Significant Isotope Effect Research Articles

Variation and fractionation of δ2H, δ18O, and δ17O stable isotopes from irrigation water to soil, grapes, and wine for the traceability of geographical origins

To investigate the variation and fractionation of stable isotopes from irrigation water to soil, grapes, and wine, δ2H, δ18O, and δ17O in different samples from 10 regions in China were determined using a water isotope analyser. The values were significantly different among regions according to the chemometric analysis. All isotopes were significantly and positively correlated with irrigation water–soil and grape–wine. A significant water isotopic fractionation effect was observed from the irrigation water to the soil, grapes, and wine. Stable isotope distribution characteristics correlated with longitude, latitude, altitude, temperature, precipitation, station pressure and wind speed. The linear discriminant analysis (LDA), random forest (RF), support vector machine (SVM), and feed-forward neural network (FNN) models 58.33–100 %, 80–100 %, 53.33–100 %, and 73.33–100 % accurate for distinguishing the geographical origins of all samples from training and test data, respectively. These findings provide a theoretical basis for authenticating the geographic origin of Chinese wines using stable isotope analysis.

Stable isotopic characterization and sources of ammonium in wet deposition at the Danjiangkou Reservoir

Ammonium nitrogen (NHx) in atmospheric wet deposition is a primary external nitrogen source for reservoir-type water sources, and identifying its sources is crucial for controlling nitrogen pollution in water. This study aimed to examine the NHx characteristics and sources in wet deposition in the Xichuan reservoir area of the Danjiangkou Reservoir, the South-to-North Water Diversion Project (SNWDP) water source, from September 2019 to August 2020. Major inorganic nitrogen concentrations in precipitation and ammonia-nitrogen isotope (δ15N–NH4+) values were measured, and the sources were analyzed using the Bayesian mixing model. The results showed that NH4+ was the main inorganic nitrogen form in wet deposition, with a concentration ranged of 0.16–5.26 mg L−1. The δ15N–NH4+ values ranged from −14.7‰ to +6.3‰, with significant isotopic effects from agricultural sources and climatic conditions, showing higher values in summer and lower values in winter. The release of ammonia (NH3) from agricultural sources was the primary NHx source (58.0%–76.6%), with fertilizer application being the most significant contributor (32.1%–46.6%). Seasonally, the relative contribution of agricultural sources to NH4+ in wet deposition was higher in autumn and winter. Spatially, agricultural activities significantly impacted atmospheric nitrogen accumulation across the reservoir area. This study quantified NHx sources in wet deposition, providing isotopic evidence and scientific references for nitrogen control measures and atmospheric nitrogen cycling studies.

Hydrogen bond symmetrisation in D2O ice observed by neutron diffraction

Hydrogen bond symmetrisation is the phenomenon where a hydrogen atom is located at the centre of a hydrogen bond. Theoretical studies predict that hydrogen bonds in ice VII eventually undergo symmetrisation upon increasing pressure, involving nuclear quantum effect with significant isotope effect and drastic changes in the elastic properties through several intermediate states with varying hydrogen distribution. Despite numerous experimental studies conducted, the location of hydrogen and hence the transition pressures reported up to date remain inconsistent. Here we report the atomic distribution of deuterium in D2O ice using neutron diffraction above 100 GPa and observe the transition from a bimodal to a unimodal distribution of deuterium at around 80 GPa. At the transition pressure, a significant narrowing of the peak widths of 110 is also observed, attributed to the structural relaxation by the change of elastic properties.

Weathering-induced Sb isotope fractionation during leaching of stibnite and formation of secondary Sb minerals

