一测多评法测定黑果小檗果实中5种花色苷含量

Determination of Five Anthocyanins in Berheris Heteropoda Schrenk Fruit by QAMS Method

建立高效液相色谱一测多评法(HPLC-QAMS)测定黑果小檗果实中5种花色苷的含量。采用Welch Topsil TM C 18色谱柱(250 mm×4.6 mm,5μm);流动相为甲醇-2.5%甲酸水溶液,梯度洗脱;检测波长为520 nm;流速为0.8 mL/min;柱温为35℃;进样量为10μL。以矢车菊素-3-O葡萄糖苷为内参物,建立飞燕草素-3-O-葡萄糖苷、矮牵牛素-3-O-葡萄糖苷、芍药素-3-O-葡萄糖苷以及锦葵素-3-O-葡萄糖苷的相对校正因子,并计算各成分含量,同时与外标法测定结果进行对比。5种花色苷分别在各自浓度范围内与峰面积的线性关系良好(0.9994≤R 2≤0.9996);平均加样回收率为99.44%~101.72%,RSD为1.32%~2.59%;6批不同产地的黑果小檗采用QAMS法与外标法所测含量无显著性差异(P>0.05)。该方法可用于黑果小檗果实中5种花色苷的含量测定。

To establish a high performance liquid chromatography-quantitative analysis of multi-component by single marker(HPLC-QAMS)method for determination of five anthocyanins in Berberis heteropoda Schrenk fruit.Welch Topsil TM C 18(250 mm×4.6 mm,5μm)was used as the column with methanol-2.5%formic acid water as the mobile phase by gradient elution.The detection wavelength was set at 520 nm,the flow rate was 0.8 mL/min,the column temperature was 35℃and the injection volume was 10μL.The relative correction factors of delphinidin-3-O-glucoside,petunidin-3-O-glucoside,peonidin-3-O-glucoside and malvidin-3-O-glucoside were established with cyanidin-3-O-glucoside as internal reference.The contents of each component were calculated and compared with the results of external standard method.Five anthocyanins showed good linear relationships within their own ranges(0.9994≤R 2≤0.9996);whose average recoveries are 99.44%~101.72%with the RSD of 1.32%~2.59%.There was no significant difference between QAMS method and external standard method in the results of 6 batches of Berberis heteropoda Schrenk fruits from different producing areas(P>0.05).The established method can be used for the determination of five anthocyanins in Berberis heteropoda Schrenk fruits.