Protonation Equilibria Research Articles

Direct Observation of Polarization in Short Hydrogen Bonds due to Proton Delocalization

Based on their relatively shallow potential energy surfaces (PESs), short hydrogen bonds with heavy atom distances about or less than 2.5 Å are believed to exhibit proton delocalization, which has been argued to be beneficial to biological catalysis1. While the degree of delocalization is usually characterized by neutron diffraction or inferred from broad bands in the IR spectra, these approaches can result in ambiguities2 and information on energetics cannot be directly obtained. Here we apply an external electric field to perturb the proton equilibrium position of a short hydrogen bond in previously characterized GFP S65T/H148D mutants with various degrees of proton affinity mismatch (ΔpKα) between the chromophore and aspartate3. The chromophore serves as a spectral probe of the proton position, allowing us to perform electronic Stark spectroscopy. The local curvature of the PES is reflected by the ease of proton polarization, and the resulting proton displacement effectively amplifies the electric field experienced by the chromophore, leading to anomalously large Stark tuning rates as readouts. These Stark tuning rates are sensitive to the presence of the short hydrogen bond, proton position, ΔpKα and deuteration, in accordance with large hydrogen bond polarizations predicted by Zundel4. The results reinforce our previous arguments on the absence of low barrier hydrogen bonds in these variants3: the general shape of PESs is double-welled with a rather high barrier in between and the proton is delocalized within each well. [1] Perrin, C. L.; Nielson, J. B. Annu. Rev. Phys. Chem. 1997, 48, 511-544. [2] Graen, T. et al. J. Am. Chem. Soc. 2016, 138, 16620-16631. [3] Oltrogge, L. M.; Boxer, S. G. ACS Cent. Sci. 2015, 1, 148-156. [4] Janoshek, R. et al. J. Am. Chem. Soc. 1972, 94, 2387-2396.

Levaquin drug complexing property towards platinum and palladium metal ions: Thermodynamic studies in aqueous solutions

The experimental protonation and complex formation equilibrium constants of levaquin (LFX) with platinum(II) and palladium(II) metal ions have been investigated at 310.15K in water solutions at ionic strength of I=0.16mol·dm−3 NaNO3 using pH-potentiometric and cyclic voltammetry techniques, and by means of Hyperquad 2008 estimation model program. Also, the dissociation constants of LFX and the equilibrium constants of its binary complexes with the studied platinum(II) and palladium(II) metal ions in I=0.16mol·dm−3 NaNO3 water solutions were observed at different temperatures such as 298.15, 310.15, 318.15 and 328.15K. The theoretical calculations of overall protonation and stability constants of the metal ion-LFX complex species in aqueous solutions were predicted as the free energy change associated with the LFX protonation, and metal ion – LFX complex formation equilibria using ab initio and density function theory calculations by applying Gaussian 09 software molecular modeling. The determination of equilibrium constants were confirmed by the spectrophotometric method with the refinement by using the HypSpec program. The usage of the experimental potentiometry/spectrophotometry techniques and theoretical predictions provides a complete picture of the microscopic equilibria of the studied systems (metal ions -LFX). Precisely, this theoretically predications could be useful to control the most real protonation constants of levaquin ligand in which the binding sites changes due to the ligand protonation/deprotonation equilibria. Also, the complexing capacities of different platinum(II) and palladium(II) metal ions towards LFX in solutions were evaluated and discussed. From the determined experimental stability constants of different metal complex species, the concentration distribution diagrams of the various metal ions - LFX complex species in solutions were estimated using HySS 2009 software.

Synthesis of cationic molybdenum-cobalt heterometallic clusters protected against hydrolysis by macrocyclic triazacyclononane complexes.

