水相中氢气浓度检测方法的建立

近年来,研究表明氢分子具有广泛的生物学效应,饮用富氢水（hydrogen-rich water,HRW）是其主要的摄取方法,但目前对于水相中氢气浓度检测方法的研究甚少。为了建立适用于测定水相中氢气浓度的检测方法,利用纯水氢气发生器制备饱和富氢水。然后,利用氢气微电极直接测定水相中的氢气浓度,结果表明,在不同氢气浓度范围内（0～1.620 0 mg·L-1和0～0.202 5 mg·L-1）,氢气浓度与微电极信号值均呈现良好的线性关系,方法检出限（method detection limit,MDL）为4.3×10-3 mg·L-1。同时,采用顶空方式将水相中的氢气转移到气相中,通过气相色谱法测定氢气的浓度,结果表明,在不同氢气浓度范围内（0～1.620 0 mg·L-1和0～0.202 5 mg·L-1）,氢气浓度与气相色谱峰面积均具有良好的线性关系,MDL为8.7×10-4 mg·L-1。研究结果表明,氢微电极法和气相色谱法均可用于水相中氢气浓度的精确定量,即成功建立了采用氢气微电极及顶空气相色谱测定水相中氢气含量的检测方法。     

超临界水处理生物质制氢技术

简要论述了工业制氢的常用方法及各自的优缺点。基于目前国际上的最新研究动向,重点就超临界水催化汽化生物质制氢技术的研究现状进行了归纳和综述。认为超临界水催化生物质制氢技术具有环境友好、资源可再生以及效率高等技术优势,具有良好开发前景,应该予以重视。重点讨论了目前研究工作所取得的成果、面临的主要技术问题和可能的解决途径,指出了未来的重点研究方向。     

Boron Nanoparticles for Room‐Temperature Hydrogen Generation from Water

Boron nanoparticles (BNPs) are of great interest for applications such as neutron capture therapy of cancer cells, hydrogen generation from water, and high energy density fuels. Boron is particularly interesting for chemical water splitting, because of its high gravimetric hydrogen generation potential of 277 g H₂ per kg B. However, only a few studies of water splitting by reaction with boron are available, and those have used high temperature steam with external heating. Room‐temperature boron hydrolysis is of great interest from both scientific and practical perspectives. The studies presented here demonstrate that high purity amorphous BNPs can be oxidized by water to produce H₂ at room temperature, without external energy input, in the presence of catalytic quantities of an alkali metal or alkali metal hydride. The BNPs are produced in a single step gas phase process via CO₂ laser‐induced pyrolysis of mixtures of B₂H₆ and SF₆. The BNPs are spherical with a primary particle diameter of 10–15 nm, narrow size distribution, and specific surface area exceeding 250 m² g^?1. This first demonstration of room‐temperature chemical splitting of liquid water using boron opens up exciting new possibilities for on‐demand hydrogen generation at high gravimetric capacity.     

Ozone produced by corona discharge in the presence of water

Significant levels of ozone have been detected in a reaction flask that was designed for studies using negative air ions generated from corona discharge. While diluting the gas in the vessel at the rate of 1.5 liters/min., more than 1.0 ppm O₃ was measured in the vessel when grounded water was present. Ozone levels were much less (15 ppb) in ambient air near the ion generator. Even in the absence of water, O₃ levels were only 7 ppb in the reaction flask. Without the corona discharge apparatus, levels of ozone in the gas phase (with water present) averaged 6 ppb. These results demonstrate that when negative air ions are generated from corona discharge in the presence of water, significant levels of ozone are produced. Therefore, O₃ could be directly responsible for many of the oxidizing effects that have been reportedly due to negative air ions.     

Photocatalytic Water Splitting for Solar Hydrogen Production Using the Carbonate Effect and the Z‐Scheme Reaction

The development of innovative technologies for solar energy conversion and storage is important for solving the global warming problem and for establishing a sustainable society. The photocatalytic water‐splitting reaction using semiconductor powders has been intensively studied as a promising technology for direct and simple solar energy conversion. However, the evolution of H₂ and O₂ gases in a stoichiometric ratio (H₂/O₂ = 2) is very difficult owing to various issues, such as an unfavorable backward reaction and mismatched band potentials. Two important findings have widened the variety of photocatalysts available for stoichiometric water‐splitting, viz. the carbonate anion effect and the Z‐scheme photocatalytic reaction using a redox mediator. The bicarbonate anion has been found to act as a redox catalyst via preferential peroxide formation and subsequent decomposition to O₂. As the Z‐scheme reaction using a redox mediator mitigates band potential mismatches, it is widely applicable for various visible‐light‐active photocatalysts. This review describes the development of photocatalytic water‐splitting for solar hydrogen production using the carbonate anion effect and the Z‐scheme reaction. Moreover, recent developments in photocatalysis–electrolysis hybrid systems, an advanced Z‐scheme reaction concept, are also reviewed for practical and economical hydrogen production.     

