Side-Chain Sulfonation to Modulate Hydroxyl Radical Generators for Efficient Suppression of Hepatoma Cells under Hypoxia.

Synthetic boron-doped diamond thin film is a new promising anode material. Because of its properties (high anodic stability under drastic conditions and wide potential window), it is widely investigated for numerous possible electrochemical applications such as electrosynthesis, preparation of powerful oxidants and electroincineration. In the first part of this work, simple charge transfer was investigated at boron-doped diamond electrode through the study of an outer sphere system in the potential region of water stability. In a second part of this work, the electrochemical oxygen transfer reaction (EOTR) was studied in more details. Hydroxyl radicals are one of the most important intermediates produced during EOTR. Their formation depends on the electrode material as well as the potential and implies different mechanisms and reactivities. At low potential, hydroxyl radicals are produced by the dissociative adsorption of water followed by the hydrogen discharge. This reaction is assumed to take place at electrocatalytic material like platinum. When the potential is higher than 1.23 V vs SHE (thermodynamic potential of water decomposition in acidic medium), the water discharge occurs, leading to the formation of hydroxyl radicals. From this, two classes of materials can be distinguished: active and non active electrodes. It is well established that at active electrodes, a strong interaction with hydroxyl radicals exists and the EOTR occurs via the formation of an higher oxide. In contrast, at non active electrodes, the substrate does not participate in the process and the oxidation is assisted by hydroxyl radicals that are weakly adsorbed at the electrode surface. Assuming that hydroxyl radicals are the main intermediates of the reaction, a model was developed to predict the organic compounds oxidation (COD-ICE model). Another part of this work deals with the validation of the theoretical models. In addition to the COD-ICE model, another model describing the oxidation reaction in terms of flux of both hydroxyl radicals and organics (γ-ν model) was developed. Both models permitted on the one hand to predict and describe the evolution of the oxidation reaction, and on the other hand to confirm the role of hydroxyl radicals. Moreover, it was possible to perform, depending on the conditions of applied current, either a partial oxidation (into intermediates) or a total incineration (into CO2) of the organic compound. The models, developed for a one-compartment electrochemical flow cell, were also validated in both a two-compartments cell and a new electrochemical cell, called turbine cell. In addition, the development of this cell allowed us to work with well established hydrodynamic conditions. The wide potential window that exists at boron-doped diamond electrode (BDD) theoretically allows the formation of free hydroxyl radicals, whose redox potential is estimated at about 2.6 V (vs SHE). The principal aim of this work was to highlight the presence of hydroxyl radicals at BDD electrode and to study their reactivity. First, we have investigated the production of hydrogen peroxide and the competitive reaction of carboxylic acids, both of which indicated the presence of hydroxyl radicals. Then, spin trapping was performed to detect hydroxyl radicals. This method consists in trapping the radical with an appropriate scavenger to produce a stable adduct, which can be analyzed by different techniques such as electron spin resonance (ESR), UV-visible and liquid chromatography (HPLC) measurements. The spin trapping at BDD electrode was performed through three experiments, viz., the electrolysis of a solution of 5,5-dimethyl- 1-pyrroline-N-oxide (DMPO) or 4-nitroso-N,N-dimethylaniline (p-nitrosoaniline or RNO) and the hydroxylation of salicylic acid using ESR, UV and HPLC analysis, respectively. These results have confirmed the presence and the key role of hydroxyl radicals during oxidative processes at BDD electrode. The hydroxylation of salicylic acid, whose oxidation mechanism is well established and yields to two dihydroxylated isomers (2,3- and 2,5-DHBA), was investigated in more details to study the reactivity of hydroxyl radicals. The results were compared to the reactivity of hydroxyl radicals chemically produced by Fenton reaction and UV-photolysis. The comparison was based on the investigation of the isomer distribution. On the basis of our results and by analogy with chemical and biological results, a mechanism for salicylic acid hydroxylation was proposed.

