| 罗恒,何燕利,韩章润,等.运用酶解及氘代PMP衍生方法对鲨鱼硫酸软骨素结构的液质联用分析[J].中国海洋药物,2020,39(3):51-57. |
| 运用酶解及氘代PMP衍生方法对鲨鱼硫酸软骨素结构的液质联用分析 |
| Structural Analysis of Chondroitin Sulfate from Shark Cartilage by LC-MS Analysis of Enzyme-digested and Deuterated PMP-labeled Products |
| 投稿时间：2019-05-28 修订日期：2019-10-27 |
| DOI： |
| 中文关键词: 硫酸软骨素,氘代PMP,液质联用 |
| English Keywords:Chondroitin sulfate, Deuterated PMP, LC-MS |
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| 中文摘要: |
| 硫酸软骨素是一种具有生物活性、由N-乙酰半乳糖胺和葡萄糖醛酸作为重复二糖单元组成的线性多糖。 硫酸软骨素广泛存在于结缔组织、皮肤组织、关节滑液中,在人和动物软骨组织中含量最高。在欧洲,硫酸软骨素是治疗关节炎的临床药物；在美国,硫酸软骨素是销量排名第二的保健品。不同品种、年龄等动物软骨来源的硫酸软骨素的含量、结构与生物活性均不完全相同。由于鲨鱼软骨有抗癌作用,来源于鲨鱼软骨的硫酸软骨素价格昂贵,鉴定鲨鱼来源的硫酸软骨素结构始终是一个重要而没有理想方法的科学问题。在本篇文章中,我们首先合成了三种氘代3-甲基-1-苯基-2-吡唑啉-5-酮 (PMP),进而运用4种不同分子量的PMPs(d0-,d3-,d5-,及d8-PMP)对酶解后的鲨鱼硫酸软骨素进行了衍生,随后的液质联用分析共鉴定了7种二糖及4种含2、3、4个硫酸根的四糖结构。质谱结果表明四种PMPs对降解的各种二糖与四糖的衍生效率相当,从而为同时分析比较不同批次及不同来源的鲨鱼硫酸软骨素的结构提供了一种新型、快速、准确的定量定性方法。 |
| English Summary: |
| Chondroitin sulfate is a biologically active linear polysaccharide consisting of N-acetylgalactosamine and glucuronic acid as repeating disaccharide units. Chondroitin sulfate is widely found in connective tissue, skin, synovial fluid, and mostly enriched in human and animal cartilage. In Europe, chondroitin sulfate is a clinical drug for the treatment of arthritis; in the United States, chondroitin sulfate is ranked number two in market share among all healthy supplements. The content, structure, and biological activities of cartilage chondroitin sulfate vary greatly depending on the source of cartilage and the ages of the animals. Because shark cartilage has anti-cancer effect, chondroitin sulfate isolated from shark cartilage is very expensive. Up to date, correctly identifying shark-derived chondroitin sulfate is still a scientifically challenging issue. In current study, we first synthesized three kinds of deuterated 3-methyl-1-phenyl-2-pyrazolin-5-one (PMP), and then used the four different molecular weight PMPs (d0-, d3 - d5-, and d8-PMP) to derive the chondroitinase ABC-digested shark chondroitin sulfate. By LC-MS analysis, the structures of seven disaccharides and four tetrasaccharides containing 2,3,4-sulfates, respectively, from shark cartilage chondroitin sulfate were identified. MS analysis further showed that the four PMPs had the comparable derivatization efficiency for all di- and tetrasaccharides. Thus, our study provided a new, rapid and accurate method that made the simultaneous analysis and comparison of the structures of shark chondroitin sulfate from different batches or sources possible. |
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