世界中医药
文章摘要
高效液相色谱法检测二仙汤中淫羊藿黄酮类成分的含量
Determination of Epimedium Flavonoids in Erxian Decoction by High Performance Liquid Chromatography
投稿时间:2019-03-13  
DOI:10.3969/j.issn.1673-7202.2019.07.006
中文关键词: 高效液相色谱法  二仙汤  淫羊藿苷  淫羊藿次苷Ⅱ  朝藿定B  朝藿定C
英文关键词: High performance liquid chromatography  Erxian decoction  Icariin  Icariin II  Chaohuding B  Chaohuding C  Content analysis
基金项目:国家自然科学基金项目(81760732);延安市科技惠民计划项目(2017HW-11-01)
作者单位
高芳芳1,刘耀龙2,李慧1,凡芳1 1 延安大学附属医院药剂科延安716000 2 陕西省延安市中医医院保健科延安716000 
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中文摘要:
      目的:通过高效液相色谱法(HPLC)对二仙汤中淫羊藿黄酮类成分进行含量检测,并采用4个不同产地的淫羊藿进行多角度分析。方法:进样体积为1 μL;色谱柱的的温度控制在40 ℃;流动相A以0.1%色谱纯甲酸和5 mmol/L的色谱纯乙酸铵均匀混合而成;流动相B以色谱纯甲醇和超纯水均匀混合而成,研究中进行所有高效液相色谱线性梯度洗脱程序。检测波长设置在270 nm时可以检测到较多的吸收峰。前期对HPLC进行常规的可靠性方法学考察,包括专属性、精密度、稳定性等,然后进一步对淫羊藿黄酮类成分淫羊藿苷、淫羊藿次苷Ⅱ、朝藿定B、朝藿定C的含量进行检测分析。结果:二仙汤中淫羊藿黄酮类成分淫羊藿苷对照品、淫羊藿次苷Ⅱ对照品、朝藿定B对照品和朝藿定C对照品均可见良好的分离度,且不存在任何内源性的干扰物质,每一个黄酮类成分专属性均较好。淫羊藿苷线性回归方程:Y=34.012X-12.99;淫羊藿次苷Ⅱ线性回归方程:Y=39.875X-7.134;朝藿定B线性回归方程:Y=23.997X-8.416;朝藿定C线性回归方程:Y=29.896X-15.38。淫羊藿苷、淫羊藿次苷Ⅱ、朝藿定B和朝藿定C精密度试验的RSD值分别为0.33%、0.64%、0.57%、0.52%;稳定性试验RSD值分别为1.09%、1.62%、1.78%、0.41%;重复性试验RSD值分别为2.61%、2.75%、2.48%、1.22%;二仙汤供试品溶液中淫羊藿黄酮类成分淫羊藿苷、淫羊藿次苷Ⅱ、朝藿定B和朝藿定C的平均回收率分别为(96.87±2.15)%、(99.46±1.52)%、(101.64±2.43)%、(99.79±2.31)%。二仙汤供试品溶液中淫羊藿黄酮类成分的含量高于淫羊藿单味药材供试品溶液;从淫羊藿不同产地中黄酮类成分含量的角度比较,发现四川产地的淫羊藿中测得的黄酮类成分高于其他3个产地,陕西产地淫羊藿次之,其后为湖北产地淫羊藿,而湖南产地淫羊藿黄酮类成分含量最低。结论:采用HPLC检测二仙汤中淫羊藿黄酮类成分含量具有良好的可靠性,且二仙汤供试品溶液中淫羊藿黄酮类成分的含量高于淫羊藿单味药材供试品溶液。
英文摘要:
      To determine the content of epimedium flavonoids in Erxian Decoction by high performance liquid chromatography(HPLC),and to analyze Herba Epimedii from 4 different habitats.Methods:Sample volume was 1 μL; column temperature was controlled at 40 ℃; mobile phase A was homogeneously mixed with 0.1% pure formic acid and 5 mmol/L pure ammonium acetate; mobile phase B was homogeneously mixed with pure methanol and ultra-pure water.Linear gradient elution procedures of high performance liquid chromatography were studied.When the detection wavelength was set at 270 nm,more absorption peaks can be detected.In the early stage,the routine reliability methodologies of HPLC were investigated,including specificity,precision,stability,etc.Then the contents of icariin,icariin II,Icariin B,Icariin C were further determined and analyzed.Results:1)Icariin,Icariin II,Chaohoding B and Chaohoding C in Erxian Decoction showed good isolation,and there was no endogenous interfering substance.Each flavonoid had good specificity. 2)The results of linear regression analysis showed that the linear regression equation of icariin was Y=34.012X-12.99; the linear regression equation of icariin II was Y=39.875X-7.134; the linear regression equation of Chaohuoding B was Y=23.997X-8.416; and the linear regression equation of Chaohuoding C was Y=29.896X-15.38. 3)The RSD values of precision tests of icariin,icariin II,Chaixiding B,Chaixiding B,and Chaixiding C were 0.33%,0.64%,0.57%,0.52%.RSD values of stability test were 1.09%,1.62%,1.78%,0.41%.RSD values of repetitive test were 2.61%,2.75%,2.48%,1.22%. 4)The average recoveries of icariin,icariin II,Chaohoding B and Chaohoding C were(96.87±2.15)%,(99.46±1.52)%,(101.64±2.43)%,(99.79±2.31)%,respectively. 5)The content of epimedium flavonoids in Erxian Decoction was higher than that of sample solution of single herb of Herba Epimedii.From the point of view of the content of flavonoids in different habitats of Herba Epimedii,it was found that the content of flavonoids in Herba Epimedii from Sichuan was higher than that in the other 3 habitats,followed by Herba Epimedii from Shaanxi,Herba Epimedii from Hubei,and Herba Epimedii from Hunan.The content of flavonoids was the lowest.Conclusion:The determination of flavonoids in Erxian Decoction by HPLC has good reliability,and the content of flavonoids in Erxian Decoction is higher than that of single herb of Herba Epimedii.
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