Four kinds of cobalt-polyoxomolybdenum clusters were synthesized by a combination of polyoxomolybdates and a Co-tacn (tacn = 1,4,7-triazacyclononane) complex. The heterometallic polynuclear clusters were obtained by the reaction of Na2MoO4 and [Co(tacn)(H2O)3](CF3SO3)3·H2O in water. The peripherals of polyoxomolybdates are capped by thermodynamically stable [Co(tacn)]3+ protecting groups to prevent further hydrolysis and condensation reactions. Intra-molecular hydrogen bonds between N-H groups on tacn and oxygen atoms on the molybdate core contribute to the stability of the complexes in water. Under the optimized synthetic conditions, the molar ratios of Na2MoO4 and [Co(tacn)(H2O)3](CF3SO3)3·H2O were adjusted to 1 : 0.3, 1 : 0.9, or 1 : 1.5, producing [{Co(tacn)}2Mo3O12]·2NaCF3SO3·7H2O (1), [{Co(tacn)}4H2Mo7O27](CF3SO3)2·13H2O (2), and [{Co(tacn)}4H3Mo4O17](CF3SO3)5·6H2O (3), respectively. As one of the most important factors, the adjustments of the pH values by changing the molar ratio enabled us to isolate these clusters. The presence of pyridine produces a neutral complex, [Co2(tacn)2(pyridine)Mo5O18]·4.5H2O (4) containing a lacunary Lindqvist-type polyoxomolybdate. The synthesis of complexes 2-4 is also possible starting from cluster 1, implying that 1 is a precursor for the formation of 2-4 in aqueous solution. Complexes 2-4 have a common structural building block, [{Co(tacn)}2Mo3O13], in each of the cluster units. The 1H NMR spectra of 2 and 3 in aqueous solution show multiple splitting patterns of methylene signals on tacn ligands according to the chemical environments indicated by the crystallographic structure. The 59Co NMR spectrum of cluster 3 in CH3NO2 has two signals from two different Co units with a 3 : 1 integration ratio ensuring the structural integrity in solution. The broadening of one of the signals for the 59Co NMR spectrum of cluster 3 in water as a protic solvent reveals the presence of the deprotonation equilibrium of the acidic protons on the cluster framework in water.

A headgroup linker perturbs pKavia acyl chain migration: designing base-labile supramolecular assemblies.

Acyl chain transfer, which perturbs the protonation equilibrium of amine and reduces the apparent pKa by 2.0-2.5 units, is used to develop a liposome-based drug delivery system.

Thermodynamic studies on metal ions – Ninhydrin – Glycine interactions in aqueous solutions

The experimental protonation and complex formation equilibrium constants of ligands (ninhydrin (NHN) and glycine (Gly)) with trivalent (iron(III), aluminium(III), and chromium (III)), divalent (copper(II), cobalt(II), nickel(II), strontium(II), manganese(II), and mercury(II)) and monovalent silver(I) metal ions have been investigated at 310.15 K in water solutions at ionic strength of 13.60 g·dm−3 NaNO3 using pH-potentiometric, UV–visible spectrophotometric, and cyclic voltammetry techniques, and by means of Hyperquad 2008 estimation model program. Also, the dissociation constants of NHN and the equilibrium constants of its binary complexes and ternary complexes with the studied metal ions in 13.60 g·dm−3 NaNO3 water solutions were observed at different temperatures such as (298.15, 310.15, 318.15 and 328.15) K. The theoretical calculations of overall protonation and stability constants of the metal ion-ninhydrin complex species in aqueous solutions were predicted as the free energy change associated with the ninhydrin protonation, and metal ion – ninhydrin – glycine complex formation equilibria using ab initio and density function theory calculations by applying Gaussian 09 software molecular modelling. The convention of the experimental Potentiometry/Spectrophotometry techniques and theoretical calculations provide a complete picture of the microscopic equilibria of the studied metal ions – ninhydrin – glycine systems. Precisely, this theoretical predication could be useful to control the most real protonation constants of ninhydrin and glycine ligands in which the binding sites change due to the ligand protonation/deprotonation equilibria. Also, the complexing capacities of different metal ions towards ninhydrin and glycine in solutions were evaluated and discussed. From the determined experimental stability constants of different metal complex species, the concentration distribution diagrams of the various metal ions – ninhydrin – glycine complex species in solutions were estimated using HySS 2009 software.

EFFECT OF CATIONIC MICELLES OF CETYLTRIMETHYLAMMONIUM BROMIDE ON PROTONATION EQUILIBRIA OF SALICYLIC ACID DERIVATIVES

To gain more information about the effect of surfactant on salicylic acid derivatives, the stoichiometric protonation constants of 5-Sulphosalicylic acid and 5-Hydroxysalicylic acid in 0.0%-2.5% (w/v) cetyltrimethylammonium bromide (CTAB) - water mixtures were determined at an ionic strength of 0.16 mol dm-3 and at 303 K. A potentiometric method was used and the calculation of constants was carried out using the computer program MINIQUAD75. These protonation constants values have been found to shift in micellar media as compared to those in pure water. The differences in the values have been attributed to the solvent properties of the interfacial and bulk phases involving contribution from the micellar surface potential in the case of charged micelles. The trend of log values of step-wise protonation constants with mole fraction of the medium have been explained based on specific solute-solvent interactions. In this study Distributions of species, percentage of species composition, protonation equilibria and effect of influential parameters on the protonation were also discussed.

