采用简单、有效、绿色的三相萃取体系分离富集螺旋藻多糖,分别研究了(NH4)2SO4质量分数、叔丁醇加入量、pH、萃取温度、萃取时间对多糖得率的影响,并对萃取后多糖结构特征进行了分析;结果表明,(NH4)2SO4质量分数为20%,叔丁醇加入量为15 mL,体系pH为7.0,在35 ℃条件下,萃取30 min后,螺旋藻多糖得率为(9.25±0.42)%;红外光谱表明其具有多糖的特征吸收峰,紫外-可见波长扫描显示其不含蛋白质和核酸,采用凝胶渗透色谱测定多糖重均分子质量为3.597×104,其单糖物质量组成为n(鼠李糖):n(甘露糖):n(葡萄糖)=5.14:5.78:81.37;该法可为天然产物活性成分提取和纯化提供借鉴作用。
Three-phase partitioning (TPP), which is a simple, efficient, and green bioseparation technique, was used to extract and separate polysaccharides from Spriulina platensis (PSP). The effect of mass fraction of (NH4)2SO4, amount of t-butanol, pH, temperature, and extraction time on the yield of PSP were studied. The structural feature of extracted PSP were also analyzed. The optimal parameters with a high yield of (9.25±0.42)% were as follows: mass fraction of (NH4)2SO4, 20.0%; amount of t-butanol, 15 mL; pH 7.0; temperature, 35 ℃; and extraction time, 30 min. The results of IR showed that PSP had the characteristic absorption peaks of polysaccharide, UV wavelength scanning result showed that PSP do not contain protein and nucleic acids. The weight average molecular of PSP determined by gel permeation chromatography can reach 3.597×104, which is composed of Rhamnose, mannose, and glucose in a molar ratio of 5.14:5.78:81.37, respectively. Hence, TPP was an excellent efficient to concurrent extraction and purification polysaccharides.
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