In this work, we investigated the extent of antimony (Sb) isotopic fractionation during weathering of stibnite at supergene conditions. Antimony isotope data have been obtained from secondary Sb minerals collected from Pezinok, Dobšiná (both Slovakia) and Allchar (North Macedonia) deposits and mine tailings. The Sb isotope compositions of sulfides and secondary Sb minerals formed on the primary stibnite [Sb2S3] or in mine tailings grains were compared with each other. Furthermore, we experimentally investigated Sb isotope fractionation during stibnite leaching with different acids. Our study reveals a large isotopic range for δ123Sb (from −0.50 to +0.69 ‰) for secondary Sb minerals. They are either isotopically indistinguishable or isotopically lighter than the primary stibnite. Isotopically indistinguishable weathering products likely formed by quantitative Sb transfer from stibnite to the secondary minerals, such as brandholzite [Mg(H2O)6[Sb(OH)6]2] from Pezinok. Isotopic fractionation towards lighter δ123Sb was observed for adsorption of Sb onto iron oxides. Distinctly isotopically lighter δ123Sb was observed in secondary Sb minerals tripuhyite [FeSbO4], chapmanite [Fe3+2Sb3+(Si2O5)O3(OH)], hydroxyferroromeite [Fe2Sb2O6(OH)], and stibiconite [Sb3O6OH] that either replace stibnite or formed in mine tailings from the pore solutions. These secondary minerals were likely generated by partial precipitation of Sb from aqueous solutions produced by dissolution of stibnite. In the leaching experiments with HCl and oxalic acid, Sb was leached without significant isotope effects during the first 2–3 days, followed by a drop of the dissolved Sb concentration associated with Sb isotope fractionation towards high δ123Sb in the leachate (by up to 0.5 ‰) after 4–7 days. We interpret these observations to be related to the precipitation of secondary Sb oxides with low δ123Sb, resulting in an isotopically heavy dissolved Sb pool. These findings are in agreement with previous results of isotopically heavy groundwater and mine drainage water with δ123Sb > +0.36 ‰ that may suggest that the ‘truly’ dissolved (operationally defined as <0.45 μm) Sb fraction in general may be isotopically heavy.

Undetected perovskite phase interference with zirconolite dissolution measurements

Zirconolite ceramics present a chemically durable host matrix for waste actinides, but zirconolite dissolution rates reported in the literature often vary significantly. Here, the release of Ca and Al from a hot isostatically pressed zirconolite (Ca0.8Zr0.9Ce0.3Ti1.6Al0.4O7.0) was shown to be predominantly driven by preferential dissolution of minor perovskite and alumina phases. Both phases were undetectable by XRD, and the perovskite was difficult to detect by SEM-EDS. Whilst the zirconolite phase exhibited no signs of alteration, dissolution of the perovskite proceeded congruently without forming a hydrated altered layer or diffusion of protons into the solid that would be indicative of an ion-exchange mechanism. The weak temperature dependence of dissolution (40, 90 and 150 °C) showed that kinetics were limited by transport and a mixed transport-surface controlled reaction for Ca and Al, respectively. A significant H2O-D2O isotope effect on dissolution was observed for Ca but not for Al at all temperatures. The former was consistent with an abated rate of hydrolysis in the absence of a contribution from diffusion, whilst the latter could be attributed to differences in the activated complex for Ca and Al release through hydrolysis. These results demonstrate the role of a secondary phase perovskite in the dissolution kinetics of zirconolite even when perovskite occurs at low concentration and evades detection by bulk techniques such as XRD. This study provides a potential explanation of variations in zirconolite ceramic dissolution rates present in the literature and provides a null result to tests of an incongruent Ca release mechanism from zirconolite.

Evolution of hydrogen isotopes retention behavior of in-situ boronization films in EAST

Effective management of hydrogen isotopes retention in plasma-facing materials (PFMs) is crucial, particularly when utilizing tritium (T) as fuel, for the success of burning plasma operations. Boronization, a widely employed technique for controlling fuel recycling and mitigating impurity influx from plasma-surface interactions into the core of burning plasma, significantly influences hydrogen isotopes retention in PFMs. In this study, boronization films were generated by ion cyclotron range of frequency (ICRF) discharge assisted boronization with C2B10H12 as boron source on tungsten substrates at Experimental Advanced Superconducting Tokamak (EAST) which employing ITER-like water-cooled W monoblock PFMs and components (PFCs), followed by in-situ glow discharge (GD) cleaning for 2 h and 20 shots (180 s) edge-plasma exposure. Employing the Material and Plasma Evaluation System (MAPES), representative samples were analyzed after each process. The resultant carbon–boron films, dense and continuous, exhibited thickness up to 120 nm and were identified as amorphous in structure. It was observed that the D2-GD cleaning resulted in a significant isotopic exchange effect, effectively reducing the hydrogen (H) retention in the carbon–boron films. This hydrogen isotope replacement efficiency was found to be influenced by the thickness of the films. Notably, after boundary plasma exposure, samples with thicker films demonstrated an enhanced capacity to capture deuterium (D), adsorbing 10 times more D than bare tungsten (W). Our findings offer transformative insights for T recycling analysis and the plasma operation of devices like International Thermonuclear Experimental Reactor (ITER), highlighting the impact of boronization and subsequent treatments on hydrogen isotope retention behavior in PFMs.

