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At least 163 records · Page 9

A colorimetric method to measure in vitro nitrogenase functionality for engineering nitrogen fixation

Biological nitrogen fixation (BNF) is the reduction of N 2 into NH 3 in a group of prokaryotes by an extremely O 2 -sensitive protein complex called nitrogenase. Transfer of the BNF pathway directly into plants, rather than by association with microorganisms, could generate crops that are less dependent on synthetic nitrogen fertilizers and increase agricultural productivity and sustainability. In the laboratory, nitrogenase activity is commonly determined by measuring ethylene produced from the nitrogenase-dependent reduction of acetylene (ARA) using a gas chromatograph. The ARA is not well suited for analysis of large sample sets nor easily adapted to automated robotic determination of nitrogenase activities. Here, we show that a reduced sulfonated viologen derivative (S 2 V red ) assay can replace the ARA for simultaneous analysis of isolated nitrogenase proteins using a microplate reader. We used the S 2 V red to screen a library of NifH nitrogenase components targeted to mitochondria in yeast. Two NifH proteins presented properties of great interest for engineering of nitrogen fixation in plants, namely NifM independency, to reduce the number of genes to be transferred to the eukaryotic host; and O 2 resistance, to expand the half-life of NifH iron-sulfur cluster in a eukaryotic cell. This study established that NifH from Dehalococcoides ethenogenes did not require NifM for solubility, [Fe-S] cluster occupancy or functionality, and that NifH from Geobacter sulfurreducens was more resistant to O 2 exposure than the other NifH proteins tested. It demonstrates that nitrogenase components with specific biochemical properties such as a wider range of O 2 tolerance exist in Nature, and that their identification should be an area of focus for the engineering of nitrogen-fixing crops.

59 BASIC BIOLOGICAL SCIENCES↗

Control of chloroplast degradation and cell death in response to stress

Chloroplasts are the sites of photosynthesis in plants and algae and, by extension, are essential for most life on Earth. Their maintenance is costly and complex due to the inherent photo-oxidative damage incurred by photosynthetic chemistry. Chloroplast degradation and cell death are mechanisms by which plants acclimate to such stress and serve a dual purpose: protecting cells and organs by removing reactive oxygen species-producing chloroplasts and redistributing nutrients to other tissues. Here I review recent progress in understanding the molecular mechanisms initiating and facilitating such degradation and show these are complex processes involving multiple pathways. Furthermore, due to the links to photosynthesis and nitrogen metabolism, there is great potential to manipulate these pathways to increase crop yield and quality under stressful environments.

59 BASIC BIOLOGICAL SCIENCES↗

Alternant Hydrocarbon Diradicals as Optically Addressable Molecular Qubits

High-spin molecules allow for bottom-up qubit design and are promising platforms for magnetic sensing and quantum information science. Optical addressability of molecular electron spins has also been proposed in first-row transition-metal complexes via optically detected magnetic resonance (ODMR) mechanisms analogous to the diamond-nitrogen-vacancy color center. However, significantly less progress has been made on the front of metal-free molecules, which can deliver lower costs and milder environmental impacts. At present, most luminescent open-shell organic molecules are π-diradicals, but such systems often suffer from poor ground-state open-shell characters necessary to realize a stable ground-state molecular qubit. In this work, we use alternancy symmetry to selectively minimize radical–radical interactions in the ground state, generating π-systems with high diradical characters. We call them m-dimers, referencing the need to covalently link two benzylic radicals at their meta carbon atoms for the desired symmetry. Through a detailed electronic structure analysis, we find that the excited states of alternant hydrocarbon m-diradicals contain important symmetries that can be used to construct ODMR mechanisms leading to ground-state spin polarization. Furthermore, the molecular parameters are set in the context of a tris(2,4,6-trichlorophenyl)methyl (TTM) radical dimer covalently tethered at the meta position, demonstrating the feasibility of alternant m-diradicals as molecular color centers.

