IN SITU MASS SPECTRAL ANALYSIS OF ELECTROPHORETOGRAMS

生化研究广泛使用静态层析，如 高分辨率二维纸层析和薄层层析 电泳法。从这些技术中获得的信息是有限的。 由于传统的检测系统的性能相对较差 使用。这项提议支持质谱学的发展 一种可以对分离的生物分子进行原位分析的系统，具有特定的 重点分析高效两相分离的多肽混合物 空间电泳法。从群众中获得的信息 光谱，分离的成分的同一性和均匀性可以是 在不损失色谱图完整性的情况下建立。在……里面 有利的情况下，化合物可以在分析后被回收 进一步的表征或加工。在这台新仪器里，完好无损 色谱图放置在真空中，成分封装在 通过色谱图在XY方向上的移动使基质成为焦点 开发平面。一种高聚焦二次离子质谱仪 初级离子束将物质直接从色谱图溅射到 四极质谱仪。在x和x方向的空间分辨率均为10微米 Y可以通过以下方式提高对分离不良化合物的有效分辨率 质谱学数据的处理。一种自动反馈系统 通过增加空间数据来最大化数据收集的效率 当化合物被带入仪器焦点时，分辨率。首字母 将应用于分离的肉碱样品的分析 用薄层层析法分离镰状细胞血红蛋白多肽 通过电泳法，以及一项新的实验，选择序列监测， 它精确定位包含特定氨基酸序列的多肽 在用二维层析法分离的混合物中。这些 初步实验证明了新系统的性能存在问题 它们具有显著的生物化学意义。在拟议的 在此期间，将开发一种自动分析系统，该系统可以 用于二维分离的复杂混合物的一般分析 电泳法。

Biochemical studies make extensive use of static chromatographies such as high resolution two dimensional paper and thin layer chromatography and electrophoresis. The information available from such techniques is limited by the relatively poor performance of the detection systems traditionally used. This proposal supports the development of a mass spectrometric system which can analyze separated biomolecules in situ, with a particular focus on the analysis of peptide mixtures separated by high performance two dimensional electrophoresis. With information available from the mass spectrum, the identity and homogeneity of the separated components can be established without loss of the integrity of the chromatogram. In favorable instances, the compounds can be recovered after analysis for further characterization or processing. In this new instrument, intact chromatograms are placed under vacuum, and components encapsulated in the matrix are brought into focus by movement of the chromatogram in the xy development plane. A secondary ion mass spectrometer with a highly focused primary ion beam sputters material directly from the chromatogram into a quadrupole mass analyzer. A spatial resolution of 10 microns in both x and y can increase the effective resolution of poorly separated compounds by processing of the mass spectral data. A feedback system automatically maximizes the efficiency of data collection by increasing spatial resolution as compounds are brought into the instrument focus. Initial applications will be to the analysis of carnitine-based samples separated by thin layer chromatography, to sickle cell hemoglobin peptides separated by electrophoresis, and to a new experiment, selected sequence monitoring, which pinpoints peptides containing a specific sequence of amino acids from within a mixture separated by two dimensional chromatography. These initial experiments document the performance of the new system in problems which have significant biochemical interest. By the end of the proposed period, an automated analysis system will have been developed which can be used for general analysis of complex mixtures separated by two dimensional electrophoresis.