树皮藻岩藻聚糖硫酸酯的纯化及其化学结构研究

Purification and chemical structure of fucoidan extracted from brown seaweed Ecklonia maxima

为研究树皮藻Ecklonia maxima岩藻聚糖硫酸酯的单糖组成及其化学结构,以产自南非的树皮藻为原料测定其基本成分,采用DEAE-Sepharose Fast Flow阴离子层析柱对树皮藻岩藻聚糖硫酸酯粗品进行分离纯化,通过柱前衍生高效液相色谱法、红外光谱法和核磁共振波谱法测定单糖组成和化学结构。结果表明:树皮藻干基中基本营养成分分别为灰分18.59%、蛋白质9.16%、脂肪0.74%;对树皮藻岩藻聚糖硫酸酯粗品进行纯化得到4个组分,分别命名为SPF1、SPF2、SPF3、SPF4,收集冻干各组分,检测各组分得率分别为18.19%、24.02%、5.61%、4.23%,硫酸根含量分别为12.07%、5.44%、15.58%、19.30%;4个组分主要由岩藻糖、半乳糖、鼠李糖、甘露糖和葡萄糖醛酸组成,还含有少量的葡萄糖和木糖,其中SPF3、SPF4主要由岩藻糖和半乳糖组成,岩藻糖含量分别为70.92%和86.20%,4个组分的红外光谱显示出典型的多糖吸收峰,SPF1、SPF2在13C核磁共振谱177×10-6附近均出现糖醛酸C6信号峰,SPF3、SPF4在18×10-6附近均出现信号峰,为α-L-吡喃岩藻糖C6信号峰。

The basic components were determined in brown seaweed Ecklonia maxima from South Africa and then crude fucoidan was subjected to a DEAE column chromatography, with four fractions named as SPF1 , SPF2 , SPF3, and SPF4. It was found that the seaweed had ash content of 18. 5 9 % , protein content of 9. 16% , and fat content of 0. 74% on the dry basis. T he four fractions were collected and freezing dried, and then the yield, carbo-hydrate content and sulphate content were measured. T he results showed that the yield and sulphate content were shown to be 18. 19% and 12. 07% in S P F1 , 24. 0 2 % and 5. 4 4 % in S P F2, 5. 6 1 % and 15. 5 8 % in S P F3 ,and 4.23% and 19.30% in SPF4. Th e monosaccharide compositions and structural features of each fraction were ana-lyzed by HPLC , FTIR , and NMR spectroscopy. T he monosaccharides S P F1 - S P F4 was primarily composed of fu- cose, galactose, rhamnose, mannose, and glucuronic acid, with small amount of glucose and xylose, in which fu- cose （70. 92%） and galactose （86. 2 0 % ） were the main constituents in S P F3 and S P F4. Th e IR spectra of the four fractions showed the typical absorption bands of polysaccharides, with signal peak of glucuronic acid C6 in SPF1 and SPF2 in the vicinity of 177×10^-6 in l3C NMR spectroscopy. Th e 18×10^-6 regions of SPF3 and S P F4 were assigned to C6 of α-L -fucopyranose.