Chemistry↗

Charge collection and trapping mechanisms in hexagonal boron nitride epilayers

Understanding charge collection and trapping mechanisms is crucial for using hexagonal boron nitride (h-BN) as active layers for many photonic and electronic devices such as deep UV detectors and emitters, neutron detectors, and single photon emitters. Charge collection and trapping mechanisms in h-BN epilayers have been investigated by probing impurity related optical emissions under an applied electrical field. Our results suggested that the existence of oxygen impurities affects the charge collection efficiency and results in an additional emission peak at 3.75 eV, corresponding to a donor-acceptor pair (DAP) recombination involving O N (oxygen residing on the nitrogen site) donors and the V B -H (boron vacancy bonded with hydrogen complex) deep level acceptors. Experimental results further revealed that the applied electric field induces an anti-correlation between the emission intensity of the DAP transition and the charge collection efficiency from which it was shown that it is possible to find an expression to quantitatively measure the maximum charge collection efficiency in h-BN. The results introduce not only a coherent picture for the relationship between common impurities in h-BN and charge collection and trapping mechanisms but also useful insights into possible approaches to improve the quality, purity, and charge collection of the h-BN epilayers.

71 CLASSICAL AND QUANTUM MECHANICS, GENERAL PHYSIC↗

Direct synthesis and chemical vapor deposition of 2D carbide and nitride MXenes

Two-dimensional transition-metal carbides and nitrides (MXenes) are a large family of materials actively studied for various applications, especially in the field of energy storage. MXenes are commonly synthesized by etching the layered ternary compounds, called MAX phases. We demonstrate a direct synthetic route for scalable and atom-economic synthesis of MXenes, including compounds that have not been synthesized from MAX phases, by the reactions of metals and metal halides with graphite, methane, or nitrogen. The direct synthesis enables chemical vapor deposition growth of MXene carpets and complex spherulite-like morphologies that form through buckling and release of MXene carpet to expose fresh surface for further reaction. The directly synthesized MXenes showed excellent energy storage capacity for lithium-ion intercalation.

Wang, Di↗

Viking to Mars - Profile of a space expedition

The Viking spacecraft, operations, and findings are reviewed and numerous pictures are presented in an attempt to capture the atmosphere of the Viking expedition. The details of the two Viking spacecraft, each consisting of an orbiter and lander combination launched a Titan III/Centaur are described and illustrated, along with the Viking ground-data and communications system. The principal conclusions of the Viking mission to date are: detection of nitrogen, argon, krypton, and xenon; determination of isotopic ratios of carbon, nitrogen, oxygen, and argon; uniform diurnal meteorological conditions; determination of major elemental abundances; complex surface chemistry; no ubiquitous organic material; 4 to 7% of the sampled surface material is magnetic; discovery of ancient extensive fluvial activity; north permanent polar cap made of water ice; and significant variations of the atmospheric water vapor, the summer hemisphere being much more humid than the winter hemisphere.

Martin, J. S., Jr.↗

Simulation and Experimental Measurements of Inductively Coupled CF4 and CF4/Ar Plasmas

The recently developed code SEMS (semiconductor equipment modeling software)is applied to the simulation of CF4 and CF4/Ar inductively coupled plasmas (ICP). This work builds upon the earlier nitrogen, transformer coupled plasma (TCP) SEMS research by demonstrating its accuracy for more complex reactive mixtures, moving closer to the realization of a virtual plasma reactor. Attention is given to the etching of and/or formation of carbonaceous films on the quartz dielectric window and diagnostic aperatures. The simulations are validated through comparisons with experimental measurements using FTIR (Fourier Transform Infrared) and UV absorption spectroscopy for CFx and SiFx neutral radicals, QMS (quadrupole mass spectrometry) for the ions, and Langmuir probe measurements of electron number density and temperature in an ICP GEC reference cell.

Hash, D. B.↗

An Overview of the Impact of Energetic Particle Precipitation on the Mesosphere and Stratosphere