Radiationless decay spectrum of O 1s double core holes in liquid water.

We present a combined experimental and theoretical investigation of the radiationless decay spectrum of an O 1s double core hole in liquid water. Our experiments were carried out using liquid-jet electron spectroscopy from cylindrical microjets of normal and deuterated water. The signal of the double-core-hole spectral fingerprints (hypersatellites) of liquid water is clearly identified, with an intensity ratio to Auger decay of singly charged O 1s of 0.0014(5). We observe a significant isotope effect between liquid H2O and D2O. For theoretical modeling, the Auger electron spectrum of the central water molecule in a water pentamer was calculated using an electronic-structure toolkit combined with molecular-dynamics simulations to capture the influence of molecular rearrangement within the ultrashort lifetime of the double core hole. We obtained the static and dynamic Auger spectra for H2O, (H2O)5, D2O, and (D2O)5, instantaneous Auger spectra at selected times after core-level ionization, and the symmetrized oxygen-hydrogen distance as a function of time after double core ionization for all four prototypical systems. We consider this observation of liquid-water double core holes as a new tool to study ultrafast nuclear dynamics.

Significant Isotope Effects from the Nonadiabatic Couplings in the Cl(2P) + HD(v = 0, j = 0) Reaction.

The impact of non-Born-Oppenheimer couplings on the isotopic effects in the reaction of the Cl(2P) atom with the HD (v = 0, j = 0) molecule is investigated with our recently developed nonadiabatic time-independent quantum scattering methods, where the full open-shell characteristics are included in the six-state model, and also with the recently developed two-state model solving by time-independent methods, where part of the open-shell characteristic is included. The same reaction is also calculated with the simple adiabatic model using the lowest adiabatic potential energy surface. Compared with the results from different models, it is found that the reactivity of the Cl + HD → HCl + D channel is significantly overestimated in the adiabatic model. In contrast, the reactivity of the other channel agrees well with the nonadiabatic models. This is due to the van der Waals well in the reactant channel being changed a lot by including the nonadiabatic couplings. These quantum dynamics calculations suggest that sometimes the adiabatic model should be used with caution; otherwise, it may result in significant deviations for some reactions.

Hyperfine excitation of NH and ND by molecular hydrogen

The NH and ND radicals are of key importance in the comprehension of nitrogen chemistry and the enhancement of deuterated molecules in the interstellar medium. Observations by space telescopes yield spectra that can resolve the fine and hyperfine structure of these radicals, a consequence of the electronic and magnetic interactions of nitrogen, hydrogen, and deuterium nuclei. Accurate rate coefficients, induced by collisions with H2, are required to interpret spectra of these radicals. We report the first rate coefficients for fine and hyperfine transitions of NH and ND in collision with both ortho- and para-H2. Based on a recent four-dimensional potential energy surface, fine-structure resolved cross sections and rate coefficients are computed with the time-independent close-coupling method over a temperature range of 5–300 K. Our calculations include the first 25 energy levels of NH and ND. Hyperfine resolved cross sections and rate coefficients are determined using the infinite-order sudden (IOS) approximation between 5 and 200 K for NH and 100 K for ND. We consider the first 71 and 105 energy levels of NH and ND, respectively. General propensity rules are discussed. We found a significant isotopic substitution effect in the rate coefficients. In addition, the rate coefficients for collisions with H2 are larger than those with He by a factor of up to 5, leading to lower critical densities for collisional excitation with H2 than He. The impact of the new set of collisional data has been investigated in simple radiative transfer models of the NH emission seen toward the Orion Bar and the ejecta of the η Carinae binary star. We observed significant differences by a factor of 5 between the presently determined column densities for NH compared to those from the literature using He as a collider.

