引用本文:	张齐,王晶,谭姣姣,张全斌,秦浩然.低硫酸化杂多糖水解单糖高效液相色谱指纹图谱研究[J].海洋科学,2017,41(12):96-103.

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低硫酸化杂多糖水解单糖高效液相色谱指纹图谱研究
张齐^1,2,3, 王晶^1,2, 谭姣姣^1,2,3, 张全斌^1,2, 秦浩然⁴
1.中国科学院海洋研究所;2.青岛海洋科学与技术国家实验室, 海洋生物学与生物技术功能实验室;3.中国科学院大学;4.青岛市实验高级中学

摘要:

低硫酸化杂多糖(UF)是从海洋褐藻中分离的一种单糖组成复杂的硫酸化杂多糖, 在细胞和动物实验中均表现出显著的神经保护活性, 对帕金森病具有显著作用。本研究采用水提-醇沉-DEAE-Sepharose Fast Flow交换层析柱制备UF, 经三氟乙酸(TFA)水解和1-苯基-3-甲基-5吡唑啉酮(1-phenyl-3-methyl-5-pyrazalone, PMP)衍生化, 通过对水解条件、衍生化试剂使用量、衍生时间的考察,确定最佳衍生条件, 通过高效液相色谱(HPLC)法测定UF的单糖组成, 并建立UF的指纹图谱。在UF水解单糖的指纹图谱中, 共标示出8个特征共有峰, 经鉴别主要色谱峰8个, 分别是D-甘露糖、D-甘露糖醛酸、L-鼠李糖、D-葡萄糖醛酸、D-葡萄糖、D-半乳糖、D-木糖及L-岩藻糖, 其中岩藻糖含量最高, 其次是半乳糖。该方法操作简便、快速、灵敏度高、重复性好, 可作为UF检查和鉴别的方法, 建立实验室UF质量控制标准, 为临床用UF质量标准的建立奠定基础。

关键词: 低硫酸化杂多糖岩藻多糖指纹图谱柱前衍生化高效液相色谱

DOI：10.11759/hykx20170627001

分类号:

基金项目:山东省重点研发计划项目(2016GSF115031); 中国科学院战略生物资源网络服务计划项目(ZSTH-0025); 国家自然科学基金(41406144); 中国科学院青年创新促进会(2016190)

HPLC fingerprints of monosaccharides from sulfated heteropolysaccharides by precolumn derivatization

ZHANG Qi,WANG Jing,TAN Jiao-jiao,ZHANG Quan-bin,QIN Hao-ran

Abstract:

Sulfated heteropolysaccharide (UF) is a novel marine polysaccharide extracted from Laminaria japonica, with a complex chemical composition and structure. UF showed a significant neuroprotective activity both in cell and animal studies. UF was extracted with hot distilled water and further fractionated by DEAE-Sepharose Fast Flow column chromatography. The optimal hydrolysis and PMP derivatization conditions were investigated. UF was hydrolyzed into monosaccharides by trifluoroacetic acid and derivated with 1-phenyl-3-methyl-5-pyrazolone and analyzed by HPLC to study the monosaccharide components and fingerprint of UF. Nine peaks were detected in the fingerprint, of which 8 were identified as D-mannose, D-Mannuronic acid, L-rhamnose, D-glucuronic acid, D-glucose, D-galactose, D-xylose, and L-fucose. Fucose content was the highest, followed by galactose content. The result shows that the method is convenient, sensitive, and rapid such that it can be applied for the quality control of UF.

Key words: Sulfated heteropolysaccharides fucoidan fingerprint precolumn HPLC