Energetic precipitating particles (EPPs) can cause significant constituent changes in the polar mesosphere and stratosphere (middle atmosphere) during certain periods. Both protons and electrons can influence the polar middle atmosphere through ionization and dissociation processes. EPPs can enhance HOx (H, OH, HO2) through the formation of positive ions followed by complex ion chemistry and NOx (N, NO, NO2) through the dissociation of molecular nitrogen. The HO, increases can lead to ozone destruction in the mesosphere and upper stratosphere via several catalytic loss cycles. Such middle atmospheric HOx,-caused ozone loss is rather short-lived due to the relatively short lifetime (hours) of the HOx constituents. The HOx,-caused ozone depletion has been observed during several solar proton events (SPEs) in the past 40 years. HOx enhancements due to SPEs were confirmed by observations in the past solar cycle. A number of modeling studies have been undertaken over this time period that show predictions of enhanced HO, accompanied by decreased ozone due to energetic particles. The NO, family has a longer lifetime than the HOx family and can also lead to catalytic ozone destruction. EPP-caused enhancements of the NOx family can affect ozone promptly, if produced in the stratosphere, or subsequently, if produced in the 1ow.er thermosphere or mesosphere and transported to the stratosphere. NOx enhancements due to auroral electrons, medium and high energy electrons, relativistic electron precipitation (REP) events, and SPEs have been measured and/or modeled for decades Only a small number of SPEs (less than 10) in each solar cycle have sufficient flux of high energy protons (greater than 30 MeV) to produce a significant amount of NOx directly in the stratosphere to cause a measurable ozone destruction. Very high energy electrons (greater than 1500 keV) during REP events can also produce NOx directly in the stratosphere, however, the frequency and intensity of these electrons is uncertain. Indirect stratospheric EPP effects result when NO, is produced in the mesosphere and lower thermosphere and transported to the stratosphere during the late fall or winter. Such EPP-produced NOx may last up to months beyond its initial production. This EPP-produced mesospheric and lower thermospheric NOx primarily results from auroral electrons (approx. 1- 30 keV), medium- and high-energy electrons (approx. 30-1500 keV), and medium energy protons (approx. 1-30 MeV). The EPP-caused direct and indirect effects on the stratosphere are important in the polar regions during particular years, especially near solar maximum. This talk will attempt to provide an overview of several of the EPP-related important processes and their impact on the mesosphere and stratosphere. Much progress has been made in this field in the past several years and it is anticipated that other workshop participants will address aspects of this topic in their presentations.

Jackman, Charles H.↗

Direct EPP Affects on the Middle Atmosphere

Energetic precipitating particles (EPPs) can cause significant direct constituent changes in the mesosphere and stratosphere (middle atmosphere) during certain periods. Both protons and electrons can influence the polar middle atmosphere through ionization and dissociation processes. EPPs can enhance HOx (H, OH, HO2) through the formation of positive ions followed by complex ion chemistry and NOx (N, NO, NO2) through the dissociation of molecular nitrogen. The HOx increases result in direct ozone destruction in the mesosphere and upper stratosphere via several catalytic loss cycles. Such middle atmospheric HOx-caused ozone loss is rather short-lived due to the relatively short lifetime (hours) of the HOx constituents. The NOx family has a considerably longer lifetime than the HOx family and can also lead to catalytic ozone destruction. EPP-caused enhancements of the NOx family can affect ozone directly, if produced in the stratosphere. Ozone decreases from the EPPs lead to a reduction in atmospheric heating and, subsequent atmospheric cooling. Conversely, EPPs can cause direct atmospheric heating through Joule heating. Measured HOx constituents OH and HO2 showed increases due to solar protons. Observed NOx constituents NO and NO2 were enhanced due to both solar protons and precipitating electrons. Other hydrogen- and nitrogen-ocntaining constituents were also measured to be directly influenced by EPPs, including N2O, HNO3, HO2NO2, N2OS, H2O2, ClONO2, HCl, and HOCl. Observed constituents ClO and CO were directly affected by EPPs as well. Many measurements indicated significant direct ozone decreases. A significant number of satellites housed instruments, which observed direct EPP-caused atmospheric effects, including Nimbus 4 (BUV), Nimbus 7 (SBUV), several NOAA platforms (SBUV/2), SME, UARS (HALOE, CLAES), SCISAT-1 (ACE-FTS), Odin (OSIRIS), Envisat-l (GOMOS, MIPAS, SCIAMACHY), and Aura (MLS). Measurements by rockets and ground-based radar also indicated EPP direct impacts. Atmospheric models have been used with some success in predicting the direct EPP impacts on the mesosphere and stratosphere. A review of the observed direct effects of EPP on the middle atmosphere will be given in this presentation.

