阀切换离子色谱-质谱联机用于热纤梭菌胞外水溶性代谢物组学分析

In the fields of metabolomics, HPLC techniques cover a wide range of compounds, however ionic and polar compounds such as organic acids, carbohydrates, nucleotides and amino acids are difficult to separate on traditional reverse phase columns or HILIC columns. Ion exchange chromatography has been proven to be a much better separation process for these compounds. Ion Chromatography Coupled with Mass Spectrum technique is an important tool for polar compounds analysis. But the peak capacity of one dimension is limited and the time of analysis is very long. In this project, an ion chromatography tandem mass spectrometry system will be set up with valve switching and electrolytic anion suppressor according to different ion rention mechanism. Simultaneous separation and identification of the polar compounds will be realized using the system,meanwhile the peak capacity and work efficiency will be increased. Clostridium thermocellum is selected as the research object and organic acids, carbohydrates, nucleotides and amino acids are targeted metabolites. We will study the extracellular metabolomics of wild type strain and ethanol tolerant strain of Clostridium thermocellum. We hope to find the otherness of metabolic footprinting and supply more scientific and direct data for reasonably design of experiments and procedures.

在生物能源领域的代谢物组学研究中，反相和亲水液相色谱可分析大范围的小分子代谢物，然而一些离子和极性化合物如有机酸、糖类化合物和氨基酸等重要代谢物则分离比较困难。离子色谱-质谱联用技术是解决复杂样品中极性化合物分离和鉴定的重要手段，一维离子色谱由于峰容量低，分析时间长，在实际应用中受到限制。针对上述制约因素，本项目拟结合多种离子分离机理，通过阀切换和微膜电解抑制技术构建离子色谱-质谱联用系统，解决一维离子色谱-质谱分离度不足、组分共淋洗和盐的干扰问题，有效提高分析准确度和效率，并实现上述极性代谢物的在线同时分析。以氨基酸、糖类、有机酸等热纤梭菌胞外靶向代谢物为研究对象，利用建立的离子色谱-质谱联用系统进行热纤梭菌乙醇耐受株和野生株的代谢足印分析，找到差异性代谢物，为设计更合理的热纤梭菌反应调控路线提供更加科学和直接的实验依据。

在生物能源领域的代谢物组学研究中，反相和亲水液相色谱可分析大范围的小分子代谢物，然而一些离子和极性化合物如有机酸、糖类化合物和氨基酸等重要代谢物则分离比较困难。离子色谱-质谱联用技术是解决复杂样品中极性化合物分离和鉴定的重要手段，一维离子色谱由于峰容量低，分析时间长，在实际应用中受到限制。针对上述制约因素，本项目结合多种离子分离机理，通过阀切换和微膜电解抑制技术构建离子色谱-质谱联用系统，解决了一维离子色谱-质谱分离度不足、组分共淋洗和盐的干扰问题，有效提高分析准确度和效率，并实现了上述极性代谢物的在线同时分析。以氨基酸、糖类、有机酸等热纤梭菌胞外靶向代谢物为研究对象，利用建立的离子色谱-质谱联用系统进行热纤梭菌胞外靶向水溶性代谢物的分析，为设计更合理的热纤梭菌反应调控路线提供了更加科学和直接的实验手段。