Since reactive oxygen species (ROS) and hydroxyl radicals (*OH) play an important role in the pathogenesis of many human degenerative diseases, much attention has focused on the development of safe and effective antioxidants. Preliminary experiments have revealed that the methanol (MeOH) extract of the stem of Prunus davidiana exerts inhibitory/scavenging activities on 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals, total ROS and peroxynitrites (ONOO-). In the present study, the antioxidant activities of this MeOH extract and the organic solvent-soluble fractions, dichloromethane (CH2Cl2), ethyl acetate (EtOAc), and n-butanol (n-BuOH), and the water layer of P. davidiana stem were evaluated for the potential to inhibit *OH and total ROS generation in kidney homogenates using 2',7'-dichlorodihydrofluorescein diacetate (DCHF-DA), and for the potential to scavenge authentic ONOO-. We also evaluated the inhibitory activity of seven flavonoids isolated from P. davidiana stem, kaempferol, kaempferol 7-O-beta-D-glucoside, (+)-catechin, dihydrokaempferol, hesperetin 5-O-beta-D-glucoside, naringenin and its 7-O-beta-D-glucoside, on the total ROS, *OH and ONOO- systems. For the further elucidation of the structure-inhibitory activity relationship of flavonoids on total ROS and *OH generation, we measured the antioxidant activity of sixteen flavonoids available, including three active flavonoids isolated from P. davidiana, on the total ROS and *OH systems. We found that the inhibitory activity on total ROS generation increases in strength with more numerous hydroxyl groups on their structures. Also, the presence of an ortho-hydroxyl group, whether on the A-ring or B-ring, and a 3-hydroxyl group on the C-ring increased the inhibitory activity on both total ROS and *OH generation.

Several studies indicate the presence of hydroxyl radical (OH.) as well as its involvement in the myocardial reperfusion injury. A transition metal-like iron is necessary for the conversion of superoxide anion (O2-) to a highly reactive and cytotoxic hydroxyl radical (OH.). In the present study, we have examined the generation of OH. and free iron in reperfused hearts following either normothermic (37 degrees C) or hypothermic ischemia (5 degrees C). Employing the Langendorff technique, isolated rat hearts were subjected to global ischemia for 30 min at 37 degrees C or 5 degrees C and were then reperfused for 15 min at 37 degrees C. The results of the study suggest that both the OH. generation in myocardium and free iron release into perfusate were significantly lower in hearts made ischemic at 5 degrees C as compared to 37 degrees C. Release of myoglobin and lactic acid dehydrogenase into perfusate also followed a similar pattern. Furthermore, in in vitro studies, chemically generated O2- at 5 degrees C caused a significantly lower rate of oxidation of oxymyoglobin as well as generation of OH. and free iron as compared to 37 degrees C. These results suggest that (1) reperfusion of hypothermic ischemic heart is associated with a reduction in the generation of OH. and cellular damage compared to that of normothermic ischemic heart, and (2) myoglobin, an intracellular protein, is a source of free iron and plays a role in the reperfusion injury mediated by free radicals.

The purpose of this study was to evaluate the oxidative effect in human lymphocytes after acute nickel (Ni) treatment for 1 h; levels of intracellular reactive oxygen species (ROS), lipid peroxidation (LPO) and hydroxyl radicals ((*)OH) were examined in isolated lymphocytes. The potential effects of antioxidants were also examined. After acute treatment, NiCl(2) (0-10 mM) significantly decreased the viability of lymphocytes. NiCl(2) appear to increase the degree of dichlorofluorescein (DCF) fluorescence and the levels of thiobarbituric acid-reactive substances (TBARS) in human lymphocytes in vitro in a concentration-dependent manner. The level of (*)OH was quantified by two main hydroxylated derivates, 2,3- and 2,5-dihydroxybenzate (DHB). Levels of 2,3- and 2,5-DHB were significantly higher in the Ni-treated group than in controls. Catalase partially reduced the NiCl(2)-induced elevation of oxidants and TBARS, whereas superoxide dismutase (SOD) enhanced the level of oxidants and TBARS. Both NiCl(2)-induced fluorescence and LPO were prevented significantly by glutathione (GSH) and mannitol. NiCl(2)-induced increase in generation of (*)OH was prevented significantly by catalase, GSH and mannitol, but not by SOD. These results suggest that NiCl(2)-induced lymphocyte toxicity may be mediated by oxygen radical intermediates, for which the accelerated generation of (*)OH may plays an important role in Ni-induced oxidative damage of human lymphocytes. Catalase, GSH and mannitol each provides protection against the oxidative stress induced by Ni.

