高效液相色谱法同时测定水辣蓼提取物中6种黄酮成分的含量

Simultaneous determination of six kinds of flavonoid in extract of Polygonum hydropiper L. by HPLC

建立了同时定量检测水辣蓼中6种黄酮类成分的高效液相色谱方法：选择Waters Symmetry shield RP18（4.6 mm×250 mm,5.0 μm）色谱柱,流动相甲醇-0.4％甲酸水溶液（梯度洗脱）,流速1 mL/min,柱温30℃,检测波长359 nm,进样量10 μL.结果表明,6种黄酮成分在50 min内达到基线分离,在0.1～20μg/mL浓度范围内,6种黄酮成分的线性相关系数R2≥0.9995,平均加样回收率为96.65％～99.54％,RSD为0.82％～ 1.38％ （n=3）.该方法准确、简便,重复性好,可用于水辣蓼中芦丁、金丝桃苷、槲皮苷、槲皮素、异鼠李素和山奈酚的含量测定,为控制水辣蓼药材的质量提供依据.

This paper established a method of quantitative detection for six kinds of flavonoid content in extract of Polygonum hydropiper L. by high performance liquid chromatography （HPLC）. The analysis was performed on a Waters Symmetry shield RP18 （4.6 mm×250 mm, 5.0μm） column. Mobile phase was methanol-0.4% formic acid aqueous solution （gradient elution）. The flow rate was set at 1.0 mL/min and the eluent was detected at 359 nm at 30℃. The calibration curves of the six kinds of flavonoid were linear（R2〉0.9995） over the concentration range of 0.1-20 μg/mL. The mean recoveries were 96.65% to 99.54%, RSD was 0.82%-1.38% （n=3）. The results indicate that the method is accurate and simple, has good repeatability, can apply to simultaneous determination of Rutin, Hyperoside, Quercitrin, Quercetin, Isorhamnetin and Kaempferol in the extract of P. hydropiper L., to provide the basis for control the quality of P. hydropiper L.