黑松露多糖提取工艺优化及体外抗氧化活性分析

Extraction Optimization for the Polysaccharides from Tuber sinense and Their in vitro Antioxidant Activities

选取云南黑松露为研究对象,对黑松露多糖优化提取工艺,纯化后单糖组成和体外抗氧化活性进行研究。应用响应曲面法优化后最佳提取工艺条件为:提取温度75.36℃,提取时间1.02 h与料液比1:31.32 g/mL,在此条件下,多糖的实验得率为11.79%,预测得率为11.86%。采用DEAE-Sepharose快速流动柱从黑松露粗多糖中分离纯化出4个新的多糖组分(TSP-1、TSP-2、TSP-3、TSP-4)。应用离子色谱法分析多糖成分,得出TSP-1的单糖组成为盐酸氨基半乳糖、葡萄糖和甘露糖,比例为2.8:77.1:20;TSP-2为鼠李糖、盐酸氨基葡萄糖、半乳糖、葡萄糖和甘露糖,比例为18.7:1.5:2:40.6:37.3;TSP-3为鼠李糖、盐酸氨基葡萄糖、葡萄糖、甘露糖、葡萄糖醛酸和半乳糖的比例为13:3.5:58.4:21.1:2.7:1.2。粗多糖(crud TSP)和TSP-1、TSP-2、TSP-3三个纯化组分多糖浓度在0.25~4 mg/mL范围内,对DPPH自由基的最大清除率分别为73.93%、36.67%、73.60%和54.10%;对ABTS自由基的最大清除率分别为60.47%、36.20%、41.87%和52.73%;金属螯合力分别为61.63%、27.00%、52.50%和43.17%;还原力吸光度值分别为0.39、0.34、0.28和0.56。该研究旨在为研究黑松露多糖具生物活性的物质基础及开发利用提供理论依据,并对其保健食品研发具有重要意义。

Tuber sinense from Yunan was used as the research object,and the extraction process for extracting polysaccharides from Tuber sinense was optimized.The monosaccharide compositions and in vitro antioxidant activities of the purified polysaccharides were analyzed.The optimal extraction conditions obtained via optimization by the response surface method were as follows:extraction temperature 75.36℃,extraction time 1.02 h and material-liquid ratio 1:31.32(g/mL).Under these conditions,the experimental yield of polysaccharides was 11.79%,whilst the predictive yield was11.86%.Four new polysaccharide fractions(TSP-1,TSP-2,TSP-3,TSP-4)were separated and purified from the crude polysaccharides from Tuber sinense by using the DEAE-sepharose fast flow column.The compositions of the polysaccharides were analyzed by ion chromatography.The monosaccharide components of TSP-1 included galactosamine hydrochloride,glucose and mannose in a ratio of 2.8:77.1:20;TSP-2 included rhamnose,glucosamine hydrochloride,galactose and mannose in a ratio of 18:1.4:44.7:35.9;TSP-3 was composed of rhamnose,glucosamine hydrochloride,glucose,mannose,glucuronic acid and galactose in a ratio of13:3.5:58.4:21.1:2.7:1.2.In the range of 0.25~4 mg/mL,the maximum rates of crude polysaccharide(crud TSP),TSP-1,TSP-2 and TSP-3 were 73.93%,36.67%,73.60%and 54.10%for scavenging DPPH radical,60.47%,36.20%,41.87%and 52.73%for scavenging ABTS radical,61.63%,27.00%,52.50%and 43.17%for chelating metal,and 0.39,0.34,0.28,0.56 for the absorbance corresponding to reducing power.This research aimed to provide a theoretical foundation for the study of the bioactive material basis and the development and utilization of Tuber sinense polysaccharides.The research results are of great significance to the research and development of wellness foods.