HX-MS2 for high performance conformational analysis of complex protein states

Water-mediated hydrogen exchange (HX) processes involving the protein main chain are sensitive to structural dynamics and molecular interactions. Measuring deuterium uptake in amide bonds provides information on conformational states, structural transitions and binding events. Increasingly, deuterium levels are measured by mass spectrometry (MS) from proteolytically generated peptide fragments of large molecular systems. However, this bottom-up method has limited spectral capacity and requires a burdensome manual validation exercise, both of which restrict analysis of protein systems to generally less than 150 kDa. In this study, we present a bottom-up HX-MS² method that improves peptide identification rates, localizes high-quality HX data and simplifies validation. The method combines a new peptide scoring algorithm (WUF, weighted unique fragment) with data-independent acquisition of peptide fragmentation data. Scoring incorporates the validation process and emphasizes identification accuracy. The HX-MS² method is illustrated using data from a conformational analysis of microtubules treated with dimeric kinesin MCAK. When compared to a conventional Mascot-driven HX-MS method, HX-MS² produces two-fold higher α/β-tubulin sequence depth at a peptide utilization rate of 74%. A Mascot approach delivers a utilization rate of 44%. The WUF score can be constrained by false utilization rate (FUR) calculations to return utilization values exceeding 90% without serious data loss, indicating that automated validation should be possible. The HX-MS² data confirm that N-terminal MCAK domains anchor kinesin force generation in kinesin-mediated depolymerization, while the C-terminal tails regulate MCAK-tubulin interactions.     

An Apparatus for Measuring Volumes of Small Objects

An apparatus for measuring volumes of small objects such as tissue blocks is described. The apparatus measures volumes by fluid displacement and consists of a micropipette adapted to fit the mouth of an Erleiuneyer flask, a Luer adaptor fused to the side of the flask, and a glass syringe. When assembled with fluid enclosed, the fluid rises to a low level in the micropipette. Withdrawal of fluid into the syringe lowers the fluid level below the mouth of the flask. The micropipette is raised, the object to be measured is placed in the flask, and the micropipette is joined to the flask again. Fluid returned to the flask from the syringe rises to a higher level in the micropipette. The difference between the two fluid levels equals the volume of the object measured.

This apparatus gives reproducible measurements and can be calibrated for absolute volume determination. It is inexpensive to construct and easy to use.     

Advances In Biotreatment of Acid Mine Drainage and Biorecovery of Metals: 2. Membrane Bioreactor System for Sulfate Reduction

Several biotreatmemt techniques for sulfate conversion by the sulfate reducing bacteria (SRB) have been proposed in the past, however few of them have been practically applied to treat sulfate containing acid mine drainage (AMD). This research deals with development of an innovative polypropylene hollow fiber membrane bioreactor system for the treatment of acid mine water from the Berkeley Pit, Butte, MT, using hydrogen consuming SRB biofilms. The advantages of using the membrane bioreactor over the conventional tall liquid phase sparged gas bioreactor systems are: large microporous membrane surface to the liquid phase; formation of hydrogen sulfide outside the membrane, preventing the mixing with the pressurized hydrogen gas inside the membrane; no requirement of gas recycle compressor; membrane surface is suitable for immobilization of active SRB, resulting in the formation of biofilms, thus preventing washout problems associated with suspended culture reactors; and lower operating costs in membrane bioreactors, eliminating gas recompression and gas recycle costs. Information is provided on sulfate reduction rate studies and on biokinetic tests with suspended SRB in anaerobic digester sludge and sediment master culture reactors and with SRB biofilms in bench-scale SRB membrane bioreactors. Biokinetic parameters have been determined using biokinetic models for the master culture and membrane bioreactor systems. Data are presented on the effect of acid mine water sulfate loading at 25, 50, 75 and 100 ml/min in scale-up SRB membrane units, under varied temperatures (25, 35 and 40 °C) to determine and optimize sulfate conversions for an effective AMD biotreatment. Pilot-scale studies have generated data on the effect of flow rates of acid mine water (MGD) and varied inlet sulfate concentrations in the influents on the resultant outlet sulfate concentration in the effluents and on the number of SRB membrane modules needed for the desired sulfate conversion in those systems. The pilot-scale data indicate that the SRB membrane bioreactors systems can be applied toward field-scale biotreatment of AMD and for recovery of high purity metals and an agriculturally usable water.     