Synthesis of Deuterated and Sulfurated Polymers by Inverse Vulcanization: Engineering Infrared Transparency via Deuteration.

The synthesis of deuterated, sulfurated, proton-free, glassy polymers offers a route to optical polymers for infrared (IR) optics, specifically for midwave IR (MWIR) photonic devices. Deuterated polymers have been utilized to enhance neutron cross-sectional contrast with proteo polymers for morphological neutron scattering measurements but have found limited utility for other applications. We report the synthesis of perdeuterated d14-(1,3-diisopropenylbenzene) with over 99% levels of deuteration and the preparation of proton-free, perdeuterated poly(sulfur-random-d14-(1,3-diisopropenylbenzene)) (poly(S-r-d14-DIB)) via inverse vulcanization with elemental sulfur. Detailed structural analysis and quantum computational calculations of these reactions demonstrate significant kinetic isotope effects, which alter mechanistic pathways to form different copolymer microstructures for deutero vs proteo poly(S-r-DIB). This design also allows for molecular engineering of MWIR transparency by shifting C-H bond vibrations around 3.3 μm/3000 cm-1 observed in proteo poly(S-r-DIB) to 4.2 μm/2200 cm-1. Furthermore, the fabrication of thin-film MWIR optical gratings made from molding of deuterated-sulfurated, proton-free poly(S-r-d14-DIB) is demonstrated; operation of these gratings at 3.39 μm is achieved successfully, while the proteo poly(S-r-DIB) gratings are opaque at these wavelengths, highlighting the promise of MWIR sensors and compact spectrometers from these materials.

Determination of intermolecular and intramolecular isotopic compositions of low-abundance gaseous hydrocarbons using an online hydrocarbon gas concentration method

The stable isotopic composition of natural gas can be used to identify its origin and source. However, low concentrations of gaseous hydrocarbons in high-mature natural and shale gases hinder accurate determination of their compound- and position-specific isotopic compositions. In this study, an online C2+ hydrocarbon gas concentration system combined with gas chromatography–isotope ratio mass spectrometry (GC–IRMS) or gas chromatography–pyrolysis–gas chromatography–isotope ratio mass spectrometry (GC–Py–GC–IRMS) was developed to determine compound- and position-specific isotopic compositions of low-abundance gaseous hydrocarbons. The lower limit of the gas concentration required for isotope ratio determination using the online concentration system is 0.001% (0.003%) for compound-specific carbon (hydrogen) isotopes and 0.005% for position-specific carbon isotopes and is thus applicable to most natural gas samples. The online concentration technique does not cause significant isotopic fractionation effects, and the combination with GC–IRMS and GC–Py–GC–IRMS can accurately and precisely determine the compound-specific δ13C and δD values of low-content C2+ gaseous hydrocarbons and the position-specific δ13C values (δ13Ca, δ13Cb, and SP values) of propane in low-content propane samples, respectively. The application of our method to two natural gas samples from the Ordos and Sichuan basins further confirms that the online concentration method allows simple and rapid determination of the compound- and position-specific isotopic compositions of low-abundance gaseous hydrocarbons.

Hyperfine excitation of 13CCH and C13CH by collisions with para-H2

ABSTRACT The computation of hyperfine resolved cross sections and rate coefficients for open-shell molecules in collision with H2 is a true methodological and numerical challenge. Such collisional data are however required to interpret astrophysical observations. We report the first hyperfine resolved rate coefficients for (de-)excitation of 13CCH and C13CH isotopologues induced by collisions with para-H2. These calculations have been performed using a recently published C2H–H2 potential energy surface. Hyperfine resolved cross sections and rate coefficients between the first 98 energy levels of the two isotopologues were determined using a recoupling technique for temperatures ranging from 5 to 100 K. Significant isotopic substitution effects were found, showing the necessity of computing isotopologue specific collisional data. These rate coefficents have then been used in a simple radiative transfer modelling for typical molecular cloud conditions.

Deuterium Solvent Kinetic Isotope Effect on Enzymatic Methyl Transfer Catalyzed by Catechol O-methyltransferase.

