超声辅助酶提取-离子色谱法测定六堡茶多糖中单糖组分及含量

Determination of monosaccharide components and content in polysaccharides from Liubao tea by ultrasonic-assisted enzymatic extraction-ion chromatography

中文关键词:  超声辅助酶提取  离子色谱法  六堡茶  茶多糖  单糖

英文关键词:ultrasonic-assisted enzymatic extraction  ion chromatography  Liubao tea  tea polysaccharide  monosaccharide

中文摘要:

      目的 建立超声辅助酶提取-离子色谱法测定六堡茶多糖中单糖组分及含量的方法。方法 采用单因素实验和正交实验对六堡茶多糖中单糖组分及含量的提取条件进行优化, 同时优化了离子色谱法检测六堡茶多糖中单糖的色谱条件, 并对所建立的方法进行方法学评价。结果 通过极差和方差分析, 六堡茶多糖中单糖组分及含量最佳提取工艺条件为m纤维素酶:m果胶酶(1:5)、酶添加量4.0%、酶解pH 5.0、料液比1:25 (g:mL)、酶解-超声时间40 min、酶解-超声温度50 ℃; 经方法学评价, 7种单糖均具有良好的线性关系, 相关系数均大于0.995, 相对标准偏差小于等于3.2%, 定量限良好, 回收率为83.33%~101.74%。通过对六堡茶多糖中单糖组分的分析, 表明六堡茶中组成茶多糖的有鼠李糖、阿拉伯糖、半乳糖、葡萄糖、D-木糖、D-甘露糖、半乳糖醛酸。结论 超声辅助酶提取方法可以大量缩短样品提取时间, 可以用于快速测定六堡茶多糖中单糖的组分及含量测定。

英文摘要:

      Objective To establish a method for the determination of the monosaccharide components and content in polysaccharides from Liubao tea by ultrasonic-assisted enzymatic extraction-ion chromatography. Methods Optimization of extraction conditions for monosaccharide components and content in Liubao tea polysaccharides by single-factor experiments and orthogonal experiments. The chromatographic conditions for the determination monosaccharides in Liubao tea polysaccharides by ion chromatography were optimized simultaneously, and the established method was evaluated methodologically. Results Through analysis of range and variance, the optimal extraction process conditions for monosaccharide components and content in Liubao tea polysaccharides were that mcellulase to mpectinase ratio of 1:5, enzyme addition level of 4.0%, enzymatic hydrolysis pH of 5.0, solid-to-liquid ratio of 1:25 (g:mL), enzymatic hydrolysis-ultrasonic treatment duration of 40 minutes and enzymatic hydrolysis-ultrasonic temperature at 50 ℃; after methodological evaluation, all 7 kinds of monosaccharides exhibited good linear relationships, with correlation coefficients greater than 0.995, the relative standard deviation of precision was less than 3.2%, the limit of quantitation were satisfactory, and the recovery rates ranged from 83.33% to 101.74%. Through the analysis of monosaccharide components in the polysaccharides of Liubao tea showed that the tea polysaccharides in Liubao tea were composed of rhamnose, arabinose, galactose, glucose, D-xylose, D-mannose and galacturonic acid. Conclusion The ultrasonic-assisted enzymatic extraction method can significantly shorten the extraction time of sample, and can be utilized for the rapid determination of monosaccharide components and content in the polysaccharides of Liubao tea.