PH Dependence of the Imidazole-2-carboxaldehyde Hydration Equilibrium: Implications for Atmospheric Light Absorbance

Imidazole-2-carboxaldehyde (IC) has been identified as an aqueous “brown carbon” absorber and a possible atmospheric photosensitizer. IC exists in a pH-dependent equilibrium between its aldehyde and geminal diol form; the diol form is the dominant species in solution at pH <5. Calculated molar absorptivity coefficients are 13700 ± 200 cm–1 M–1 at 287 nm for the aldehyde and 7800 ± 100 cm–1 M–1 at 212 nm for the diol. This shift from aldehyde to diol changes the peak light absorption of the aqueous solution from 287 to 212 nm, which is beyond the actinic range and may have implications for radiative forcing. The observed pH-dependent shift in the hydration equilibrium of IC is driven by the interaction between its hydration and protonation equilibria. Calculated pKa values are 2.5 ± 0.4 and 5.94 ± 0.05 for the aldehyde and diol, respectively, and are consistent with the trend toward increasing diol as the pH decreases. The acid–base equilibrium affects both the solubility and the major species in solution ...

Understanding the Regioselectivity of Aromatic Hydroxylation over Divanadium-Substituted γ-Keggin Polyoxotungstate

The aromatic hydroxylation of pseudocumene (PC) with aqueous hydrogen peroxide catalyzed by the divanadium-substituted γ-Keggin polyoxotungstate TBA4[γ-PW10O38V2(μ-O)(μ-OH)] (TBA-1H, TBA = tetrabutylammonium) has been studied using kinetic modeling and DFT calculations. This reaction features high chemoselectivity and unusual regioselectivity, affording 2,4,5-trimethylphenol (TMP) as the main product. Then the computational study was extended to the analysis of the regioselectivity for other alkoxy- and alkylarene substrates. The protonation/deprotonation of TBA-1H in MeCN/tBuOH (1:1) was investigated by 31P NMR spectroscopy. Forms with different protonation states, [γ-PV2W10O40]5– (1), [γ-HPV2W10O40]4– (1H), and [γ-H2PV2W10O40]3– (1H2), have been identified, and the protonation equilibrium constants were estimated on the basis of the 31P NMR data. DFT calculations were used to investigate the oxygen transfer process from hydroperoxo species, [γ-PW10O38V2(μ-O)(μ-OOH)]4– (2) and [γ-PW10O38V2(μ-OH)(μ-OOH)]3...

Measurement and thermodynamic analysis of the acid-base equilibria of two bioactive flavanone glycosides in different temperatures and non-aqueous mixtures

In this research, the acid-base properties of two glycosilated flavanones namely naringoside and hesperidin were thermodynamically studied using spectrophotometric and potentiometric techniques at various temperatures (293.15–313.15) K. Because of very low solubility of these flavanones in aqueous solution, the protonation constants were obtained in ethanol-water mixtures (60–90% v/v) containing 0.10moldm−3 tetra-n-butylammonium perchlorate (TBAP) as supporting electrolyte for naringoside and DMSO-water mixed solvents (50–95% v/v) containing 0.10moldm−3 NaCl for hesperidin. Data process for the calculation of the protonation constants was done using the computer program STAR. The thermodynamic quantities (ΔG°, ΔH°, and ΔS°) of the protonation process were obtained by means of modified van't Hoff equation. It was found that the acid dissociation reaction is exothermic and the entropic contribution is low for both the flavanones. Finally, the influence of solute-solvent interactions on the protonation equilibria in various temperatures was modelled using Kamlet, Abboud and Taft (KAT) solvatochromic parameters and Hildebrand's solubility index.

Protonation equilibria and pore-opening structure of the dual-histidine influenza B virus M2 transmembrane proton channel from solid-state NMR

