普乐安胶囊中槲皮素和山柰素的HPLC法含量测定研究

Optimize the HPLC Test Condition of Quercetin and Kaempferol in Pule'an Capsules

目的：采用高效液相色谱（HPLC）法测定普乐安胶囊中槲皮素和山柰素的含量并进行条件优化。方法：高效液相色谱柱为Waters C18色谱柱（4.6 mm×250 mm,5μm）;最佳流动相为甲醇：0.4%磷酸溶液（48∶52）;流速为1.0 m L·min-1;检测波长为360 nm;柱温为35℃;进样量10μL。结果：普乐安胶囊中槲皮素和山柰素分别在0.01248~0.3120μg和0.03197~0.3996μg质量浓度范围内与峰面积呈现良好线性关系（r=1.000;r=1.000）,平均回收率分别为99.05%和98.82%,RSD分别为0.88%和1.03%。结论：该方法操作简便、灵敏、准确,重现性好,能有效测定普乐安胶囊中槲皮素和山柰素的含量。特别是流动相组成比例优化条件下,样品普乐安胶囊中槲皮素与有关杂质实现了很好的分离。

Objective： To optimize the high performance liquid chromatography（ HPLC） method test condition of quercetin and kaempferol in pule＇an capsules. Methods： The HPLC column was Waters C18column（ 4. 6 mm × 250 mm,5 μm）; optimal mobile phase was methanol： 0. 4% phosphoric acid（ 48∶ 52）; flow rate was 1. 0 m L · min- 1; detection wavelength 360 nm; column temperature was 35 ℃; the injection volume was 10μL. Results： Pule＇an capsules quercetin and kaempferol were within range 0. 01248 ~ 0. 3120μg and 0. 03197 ~ 0. 3996 μg concentration showed a good linear relationship（ r = 1. 000; r = 1. 000） and the peak area,the average recoveries were 99. 05% and 98. 82%,RSD were 0. 88% and 1. 03% respectively. Conclusion： The method is simple,sensitive,accurate,reproducible and effective determination of pule＇an capsules quercetin and kaempferol. Especially the mobile phase composition ratio of optimum conditions,the sample pule＇an capsules quercetin with the impurities to achieve a good separation.