HPLC测定不同树龄杜仲叶中黄酮类成分的含量

目的]建立高效液相色谱同时测定不同树龄杜仲叶中绿原酸、芦丁、槲皮素和山萘酚含量的方法。方法]采用Shim—packVP—ODS（4．6×150mm，5μm）色谱柱，以甲醇-0．4％磷酸水溶液为流动相进行梯度洗脱，流量1．0ml/min，用二极管阵列检测器扫描检测，以保留时间结合待测物质的紫外吸收光谱进行定性分析。采用外标法进行定量分析，检测波长为360nm，柱箱温度为40℃。结果]绿原酸的线性范围为4．63～92．65μg，r=0．9998，回收率为98％～101．5％；芦丁的线性范围为0．714～14．285μg／ml，r=0．9999，回收率为96％-98．4％；槲皮素的线性范围为0．101～2．066μg／ml，r=0．9970，回收率为94％～101％；山萘酚的线性范围为0．043～0．886μg／ml，r=0．9980，回收率92％～96％。结论]对不同树龄杜仲叶中4种有效成分含量用方差分析进行多重比较，从绿原酸含量方面考虑．三年树龄杜仲叶采收最佳。

Determination of Flavonoids in Leaves of Eucommia ulmoides Oliver at Different Tree Age by HPLC

Objective]To simultaneously determine contents of chlorogenic acid,rutin,quercetin and kaempferol in leaves of Eucommia ulmoides Oliver at different tree age by high performance liquid chromatography method(HPLC).Method]Mixed solution of methanol and 0.4% phosphoric acid solution in different volume ratio were used as mobile phase in the gradient elution,whose flow rate was 1.0 ml/min.Using Shim-pack VP-ODS(4.6 mm×150 mm,5 μm) as chromatographic column,the bioactive constituents were analyzed qualitatively base on their retention time combination with ultraviolet absorption spectra.The quantization analysis was carried out by external standard method with the detection wavelength at 360 nm and the column temperature at 40 ℃.Result]The linear ranges of chlorogenic acid,rutin,quercetin and kaempferol were 4.63-92.65(r=0.999 8),0.714-14.285(r=0.999 9),0.101-2.066(r=0.997 0) and 0.043-0.886 μg/ml(r=0.998 0),respectively.The recoveries of bioactive constituents were 98%-101.5%,96%-98.4%,94%-101% and 92%-96% respectively.Conclusion]The contents of four kind's bioactive constituents in leaves of Eucommia ulmoides Oliver at different tree age were analyzed by way of variance analysis and multiple comparisons.Considering the content of chlorogenic acid,leaves of Eucommia ulmoides Oliver about three-year-old tree age was the best.