The formation of hydrogen peroxide during the oxidation of reduced nicotinamide adenine dinucleotide by cytochrome o from Vitreoscilla.

The formation of hydrogen peroxide during the oxidation of NADH by purified preparations of cytochrome o has been demonstrated by employing three independent methods: polarographic, colorimetric, and fluorometric. The first two methods were used to assay for the accumulation of hydrogen peroxide and showed that hydrogen peroxide did accumulate as a product, but only about 30% of the oxygen consumed or 15 to 20% of the NADH oxidized was recoverable as hydrogen peroxide. This lack of 1:1 stoichiometry was not due to residual catalase activity in these preparations which could be eliminated by freeze-thawing. Thus, hydrogen peroxide may not be the sole or primary product of the NADH-cytochrome o oxidase reaction. The fluorometric assay could be coupled directly to the NADH-cytochrome o oxidase reaction in one medium, and this method showed that hydrogen peroxide was generated continuously from the beginning of the reaction in a 1:1 stoichiometry, hydrogen peroxide generated to NADH oxidized. This result suggests that hydrogen peroxide is an intermediate that can be trapped efficiently under the conditions of the fluorometric assay, whereas under the conditions of the first two assays most of the hydrogen peroxide generated undergoes further reaction. Exogenously added FAD or FMN increased the percentage of hydrogen peroxide that accumulated in the NADHcytochrome o oxidase reaction. Flavin is believed to act on the reductase side of cytochrome o so the increased percentage of hydrogen peroxide is not likely to result from the direct reaction of reduced flavin with oxygen.     

Single Atoms and Clusters Based Nanomaterials for Hydrogen Evolution,Oxygen Evolution Reactions,and Full Water Splitting

The sustainable and scalable production of hydrogen through hydrogen evolution reaction (HER) and oxygen through oxygen evolution reaction (OER) in water splitting demands efficient and robust electrocatalysts. Currently, state‐of‐the‐art electrocatalysts of Pt and IrO₂/RuO₂ exhibit the benchmark catalytic activity toward HER and OER, respectively. However, expanding their practical application is hindered by their exorbitant price and scarcity. Therefore, the development of alternative effective electrocatalysts for water splitting is crucial. In the last few decades, substantial effort has been devoted to the development of alternative HER/OER and water splitting catalysts based on various transition metals (including Fe, Co, Ni, Mo, and atomic Pt) which show promising catalytic activities and durability. In this review, after a brief introduction and basic mechanism of HER/OER, the authors systematically discuss the recent progress in design, synthesis, and application of single atom and cluster‐based HER/OER and water splitting catalysts. Moreover, the crucial factors that can tune the activity of catalysts toward HER/OER and water splitting such as morphology, crystal defects, hybridization of metals with nonmetals, heteroatom doping, alloying, and formation of metals inside graphitic layered materials are discussed. Finally, the existing challenges and future perspectives for improving the performance of electrocatalysts for water splitting are addressed.     

Is there a weak H‐bond → LBHB transition on tetrahedral complex formation in serine proteases?

The transformation of a weak hydrogen bond in the free enzyme into a low-barrier hydrogen bond (LBHB) in the tetrahedral intermediate has been suggested as an important factor facilitating catalysis in serine proteases. In this work, we examine the structure of the H-bond in the Asp102-His57 diad of serine proteases in the free enzyme and in a covalent tetrahedral complex (TC) with a trifluoromethylketone inhibitor. We apply ab initio quantum mechanical calculations to models consisting of a large molecular fragment of the enzyme active site, and the combined effect of the rest of the protein body and the solvation by surrounding bulk water was simulated by a self-consistent reaction field method in our novel QM/SCRF(VS) approach. Potential profiles of adiabatic proton transfer in the Asp102-His57 diad in these model systems were calculated. We conclude that the hydrogen bond in both the free enzyme and in the enzyme-inhibitor TC is a strong ionic asymmetric one-well hydrogen bond, in contrast to a previous suggestion that it is a weak H-bond in the former and a double-well LBHB in the latter.     

