基于微流控芯片－质谱的肺癌代谢组分析方法研究

项目名称： 基于微流控芯片－质谱的肺癌代谢组分析方法研究

项目编号： No.20805017

项目类型： 青年科学基金项目

立项/批准年度： 2009

项目学科： 化学工业

项目作者： 徐晖

作者单位： 华中师范大学

项目金额： 23万元

中文摘要： 本项目充分地发挥毛细管电泳和液相色谱的高效分离的优势，和高灵敏度的质谱检测器相结合，建立高效、快速、灵敏、集成化的肺癌代谢标志物的毛细管电泳－质谱、液相色谱-质谱分析新方法。采用补充鞘流液和适当增加背压的方法，有效地解决了毛细管电泳与质谱接口所遇到的分离电压施加和喷雾离子化效率等问题。采用毛细管内原位光引发合成聚甲基丙烯酸-乙二醇二甲基丙烯酸酯整体柱，在管内进行在线衍生化和在线预富集，实现痕量醛类代谢物的再次富集，提高分析灵敏度。同时我们采用衍生化技术，将质谱响应信号弱的醛类以2, 4-二硝基苯肼为衍生化试剂进行衍生化反应,实现代谢物的选择性、高灵敏度的测定。除此之外，我们还采用5种离线样品前处理方法，分离提纯样品，减小基质效应，进一步提高质谱分析灵敏度和重现性。实验结果显示肺癌患者血清、尿液中己醛和庚醛的含量明显高于正常对照组，这两种醛可以作为潜在的肺癌生物标志物，用于肺癌的临床早期诊断和筛查。所建立的分析方法简单、快速、灵敏、选择性高，非常适合于复杂生物样品中挥发性化合物的分析测定，在临床醛类疾病标志物分析中具有潜在应用前景。

中文关键词： 毛细管电泳－质谱联用；代谢组学；前处理技术；肺癌;醛类物质

英文摘要： In this work, we are currently interested in the applying of capillary electrophorsis-mass spectrometry system as an alternative tool for the metabonomics analysis of lung cancer. An effective, rapid, sensitive, integrated method for the metabonomics analysis of lung cancer is developed by combining the home-made capillary electrophoresis analysis system with the mass spectmetry detector. First, capillary electrophoresis was hyphenated with ESI mass spectrometer (CE/MS) with the assistant of external supplement of sheath liquid. A plug of poly(metharylic acid-co-ethlyene dimethacrylate) monolith was in-situ synthesized inside a fused-silica capillary for online derivatization and online preconcentration.2, 4-Dinitrophenylhydrazine (DNPH) as derivatizing reagent was ultilized to enhance the sensitivity of aldehydes. Besides, the interferences of sample matrix were removed and the target analytes were preconcentrated through off-line sample pretreatment techniques, such as liquid-phase microextraction, magnetic solid-phase extraction and monolith frit-based solid-phase microextraction.The experimental results showed that the concentrations of hexanal and heptanal determined in the urine and serum samples of lung cancer patients are obviously higher than those in normal people.The results suggested that hexanal and heptanal in human urine and serum can be used as the biomarker candidates of lung cancer in the early diagnosis. Furthermore, these methods may provide convenient and efficient tools for the determination of aldehydes in complex biological matrixes.

英文关键词： capillary electrophoresis-mass spectrometry; metabonomics; sample preparation; lung cancer;aldehydes