基于硫酸-苯酚法的可溶性糖高通量测定方法及其应用

High-throughput method for determining soluble sugar based on sulfuric acid-phenol method and its application

中文关键词:  硫酸-苯酚法  高通量  可溶性糖  五味子  枸杞子  梨

英文关键词:sulfuric acid-phenol method  high-throughput  soluble sugar  Schisandra chinensis fruits  Lycium barbarum fruits  pears

中文摘要:

      目的 基于硫酸-苯酚法建立一种测定可溶性糖含量的高通量方法, 并应用于五味子、枸杞子和梨的可溶性糖含量测定。方法 以硫酸-苯酚比色法为基础, 在微孔板中进行反应和测定, 对苯酚加入顺序、反应温度、浓硫酸用量和苯酚浓度进行考察和优化。以五味子、枸杞子和梨为研究对象, 进行方法学考察, 并与传统苯酚-硫酸法的含量测定结果进行比较。结果 先加入浓硫酸后加入苯酚的反应顺序能获得较高吸光度值, 葡萄糖-果糖标准曲线在1.515~30.285 μg/孔范围内线性关系良好, 方法的精密度、重复性和稳定性较好, 测定五味子、枸杞子、梨样品中葡萄糖-果糖的加样回收率分别为98.34%、103.88%、101.26%, 相对标准偏差均小于5%。结论 本研究方法与传统方法相比操作简单、试剂用量小、线性范围广, 可以用作植物样品中可溶性糖的高通量测定。

英文摘要:

      Objective To establish a high-throughput method based on sulfuric acid-phenol method for determining soluble sugar, and apply for the determination of soluble sugar of Schisandra chinensis fruits, Lycium barbarum fruits and pears. Methods The reaction and assay were carried out in the microplate based on the sulfuric acid-phenol colorimetric method, and the influences of phenol addition order, reaction temperature, volume of concentrated sulfuric acid and phenol concentration on the determination of soluble sugar were investigated and the reaction conditions were optimized. The present method was validated with and applied for samples of Schisandra chinensis fruits, Lycium barbarum fruits and pears, and the content of soluble sugar in the 3 samples determined by the present method and the traditional phenol-sulfuric acid method were compared. Results The higher absorbance value can be obtained by adding concentrated sulfuric acid first and then phenol. The standard curve of glucose-fructose had a good linear relationship in the range of 1.515 to 30.285 μg/well, the method had good precision, repeatability and stability, and the recoveries of glucose-fructose spiked in samples of Schisandra chinensis fruits, Lycium barbarum fruits and pears were 98.34%, 103.88%, 101.26%, with the relative standard deviations value less than 5%. Conclusion Compared with the traditional method, the present method is of simple operation, small reagent consumption and has wide linear range, and can be used as a high-throughput method for determining soluble sugar in plant samples.