真鱿软骨硫酸软骨素的提取、鉴定及结构分析  

Extraction and structural characterization of chondroitin sulfate from squid cartilage(Loligo chinesis)

作　　者：胡靖[1] 陶圣男 李八方[1] 候虎 张朝辉[1] 赵雪[1] 

机构地区：[1]中国海洋大学食品科学与工程学院,山东青岛266000

出　　处：《中国海洋药物》2016年第3期37-42,共6页Chinese Journal of Marine Drugs

基　　金：国家自然科学基金项目(30800858);国家自然科学基金-山东省联合基金专项(U1406402)资助

摘　　要：目的采用酶解提取法,从真鱿软骨中提取1种新型的硫酸软骨素,对其化学结构进行研究。方法采用酶解法、氯化十六烷基吡啶(CPC)沉淀法从鱿鱼软骨中提取纯化硫酸软骨素,通过PMP柱前衍生液相色谱法、聚丙烯酰氨凝胶电泳(PAGE)、核磁共振波谱法(NMR)和液质联用分析(HPLC-MS),对硫酸软骨素的化学结构进行分析。结果使用木瓜蛋白酶酶解鱿鱼软骨,乙酸缓冲液在料液比为1∶30(g/mL)条件下,硫酸软骨素的提取率最高,蛋白质残留最低,经过CPC纯化,得到了纯度高达99.5%的硫酸软骨素。由PAGE电泳可以看出其分子量很大且能够被硫酸软骨素ABC酶酶解。单糖分析表明,鱿鱼硫酸软骨素主要由乙酰氨基半乳糖、葡萄糖醛酸以及少量的葡萄糖组成。硫酸软骨素ABC酶将其水解为聚合度2-18的偶数糖,采用HPLC-MS分析,可以得到所有寡糖的全指纹谱图。结合1H-NMR分析可知,从真鱿软骨提取出来的硫酸软骨素主要含有6-单硫取代的硫酸软骨素C二糖单元和4,6-双硫取代硫酸软骨素E二糖单元,其所占比例分别为60.3%和39.7%,硫酸根含量为22.3%,是属于高硫的硫酸软骨素。结论从真鱿软骨中提取出了具有高硫酸根含量的硫酸软骨素组分,可用于开发保健食品和海洋药物。Objective The extraction and chemical character of chondroitin sulfate from squid cartilage were studied.Methods The chondroitin sulfate was extracted after enzymatic hydrolysis and purified with Cetylpyridinium Chloride（CPC）.The purity and molecular weight of chondroitin sulfate were examined by carbazole method and PAGE.The chemical structure of chondroitin sulfate was analyzed by PMP reverse-phased HPLC,NMR and HPLC-MS.Results The chondroitin sulfate was extracted from squid cartilage with acetate buffer at ratio of 1∶30（g/mL）after hydrolyzed by papain.After CPC purification,the purity of chondroitin sulfate reached 99.5%.PAGE demonstrated the chondroitin sulfate＇s molecular weight is large,and the chondroitin sulfate could be completely digested by chondroitin sulfate ABC enzyme.Monosaccharide composition analysis demonstrated that the chondroitin sulfate was composed of acetyl galactosamine,glucuronic acid and a small amount of glucose.HPLC-MS analysis showed chondroitin sulfate was hydrolyzed by chondrotinase ABC to even-numbered oligosaccharides from dp2-dp18 and showed the mapping of all oligosaccharides.LC-MS and NMR analysis showed that chondroitin sulfate mainly contained 6-monosulfated chondroitin sulfate C disaccharide（60.3%）and 4,6-disulfated chondroitin sulfate disaccharide（39.7%）,and sulfate content was 22.3%.Conclusion The chondroitin sulfate from squid cartilage was a kind of highly sulfated polysaccharide with good potential as functional food and drug.

关 键 词：真鱿 硫酸软骨素 提取 结构 

分 类 号：R931[医药卫生—生药学]