基于HPLC梯度洗脱法同时测定止咳喘颗粒中7种成分的含量

Study on Quantitative Control of Seven Constituents in Zhikechuan Granules by HPLC with Gradient Elution

目的:建立高效液相梯度洗脱法(HPLC-GEM)同时测定止咳喘颗粒中去芹糖桔梗皂苷E、桔梗皂苷E、桔梗皂苷D3、甘草苷、甘草酸、金丝桃苷和槲皮素的含量。方法:色谱柱为Agilent TC-C18柱(250 mm×4.6 mm,5μm),柱温:30℃;流动相为乙腈-0.5%冰醋酸溶液,梯度洗脱,体积流量:1.0 ml·min^-1;检测波长分别为210 nm(检测去芹糖桔梗皂苷E、桔梗皂苷E和桔梗皂苷D3)、237 nm(检测甘草苷和甘草酸)和360 nm(检测金丝桃苷和槲皮素)。结果:去芹糖桔梗皂苷E、桔梗皂苷E、桔梗皂苷D3、甘草苷、甘草酸、金丝桃苷和槲皮素质量浓度分别在0.74~14.80μg·ml^-1(r=0.9999)、1.16~23.20μg·ml^-1(r=0.9996)、0.53~10.60μg·ml^-1(r=0.9995)、1.39~27.80μg·ml^-1(r=0.9993)、6.47~129.40μg·ml^-1(r=0.9997)、4.18~83.60μg·ml^-1(r=0.9994)、1.06~21.20μg·ml^-1(r=0.9998)范围内与峰面积线性关系良好;平均加样回收率分别为97.70%、98.69%、96.98%、99.23%、100.09%、99.47%和98.24%,RSD分别为1.33%、1.20%、1.03%、0.97%、0.69%、0.72%和1.49%(n=9)。结论:该方法操作简便、重复性好,可作为评价止咳喘颗粒的质量控制方法。

Objective: To establish an HPLC with gradient elution method( HPLC-GEM) for the simultaneous determination of deapi-platycoside E,platycoside E,platycodin D3,liquiritin,glycyrrhizic acid,hyperoside and quercetin in Zhikechuan granules. Methods: The method was performed on an Agilent TC-C18 column( 250 mm × 4.6 mm,5 μm) with the mobile phase consisting of acetonitrile-0.5% acetic acid solution with gradient elution. The flow rate was 1.0 ml · min^-1 and the column temperature was set at 30 ℃ . The detection wavelength was set at 210 nm for deapi-platycoside E,platycoside E and platycodin D3,237 nm for liquiritin and glycyrrhizic acid,and 360 nm for hyperoside and quercetin. Results: Deapi-platycoside E,platycoside E,platycodin D3,liquiritin,glycyrrhizic acid,hyperoside and quercetin showed good linear relationship within the range of 0.74^-14.80 μg·ml^-1( r = 0.999 9),1.16-23.20 μg·ml^-1( r = 0.999 6),0.53^-10.60 μg·ml^-1( r = 0.999 5),1.39-27.80μg·ml^-1( r = 0.999 3),6.47^-129.40 μg·ml^-1( r = 0.999 7),4.18-83.60 μg·ml^-1( r = 0.999 4) and 1.06-21.20 μg·ml^-1( r= 0.999 8),respectively. The average recovery was 97.70%,98.69%,96.98%,99.23%,100.09%,99.47% and 98.24% with the RSDs of 1.33%,1.20%,1.03%,0.97%,0.69%,0.72% and 1.49%( n = 9),respectively.Conclusion: The method is easy to be operated and repeatable,and valuable as a quality control method for Zhikechuan granules.