Emily A. Kurfman, Maria F. Mora, Peter A. Willis, Susan M. Lunte
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引用次数: 0
Abstract
The search for chemical indicators of life is a fundamental component of potential future spaceflight missions to ocean worlds. Capillary electrophoresis (CE) is a useful separation method for the determination of the small organic molecules, such as amino acids and nucleobases, that could be used to help determine whether or not life is present in a sample collected during such missions. CE is under development for spaceflight applications using multiple detection systems, such as laser induced fluorescence (LIF) and mass spectrometry (MS). Here we report CE-based methods for separation and detection of major polar metabolites in cells, such as amino acids, nucleobases/sides, and oxidized and reduced glutathione using detectors that are less expensive alternatives to LIF and MS. Direct UV detection, indirect UV detection, and capacitvely coupled contactless conductivity detection (C4D) were tested with CE, and a combination of direct UV and C4D allowed the detection of the widest variety of metabolites. The optimized method was used to profile metabolites found in samples of Escherichia coli and Pseudoalteromonas haloplanktis and showed distinct differences between the species.
寻找生命的化学指标是未来潜在的海洋世界太空飞行任务的基本组成部分。毛细管电泳(CE)是一种有用的分离方法,可用于测定氨基酸和核碱基等小分子有机物,从而帮助确定在此类飞行任务中收集的样本中是否存在生命。目前正在开发使用激光诱导荧光(LIF)和质谱(MS)等多种检测系统的 CE 航天应用。在此,我们报告了基于 CE 的细胞内主要极性代谢物的分离和检测方法,如氨基酸、核碱基/核苷酸、氧化和还原谷胱甘肽,使用的检测器是 LIF 和 MS 的廉价替代品。直接紫外检测法、间接紫外检测法和电容耦合非接触电导检测法(C4D)与 CE 一起进行了测试,直接紫外检测法和 C4D 的组合可以检测到最多种代谢物。优化后的方法用于分析大肠杆菌和卤代假交替单胞菌样本中发现的代谢物,结果显示不同物种之间存在明显差异。
期刊介绍:
ELECTROPHORESIS is an international journal that publishes original manuscripts on all aspects of electrophoresis, and liquid phase separations (e.g., HPLC, micro- and nano-LC, UHPLC, micro- and nano-fluidics, liquid-phase micro-extractions, etc.).
Topics include new or improved analytical and preparative methods, sample preparation, development of theory, and innovative applications of electrophoretic and liquid phase separations methods in the study of nucleic acids, proteins, carbohydrates natural products, pharmaceuticals, food analysis, environmental species and other compounds of importance to the life sciences.
Papers in the areas of microfluidics and proteomics, which are not limited to electrophoresis-based methods, will also be accepted for publication. Contributions focused on hyphenated and omics techniques are also of interest. Proteomics is within the scope, if related to its fundamentals and new technical approaches. Proteomics applications are only considered in particular cases.
Papers describing the application of standard electrophoretic methods will not be considered.
Papers on nanoanalysis intended for publication in ELECTROPHORESIS should focus on one or more of the following topics:
• Nanoscale electrokinetics and phenomena related to electric double layer and/or confinement in nano-sized geometry
• Single cell and subcellular analysis
• Nanosensors and ultrasensitive detection aspects (e.g., involving quantum dots, "nanoelectrodes" or nanospray MS)
• Nanoscale/nanopore DNA sequencing (next generation sequencing)
• Micro- and nanoscale sample preparation
• Nanoparticles and cells analyses by dielectrophoresis
• Separation-based analysis using nanoparticles, nanotubes and nanowires.