过氧化氢抗坏血酸法制备小分子量鲍鱼脏器多糖工艺研究

Study on preparation processing of low molecular weight abalone viscera polysaccharide by hydrogen peroxide combined with ascorbic acid method

目的 采用过氧化氢抗坏血酸法制备小分子量鲍鱼脏器多糖。方法 利用过氧化氢抗坏血酸法降解鲍鱼脏器多糖,通过单因素实验考察反应体系中过氧化氢浓度、反应时间和反应温度对多糖水解率的影响,在单因素实验基础上,结合Box-Behnken响应面分析法对影响多糖水解率的3个主要因素:过氧化氢浓度、反应时间和反应温度进行优化;利用葡聚糖凝胶层析法分离纯化降解后的多糖组分,并通过高效液相色谱法测定多糖分子量。结果 各因素对多糖水解的影响大小为:过氧化氢浓度>温度>时间。在过氧化氢浓度0.2%、反应时间3h、反应温度60℃条件下,多糖水解率最高,为69.85%。分离纯化后的小分子量鲍鱼脏器多糖为单组分,通过高效液相色谱法测得该多糖分子量为4011 Da。结论 采用过氧化氢抗坏血酸法降解鲍鱼脏器多糖,可进一步为鲍鱼脏器深加工提供参考。

Objective To prepare low molecular weight abalone viscera polysaccharide by hydrogen peroxide combined with ascorbic acid method. Methods The polysaccharide in abalone viscera was degraded by hydrogen peroxide combined with ascorbic acid method. The effects of hydrogen peroxide concentration, reaction time and reaction temperature on the hydrolysis rate of polysaccharide in the reaction system were investigated by single factor experiment. On the basis of single factor experiment, the 3 main factors affecting the hydrolysis rate of polysaccharide were optimized by Box-Behnken response surface analysis method: Hydrogen peroxide concentration, reaction time and reaction temperature;the degraded polysaccharides were separated and purified by sephadex gel chromatography, and the molecular weight of polysaccharides was determined by high performance liquid chromatography. Results The influence of various factors on the hydrolysis of polysaccharides were hydrogen peroxide concentration, temperature and time in turns. The best hydrolysis rate of polysaccharide was 69.85% under the concentration of hydrogen peroxide of 0.2%, the reaction time of 3 h, and the reaction temperature of 60℃. The low molecular weight polysaccharide was single component after separation and purification. The molecular weight was determined by high performance liquid chromatography to be 4011 Da. Conclusion The abalone viscera polysaccharide can be degraded by hydrogen peroxide ascorbic acid method, which provides reference for further processing of abalone viscera.