微量苯酚-硫酸法快速测定西瓜翠衣多糖含量

Rapid determination of the polysaccharide content from Citrullus lanatus peel by micro-phenol-sulfuric acid method

中文关键词:  微量  苯酚-硫酸法  西瓜翠衣  多糖

英文关键词:microplate format  phenol-sulfuric acid method  Citrullus lanatus peel  polysaccharide

中文摘要:

      目的 建立微量苯酚-硫酸法对西瓜翠衣多糖进行快速测定的方法。方法 利用微量苯酚-硫酸法对西瓜翠衣多糖含量进行测定, 葡萄糖为标准品, 直接在96孔板进行加样操作, 探讨检测波长、试剂添加顺序及用量、显色时间等因素对吸光值的影响, 并测定了精密度、稳定性、重复性和回收率。结果 微量苯酚-硫酸法测定西瓜翠衣多糖条件优化结果为: 检测波长为490 nm, 6%苯酚溶液25 μL, 浓硫酸125 μL, 室温, 反应时间10 min。用该方法测定西瓜翠衣多糖方法稳定, 精密度高, 在1~5 μg范围内吸光度与被测物含量呈良好线性关系, r2=0.999, 平均回收率为104.68%, 相对标准偏差为4.29%。结论 优化后的微量苯酚-硫酸法测定西瓜翠衣多糖具有高通量, 精密度高, 稳定性和重复性好的特点, 同时操作方便, 快速, 节约试剂, 为西瓜翠衣多糖的快速检测奠定了基础, 对其他多糖快速测定具有参考价值。

英文摘要:

      Objective To establish a rapid determination method for polysaccharides in Citrullus lanatus peel by the micro-phenol-sulfuric acid method. Methods Glucose was used as the standard substance, the sample was directly added into a 96-well plate. The effects of factors such as detection wavelength, reagent addition sequence and amount, color development time on absorbance were explored, the precision, stability, repeatability and recovery rate were also measured. Results The detection wavelength was 490 nm, 25 μL of 6% phenol solution, 125 μL of concentrated sulfuric acid, at room temperature, and the reaction time was 10 minutes. There was a good linear relationship between the absorbance and the content of the measured substance within the range of 1 to 5 μg, with r2=0.999 and an average recovery rate of 104.68%, relative standard deviation 4.29%. Conclusion The optimized micro-phenol-sulfuric acid method for determining the polysaccharides has the characteristics of high throughput, high precision, good stability and repeatability. Meanwhile, it is convenient to operate, rapid, and saves reagents, laying a foundation for the rapid detection of polysaccharides in Citrullus lanatus peel and having reference value for the rapid determination of other polysaccharides.