Formation of Hydroxyl Radicals on Reaction of Hypochlorous Acid with Ferrocyanide, a Model IRON(II) Complex

Hypochlorous acid reacts with the model iron(II) complex, ferrocyanide (Fe(CN)₆^4-) in aqueous solution with the rate constant 220 ± 15 dm³ mol^-1 s^-1. Free hydroxyl radicals are formed in this reaction in 27% yield as shown by the hydroxylation of benzoate to give a product distribution identical to that of free (radiolytically generated) hydroxyl radicals. This reaction is three orders of magnitude faster than the analogous reaction involving hydrogen peroxide (the Fenton reaction), suggesting that the hypochlorous acid generated by activated neutrophils may be a source of hydroxyl radicals.     

Avoiding high-valent iron intermediates: superoxide reductase and rubrerythrin

The Fenton or Fenton-type reaction between aqueous ferrous ion and hydrogen peroxide generates a highly oxidizing species, most often formulated as hydroxyl radical or ferryl ([Fe(IV)O](2+)). Intracellular Fenton-type chemistry can be lethal if not controlled. Nature has, therefore, evolved enzymes to scavenge superoxide and hydrogen peroxide, the reduced dioxygen species that initiate intracellular Fenton-type chemistry. Two such enzymes found predominantly in air-sensitive bacteria and archaea, superoxide reductase (SOR) and rubrerythrin (Rbr), functioning as a peroxidase (hydrogen peroxide reductase), contain non-heme iron. The iron coordination spheres in these enzymes contain five or six protein ligands from His and Glu residues, and, in the case of SOR, a Cys residue. SOR contains a mononuclear active site that is designed to protonate and rapidly expel peroxide generated as a product of the enzymatic reaction. The ferrous SOR reacts adventitiously but relatively slowly (several seconds to a few minutes) with exogenous hydrogen peroxide, presumably in a Fenton-type reaction. The diferrous active site of Rbr reacts more rapidly with hydrogen peroxide but can divert Fenton-type reactions towards the two-electron reduction of hydrogen peroxide to water. Proximal aromatic residues may function as radical sinks for Fenton-generated oxidants. Fenton-initiated damage to these iron active sites may become apparent only under extremely oxidizing intracellular conditions.     

Relaxation of liquid water states with altered stoichiometry

The integral parameter of the physical state of liquid water, its stoichiometric composition, is considered. This factor takes into account that the substance of liquid water contains hydrogen and oxygen atoms in the proportion other than the ideal 2: 1 ratio characteristic of a separate water molecule. The stoichiometry index x of water — the deviation from the ideal ratio — is shown to be an independent metastable variable that determines equilibrium concentrations of the oxygen-hydrogen molecules. The optical method for measuring x is suggested, and a laser interferometer is described that is capable of measuring the stoichiometry index to an accuracy of 10^?5. Possible applications of the instrument in biophysics of water systems are discussed.     

Multiple controls for the variability of hydrogen isotopic compositions in higher plant n-alkanes from modern ecosystems

We performed a global scale analysis of available leaf wax n -alkane δ D data compiled from our new results, as well as from the literature and expressed as average values of D/H ratios from three common lipids of n -alkanes with odd carbon numbers ( n -C₂₇, n -C₂₉, and n -C₃₁) from living higher plants. Our results clearly indicate multiple controls of hydrogen isotope composition and its variability in plants leaf wax. (1) At the global scale, precipitation δ D values play a dominating factor that exercises the first order of control for hydrogen isotopic compositions in plant leaf wax. The hydrogen isotopic composition of plant leaf wax tracks the decreasing trend of precipitation δ D with increasing latitude. (2) Because of different water acquisition systems, plant life form influences the hydrogen isotopic composition of leaf wax n -alkanes with woody plants and grasses having different responses to the change of global precipitation δ D. (3) Physiological difference, due to different photosynthesis pathways or different water usage strategies, can leave an imprint on δ D patterns of plant leaf waxes, causing δ D variations among plants using the same source water. While these results better explain the variability of hydrogen isotope composition in leaf wax, they also have important implications for the interpretation of n -alkane δ D data from fossils and ancient sediments.     