Catechol o-methyltransferase plays a key role in the metabolism of catecholamine neurotransmitters. At present, its catalytic mechanism, overall structure, and kinetic characteristics have been basically clarified, but few people have paid attention to the function of solvents on enzymatic methyl transfer reactions. The influence of solvents on enzymatic reactions has always been a fuzzy hot topic. In addition, as a well-studied typical methyltransferase, COMT is a good test bed for exploring the source of the solvent isotope effect, which is a powerful tool in enzymatic mechanism research. We have measured the kinetic parameters of methyl transfer catalyzed by COMT in both normal water (H2O) and heavy water (D2O) by high-performance liquid chromatography (HPLC) in the range of pL 6 ~ 11. The kinetic characteristics of COMT in H2O and D2O were significantly different under different pH/pD conditions. Significant solvent kinetic isotope effects (SKIE) were obtained, especially inverse solvent kinetic isotope effects (SKIE < 1) were observed in this methyl transfer reaction for the first time. Traditional factors which could interpret the solvent isotope effect were ruled out. It's suggested that the solvent might affect the overall conformation as well as the flexibility of protein through non-covalent forces, thus altering the catalytic activity of COMT and leading to the solvent isotope effect.

Hydrogen isotope effects: A new path to high-energy aqueous rechargeable Li/Na-ion batteries

Aqueous rechargeable Li/Na-ion batteries have shown promise for sustainable large-scale energy storage due to their safety, low cost, and environmental benignity. However, practical applications of aqueous batteries are plagued by water's intrinsically narrow electrochemical stability window, which results in low energy density. In this perspective article, we review several strategies to broaden the electrochemical window of aqueous electrolytes and realize high-energy aqueous batteries. Specifically, we highlight our recent findings on stabilizing aqueous Li storage electrochemistry using a deuterium dioxide-based aqueous electrolyte, which shows significant hydrogen isotope effects that trigger a wider electrochemical window and inhibit detrimental parasitic processes.

Effects of hydrogen bonding on intramolecular/intermolecular proton-coupled electron transfer using a Ruthenium-anthraquinone dyad in ultrafast time domain

A ruthenium-anthraquinone dyad consisting of the bi-imidazole spacer in which one imidazole proton acts as a hydrogen bonding agent is employed to investigate the effects of H-bonding on both photo-induced electron transfer (PET) and proton-coupled electron transfer process (PCET). The H-bonding of the anthraquinone radical anion (AQ•-) with a bi-imidazole spacer in complex 1 generated by the electron transfer reduction of complex 1 is indicated by the symbolic anodic shift of the reduction potential of anthraquinone. The present investigation has established that the arrangement of an intramolecular H-bond between the attached AQ and the imidazole proton consequences in a notable increase in the rate of intramolecular electron transfer from Ru(II) to AQ. Further, we have also demonstrated the ultrafast electron transfer and PCET dynamics in presence of water. We have successfully detected the transient absorption photoproduct Ru(III) by which we could determine the time constants of each step which is rare in literature. Most importantly, our study demonstrates electron transfer and intra-molecular PCET dynamics in the femtosecond time domain. The results of the steady-state and transient absorption spectroscopic studies suggest that bi-imidazole spacer and water molecules have a significant role in both PET and PCET dynamics as a proton source in the excited state. We have demonstrated PCET reaction in ultrafast time scale using Ru-polypyridyl complex, with significant kinetic isotope effect.

The role of immiscible sulfides for sulfur isotope fractionation in arc magmas: Insights from the Talkeetna island arc crustal section, south-central Alaska

