酸降解-液相色谱-质谱分析技术鉴别硫酸软骨素
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- 英文论文题名：Identification of chondroitin sulfate using acid hydrolysis and HPLC-MS~n analysis
- 论文作者：鲁姣姣 ; 刘斌 ; 徐鑫 ; 郭丽 ; 艾春青 ; 宋爽
- 英文论文作者：Lu Jiaojiao ; Liu Bin ; Xu Xin ; Guo Li ; Ai Chunqing ; Song Shuang ; School of Food Science and Technology ; Dalian Polytechnic University ; National Engineering Research Center of Seafood ; Collage of food and pharmaceutical engineering ; Suihua University
- 年：2016
- 作者机构：大连工业大学食品学院;国家海洋食品工程技术研究中心;绥化学院食品与制药工程学院;
- 论文关键词：硫酸软骨素 ; 糖胺聚糖 ; 酸降解 ; 寡糖 ; 液相色谱质谱法
- 英文论文关键词：chondroitin sulfate ; glycosaminoglycan ; acid hydrolysis ; oligosaccharide ; HPLC-MS~n
- 会议召开时间：2016-11-09
- 会议录名称：中国食品科学技术学会第十三届年会论文摘要集
- 英文会议录名称：Abstracts of the 13th Annual Meeting of CIFST
- 语种：中文
- 分类号：TS207.3;O657.63
- 学会代码：ZGSP
- 会议名称：中国食品科学技术学会第十三届年会
- 会议地点：中国北京
- 主办单位：中国食品科学技术学会
- 学会名称：中国食品科学技术学会
- 页数：2
- 文件大小：113k
- 原文格式：O
- 会议级别：全国
摘要
鉴定复杂生物基质中的硫酸软骨素(chondroitin sulfate)较为困难,不仅操作过程繁琐,耗时长,且对样品的纯度要求高。本研究建立了一种酸降解结合1-苯基-3-甲基-5-吡唑啉酮(1-phenyl-3-methyl-5-pyrazolone,PMP)衍生化的液相色谱串联质谱技术(high-performance liquid chromatography-mass spectrometry,HPLC-MS~n)分析硫酸软骨素的方法。首先,对猪骨和鲨鱼硫酸软骨素标准品进行酸水解,降解产物经PMP衍生化后,利用HPLC-MS~n进行检测。经过多级质谱分析,鉴定出了包括二糖、三糖和四糖在内的10种寡糖,其中一些寡糖还带有硫酸基或乙酰基。同时,我们考察了三氟乙酸浓度(0.1~2.0M)对这些寡糖产率的影响,结果表明三氟乙酸的最优浓度为0.2mol/L。然后,将该分析方法应用于经过简单提取的鲟鱼脊骨和猪喉管中硫酸软骨素的检测,以这10种寡糖为依据,实现了这两种生物样品中硫酸软骨素的快速鉴定,证明该方法能在短时间内有效分析基质复杂的生物样品中的硫酸软骨素。
It is challenging to identify chondroitin sulfate(CS) from a complex biological matrix,and the process is typically time-consuming and labor intensive,requiring isolation and multistep purification.Therefore,a method based on acid hydrolysis and high performance liquid chromatography-mass spectrometry(HPLC-MS ") analysis upon l-phenyl-3-methyl-5-pyrazolone(PMP) derivatization has been explored.Firstly,acid hydrolysates of commercial CS reference standards from shark cartilage and porcine bone were elucidated using HPLC-MS" after PMP derivatization,and altogether 10 di-,tri- and tetra- saccharides with or without sulfate/acetyl groups were identified by their multi-stage mass spectra.Then the trends of reaction yields of these oligosaccharides alone with trifluoroacetic acid(TFA) concentrations(0.1 ~2.0 M) were investigated,and 0.2 M TFA was recommended.Finally,using these 10 oligosaccharide derivatives as markers,the present method accomplished the identification of CS in sturgeon backbone and porcine trachea after a simple isolation procedure,proving its powerful potent to profile CS in a complex biological matrix within short time.
It is challenging to identify chondroitin sulfate(CS) from a complex biological matrix,and the process is typically time-consuming and labor intensive,requiring isolation and multistep purification.Therefore,a method based on acid hydrolysis and high performance liquid chromatography-mass spectrometry(HPLC-MS ") analysis upon l-phenyl-3-methyl-5-pyrazolone(PMP) derivatization has been explored.Firstly,acid hydrolysates of commercial CS reference standards from shark cartilage and porcine bone were elucidated using HPLC-MS" after PMP derivatization,and altogether 10 di-,tri- and tetra- saccharides with or without sulfate/acetyl groups were identified by their multi-stage mass spectra.Then the trends of reaction yields of these oligosaccharides alone with trifluoroacetic acid(TFA) concentrations(0.1 ~2.0 M) were investigated,and 0.2 M TFA was recommended.Finally,using these 10 oligosaccharide derivatives as markers,the present method accomplished the identification of CS in sturgeon backbone and porcine trachea after a simple isolation procedure,proving its powerful potent to profile CS in a complex biological matrix within short time.
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