Jackman, Charles H.↗

An Overview of Energetic Particle Precipitation Effects on the Earth's Atmosphere and (Potentially) Climate

Energetic precipitating particles (EPPs) can cause significant constituent changes in the polar mesosphere and stratosphere (middle atmosphere) during certain periods. Both protons and electrons can influence the polar middle atmosphere through ionization and dissociation processes. EPPs can enhance HOx (H, OH, HO2) through the formation of positive ions followed by complex ion chemistry and NOx (N, NO, NO2) through the dissociation of molecular nitrogen. The solar EPP-created HOx increases can lead to ozone destruction in the mesosphere and upper stratosphere via several catalytic loss cycles. Such middle atmospheric HOx-caused ozone loss is rather short-lived due to the relatively short lifetime (hours) of the HOx constituents. The HOx-caused ozone depletion of greater than 30% has been observed during several large solar proton events (SPEs) in the past 50 years. HOx enhancements due to SPEs were confirmed by observations in solar cycle 23. A number of modeling studies have been undertaken over this time period that show predictions of enhanced HOx accompanied by decreased ozone due to energetic particles. The solar EPP-created NOx family has a longer lifetime than the HOx family and can also lead to catalytic ozone destruction. EPP-caused enhancements of the NOx family can affect ozone promptly, if produced in the stratosphere, or subsequently, if produced in the lower thermosphere or mesosphere and transported to the stratosphere. NOx enhancements due to auroral electrons, medium and high energy electrons, relativistic electron precipitation (REP) events, and SPEs have been measured and/or modeled for decades. Model predictions and measurements show that certain years have significant winter-time meteorological events, which result in the transport of EPP-caused NOx enhancements in the upper mesosphere and lower thermosphere to lower altitudes. The NOx-caused ozone depletion has also been observed during several solar proton events (SPEs) in the past 50 years. Model predictions indicate that the longer-lived SPE-caused polar stratospheric and mesospheric ozone decrease can be >10% for up to five months past the largest events and is statistically significant; however, total ozone measurements do not indicate any long-term SPE impact.

Jackman, Charles H.↗

Insights into Sustainable Nitrogen Fixation by Gas-phase Spectroscopic Measurements and Global Modeling of Reaction Intermediates in Humid Nitrogen Plasma

In this article, there is growing interest in reacting molecular nitrogen and water to sustainably synthesize fixed forms of nitrogen such as ammonia. In particular, low-temperature plasmas can activate these relatively inert feedstocks at or near room temperature without a catalyst. However, because of the enhanced reactivity and nonequilibrium chemistry, a diverse range of products is formed, and the underlying reaction mechanisms are exceedingly complex. In this work, we studied a simplified reactor consisting of a gaseous plasma containing controlled mixtures of nitrogen gas and water vapor. Densities of key chemical species such as N, H, OH, NH, and NO were measured by emission and laser-based spectroscopy as a function of the relative humidity. A global model was constructed and the reaction network was validated by comparing calculated species densities with experiments. We discover that N, a key initial intermediate for ammonia, strongly decreases in the presence of water vapor, and as a result, ammonia formation becomes limited at high relative humidity. This decrease is surprisingly not because N itself reacts, but because one of its main sources, an excited molecular nitrogen state, is reacted away. In addition, oxidation pathways for nitrogen, which lead to NO and related products, are found to be favored over reduction pathways because the corresponding reverse reactions are less significant. Together, this understanding helps explain previously reported observations of selectivity toward nitrogen oxides over ammonia, particularly at higher relative humidities.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Dominant heterocyclic composition of dissolved organic nitrogen in the ocean: A new paradigm for cycling and persistence

Marine dissolved organic nitrogen (DON) is one of the planet’s largest reservoirs of fixed N, which persists even in the N-limited oligotrophic surface ocean. The vast majority of the ocean’s total DON reservoir is refractory (RDON), primarily composed of low molecular weight (LMW) compounds in the subsurface and deep sea. However, the composition of this major N pool, as well as the reasons for its accumulation and persistence, are not understood. Past characterization of the analytically more tractable, but quantitatively minor, high molecular weight (HMW) DON fraction revealed a functionally simple amide-dominated composition. While extensive work in the past two decades has revealed enormous complexity and structural diversity in LMW dissolved organic carbon, no efforts have specifically targeted LMW nitrogenous molecules. Here, we report the first coupled isotopic and solid-state NMR structural analysis of LMW DON isolated throughout the water column in two ocean basins. Together these results provide a first view into the composition, potential sources, and cycling of this dominant portion of marine DON. Our data indicate that RDON is dominated by 15N-depleted heterocyclic-N structures, entirely distinct from previously characterized HMW material. This fundamentally new view of marine DON composition suggests an important structural control for RDON accumulation and persistence in the ocean. The mechanisms of production, cycling, and removal of these heterocyclic-N-containing compounds now represents a central challenge in our understanding of the ocean’s DON reservoir.

