. 27 mutual peaks were labeled and the similarity was more than 0.808. Conclusion The established method is accurate, simple and reproducible, which can provide reference for quality control of Radix Astragali.
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[基金项目]
国家自然科学基金资助项目(21502142)