反相高效液相色谱法测定青荚叶中表儿茶素含量

Determination of epicatechin in Helwingia japonica using reversed phase high-performance liquid chromatrography

目的:采用反相高效液相色谱法建立青荚叶中表儿茶素含量的测定方法。方法:用甲醇-水(体积比1:1)为溶剂,超声波法提取青荚叶中的表儿茶素。采用反相高效液相色谱法。色谱条件为:色谱柱:ZorbaxXDB-C18柱(4.6mm×250mm,5μm);流动相组成:流动相A,400mL0.04mol/L柠檬酸+80mLN,N-二甲基甲酰胺+20mL四氢呋喃;流动相B,甲醇;流动相的洗脱方式为等强度洗脱:流动相A+B(体积比85:15);流速1.0mL/min;柱温:35℃;检测器为二极管阵列检测器,检测波长280nm。采用外标法定量。结果:表儿茶素线性范围为0～364.00mg/L(r=0.9999),平均回收率为98.4%,RSD为2.08%(n=3)。青荚叶中表儿茶素含量为(5.334±0.111)mg/g。结论:该方法简便、准确且灵敏,适用于青荚叶中表儿茶素含量测定。

Aim：To establish a method of reversed phase high-performance liquid chromatrography（RP-HPLC） to determine the content of epicatechin in Helwingia japonica.Methods：RP-HPLC was adopted,on a column of Zorbax XDB-C18（4.6 mm×250 mm,5 μm）,using the mobile phase of A（400 mL 0.04 mol/L citric acid＋80 mL N,N-dimethylformamide＋20 mL tetra-hydrofuran）＋ B（methanol）（85：15）,at a flow rate of 1.0 mL/min,and the detection wavelength of 280 nm.Results：The calibration curve of epicatechin was linear over the range of 0～364.00 mg/L（r=0.999 9）.The average recovery rate was 98.4%,and RSD was 2.08%（n=3）.The content of epicatechin in Helwingia japonica was（5.334±0.111） mg/g.Conclusion：The method is simple,accurate,and sensible,and is suitable for the determination of epicatechin in Helwingia japonica.