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Separation of isobaric Amino Acids and Small Molecule Metabolites Using Multipass Ion Mobility Analysis

Amino acids along with small molecule metabolites are important biomarkers for the study and detection of diseases that are initially analyzed in untargeted omics fashion. Amino acids and many metabolites are isomeric, and their specific form can have a significant impact on biological function. Chromatographic separation of isomers is challenging, and they cannot be resolved by mass spectrometry alone. Ion mobility is a technique that allows the separation of ions based on their size, shape, and charge. Here we present the results of the separation of isobaric amino acids and small molecule metabolites using a system that allows for multi-pass ion mobility separation which, in turn, enhances the ion mobility resolution of the separation. Amino acid standards and small molecule metabolites commercially available were infused directly into a SELECT SERIES™ Cyclic™ IMS system. Solutions of individual and mixture of the isobaric species were used and the mobility conditions were optimized for multiple passes for each corresponding set of isobaric species. Both ionization polarities and various solvent adducts were tested to provide the best signal intensity and separation. Amino acids such as leucine and isoleucine have previously been separated with ion mobility in a system with lower ion mobility resolution (SYNAPT™ G2 mass spectrometer) rendering about 90% of valley and mobility resolution of around 40 Ω/ΔΩ. With the enhanced ion mobility resolution using the cyclic IMS technique we have obtained almost complete separation of those amino acids rendering a valley of about 10% after 15 passes. For this case, the mobility resolution is around 250 Ω/ΔΩ. Another example is the separation of glucose-6-phosphate from glucose-1-phosphate, and fructose-6phosphate which were separated after 10 passes. For this case, the mobility resolution is around 205 Ω/ΔΩ.

Hernando J Olivos↗

Microchrons - The Rb-87/Sr-87 dating of microscopic samples

The present investigation is concerned with the extension of the Rb-Sr dating technique to microscopic samples of individual mineral grains as small as approximately 30 micrometers in size. The measurement of the isotopic composition and the relative number of atoms of Rb and Sr has presented difficulties because of the need to separate Rb from Sr due to the isobaric interference at mass 87. This separation is critical since the changes in Sr-87/Sr-86 are directly related to the age of the sample. The approach used in the considered investigation is a modified version of the direct loading technique reported by Wasserburg et al. (1977). It is shown that measurements can be conducted of the Rb and Sr isotopic compositions and Rb/Sr atom ratios in microsamples containing 10 to the 11th to 10 to the 12th atoms of Sr. Rb-Sr model ages may prove to be of considerable importance in identifying and studying interplanetary dust particles.

Papanastassiou, D. A.↗

Elucidating the Gravome: Quantitative Proteomic Profiling of the Response to Chronic Hypergravity in Drosophila

Altered gravity conditions, such as experienced by organisms during spaceflight, is known to cause transcriptomic and proteomic changes. We describe the proteomic changes in the whole body of adult Drosophila melanogaster (fruit fly), but focus specifically on the localized changes in the adult head in response to chronic hypergravity (3G) treatment. Canton S adult female flies (2-3 days old) were exposed to chronic hypergravity for 9 days and compared with parallel 1G controls. After hypergravity treatment, whole flies and fly heads were separated, and evaluated for quantitative comparison of the two gravity conditions using an isobaric tagging liquid chromatography-tandem mass spectrometry approach. Data revealed a total of 1948 (whole flies) and 1480 (head) proteins to be differentially present in hypergravity-treated flies. Gene Ontology analysis of head specific proteomics revealed host immune response and humoral stress proteins were significantly upregulated. Proteins related to calcium signaling, ion transport and ATPase were decreased. Enhanced expression of cuticular proteins may suggest an alteration in chitin metabolism and in chitin-based cuticle development. We therefore present a comprehensive quantitative survey of proteomic changes in response to chronic hypergravity in Drosophila, which will help elucidate the underlying molecular mechanisms associated with altered gravity environments.

