Enabling technologies for high performance mass spectrometry applications

质谱（MS）由于其上级灵敏度和分辨率以及其提供的结构信息而成为最优选的分析工具。在目前可用的MS方法中，液相和气相色谱法（LC和GC）与MS结合使用最为广泛。然而，LC/GC-MS在许多情况下遭受长的分析时间和低灵敏度，这增加了分析成本，并且禁止在许多情况下使用MS，这些情况可以通过MS技术来帮助，例如食品，环境和药物分析，以及其在临床和护理环境中的应用。提出的研究计划旨在解决阻碍MS方法应用的一些主要问题，并使一些目前耗时的方法具有高通量，并提高其他MS方法的性能。提出了新方法在蛋白质结构和结构动力学研究中的应用。样品提取和样品富集通常决定分析方法的质量及其成功实施。我们将探索使用新的溶剂系统，如低共熔溶剂与机械化学方法进行溶质特异性提取和富集。将开发新的液体和固体萃取方法用于环境电离质谱（AIMS）检测，以加快分析过程，同时保持灵敏度和特异性。我们将重点介绍AIMS技术之一，称为真实的时间质谱直接分析（DART-MS）。萃取系统与AIMS的结合将使我们能够在比目前使用的LC-MS方法短得多的时间内分析环境和生物样品。将研究溶剂和固体的物理化学性质及其与分析物的相互作用，以及这些性质对AIMS的电离和检测效率的影响。对于更复杂的系统，在分析物被引入MS之前必须添加分离步骤。我们将继续开发新的方法来使用毛细管电泳质谱（CEMS）。特别感兴趣的是我们的努力，在毛细管等电聚焦（cIEF）MS。我们已经证明，这种方法是可行的，我们的下一步是探索的可能性相结合的IEF与串联质谱的微小结构差异的研究蛋白质异构体。CE-MS是研究蛋白质结构动力学和构象变化的有力工具。我们将开发新的方法来监测氢-氘交换（HDX），以表征蛋白质分子中暴露于溶液的部分、受其三级结构保护的部分以及它们的构象变化和二硫键在不同类型的应激条件下的争夺。

Mass spectrometry (MS) has become the most preferred analytical tool because of its superior sensitivity and resolution, as well as the structural information it provides. Of the currently available MS methods, liquid and gas chromatography (LC and GC) coupled with MS have been the most widely used. However, LC/GC-MS in many cases suffer from long analysis time and low sensitivity, that increase the cost of analyses and prohibit the use of MS in many situations that could have helped by the MS technology, such as food, environmental and pharmaceutical analysis, as well as its application in clinical and point of care settings. The proposed research program aims to address some major issues that are impeding the application of MS methods and make some of the currently time-consuming methods high throughput, and improve the performance other MS methods. Applications of the new methods for the study of protein structure and structure dynamics are proposed. Sample extraction and sample enrichment often determine the quality of the analytical methods and their successful implementation. We will explore the use of new solvent systems such as deep eutectic solvents with mechanochemical method for solute specific extraction and enrichment. New liquid and solid extraction methods will be developed for ambient ionization mass spectrometry (AIMS) detection to speed up the analytical process while maintaining sensitivity and specificity. We will focus on one of the AIMS technologies called direct analysis in real time mass spectrometry (DART-MS). The combination of the extraction systems with AIMS will allow us to analyze environmental and biological samples in much shorter times than the currently used LC-MS methods. Physicochemical properties of the solvents and the solids, and their interaction with the analytes will be studied, as well as the effect of these properties on the ionization and detection efficiency of AIMS. For more complex systems a separation step must be added before the analytes are introduced into the MS. We will continue to develop novel methods to use capillary electrophoresis mass spectrometry (CEMS). Of particular interest is our efforts in capillary isoelectric focusing (cIEF) MS. We have demonstrated that this method is feasible, and our next step is to explore the possibility of combining IEF with tandem mass spectrometry for the study of minute structural differences in protein isomers. CE-MS can be a powerful tool for the study of protein structural dynamics and conformation changes in solution in their native states. We will develop new method to monitor the hydrogen-deuterium exchange (HDX) to characterize the part of the protein molecules that are exposed to the solution, and the parts of that are protected by their tertiary structures, as well as their conformational changes and disulfide bonds scramble in different kind of stress conditions.