While sulfur isotopes have proven powerful tracers of sulfur (re)cycling within subduction zones, the origin of the 34S-enrichment seen in arc magmas remains a subject of debate, with competing hypotheses implicating both mantle and crustal processes. Herein, we investigate these competing models through the study of the Early–Middle Jurassic Talkeetna Arc section exposed in the Chugach Mountains, south-central Alaska, reporting the sulfur isotope systematics of rocks spanning a depth transect that extends from the upper mantle (∼0.9–1.2 GPa, ∼30–35 km) up into the mid-crust (∼0.2–0.5 GPa, ∼5–15 km). Marked by an increase in δ34S values from −1.28 to +5.61‰, these data reveal a significant isotope effect associated with melt differentiation, with 32S being preferentially sequestered from primitive basaltic melts into ultramafic and mafic cumulates, yielding progressively more evolved melts enriched in 34S. Leveraging a quantitative petrological model that constrains the liquid line of descent and sulfide saturation as a function of sulfur content, oxygen fugacity, pressure and temperature, we then demonstrate that, in the presence of dissolved oxidized sulfur species, the saturation of immiscible magmatic sulfides is capable of generating the observed sulfur isotope fractionation. While fluid saturation is unlikely to occur at lower crustal depths, ascending melts will almost certainly experience fluid saturation and degassing. As a result, sulfide immiscibility and magma degassing are not mutually exclusive and may, instead, represent complementary processes that combine to explain the observed range of positive δ34S compositions. In Talkeetna, crustal assimilation is unlikely to have played a role to increase δ34S values as there was little to no assimilation of pre-existing oceanic crust containing seawater sulfate during the formation of the Talkeetna Arc section. Furthermore, our model suggests that primitive melt in isotopic equilibrium with the most primitive ultramafic cumulates have mantle-like δ34S values between ∼0 and +1.1‰, which suggest limited input of 34S-enriched slab-derived sulfur into the sub-arc mantle. Collectively, these findings emphasize that mantle-like δ34S values for primitive arc melt may be common, while revealing the underappreciated importance of deep crustal crystallization of immiscible magmatic sulfides to generate positive δ34S compositions in erupted arc magmas.

Enrichment of hepatic glycogen and plasma glucose from H₂18 O informs gluconeogenic and indirect pathway fluxes in naturally feeding mice.

Deuterated water (2 H2 O) is a widely used tracer of carbohydrate biosynthesis in both preclinical and clinical settings, but the significant kinetic isotope effects (KIE) of 2 H can distort metabolic information and mediate toxicity. 18 O-water (H2 18 O) has no significant KIE and is incorporated into specific carbohydrate oxygens via well-defined mechanisms, but to date it has not been evaluated in any animal model. Mice were given H2 18 O during overnight feeding and 18 O-enrichments of liver glycogen, triglyceride glycerol (TG), and blood glucose were quantified by 13 C NMR and mass spectrometry (MS). Enrichment of oxygens 5 and 6 relative to body water informed indirect pathway contributions from the Krebs cycle and triose phosphate sources. Compared with mice fed normal chow (NC), mice whose NC was supplemented with a fructose/glucose mix (i.e., a high sugar [HS] diet) had significantly higher indirect pathway contributions from triose phosphate sources, consistent with fructose glycogenesis. Blood glucose and liver TG 18 O-enrichments were quantified by MS. Blood glucose 18 O-enrichment was significantly higher for HS versus NC mice and was consistent with gluconeogenic fructose metabolism. TG 18 O-enrichment was extensive for both NC and HS mice, indicating a high turnover of liver triglyceride, independent of diet. Thus H2 18 O informs hepatic carbohydrate biosynthesis in similar detail to 2 H2 O but without KIE-associated risks.

Ultrafast excited state dynamics of pyridine N-oxide derivative in solution; femtosecond fluorescence up-conversion and theoretical calculations

