苷苔岩藻聚糖硫酸酯分离纯化及结构研究

摘要

Crude polysaccharide was separated from fucoidan ( EC-FUC ) extracted from brwn seaweed Ecklonia cava sampled from coast of Cheju Island in R. O. Korea by DEAE-Sepharose fast flow anion exchange chromatog-raphy, and future purified by Sephacryl S-300, S-400 gel column chromatography. The molecular weight and con-tent of polysaccharide and sulfate of the EC-FUC were determined by phenol sulfuric acid method and BaCl2 gelatin method, respectively. High performance liquid chromatography, infrared spectroscopy and nuclear magnetic reso-nance spectroscopy were used to analyze the structural characterization of purified components. The results showed that the purified EC-FUC was comprised of three peaks including ECF-1 with total polysaccharide of 41.7% and sulfate content of 3.68%, ECF-2 with total polysaccharide of 57.79% and sulfate content of 3.41%, and ECF-3 with total polysaccharide of 75.78%, and sulfate content of 18.03%. The yields were 6.27% in ECF-1, 8.77% in ECF-2 and 16.19% in ECF-3, with molecular weight of 16430 in ECF-1, 16430 in ECF-2 and 222460 in ECF-3. The primary monosaccharide was shown to be glucose in ECF-1, mannuronic acid, glucuronic acid and fucose in ECF-2, and glucuronic acid and fucose in ECF-3. Infrared spectrum analysis revealed that the sulfated group was combined to C2 or C3 in the three fractions and 13 C nuclear magnetic resonance ( NMR) analysis indica-ted that there was mannuronic acid in ECF-2 and that 17.97×10-6 regions of ECF-3 was assigned to C6α-L-fuco-pyranose.%为探究苷苔Ecklonia cava岩藻聚糖硫酸酯粗品(EC-FUC)的组成结构,以产自韩国济州岛海岸的苷苔Ecklonia cava中提取的EC-FUC为原料,利用DEAE-Sepharose fast flow阴离子交换层析和Sephacryl S-300、S-400凝胶柱层析对EC-FUC进行分离纯化及分子量的测定,采用苯酚-硫酸法和明胶-BaCl2法分别测定纯化各组分的总糖含量及硫酸根含量,采用高效液相色谱法、 红外光谱法、 核磁共振波谱法分别测定各个纯化组分的单糖组成并分析多糖结构.结果表明:EC-FUC纯化后得到ECF-1、ECF-2和ECF-3组分,其总糖含量分别为41.7%、57.79%、75.78%,硫酸根含量分别为3.68%、3.41%、18.03%,回收率分别为6.27%、8.77%、16.19%,其相对分子质量分别为16430、16430、222460;ECF-1单糖组成主要为葡萄糖,ECF-2单糖组成主要为甘露糖醛酸、 葡萄糖醛酸和岩藻糖,ECF-3单糖组成主要为岩藻糖和葡萄糖醛酸;红外光谱分析显示,ECF-1、ECF-2和ECF-3硫酸基主要连接在岩藻糖的C2、C3位置;13 C核磁共振分析显示,ECF-2中存在甘露糖醛酸,ECF-3在17.97×10-6处出现α-L-吡喃岩藻糖C6信号峰.本研究结果可为苷苔藻聚糖硫酸酯组成结构提供丰富的信息.