The influenza A and B viruses are the primary cause of seasonal flu epidemics. Common to both viruses is the M2 protein, a homotetrameric transmembrane proton channel that acidifies the virion after endocytosis. Although influenza A M2 (AM2) and B M2 (BM2) are functional analogs, they have little sequence homology, except for a conserved HXXXW motif, which is responsible for proton selectivity and channel gating. Importantly, BM2 contains a second titratable histidine, His-27, in the tetrameric transmembrane domain that forms a reverse WXXXH motif with the gating tryptophan. To understand how His-27 affects the proton conduction property of BM2, we have used solid-state NMR to characterize the pH-dependent structure and dynamics of His-27. In cholesterol-containing lipid membranes mimicking the virus envelope, 15N NMR spectra show that the His-27 tetrad protonates with higher pKa values than His-19, indicating that the solvent-accessible His-27 facilitates proton conduction of the channel by increasing the proton dissociation rates of His-19. AM2 is inhibited by the amantadine class of antiviral drugs, whereas BM2 has no known inhibitors. We measured the N-terminal interhelical separation of the BM2 channel using fluorinated Phe-5. The interhelical 19F-19F distances show a bimodal distribution of a short distance of 7 Å and a long distance of 15-20 Å, indicating that the phenylene rings do not block small-molecule entry into the channel pore. These results give insights into the lack of amantadine inhibition of BM2 and reveal structural diversities in this family of viral proton channels.

1H NMR studies of molecular interaction of D-glucosamine and N-acetyl-D-glucosamine with capsaicin in aqueous and non-aqueous media

Complex formation of D-glucosamine (Gl) and N-acetyl-D-glucosamine (AGl) with capsaicin (Cp) were studied by 1H NMR titrations in H2O-d2 and DMSO-d6; capsaicin is the major bioactive component of chili peppers. Every titration curve has been interpreted by formulating a suitable model for the reaction equilibrium, to elucidate intermolecular interactions. In DMSO, glucosamine cations associate with each other to yield linear aggregates, and undergo pseudo-1:1-complexation with capsaicin, the formation constant being ca. 30 M−1. N-Acetylglucosamine, without self-association, forms a 2:1-complex AGl2Cp with the stability of ca. 70 M−2. These complexations are achieved by intermolecular hydrogen bonds. In D2O, glucosamine undergoes reversible protonation equilibrium between Gl0 and GlH+ with the logarithmic protonation constants log KD = 8.63 for α-glucosamine and 8.20 for β-isomer. Both anomeric isomers of deprotonated glucosamine form Gl0Cp-type complexes of capsaicin, in a competitive manner, with a formation constant of 1040 M−1 for the α-glucosamine complex and 830 M−1 for the β-complex; the anomeric carbons result in the difference in thermodynamic stability. The reactant molecules are closed up by the solvent-exclusion effect and/or the van der Waals interaction; the resulting pair is stabilized by intermolecular hydrogen bonding within a local water-free space between the component molecules. By contrast, neither protonated glucosamine (GlH+) nor N-acetylglucosamine yields a capsaicin complex with the definite stoichiometry. The monosaccharides recognize capsaicin under only a controlled condition; the same phenomena are predicted for biological systems and nanocarriers based on polysaccharides such as chitosan.

Reversible Modulation of the Electrostatic Potential of a Colloidal Quantum Dot through the Protonation Equilibrium of Its Ligands.

This Letter describes the reversible modulation of the electrostatic potential at the interface between a colloidal PbS quantum dot (QD) and solvent, through the protonation equilibrium of the QD's histamine-derivatized dihydrolipoic acid (DHLA) ligand shell. The electrostatic potential is sensitively monitored by the yield of photoinduced electron transfer from the QD to a charged electron acceptor, 9,10-anthraquinone-2-sulfonate (AQ). The permeability of the DHLA coating to the AQ progressively increases as the average degree of protonation of the ligand shell increases from 0 to 92%, as quantified by 1H NMR, upon successive additions of p-toluenesulfonic acid; this increase results in a decrease in the photoluminescence (PL) intensity of the QDs by a factor of 6.7. The increase in permeability is attributable to favorable electrostatic interactions between the ligands and AQ. This work suggests the potential of the combination of near-IR-emitting QDs and molecular quenchers as robust local H+ sensors.

Synthesis of a tetrazine-based catecholamide derivative and its evaluation as a chelating agent for removal of Cd(II), Co(II), and Cu(II)

The synthesis and structural characterization of a tetrazine-based catecholamide (CAM) ligand, N,N′-bis(N″-(aminoethyl)-2,3-bis(hydroxy)benzamide)-1,2,4,5-tetrazine-3,6-diamine (5), were investigated. All compounds were characterized by 1H NMR spectroscopy, 13C NMR spectroscopy, and FTIR spectroscopy. The protonation equilibria of 5 and complexation capacities (log βpqr) of Cd2+, Co2+, and Cu2+ complexes of 5 were evaluated through potentiometric titration and spectrophotometric titration, respectively. Species independent pM value (=−log [M]free) was used to compare metal affinities with the final sequence Cu2+ > Cd2+ > Co2+. Results show that 5 has potential for heavy metal removal.

