Amino acids are fundamental building blocks of terrestrial life as well as ubiquitous byproducts of abiotic reactions. In order to distinguish between amino acids formed by abiotic versus biotic processes it is possible to use chemical distributions to identify patterns unique to life. This article describes two capillary electrophoresis methods capable of resolving 17 amino acids found in high abundance in both biotic and abiotic samples (seven enantiomer pairs d/l-Ala, -Asp, -Glu, -His, -Leu, -Ser, -Val and the three achiral amino acids Gly, β-Ala, and GABA). To resolve the 13 neutral amino acids one method utilizes a background electrolyte containing γ-cyclodextrin and sodium taurocholate micelles. The acidic amino acid enantiomers were resolved with γ-cyclodextrin alone. These methods allow detection limits down to 5 nM for the neutral amino acids and 500 nM for acidic amino acids and were used to analyze samples collected from Mono Lake with minimal sample preparation.
We present a comprehensive discussion of the role that microchip electrophoresis (ME) instrumentation could play in future NASA missions of exploration, as well as the current barriers that must be overcome to make this type of chemical investigation possible. We describe how ME would be able to fill fundamental gaps in our knowledge of the potential for past, present, or future life beyond Earth. Despite the great promise of ME for ultrasensitive portable chemical analysis, to date, it has never been used on a robotic mission of exploration to another world. We provide a current snapshot of the technology readiness level (TRL) of ME instrumentation, where the TRL is the NASA systems engineering metric used to evaluate the maturity of technology, and its fitness for implementation on missions. We explain how the NASA flight implementation process would apply specifically to ME instrumentation, and outline the scientific and technology development issues that must be addressed for ME analyses to be performed successfully on another world. We also outline research demonstrations that could be accomplished by independent researchers to help advance the TRL of ME instrumentation for future exploration missions. The overall approach described here for system development could be readily applied to a wide range of other instrumentation development efforts having broad societal and commercial impact.
The development of therapeutic proteins and peptides is an expensive and time-intensive process. Biologics, which have become a multi-billion dollar industry, are chemically complex products that require constant observation during each stage of development and production. Post-translational modifications along with chemical and physical degradation from oxidation, deamidation, and aggregation, lead to high levels of heterogeneity that affect drug quality and efficacy. The various separation modes of capillary electrophoresis (CE) are commonly utilized to perform quality control and assess protein heterogeneity. This review attempts to highlight the most recent developments and applications of CE separation techniques for the characterization of protein and peptide therapeutics by focusing on papers accepted for publication in the in the two-year period between January 2012 and December 2013. The separation principles and technological advances of CE, capillary gel electrophoresis, capillary isoelectric focusing, capillary electrochromatography and CE-mass spectrometry are discussed, along with exciting new applications of these techniques to relevant pharmaceutical issues. Also included is a small selection of papers on microchip electrophoresis to show the direction this field is moving with regards to the development of inexpensive and portable analysis systems for on-site, high-throughput analysis.
A top-level NASA exploration goal is the search for signs of life beyond Earth. Molecular biosignatures can be sought via in situ organic chemical analyses of samples collected from planetary bodies. Current spaceflight-ready technologies lack the required sensitivity to perform these analyses on key polar organic species primarily due to the low efficiency of transferring these molecules from natural samples into chemical instrumentation. One promising approach to improve the liberation of these molecules from sample matrices prior to analysis is through the use of liquid extraction, in which organic molecules are dissolved in heated, pressurized water. This process has never been performed on a spaceflight mission. As part of instrument maturation efforts, we describe the development of the first spaceflight prototype of a fully automated subcritical water extractor designed to provide this essential functionality on a potential future planetary mission. The prototype extractor was mounted on a Mars test rover platform and successfully operated remotely in the Atacama Desert, Chile. Samples acquired by the rover's drill and sample acquisition/delivery system were remotely transferred to the extractor inlet funnel, and all subsequent extractor operations were performed automatically. To validate the instrument and demonstrate its suitability as a front-end unit for an organic analyzer, we tested low-bioload Atacama Desert soil extracts for amino acid content using capillary electrophoresis coupled to laser-induced fluorescence. We show that hot extraction under subcritical conditions is required to liberate amino acids from the sample, as no amino acids were found in the extract produced at room temperature. Plain Language SummaryThe search for signs of life beyond Earth is one of NASA's highest priorities. One powerful way to look for extraterrestrial life is to perform chemical analyses on planetary samples in order to characterize any organic molecules present. Past and current spaceflight instruments analyze various components in gases collected or gases produced from solid samples; however, the instruments either lack the sensitivity to detect organic molecules or the instruments destroy many of the organic molecules during the preparation required to perform a gas-phase-based measurement. This work presents an automated pressurized hot water extractor that can be used to release organic molecules from a solid sample. Pressurized hot water is a powerful and easy-to-handle solvent for a wide range of different compounds. The extractor acts as a front-end instrument and prepares the sample for highly sensitive, liquid-based analysis. This automated and remotely controlled prototype has been successfully tested on a simulated Mars rover mission in the Atacama Desert in Chile. Extracts of the Atacama soil samples were then analyzed by capillary electrophoresis coupled to laser-induced fluorescence to determine the extracts' amino acid content, as amino acids are an auspicious class of molecules in the search fo...
The search for biosignatures on spaceflight missions requires in situ instrumentation capable of highly selective and sensitive organic analyses. To this end, CE-LIF is a uniquely promising technique, capable of determining the type, abundance, and chirality of amino acids present in environmental samples at nanomolar concentrations. However, this type of assay requires several reagents that have not yet been used on spaceflight missions. A key concern, particularly for future missions to Europa, is the survivability of these critical components for CE separation and LIF detection under high levels of radiation. Here we present an investigation of the chemical stability of the reagents and associated fused silica capillary after a total ionizing dose of 300 krad, exceeding the predicted total ionizing dose for the potential Europa Lander Mission payload by two-fold. Neither the fused silica capillary nor the fluorescent dye (5-carboxyfluorescein succinimidyl ester) showed significant change in performance following irradiation. Following the irradiation of the pre-mixed background electrolyte, both migration time and resolution were affected. However, when the reagents (sodium tetraborate, sodium taurocholate, and γ-cyclodextrin) and the acetonitrile solution were irradiated separately and mixed afterwards, there was no change in the separation performance.
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