Hypergravity↗

In-Situ Reaction Monitoring for Lunar Applications Utilizing a Single Quadrupole Residual Gas Analyzer

With a revived focus to create a permeant presence on the moon and in preparation for future Mars exploration, it is imperative that all re-sources are utilized to their fullest potential. In-Situ Resource Utilization (ISRU) will be critical for future mission success as it would enable independent operation of missions while minimally relying on the complex supply chain created between the Earth, moon and Mars. One of the most critical resources that have been identified for ISRU is the creation of liquid oxygen (O2) for not only breathable air, but also for rocket fuel. On the moon and potentially Mars, one of the most O2 rich resources is in the presence of regolith. Although there are numerous minerals within regolith and various processes to extract O2, this is not the scope of this paper. Since the O2 extraction method can vary, a critical standardized analytical method is needed to verify O2 (or oxygenated precursor compounds) for extraction efficiency and purity. Our group utilizes a modified consumer off-the-shelf (COTS), Residual Gas Analyzer (RGA) Single Quadrupole Mass Spectrometer (QMS). Due to the current field mission requirements, the gas analysis module will need to rely only on mass-to-charge ratios (m/z) and peak intensities to differentiate and quantitate targeted gas-phase reactants or products, i.e. carbon monoxide (CO), carbon dioxide (CO2), methane (CH4) and hydrogen (H2). However, due to solar wind implanted volatiles and various other compounds present in lunar regolith, such as nitrogen (N2), helium (He), and deuterium (D2), deconvolution and quantification of isobaric compounds becomes quite difficult. Specifically, the presence of N2 makes quantification of CO, an oxygenated precursor, difficult due to both compounds having a peak at m/z 28. Approaches to differentiate isobaric components: Initially, the solution to overcome isobaric overlap was to use isotopic ratios, particularly be-tween carbon-13 and nitrogen-15. Although limited literature discuss isotopic composition and attempt to quantify the ratios, variation is particularly high and limited to a few lunar regolith samples. The following were identified as potential approaches to quantify isobaric species, N2 and CO, using our RGA. Threshold Ionization. CO and N2 both have peaks at m/z 28, but only N2 is capable of producing a peak at m/z 7. While a N2 peak at m/z 7 is generally rare, instrument parameters can be optimized in order to maximize the peak intensity. Typically a standard ionization energy of 70 eV is used to monitor a batch process and would produce reliable results for a majority of the reactants or products that we are interested in quantifying. However, to analyze the rare molecular fragment of N2 (N2+ at m/z 7), a different ionization energy may be beneficial. Threshold Ionization Mass Spectrometry (TIMS) will be explored to optimize the N2+ peak at m/z 7 by varying parameters such as ionization energy and pressure. Therefore, a quantitative value at m/z 7 will be used to determine the product gas concentration of N2, which will be subtracted from the total concentration determined at m/z 28 enabling us to approximate the amount of CO. Secondary Detector (infrared gas detector, IR). Another approach to quantitate and differentiate be-tween isobaric components is to leverage alternative modes of detection. Unlike mass spectrometry, which generally uses a separation step prior to analysis (i.e. gas chromatography), IR produces energy that is specifically absorbed depending on the bond and configuration of the molecule. This technique could be leveraged since it is generally only active on non-symmetric molecules, i.e. CO. Utilizing concentrations of CO determined from another detector could help correct concentrations of CO obtained from the QMS. However, the inclusion of another detector adds additional mission requirements, such as power, software changes and avionics updates that add additional cost and schedule requirements.

mass spectrometry↗

VTOL in ground effect flows for closely spaced jets

Results of a series of in ground effect twin jet tests are presented along with flow models for closely spaced jets to help predict pressure and upwash forces on simulated aircraft surfaces. The isolated twin jet tests revealed unstable fountains over a range of spacings and jet heights, regions of below ambient pressure on the ground, and negative pressure differential in the upwash flow field. A separate computer code was developed for vertically oriented, incompressible jets. This model more accurately reflects fountain behavior without fully formed wall jets, and adequately predicts ground isobars, upwash dynamic pressure decay, and fountain lift force variation with height above ground.