In this study we have investigated 2-ethylamino-4-nitro-6-methyl pyridine N-oxide (2E6M) molecule that belongs to important group of Proton Coupled Electron Transfer (PCET) compounds where both the charge transfer (CT) and proton transfer processes in excited states may proceed. In this case, this is possible due to the donors and acceptors of electrons and protons in this system, as well as due to the presence of intramolecular {N-H… O [2,566(3) Å}, hydrogen bond.Using stationary and time-resolved spectroscopy, as well as quantum chemical calculations on the DFT and TD DFT B3LYP/6-31G (d,p) level of theory, a partial CT nature of the S0 → S1 transition in both tautomeric forms (N and T) has been revealed. Additionally, the excited state intramolecular proton transfer (ESIPT) process shown to be more favorable in apolar and weakly polar solvents than in strongly polar acetonitrile (EN(S1) > ET(S1). The displacement of charge from the amine group and the ring to the nitro group has been observed on the changing shapes of the HOMO and LUMO orbitals involved in this transition what further quantitatively allowed to realize the increase in the dipole moment of both forms in the electronic excited state. The calculations show that in two solvents with radically different polarity (heptane, acetonitrile), dipole moments of both forms are very similar [in acetonitrile uN(S1) and uT(S1) are 11.0 D and 11.5 D, respectively]. Hence, in polar media both forms can be stabilized in a comparable manner. This made it difficult for us to assign a single fluorescent band in acetonitrile to one of the tautomeric forms. However, it seems that due to application of time-resolved spectroscopy, this problem has been clarified. The TCSPC decay curve in acetonitrile with an ultrafast lifetime assigned to the (N) form, along with the femtosecond up-conversion signals that demonstrated only an ultrafast decay without any rise-time of a new excited (T) species, allowed us to conclude that in 2E6M in strongly polar solvent the ESIPT does not occur.The unique fluorescence band origins from the (N) form. In protic solvents, the significant kinetic isotopic effects have provided us with conclusive evidence for the presence of the solvent-assisted ESIPT process. Furthermore, it was noticed that the fluorescence lifetime in D2O (100–120 fs) estimated from the up-conversion signals is about 40 times shorter relative to methanol. This may suggest that the sine qua non for the ESIPT process in 2E6M in protic solvents is the formation of a complex with a solvent molecule in the hydrogen bridge between the proton donor and proton acceptor, respectively.

Selective Oxidations Using a Cytochrome P450 Enzyme Variant Driven with Surrogate Oxygen Donors and Light.

Cytochrome P450 monooxygenase enzymes are versatile catalysts, which have been adapted for multiple applications in chemical synthesis. Mutation of a highly conserved active site threonine to a glutamate can convert these enzymes into peroxygenases that utilise hydrogen peroxide (H2O2). Here, we use the T252E‐CYP199A4 variant to study peroxide‐driven oxidation activity by using H2O2 and urea‐hydrogen peroxide (UHP). We demonstrate that the T252E variant has a higher stability to H2O2 in the presence of substrate that can undergo carbon‐hydrogen abstraction. This peroxygenase variant could efficiently catalyse O‐demethylation and an enantioselective epoxidation reaction (94 % ee). Neither the monooxygenase nor peroxygenase pathways of the P450 demonstrated a significant kinetic isotope effect (KIE) for the oxidation of deuterated substrates. These new peroxygenase variants offer the possibility of simpler cytochrome P450 systems for selective oxidations. To demonstrate this, a light driven H2O2 generating system was used to support efficient product formation with this peroxygenase enzyme.

Mitigating Matrix Effects in LC-ESI-MS-MS Analysis of a Urinary Biomarker of Xylenes Exposure.

Liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS-MS) with stable isotope-labeled internal standards (SIL-ISs) is the gold standard for quantitative analysis of drugs and metabolites in complex biological samples. Significant isotopic effects associated with deuterium labeling often cause the deuterated IS to elute at a different retention time from the target analyte, diminishing its capability to compensate for matrix effects. In this study, we systematically compared the analytical performance of deuterated (2H) SIL-IS to non-deuterated (13C and 15N) SIL-ISs for quantifying urinary 2-methylhippuric acid (2MHA) and 4-methylhippuric acid (4MHA), biomarkers of xylenes exposure, with an LC-ESI-MS-MS assay. Analytical method comparison between ISs demonstrated a quantitative bias for urinary 2MHA results, with concentrations generated with 2MHA-[2H7] on average 59.2% lower than concentrations generated with 2MHA-[13C6]. Spike accuracy, measured by quantifying the analyte-spiked urine matrix and comparing the result to the known spike concentration, determined that 2MHA-[2H7] generated negatively biased urinary results of -38.4%, whereas no significant bias was observed for 2MHA-[13C6]. Post-column infusion demonstrated that ion suppression experienced by 2MHA and 2MHA-[13C6] was not equally experienced by 2MHA-[2H7], explaining the negatively biased 2MHA results. The quantitation of urinary 4MHA results between ISs exhibited no significant quantitative bias. These results underscore the importance of the careful selection of ISs for targeted quantitative analysis in complex biological samples.