HPLC法测定天山花楸叶提取物黄酮苷的含量

为了建立天山花楸叶中总黄酮苷含量的测定方法,对样品酸水解处理再进行HPLC分析,色谱柱为Waters-C18(150 mm×4.6 mm,5μm),柱温25℃,使用可变波长检测器;流动相为甲醇-乙腈-0.4%磷酸水溶液,梯度洗脱程序为0 min(12∶20∶68)-10 min(52∶0∶48)-18 min(72∶0∶28);流速1.0 mL/min,检测波长为360 nm。结果表明:槲皮素浓度在5.98～47.84μg/mL之间与色谱峰面积呈良好的线性关系,回归方程A=66.96C-5.9964,r=0.9995。平均回收率94.86%,RSD=1.9%;山柰素浓度在4.04～28.28μg/mL之间与色谱峰面积呈良好的线性关系,回归方程A=50.114C-19.529,r=0.9999,平均回收率93.89%,RSD=2.7%;其黄酮苷百分含量为(12.62±0.40)%。该方法干扰少,灵敏,简便,重现性好,可为天山花楸叶的药用开发利用提供质量控制方法。

Determination of the Content of Flavonoid Glycosides Extracted from Sorbus tianschanica Leaves by HPLC

An method for determination of the content of flavonoid glycosides extracted from Sorbus tianschnica was established.The samples,which were hydrolyzed with acid,were analyzed by RP-HPLC.The analysis was carried out on a waters-C18（4.6 mm×150 mm,5 μm） column,The mobile phase was composed of A（methanol）-phase B（acetonitrile）-phase C（0.4% phosphoric acid water solution） in gradient elution 0 min（12：20：68）-10 min（52：0：48）-18 min（72：0：28） by flow rate of 1.0 mL/min in this method,the detection wave length was set at 360 nm,the VWD was used and the column temperature was 25 ℃.The linear relationship of quercetin was good 5.98~47.84 μg/mL,A=66.96C-5.9964,and r=0.9995.The average recovery was 94.86%,（RSD=1.9%）,the linear relationship of kaempferol was good 4.04~28.28 μg/mL,A=50.114C-19.529,and r=0.9999.The average recovery was 93.89%,（RSD=2.7%）.The content of flavonoid glycosides was（12.62±0.40）%.This method was simple,sensitive and less interference,reproducible,the utilization of quality control method with the leaf of Sorbus tianschanica.