Hydrogen exchange and ligand binding: ligand-dependent and ligand-independent protection in the Src SH3 domain

Amide hydrogen-deuterium exchange has proven to be a powerful tool for detecting and characterizing high-energy conformations in protein ensembles. Since interactions with ligands can modulate these high-energy conformations, hydrogen exchange appears to be an ideal experimental probe of the physical mechanisms underlying processes like allosteric regulation. The chemical mechanism of hydrogen exchange, however, can complicate such studies. Here, we examine hydrogen exchange rates in a simple model system, the c-Src SH3 domain interacting with a short peptide ligand. Addition of ligand slows the rates of hydrogen exchange at nearly every amide for which we can obtain data. Careful analysis, however, reveals that this slowing is due primarily to a reduction in the population of free protein in the system, and not to any specific property of the complex. We present a method to separate the contributions of free and bound protein to the exchange kinetics that has allowed us to identify the subset of amides where exchange arises directly from the complex. These results demonstrate that the slowing of hydrogen exchange induced by ligand interactions should be interpreted with caution, and more extensive experiments are required to correlate changes in hydrogen exchange with changes in structure or internal dynamics.     

Monitoring the biochemical hydrogen and methane potential of the two-stage dark-fermentative process

A two-step process has been recently proposed whereby the products of biological hydrogen production processes are used as substrates for biological methane production. The aim of the present study is to evaluate a simple bench-scale batch procedure for measuring the biochemical hydrogen and methane potential of organic substances as a two-step simulated process. Glucose fermentation showed an hydrogen and methane recovery (measured as the ratio of electron equivalents recovered as hydrogen and methane and electron equivalents of the initial substrate added) from the initial substrate of 13.3% and 75.5%, respectively, that approximates mass balance closure. On the contrary, gas recoveries ranging from 61% to 75% were measured from wastes originating from the food-industry. Moreover, the results demonstrate that the substrate origins significantly influence the ratio of H₂ and CH₄ recovery.     

稳定同位素技术在植物水分利用研究中的应用

近20a稳定同位素技术在植物生态学研究中的应用得到了长足发展,使得对植物与水分关系也有了更深一步的了解。介绍稳定同位素性碳、氢、氧同位素在研究植物水分关系中的应用及进展,以期能为国内植物水分利用研究提供参考。由于植物根系从土壤中吸收水分时并不发生同位素分馏,对木质部水分同位素分析有助于对植物利用水分来源,生态系统中植物对水分的竞争和利用策略的研究,更好地了解生态系统结构与功能。稳定碳同位素作为植物水分利用效率的一个间接指标,在不同水分梯度环境中,及植物不同代谢产物与水分关系中有着广泛的应用。同位素在土壤-植被-大气连续体水分中的应用,有助于了解生态系统的水分平衡。随着稳定同位素方法的使用,植物与水分关系的研究将取得更大的进展。     

Mathematical model for the luminol chemiluminescence reaction catalyzed by peroxidase

A kinetic model that accurately describes intensity vs. time reaction profiles for the chemiluminescence reaction between luminol and hydrogen peroxide, as catalyzed by horseradish perioxdase, is derived and evaluated. A set of three differential equations is derived and solved to provide intensity time information for the first 200 seconds of the reaction. The model accurately predicts intensity-time profiles when literature values are used for all but one of the reaction rate constants. Furthermore, the model predicts a nonlinear curve for plots of light intensity versus the initial hydrogen peroxide concentration. Experimental data confirm that such plots are nonlinear. Finally, a linear double-reciprocal plot is predicted by the model and the experimental data verify this relationship. (c) 1993 Wiley & Sons, Inc.     

A nonenzymatic method for determination of hydrogen peroxide and organic peroxides

Reduction of hydrogen peroxide and organic peroxides (t-butyl hydroperoxide and linoleic acid hydroperoxide) was achieved with homovanillic acid as hydrogen donor in the presence of the triethylenetetramine-Fe3+ complex. By the catalytic action of this complex, homovanillic acid is oxidized to its fluorescent dimer. Based on this reaction a fluorometric method for the measurement of the hydroperoxides mentioned above is described. The method can be extended to the determination of substrate-enzyme systems that produce hydrogen peroxide, e.g., glucose-glucose oxidase. The method allows the determination of substances such as hydrogen peroxide and t-butyl hydroperoxide with an accuracy and precision of less than 3%. Glucose can be determined with similar precision and an accuracy of 4.7%.