Migdal, D.↗

Utilization of FEP energetics

The research and development work on Fountain Effect Pump Systems (FEP systems) has been of interest in the competition between mechanical pumps for He II and FEP units. The latter do not have moving parts. In the course of the work, the energetics have been addressed using one part of a simple four-changes-of-state cycle. One option is the FEP ideal change of state at constant chemical potential (mu). The other option is the two-state sequence mu-P with a d mu=0 state change followed by an isobar. Questions of pump behavior, of flow rate response to temperature difference at the hot end, and related questions of thermodynamic cycle completion and heat transfer have been addressed. Porous media data obtained elucidate differences between vapor-liquid phase separation (VLPS) and Zero Net Mass Transfer (ZNMF).

Frederking, T. H. K.↗

Simulated sudden stratospheric warming - Synoptic evolution

An analysis is presented of a sudden stratospheric warming event which occurred spontaneously during a general circulation model simulation of the global atmospheric circulation. Two separate warming pulses exhibit the same dynamical evolution with a 'cycle' of about two weeks. Two distinct phases of the warming cycle are apparent: (1) the generation of an intense localized warm cell in conjunction with significant adiabatic heating associated with cross-isobar flow which has been induced by vertically propagating long wave disturbances; and (2) the northward transport of that warm cell via advection by the essentially geostrophic windfield corresponding to an intense, offset polar cyclone, in conjunction with a strong Aleutian anticyclone. During the first warming pulse in January, a moderate Aleutian anticyclone was in place prior to the warming cycle and was intensified by interaction with an eastward traveling anticyclone induced by the differential advection of the warm cell. The second warming pulse occurred in early February with a strong Aleutian anticyclone already established. In contrast to the January event, the warming in February culminated with reversal of the zonal westerlies to easterlies over a significant depth of the stratosphere.

Blackshear, W. T.↗

The heavy element yields of neutron capture nucleosynthesis

Consideration of the contribution made to the abundances of the heavy element isotopes by the S- and R-processes of nucleosynthesis has led to the determination that the previous assumption concerning the exclusive alignment of isobars to one or the other of these processes is probably in error. If the relatively small odd and even mass number abundance fluctuations characterizing R-process abundances are always the case, as assumed by this study, S-process contributions to the abundances of R-process isobars are substantial, consistent with transient flashing episodes in the S-process neutron production processes. A smooth and monotonically-decreasing curve of the abundance of the S-process yields times the neutron capture cross-section versus mass number is therefore the primary tool for the separation of the abundances due to the two processes.

Cameron, A. G. W.↗

Revised Atmospheric Angular Momentum Series Related to Earth's Variable Rotation under Consideration of Surface Topography

The atmospheric angular momentum is closely related to variations in the Earth rotation. The atmospheric excitation function (AEF), or namely atmospheric effective angular momentum function, is introduced in studying the atmospheric excitation of the Earth's variable rotation. It may be separated into two portions, i.e, the "wind" terms due to the atmospheric motion relative to the mantle and the "pressure" terms due to the variations of atmospheric mass distribution evident through surface pressure changes. The AEF wind terms during the period of 1948-2004 are re-processed from the NCEP/NCAR (National Centers for Environmental Prediction-National Center for Atmospheric Research) reanalysis 6-hourly wind and pressure fields. Some previous calculations were approximate, in that the wind terms were integrated from an isobaric lower boundary of 1000 hPa. To consider the surface topography effect, however, the AEF is computed by integration using the winds from the Earth's surface to 10 hPa, the top atmospheric model level, instead of from 1000 hPa. For these two cases, only a minor difference, equivalent to approx. 0.004 milliseconds in length-of-day variation, exists with respect to the axial wind term. However, considerable differences, equivalent to 5-6 milliarcseconds in polar motion, are found regarding equatorial wind terms. We further compare the total equatorial AEF (with and without the topographic effect) with the polar motion excitation function (PMEF) during the period of 1980-2003. The equatorial AEF gets generally closer to the PMEF, and improved coherences are found between them when the topography effect is included. Keywords: Atmospheric angular momentum, Atmospheric excitation function, Earth rotation, Topography, Wind, Pressure.

Zhou, Y. H.↗