酶法辅助提取巴戟天中多糖及耐斯糖工艺研究

目的：筛选生物酶辅助提取巴戟天多糖及耐斯糖的最佳水提工艺。方法：在单因素试验基础上，采用二次通用旋转组合设计优化工艺。测定多糖含量采用硫酸比色法，对照品为0.1 mg·mL^-1无水葡萄糖标准溶液，显色剂为5%苯酚溶液，489 nm波长处紫外分光光度仪下测定吸收度。测定耐斯糖含量采用高效液相色谱法，色谱柱为Agilent ZORBAX SB-C₁₈（5 μm，4.6 mm×150 mm），流动相为甲醇-水（3：97），流速为0.8 mL·min^-1，漂移管及雾化温度为40和80℃。结果：生物酶辅助提取巴戟天多糖及耐斯糖的最佳水提工艺为：pH 5.0、加入0.18%的纤维素酶，55℃酶解2.0 h，20倍量水提取3.5 h，提取3次，巴戟天多糖得率为10.44%，耐斯糖得率为0.083 g·g^-1。结论：二次通用旋转组合设计所得的优化工艺可靠、稳定，对巴戟天的开发利用具有一定意义。

Objective: To screen the optimum water extraction technique for the extraction of polysaccharide in Morindae Officinalis Radix and Nisi sugar by biological enzyme. Methods: On the basis of single factor test,the second general rotation combination design was used to optimize the extract process. Polysaccharides were determined at 489 nm by phenol-sulfuric acid method with 0.1 mg·mL^-1 glucose solution as standard. Nystose was determined by HPLC with Agilent ZORBAX SB-C₁₈(5 μm,4.6 mm×150 mm) as the column and methanol-water(3:97) as the mobile phase. The flow rate was 0.8 mL·min^-1,and the drift tube and atomization temperature were at 40℃ and 80℃,respectively. Results: The extraction conditions were as follows:pH 5.0,adding 0.18% cellulase, digested at 55℃ for 2.0 h,20 times of water for 3.5 h,extracted 3 times. The yield rate of Morinda polysaccharide was 10.44% and the ield rate of Nisi sugar was 0.083 g.g-1. Conclusion: The optimized technology is reliable and stable,which is significant for development and utilization of polysaccharide in Morindae Officinalis Radix.