Structural Basis for Asymmetric Conductance of the Influenza M2 Proton Channel Investigated by Solid-State NMR Spectroscopy

The influenza M2 protein forms an acid-activated proton channel that is essential for virus replication. The transmembrane H37 selects for protons under low external pH while W41 ensures proton conduction only from the N terminus to the C terminus and prevents reverse current under low internal pH. Here, we address the molecular basis for this asymmetric conduction by investigating the structure and dynamics of a mutant channel, W41F, which permits reverse current under low internal pH. Solid-state NMR experiments show that W41F M2 retains the pH-dependent α-helical conformations and tetrameric structure of the wild-type (WT) channel but has significantly altered protonation and tautomeric equilibria at H37. At high pH, the H37 structure is shifted toward the π tautomer and less cationic tetrads, consistent with faster forward deprotonation to the C terminus. At low pH, the mutant channel contains more cationic tetrads than the WT channel, consistent with faster reverse protonation from the C terminus. 15N NMR spectra allow the extraction of four H37 pKas and show that the pKas are more clustered in the mutant channel compared to WT M2. Moreover, binding of the antiviral drug, amantadine, at the N-terminal pore at low pH did not convert all histidines to the neutral state, as seen in WT M2, but left half of all histidines cationic, unambiguously demonstrating C-terminal protonation of H37 in the mutant. These results indicate that asymmetric conduction in WT M2 is due to W41 inhibition of C-terminal acid activation by H37. When Trp is replaced by Phe, protons can be transferred to H37 bidirectionally with distinct rate constants.

Nicotine – Metal ion interactions in solutions: Potentiometric, cyclic voltammetry investigations and quantum chemical calculations

The experimental protonation and complex formation equilibrium constants of nicotine (NIC) with trivalent (iron (III), and chromium (III)) and divalent (copper (II), and nickel (II)) metal ions have been investigated at 310.15 oK in water solutions at ionic strength of 13.60g·dm−3 NaNO3 using pH-potentiometric and cyclic voltammetry techniques, and by means of Hyperquad 2008 estimation model program. Also, the dissociation constants of NIC and the equilibrium constants of its binary complexes with the studied metal ions in 13.60g·dm−3 NaNO3 water solutions were observed at different temperatures such as (298.15, 310.15, 318.15 and 328.15) K. The theoretical calculations of overall protonation and stability constants of the metal ion-nicotine complex species in aqueous solutions were predicted as the Gibbs energy change associated with the nicotine protonation, and metal ion – nicotine complex formation equilibria using ab initio and density function theory calculations by applying Gaussian 09 software molecular modeling. The usage of the experimental potentiometry/spectrophotometry techniques and theoretical predictions provides a complete picture of the microscopic equilibria of the studied systems (metal ions -nicotinate). Precisely, this theoretically predications could be useful to control the most real protonation constants of nicotine ligand in which the binding sites changes due to the ligand protonation/deprotonation equilibria. Also, the complexing capacities of different metal ions towards nicotine in solutions were evaluated and discussed. From the determined experimental stability constants of different metal complex species, the concentration distribution diagrams of the various metal ions - nicotine complex species in solutions were estimated using HySS 2009 software.

From Master-Slave to Peer-to-Peer Coupling in Chemical Reaction Networks.

Design strategy through linking a driving pH oscillator (master system) to a pH sensitive complexation, precipitation, or protonation equilibrium (slave system) has been widely used to create and control concentration oscillations of chemical entities (e.g., monovalent cations, DNA, nanoparticles) not participating in the pH oscillatory system. No systematic investigation has been carried out on how the components of these equilibria affect the characteristics of the driving pH oscillators, and this feedback effect has been often neglected in previous studies. Here we show that pH sensitive species (hydrogen carbonate, EDTA) through a pH-dependent equilibrium could significantly affect the characteristics (time period and amplitude) of the driving pH oscillators. By varying the concentration of those species we are able to control the strength of the chemical feedback from slave system to master system thus introducing a transition from master-slave coupling to peer-to-peer coupling in linked chemical systems. To illustrate this transition and coupling strategies we investigate two coupled chemical systems, namely, the bromate-sulfite pH oscillator and carbonate-carbon dioxide equilibrium and the hydrogen peroxide-thiosulfate-copper(II) and EDTA complexation equilibrium. As a sign of the peer-to-peer coupling the characteristics of the driving oscillatory systems can be tuned by controlling the feedback strength, and the oscillations can be canceled above a critical value of this parameter.

