新型香豆素荧光试剂的寡糖标记研究
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- 英文论文题名：Application of a novel coumarin reagent for the analysis of oligosacchrides
- 论文作者：丁慢慢 ; 林亚维
- 英文论文作者：Manman Ding ; Yawei Lin ; Department of Chemistry School of Chemistry ; Chemical engineering and Life Sciences ; Wuhan University of Technology
- 年：2016
- 作者机构：武汉理工大学化学化工与生命科学学院化学系湖北省武汉市武汉理工大学南湖校区;
- 论文关键词：柱前衍生 ; 寡糖 ; 肟化反应 ; 高效液相色谱-荧光检测法 ; 香豆素
- 会议召开时间：2016-07-01
- 会议录名称：中国化学会第30届学术年会摘要集-第二十三分会：复杂样品分离分析
- 语种：中文
- 分类号：O657.3;O629.1
- 学会代码：ZGHY
- 会议名称：中国化学会第30届学术年会-第二十三分会 ; 复杂样品分离分析
- 会议地点：中国辽宁大连
- 主办单位：中国化学会
- 学会名称：中国化学会
- 页数：1
- 文件大小：206k
- 原文格式：D
- 会议级别：全国
摘要
柱前衍生高效液相色谱荧光检测法(HPLC-FD)是糖分析的重要方法之一。不同于常规的还原胺化反应,本研究采用新型荧光衍生试剂(4-胺氧基亚甲基-6-氯-7-羟基-香豆素),通过聚糖还原端与胺氧基的缩合反应(肟化反应)对麦芽七糖进行了成功的标记。采用ODS反相柱,HPLC-FD监控(λ_(ex)=330 nm,λ_(em)=470nm)七糖与胺氧基香豆素反应体系,观察到三个荧光衍生产物峰在10分钟内(等度洗脱,乙腈:水=15:85,v/v%)达到很好的分离。由电喷雾质谱分析,这三种衍生产物互为异构体,分别是肟式产物和五元吡喃环与六元呋喃环结构产物。产物峰峰面积在优化后的衍生条件下(反应时间3.5 h,反应温度75℃,乙酸/乙酸钠缓冲液(pH=3.0,0.2 M),香豆素与七糖的摩尔比为50/1,苯胺与七糖的摩尔比为100/1)达到最大。该研究通过采用新型荧光衍生试剂与肟化反应,为糖的定量检测奠定了良好的基础。
Oxime formation has been investigated as a simple and alternative labeling procedure to conventional reductive animation,using a novel fluorescent reagent 4-((aminooxy) methyl)-6-chloro-7-hydroxy-coumarin.The coumarin derivative was successfully applied to tag the reducing end of a model oligosaccharide,maltoheptaose.The reaction was monitored by HPLC-FD at 470 nm after excitation at 330 nm.Three fluorescent derivatization products were well separated in 10 mins on a ODS reversed-phase column with isocratic elution with ACN:H_2O(15:85,v/v%).ESI-MS analysis shows the derivatization products are interconvertible isomers,including acyclic oxime,cyclic pyranose and furanose products.Under the optimal reaction conditions,the products peak areas reach maximum.The research used a novel fluorescent reagent,investigated oxime formation derivatization procedure,and indicates an alternative labeling procedure for further analysis of oligosaccharides.
Oxime formation has been investigated as a simple and alternative labeling procedure to conventional reductive animation,using a novel fluorescent reagent 4-((aminooxy) methyl)-6-chloro-7-hydroxy-coumarin.The coumarin derivative was successfully applied to tag the reducing end of a model oligosaccharide,maltoheptaose.The reaction was monitored by HPLC-FD at 470 nm after excitation at 330 nm.Three fluorescent derivatization products were well separated in 10 mins on a ODS reversed-phase column with isocratic elution with ACN:H_2O(15:85,v/v%).ESI-MS analysis shows the derivatization products are interconvertible isomers,including acyclic oxime,cyclic pyranose and furanose products.Under the optimal reaction conditions,the products peak areas reach maximum.The research used a novel fluorescent reagent,investigated oxime formation derivatization procedure,and indicates an alternative labeling procedure for further analysis of oligosaccharides.
引文
[1]Ramsay S.,F.C.,Grace P.,et al.,Carbohyd.Res.,2001.333:59
[2]X.Zhou.,X.C.Weng.,et al.,American Chemical Society.,2013.15:3266
[2]X.Zhou.,X.C.Weng.,et al.,American Chemical Society.,2013.15:3266