紫外分光光度法测定三叶青胶囊总黄酮含量

Determination of Contents of Total Flavonoids in Tetrastigma hemsleyanum Capsules by UV

目的建立紫外分光光度法测定三叶青胶囊中的总黄酮含量的方法。方法对不同的提取方法进行考察,在单因素试验的基础上,采用正交试验考察提取温度、乙醇比例、料液比、回流时间对总黄酮提取率的影响;筛选显色系统,对Al（NO3）3溶液浓度、Na OH溶液浓度和体积、反应温度及空白溶液进行考察;采用Na NO2-Al（NO3）3-Na OH显色反应体系测定三叶青胶囊中总黄酮含量。结果回流法为最佳的提取方法,条件为加400倍40%乙醇于90℃水浴提取1.5h;Na NO2-Al（NO3）3-Na OH为最佳的显色系统,条件为5%Na NO2溶液1m L、5%Al（NO3）3溶液1m L、8%Na OH溶液10m L、反应温度40℃并以续滤液为空白;以芦丁为对照,测定波长为505nm,在14.0~84.0g·L-1范围内线性关系良好（r=1.0000）,平均回收率为99.00%（RSD1.30%,n=6）。结论该方法简便、快捷、准确,可作为三叶青胶囊中总黄酮含量测定的方法。

OBJECTIVE To establish a method of determination on the content of total flavonoids in Tetrastigma hemsleyanum Capsule by UV. METHODS Different extraction methods were investigated. Based on single-factor tests,orthogonal design was used to adopted to investigate effects of extraction temperature,ethanol concentration,solid-liquid ratio and refluxing time with extraction rate of total flavonoids as index. Different colour reaction systems were screened. Al（ NO3）3and Na OH solution concentration,Na OH solution volumn,reaction temperature and blank solution were investigated. Na NO2-Al（ NO3）3-Na OH colour reaction system was used for the determination of total flavonoids in Tetrastigma hemsleyanum Capsule. RESULTS Reflux extraction was the optimum method and the optimum conditions were as follows： extracted with 400 times the amount of 40% ethanol for 90 min in 90℃ water bath. Na NO2-Al（ NO3）3-Na OH was the optimum colour reaction system and the optimum conditions were as follows：with subsequent filtrate solution as blank,reaction temperature was 40℃ and added with 5% Na NO2 solution 1m L,5% Al（ NO3）3solution 1m L,8% Na OH solution 10 m L. With Rutin as reference and the detection wave lengeh was set at 505 nm. There was good linear relationship during 14. 0 ~ 84. 0g·L- 1（ r = 1. 0000）. The average recovery rate was 99. 00%（ RSD1. 30%,n = 6）. CONCLUSION The mothod is simple,rapid,accurate and can be used for the quantitative method of Tetrastigma hemsleyanum Capsule.