Mechanism of formate-nitrite transporters by dielectric shift of substrate acidity.

Bacterial formate-nitrite transporters (FNTs) regulate the metabolic flow of small, weak mono-acids. Recently, the eukaryotic PfFNT was identified as the malaria parasite's lactate transporter and novel drug target. Despite crystal data, central mechanisms of FNT gating and transport remained unclear. Here, we show elucidation of the FNT transport mechanism by single-step substrate protonation involving an invariant lysine in the periplasmic vestibule. Opposing earlier gating hypotheses and electrophysiology reports, quantification of total uptake by radiolabeled substrate indicates a permanently open conformation of the bacterial formate transporter, FocA, irrespective of the pH Site-directed mutagenesis, heavy water effects, mathematical modeling, and simulations of solvation imply a general, proton motive force-driven FNT transport mechanism: Electrostatic attraction of the acid anion into a hydrophobic vestibule decreases substrate acidity and facilitates protonation by the bulk solvent. We define substrate neutralization by proton transfer for transport via a hydrophobic transport path as a general theme of the Amt/Mep/Rh ammonium and formate-nitrite transporters.

Benzimidazole acrylonitriles as multifunctional push-pull chromophores: Spectral characterisation, protonation equilibria and nanoaggregation in aqueous solutions

Heterocyclic donor-π-acceptor molecular systems based on an N,N-dimethylamino phenylacrylonitrile benzimidazole skeleton have been characterised and are proposed for potential use in sensing applications. The benzimidazole moiety introduces a broad spectrum of useful multifunctional properties to the system including electron accepting ability, pH sensitivity and compatibility with biomolecules. The photophysical characterisation of the prototropic forms of these chromophores has been carried out in both solution and on immobilisation in polymer films. The experimental results are further supported by computational determination of pKa values. It is noticed that compound 3 forms nanoaggregates in aqueous solutions with aggregation-induced emission (AIE) at 600nm. All the systems demonstrate spectral pH sensitivity in acidic media which shifts towards near-neutral values upon immobilisation in polymer films or upon aggregation in an aqueous environment (compound 3). The structure-property relationships of these functional chromophores, involving their spectral characteristics, acid-base equilibria, pKa values and aggregation effects have been determined. Potential applications of the molecules as pH and biomolecular sensors are proposed based on their pH sensitivity and AIE properties.

Dangerous liaisons: anion-induced protonation in phosphate-polyamine interactions and their implications for the charge states of biologically relevant surfaces.

Although not always considered a preponderant interaction, amine-phosphate interactions are omnipresent in multiple chemical and biological systems. This study aims to answer questions that are still pending about their nature and consequences. We focus on the description of the charge state as surface charges constitute directing agents of the interaction of amine groups with either natural or synthetic counterparts. Our results allow us to quantitatively determine the relative affinities of HPO42- and H2PO4- from the analysis of the influence of phosphates on the zeta-potential of amino-functionalized surfaces in a broad pH range. We show that phosphate anions enhance the protonation of amino groups and, conversely, charged amines induce further proton dissociation of phosphates, yielding a complex dependence of the surface effective charge on the pH and phosphate concentration. We also demonstrate that phosphate-amine interaction is specific and the modulation of surface charge occurs in the physiological phosphate concentration range, emphasizing its biochemical and biotechnological relevance and the importance of considering this veiled association in both in vivo and in vitro studies.

The Key Heterolysis Selectivity of Divalent Sulfur–Sulfur Bonds for a Unified Mechanistic Scheme in the Thiosulfatolysis and Sulfitolysis of the Pentathionate Ion

The thiosulfatolysis and sulfitolysis of pentathionate were studied in the pH ranges 4.50–8.0 and 4.0–5.0 at T = (25.0 ± 0.1) °C by HPLC monitoring of the time‐dependence and distribution of different sulfur species. The two systems are first order with respect to both reactants. In contrast to tetrathionate, pentathionate and higher polythionates contain inequivalent divalent sulfur atoms; therefore, the complexity of the mechanism increases. The selective heterolytic cleavages of sulfur–sulfur bonds are essential steps for the time evolution and equilibrium distribution of the divalent sulfur species. A proposed eight‐step mechanistic model, which incorporates the proton equilibrium of sulfite and the heterolytic cleavage of divalent sulfur–sulfur bonds of hexathionate, was applied to evaluate the thiosulfatolysis and sulfitolysis of the pentathionate ion, and the kinetics of both systems are modulated by this mechanistic network.