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采用碱提醇沉法结合双酶解法对黄泥螺内脏中活性多糖进行了提取、分离和纯化。单因素试验和正交试验确定了最佳提取工艺条件:提取温度80℃、浸提时间4 h、NaOH浓度为0.5mol/L、乙醇与溶液的体积比为3∶1;最佳纯化方法为:两次醇沉结合双酶解法实现多糖的有效分离,TCA法、Sevag法结合DEAE弱阴离子型纤维素柱。复合多糖提取率为16.22%。利用G100葡聚糖凝胶过滤法对复合多糖进行筛分,成功收集到3种不同分子量的多糖:BEP1、BEP2、BEP3。并测得3种多糖的重均分子量分别为分子量为:MW(BEP1)=62185 Da,MW(BEP2)=46057 u,MW(BEP3)=21747 u;含量BEP132.15%,BEP219.48%,BEP348.37%。醋酸薄膜电泳法检测表明所得3种多糖均为单一多糖。
The alkaline polysaccharides were extracted, separated and purified by alkaline extraction and alcohol precipitation combined with double digestion method. The optimum extraction conditions were as follows: extraction temperature 80 ℃, extraction time 4 h, NaOH concentration 0.5 mol / L, volume ratio of ethanol to solution 3: 1. The best purification method was : Double alcohol precipitation combined with double enzymatic method to achieve the effective separation of polysaccharides, TCA method, Sevag method combined with DEAE weak anionic cellulose column. The polysaccharide extraction rate was 16.22%. The polysaccharide was screened by G100 dextran gel filtration method. Three polysaccharides with different molecular weights were successfully collected: BEP1, BEP2 and BEP3. The results showed that the weight average molecular weights of the three polysaccharides were as follows: MW (BEP1) = 62185 Da, MW (BEP2) = 46057 u, MW (BEP3) = 21747 u; content BEP132.15%, BEP219.48% BEP348.37%. Acetic acid film electrophoresis test showed that the three polysaccharides obtained were single polysaccharide.