高效液相色谱法测定野马追药材中槲皮素和山柰素含量

Determination of Quercetin and Kaempferol in Eupatorium Lindleyanum by HPLC

目的建立高效液相色谱(HPLC)法测定野马追中槲皮素和山柰素的含量。方法药材经提取水解后,采用HPLC法测定。色谱柱为Kromasil-C18柱(250mm×4.6mm,5μm),流动相为甲醇-0.4%磷酸(60∶40),检测波长370nm,流速0.6mL/min,柱温30℃。结果槲皮素质量浓度在6～36μg/mL范围内与峰面积线性关系良好(r=0.9999),平均回收率为97.1%,RSD=1.05%(n=6),山柰素质量浓度在4.8～28.8μg/mL范围内与峰面积线性关系良好(r=0.9995),平均回收率为97.5%,RSD=1.28%(n=6)。结论该方法快速、简便、重现性好,为野马追及其制剂提供了一种简便准确的含量测定方法。

Objective To establish a HPLC method for the determination of quercetin and kaempferol in Eupatorium Lindleyanum. Methods HPLC was used to determine the content of quereetin and kaempferol in Eupatorium lindleyanum after herb was extracted and hydrolyzed. Quercetin and kaempferol were separated completely on the Komasil-C18 column （250 mm×4. 6mm, 5 μm）, using the mixture of methanol-0.4% phosphoric acid（60：40） as the mobile phase, with UV detection at 370 nm and the flow rate of 0. 6 mL/min, the column temperature was 30 ℃. Results The calibration curve was linear in the range of 6 - 36 μg/mL of quercetin with correlation coefficient being 0. 999 9. The average recovery rate was 97.1% （RSD = 1.05%, n =6）. The calibration curve was linear in the range of 4.8-28.8 μg/mL of kaempferol with correlation coefficient being 0. 999 5. The average recovery rate was 97.5% （RSD = 1.28%, n =6）. Conclusion The method is quick and simple with good repetition. It can be used for the quality control.