58 GEOSCIENCES↗

Multisubstrate specificity shaped the complex evolution of the aminotransferase family across the tree of life

Aminotransferases (ATs) are an ancient enzyme family that play central roles in core nitrogen metabolism, essential to all organisms. However, many of the AT enzyme functions remain poorly defined, limiting our fundamental understanding of the nitrogen metabolic networks that exist in different organisms. Here, we traced the deep evolutionary history of the AT family by analyzing AT enzymes from 90 species spanning the tree of life (ToL). We found that each organism has maintained a relatively small and constant number of ATs. Mapping the distribution of ATs across the ToL uncovered that many essential AT reactions are carried out by taxon-specific AT enzymes due to wide-spread nonorthologous gene displacements. This complex evolutionary history explains the difficulty of homology-based AT functional prediction. Biochemical characterization of diverse aromatic ATs further revealed their broad substrate specificity, unlike other core metabolic enzymes that evolved to catalyze specific reactions today. Interestingly, however, we found that these AT enzymes that diverged over billion years share common signatures of multisubstrate specificity by employing different nonconserved active site residues. These findings illustrate that AT family enzymes had leveraged their inherent substrate promiscuity to maintain a small yet distinct set of multifunctional AT enzymes in different taxa. This evolutionary history of versatile ATs likely contributed to the establishment of robust and diverse nitrogen metabolic networks that exist throughout the ToL. The study provides a critical foundation to systematically determine diverse AT functions and underlying nitrogen metabolic networks across the ToL.

59 BASIC BIOLOGICAL SCIENCES↗

Line Interference Effects Using a Refined Robert-Bonamy Formalism: the Test Case of the Isotropic Raman Spectra of Autoperturbed N2

A symmetrized version of the recently developed refined Robert-Bonamy formalism [Q. Ma, C. Boulet, and R. H. Tipping, J. Chem. Phys. 139, 034305 (2013)] is proposed. This model takes into account line coupling effects and hence allows the calculation of the off-diagonal elements of the relaxation matrix, without neglecting the rotational structure of the perturbing molecule. The formalism is applied to the isotropic Raman spectra of autoperturbed N2 for which a benchmark quantum relaxation matrix has recently been proposed. The consequences of the classical path approximation are carefully analyzed. Methods correcting for effects of inelasticity are considered. While in the right direction, these corrections appear to be too crude to provide off diagonal elements which would yield, via the sum rule, diagonal elements in good agreement with the quantum results. In order to overcome this difficulty, a re-normalization procedure is applied, which ensures that the off-diagonal elements do lead to the exact quantum diagonal elements. The agreement between the (re-normalized) semi-classical and quantum relaxation matrices is excellent, at least for the Raman spectra of N2, opening the way to the analysis of more complex molecular systems.

nitrogen↗

Hyperstoichiometric Uranium Dioxides: Rapid Synthesis and Irradiation-Induced Structural Changes

