苯酚-硫酸法联合DNS法测定商陆多糖及响应面法优化超声波辅助提取研究

摘要

Objective:To find a quick and simple method for determination of polysaccharides in Radix Phytolaccae.Methods:UV spectrophotometry was used to determine the total sugar content by phenol-sulfuric acid method,DNS method was used to determine the content of reducing sugar,and the difference between the two methods was used to measure the content of polysaccharide,and the methods of repeatability,stability and recovery rate were investigated.Based on the single factor study of ultrasonic assisted extraction of polysaccharides from Radix Phytolaccae,the response rate of polysaccharide was taken as the response value,and the response surface experiment design with four factors and three levels was used to optimize the process.Results:The phenol-sulfuric acid combined with DNS method for determination of polysaccharides in Radix Phytolaccae showed a good applicability.The optimized extraction process was as follows:solid-liquid ratio 1∶40(g∶mL),ultrasonic time 30 min,ultrasonic temperature 60 ℃,ultrasonic power 350 W.Under these conditions,the average extraction rate of polysaccharides was 12.38％ ± 0.59％.Conclusion:The method is simple and reliable,and can be used for the determination of polysaccharides in Radix Phytolaccae.%目的:探寻一种商陆多糖快速、简捷的测定方法.方法:利用紫外分光光度计,分别采用苯酚-硫酸法测定商陆总糖含量,DNS法测定还原糖含量,用二者的差值作为测得的商陆多糖含量,并从重复性、稳定性、加样回收率等方面进行方法学考察;通过对商陆多糖超声波辅助提取的单因素研究,以多糖提取率为响应值,采用四因素三水平的响应面试验设计进行工艺优化.结果:苯酚-硫酸法联合DNS法测定商陆多糖具有良好的适用性;最佳提取工艺为料液比1∶40(g∶mL)、超声时间30 min、超声温度60℃、超声功率350 W,在此这条件下得到商陆多糖提取率平均为(12.38％±0.59％).结论:本实验采用的样品处理方法简单,测定方法可靠,可用于商陆多糖的测定.