寡糖的基质辅助激光解吸电离飞行时间质谱分析方法研究
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摘要
寡糖(Oligosaccharides)是生物体内重要的信息物质。寡糖的诸多生物学功能决定了它在医药领域有着广泛的应用前景。近些年来对寡糖的生物活性研究的非常多,表现在免疫调节功能、抗肿瘤、抗病毒、抗氧化、抗凝血、抗血栓、降血糖、降血脂等多种作用。寡糖作为一类新的生理活性物质,在营养与保健、疾病诊断与防治方面的应用有着极大潜力。
基质辅助激光解吸电离飞行时间质谱(matrix-assisted laser desorption ionization time of flight mass spectrometry MALDI-TOF- MS)是最近发展起来的一种软电离质谱分析技术,与传统分析技术如气相色谱技术、液相色谱技术、毛细管电泳技术及其他质谱电离源分析技术相比较, MALDI具有灵敏度高;高通量;对样品的纯度要求不高,可以直接分析未处理过的生物样品等许多优点; MALDI-TOF-MS能提供寡糖直观的分子量和结构信息,实现检测和鉴定的目的。
本研究通过对MALDI-TOF-MS分析中性寡糖和酸性寡糖的方法进行系统的研究,旨在找到合适的分析中性与酸性寡糖的基质,阐明影响MALDI分析的重要实验因素的作用,对基质作用的机理进行了探讨,并建立MALDI–TOF- MS分析寡糖的实验方法,从而实现对不同类型寡糖的分析,丰富了寡糖的分离分析技术。
根据研究目的,选择以下寡糖进行试验:中性寡糖包括还原型的中性寡糖和非还原型的中性寡糖;酸性寡糖包括含糖醛酸的酸性寡糖,含唾液酸的酸性寡糖和含硫酸基团的酸性寡糖。在试验中,以葡聚糖、棉子糖和古罗糖醛酸三糖为寡糖模型,试验了不同的基质包括单一基质和混合基质,和不同的实验因素包括基质、基质浓度、混合基质配比、点样方法,检测模式。通过MALDI-TOF-MS分析提供的各种寡糖的质量信息,应用MassLynx V4.0软件进行比较分析,确定标准的实验方法和影响分析结果的重要实验因素。在标准实验方法建立后,
Oligosaccharides is an important information matter in organism. Many biological functions of oligosaccharides make it has the widespread application prospect in the medicine domain. In recent years, the study of oligosaccharides biological activity become a hot point, shows in aspects of the immunity adjustment function, the anti-tumor, the anti-virus, the anti-oxidation, the anti-hemoglutination and the anti- thrombus activities. As a new category of biological active substances, Oligosaccharides has great potential in the nutrition and the health care, disease diagnosis and control applications.
The matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS) is one kind of“soft”ionization mass spectrometry which developed recently, compare to traditional analysis instrument like GC, HPLC, CE and other mass spectrometry ionization sources, MALDI has many advantages: high sensitivity, high flux; providing the precise molecular weight, low sample purity request. MALDI-TOF-MS can provide direct-viewing molecular weight and structure information of oligosaccharides. The aim of this research is to find out the appropriate matrix that can analyze neutral or acidic oligosaccharides, to choose experimental parameters that affecting the results when analyzing oligosaccharides by MALDI–TOF-MS, to discuss the matrix function mechanism, and to set up a general experimental method for analyze oligosaccharides. It has enriched the separation analysis technology of oligosaccharides.
According to the goal of studies, chooses oligosaccharides below to carry on the experiment: Neutral oligosaccharides including reducing neutral oligosaccharides and non-reducing neutral oligosaccharides; Acidic oligosaccharide including acidic oligosaccharides contains the uronic acid, contains acidic oligosaccharides contains the sialinic acid and acidic oligosaccharides contains the sulfur acid groups. In the
基质辅助激光解吸电离飞行时间质谱(matrix-assisted laser desorption ionization time of flight mass spectrometry MALDI-TOF- MS)是最近发展起来的一种软电离质谱分析技术,与传统分析技术如气相色谱技术、液相色谱技术、毛细管电泳技术及其他质谱电离源分析技术相比较, MALDI具有灵敏度高;高通量;对样品的纯度要求不高,可以直接分析未处理过的生物样品等许多优点; MALDI-TOF-MS能提供寡糖直观的分子量和结构信息,实现检测和鉴定的目的。
本研究通过对MALDI-TOF-MS分析中性寡糖和酸性寡糖的方法进行系统的研究,旨在找到合适的分析中性与酸性寡糖的基质,阐明影响MALDI分析的重要实验因素的作用,对基质作用的机理进行了探讨,并建立MALDI–TOF- MS分析寡糖的实验方法,从而实现对不同类型寡糖的分析,丰富了寡糖的分离分析技术。
根据研究目的,选择以下寡糖进行试验:中性寡糖包括还原型的中性寡糖和非还原型的中性寡糖;酸性寡糖包括含糖醛酸的酸性寡糖,含唾液酸的酸性寡糖和含硫酸基团的酸性寡糖。在试验中,以葡聚糖、棉子糖和古罗糖醛酸三糖为寡糖模型,试验了不同的基质包括单一基质和混合基质,和不同的实验因素包括基质、基质浓度、混合基质配比、点样方法,检测模式。通过MALDI-TOF-MS分析提供的各种寡糖的质量信息,应用MassLynx V4.0软件进行比较分析,确定标准的实验方法和影响分析结果的重要实验因素。在标准实验方法建立后,
Oligosaccharides is an important information matter in organism. Many biological functions of oligosaccharides make it has the widespread application prospect in the medicine domain. In recent years, the study of oligosaccharides biological activity become a hot point, shows in aspects of the immunity adjustment function, the anti-tumor, the anti-virus, the anti-oxidation, the anti-hemoglutination and the anti- thrombus activities. As a new category of biological active substances, Oligosaccharides has great potential in the nutrition and the health care, disease diagnosis and control applications.
The matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS) is one kind of“soft”ionization mass spectrometry which developed recently, compare to traditional analysis instrument like GC, HPLC, CE and other mass spectrometry ionization sources, MALDI has many advantages: high sensitivity, high flux; providing the precise molecular weight, low sample purity request. MALDI-TOF-MS can provide direct-viewing molecular weight and structure information of oligosaccharides. The aim of this research is to find out the appropriate matrix that can analyze neutral or acidic oligosaccharides, to choose experimental parameters that affecting the results when analyzing oligosaccharides by MALDI–TOF-MS, to discuss the matrix function mechanism, and to set up a general experimental method for analyze oligosaccharides. It has enriched the separation analysis technology of oligosaccharides.
According to the goal of studies, chooses oligosaccharides below to carry on the experiment: Neutral oligosaccharides including reducing neutral oligosaccharides and non-reducing neutral oligosaccharides; Acidic oligosaccharide including acidic oligosaccharides contains the uronic acid, contains acidic oligosaccharides contains the sialinic acid and acidic oligosaccharides contains the sulfur acid groups. In the
引文
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[3] 袁华茂. 卡拉胶寡糖与衍生物的制备及生物活性研究.博士学位论文,2005
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