Uranium dioxide (UO 2 ), the primary fuel for commercial nuclear reactors, incorporates excess oxygen forming a series of hyperstoichiometric oxides. Thin layers of these oxides, such as UO 2.12 , form readily on the fuel surface and influence its properties, performance, and potentially geologic disposal. This work reports a rapid and straightforward combustion process in uranyl nitrate–glycine–water solutions to prepare UO 2.12 nanomaterials and thin films. We also report on the investigation of the structural changes induced in the material by irradiation. Despite the simple processing aspects, the combustion synthesis of UO 2.12 has a sophisticated chemical mechanism involving several exothermic steps. Raman spectroscopy and single-crystal X-ray diffraction (XRD) measurements reveal the formation of a complex compound containing the uranyl moiety, glycine, H 2 O, and NO 3 – groups in reactive solutions and dried combustion precursors. Combustion diagnostic methods, gas-phase mass spectroscopy, differential scanning calorimetry (DSC), and extracted activation energies from DSC measurements show that the rate-limiting step of the process is the reaction of ammonia with nitrogen oxides formed from the decomposition of glycine and uranyl nitrate, respectively. However, the exothermic decomposition of the complex compound determines the maximum temperature of the process. In situ transmission electron microscopy (TEM) imaging and electron diffraction measurements show that the decomposition of the complex compound directly produces UO 2 . The incorporation of oxygen at the cooling stage of the combustion process is responsible for the formation of UO 2.12 . Spin coating of the solutions and brief annealing at 670 K allow the deposition of uniform films of UO 2.12 with thicknesses up to 300 nm on an aluminum substrate. Irradiation of films with Ar 2+ ions (1.7 MeV energy, a fluence of up to 1 × 10 17 ions/cm 2 ) shows unusual defect-simulated grain growth and enhanced chemical mixing of UO 2.12 with the substrate due to the high uranium ion diffusion in films. As a result, the method described in this work allows the preparation of actinide oxide targets for fundamental nuclear science research and studies associated with stockpile stewardship.

38 RADIATION CHEMISTRY, RADIOCHEMISTRY, AND NUCLEA↗

The photochemistry of the paleoatmosphere

Recent progress in the understanding of the chemistry and photochemistry of the paleoatmosphere is reviewed with emphasis on the application of photochemical models to the investigation of the evolution of the atmosphere. Photochemical calculations are presented which show that a primordial highly reducing atmosphere composed of methane and ammonia, if it formed at all, would be short-lived in the presence of solar ultraviolet radiation, giving way rapidly to a more mildly reducing atmosphere of carbon dioxide and nitrogen. Estimations of O2 produced from the photolysis of water vapor prior to the emergence of photosynthesis range from less than 10 to the -14th to 0.1 times the present atmospheric level, indicating the need for further research. A series of photochemical models of increasing complexity has been developed to study the evolution of atmospheric ozone taking into account reactions with O atoms, hydrogen oxides, nitrogen oxides, and chlorine as well as vertical transport, temperature and tropospheric chemistry so that the total content and vertical distribution of O3 may be determined for a specified level of paleoatmospheric O2.

Levine, J. S.↗

Ultracold cryogenic TEM with liquid helium and high stability

Cryogenic transmission electron microscopy has revolutionized structural biology and materials science. To image below liquid nitrogen temperatures, various liquid helium stages have been constructed but have proven to be complex and unstable, making high-resolution imaging challenging. This problem is even more pronounced in side-entry specimen holders common on modern transmission electron microscopes. Here, we introduce an ultracold liquid helium transmission electron microscope side-entry specimen holder, featuring continuous cryogen flow and vibration decoupling. This instrument is compatible with modern aberration-corrected microscopes and achieves sub-25 K base temperature, ±2 mK thermal stability over many hours, and atomic resolution—setting the stage for a new era of cryogenic electron microscopy.

42 ENGINEERING↗

Separation of isoprene from biologically-derived gas streams

Renewable organic precursors, including olefinic compounds such as isoprene, have attracted interest from the polymer and pharmaceutical industries. Biologically-derived processes can generate these target compounds; however, their gaseous product streams are complex mixtures of condensable organic vapors (COVs), water vapor, carbon dioxide (CO 2 ), and/or nitrogen (N 2 ). Because COVs, CO 2 and water vapor are known to alter polymer membranes, mixed gas separations data at ambient and elevated temperatures are limited. Here, this study focused on two classes of polymer membranes, glassy [polyetherimide (Ultem®)] and a rubbery [polydimethylsiloxane (PDMS)] with results indicating that isoprene separation is possible in humidified gas environment (2–4 vol% water). Gas permeabilities of these membranes did not noticeably change in the presence of humidity; however, the selectivity of these membranes was significantly lower compared to their performance under dry conditions. The role of water vapor in gas transport was derived from the energy of activation of permeation (E p ) for PDMS and Ultem® from 30–80°C in humidified mixed gas streams. For both polymers, E p data shows a slight decrease in selectivity with the other gases (hydrogen, N 2 , CO 2 , and methane) at elevated temperatures in the presence of water vapor. Thus, these COVs separations are feasible with polymer membranes in the presence of humidified gas streams, even in the case of glassy and rubbery membranes in series.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