内部萃取电喷雾电离质谱技术在动物组织分析中的应用研究

发布时间：2018-07-17 06:16

【摘要】：动物组织样品的分析是从分子层面对其本身及其代谢产物的分析,在医学、食品安全等领域的研究中具有重要意义。但是传统的分析方法大都需要复杂的研磨、萃取、分离等预处理过程,不仅冗长而耗时,更严重的是会损失组织样品内部一些重要的物质信息,从而导致分析效率低下。因此,开发出一种“绿色、快速、无损”的组织样品检测分析方法就显得非常有意义。内部萃取电喷雾电离质谱技术(iEESI-MS)是近年兴起的常压质谱技术,其独特的优势是可以在无需任何样品前处理的条件下直接获取复杂基体固形样品内部组分的化学信息,具有操作简单、分析速度快、高选择性、高灵敏度、高通量等优点。本文着重研究了iEESI-MS技术对肺癌组织与癌旁组织中氨基酸的定性定量分析、肉类中痕量瘦肉精成分的鉴定与微量肉类品质鉴定的应用。本文主要研究内容包括以下三个方面:1.采用iEESI-MS技术对肺癌组织和癌旁组织中氨基酸的分析研究。1)无需任何样品预处理,iEESI-MS技术对肺癌组织和癌旁组织中氨基酸的定性定量分析,同时所得质谱数据通过主成分分析(PCA)实现了癌症组织与癌旁组织的快速区分,为临床组织样品的快速检测提供了一种有力的技术支撑;2)将iEESI-MS与DESI-MS技术的分析性能进行对比。结果表明iEESI-MS技术具有较好的定量能力,能同时实现组织表面和内部待测物的萃取、电离过程,获得更加丰富的化学信息,并且灵敏度更高、检测限更低。2.iEESI-MS技术对肉类中非法添加成分的分析研究。开发了能直接分析肉类产品中“瘦肉精”成分的iEESI-MS技术质谱检测平台。利用该分析平台对江西省体育科学研究所从屠宰场获得的猪肉、羊肉、牛肉和超市购买牛肉样品分别进行iEESI-MS分析,实现了对肉类中痕量“瘦肉精”物质的准确、快速分析和鉴定。3.微量肉类品质的iEESI-MS分析。采用iEESI-MS质谱技术对微量瘦猪肉和肥猪肉组织样品进行质谱分析,获得其组织样品的一级指纹谱图以及实现其肉品中的营养组分的成分鉴定,并结合主成分分析(PCA)法对两种组织样品的质谱数据进行区分建立判别模型。结果表明,iEESI-MS能够直接快速地获取复杂基体动物组织样品内部丰富的物质信息,可以为食品中营养成分的分析和研究提供一种可靠的检测工具,同时也在肉类食品安全检测中具有重要的现实意义。
[Abstract]:The analysis of animal tissue samples is the analysis of itself and its metabolites at the molecular level, which is of great significance in the field of medicine, food safety and so on. However, traditional analytical methods require complex pretreatment processes, such as grinding, extraction, separation and so on, which are not only lengthy and time-consuming, but also lead to the loss of some important material information in tissue samples, which leads to low efficiency of analysis. Therefore, it is of great significance to develop a "green, fast, non-destructive" method for tissue sample detection and analysis. Internal extraction electrospray ionization mass spectrometry (iEESI-MS) is a recently developed atmospheric mass spectrometry technique. Its unique advantage is that it can directly obtain the chemical information of the internal components of complex matrix solid samples without any sample pretreatment. It has the advantages of simple operation, high analysis speed, high selectivity, high sensitivity and high throughput. In this paper, the application of iEESI-MS to the qualitative and quantitative analysis of amino acids in lung cancer tissues and adjacent tissues, the identification of trace lean components in meat and the identification of trace meat quality were studied. The main contents of this paper include the following three aspects: 1. Analysis of amino acids in lung cancer tissues and adjacent tissues by iEESI-MS. 1) the qualitative and quantitative analysis of amino acids in lung cancer tissues and adjacent tissues without any sample pretreatment. At the same time, the mass spectrum data obtained by principal component analysis (PCA) can quickly distinguish cancer tissues from adjacent tissues, which provides a powerful technical support for the rapid detection of clinical tissue samples. The analytical performance of iEESI-MS and DESI-MS is compared with that of DESI-MS. The results show that iEESI-MS has better quantitative ability, can simultaneously realize the extraction and ionization process of the substance to be tested on the surface and inside of the tissue, obtain more abundant chemical information, and have higher sensitivity. The detection limit is lower. 2. Analysis of illegally added ingredients in meat by iEESI-MS. An iEESI-MS mass spectrometry platform was developed for the direct analysis of "lean meatine" in meat products. Using the platform, the samples of pork, mutton, beef and beef purchased from slaughterhouses were analyzed by iEESI-MS in Jiangxi Institute of Sports Science, respectively, and the trace "lean meatine" in meat was accurately analyzed. Rapid analysis and identification. Analysis of trace meat quality by iEESI-MS. The tissue samples of lean pork and fat pork were analyzed by iEESI-MS mass spectrometry. The first order fingerprint of the tissue samples and the identification of nutrient components in the meat were obtained. Combined with principal component analysis (PCA), a discriminant model was established to distinguish the mass spectrum data of two kinds of tissue samples. The results show that iEESI-MS can directly and quickly obtain abundant material information from the samples of complex matrix animal tissues, and can provide a reliable tool for the analysis and research of nutritional components in food. At the same time, it has important practical significance in meat food safety detection.
【学位授予单位】：东华理工大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R73-3;TS251.7;O657.63

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