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目的:建立双波长HPLC同时测定金刚藤软胶囊中绿原酸、白藜芦醇、芦丁、槲皮素和山柰酚含量的方法。方法:采用Agilent ZORBAX SB-C18色谱柱(4.6 mm×150 mm,5μm),流动相乙腈-0.1%磷酸水溶液梯度洗脱,流速1.0 m L·min-1,柱温25℃,检测波长306 nm(绿原酸、白藜芦醇)和365 nm(芦丁、槲皮素、山柰酚)。结果:绿原酸、白藜芦醇、芦丁、槲皮素、山柰酚分别在0.058 5~2.34μg(r=0.999 8),0.025 3~1.01μg(r=0.999 9),0.042 2~0.844μg(r=0.999 6),0.018 1~0.722μg(r=0.999 9),0.0165~0.660μg(r=0.999 9)呈现良好的线性关系,平均回收率分别为98.0%,100.6%,96.0%,102.7%,98.5%,RSD分别为1.2%,2.2%,1.8%,2.9%,1.3%。结论:该方法操作简便、重复性好,可作为金刚藤软胶囊中5个活性成分的含量测定方法。
Objective: To establish a dual-wavelength HPLC method for the simultaneous determination of chlorogenic acid, resveratrol, rutin, quercetin and kaempferol in Jingangteng soft capsule. METHODS: The mobile phase was eluted with a mobile phase of acetonitrile-0.1% phosphoric acid in a gradient of 1.0 m L · min-1 using an Agilent ZORBAX SB-C18 column (4.6 mm × 150 mm, 5 μm) (Chlorogenic acid, resveratrol) and 365 nm (rutin, quercetin, kaempferol). RESULTS: The concentrations of chlorogenic acid, resveratrol, rutin, quercetin and kaempferol were respectively 0.058 5 ~ 2.34 μg (r = 0.999 8), 0.025 3 ~ 1.01 μg (r = 0.999 9), 0.042 2 ~ (R = 0.999 9), 0.0165 ~ 0.660 μg (r = 0.999 9) showed a good linear relationship with the average recoveries of 0.844 μg (r = 0.999 6), 0.018 1 ~ 0.722 μg and the average recoveries were 98.0%, 100.6% and 96.0% , 102.7% and 98.5% respectively. The RSDs were 1.2%, 2.2%, 1.8%, 2.9% and 1.3% respectively. Conclusion: The method is simple, reproducible, and can be used as a method for the determination of five active ingredients in the kumquat soft capsule.