Linear energy transfer (LET) dependence of yields of O2-dependent and O2-independent hydrogen peroxide (H2O2) in water irradiated by ionizing radiation was investigated. The radiation-induced hydroxyl radical (•OH) generation in an aqueous solution was reported to occur in two different localization densities, the milli-molar (relatively sparse) and/or molar (markedly-dense) levels. In the milli-molar-level •OH generation atmosphere, •OH generated at a molecular distance of ∼7 nm are likely unable to interact. However, in the molar-level •OH generation atmosphere, several •OH were generated with a molecular distance of 1 nm or less, and two •OH can react to directly make H2O2. An aliquot of ultra-pure water was irradiated by 290-MeV/nucleon carbon-ion beams at the Heavy-Ion Medical Accelerator in Chiba (HIMAC, NIRS/QST, Chiba, Japan). Irradiation experiments were performed under aerobic or hypoxic (<0.5% oxygen) conditions, and several LET conditions (13, 20, 40, 60, 80, or >100 keV/μm). H2O2 generation in irradiated samples was estimated by three methods. The amount of H2O2 generated per dose was estimated and compared. O2-independent H2O2 generation, i.e. H2O2 generation under hypoxic conditions, increased with increasing LET. On the other hand, the O2-dependent H2O2 generation, i.e. subtraction of H2O2 generation under hypoxic conditions from H2O2 generation under aerobic conditions, decreased with increasing LET. This suggests that the markedly-dense •OH generation is positively correlated with LET. High-LET beams generate H2O2 in an oxygen-independent manner.

In this study, steady-state and time-resolved radiolysis methods were used to determine the primary reaction pathways and kinetic parameters for the reactions of hydroxyl radical with microcystin-LR (MC-LR). The fundamental kinetic data is critical for the accurate evaluation of hydroxyl-radical based technologies for the destruction of this problematic class of cyanotoxins. The bimolecular rate constant for the reaction of hydroxyl radical with MC-LR is 2.3 (+/-0.1) x 10(10) M(-1)s(-1) based on time-resolved competition kinetics with SCN-at low conversions using pulsed radiolysis experiments. The reaction of hydroxyl radical with MC-LR can occur via a number of competing reaction pathways, including addition to the benzene ring and diene and abstraction of aliphatic hydrogen atoms. LC-MS analyses indicate the major products from the reaction of hydroxyl radicals with MC-LR involve addition of hydroxyl radical to the benzene ring and diene moieties of the Adda side chain. Transient absorption spectroscopy monitored between 260-500 nm, following pulsed hydroxyl radical generation, indicate the formation of a transient species with absorption maxima at 270 and 310 nm. The absorption maxima and lifetime of the transient species are characteristic of hydroxycyclohexadienyl radicals resulting from the addition of hydroxyl radical to the benzene ring. The rate constant for the formation of hydroxycyclohexadienyl radical is 1.0 (+/-0.1) x 10(10) M(-1)s(-1) accounting for approximately 40% of the primary reaction pathways. Representative rate constants and partitioning of hydroxyl radical reactions were assessed based on the reactivities of surrogate substrates and individual amino acids. Summation of the individual reactivities of hydroxyl radical at the different reactive sites (amino acids) leads to a rate constant of 2.1 x 10(10) M(-1) s(-1) in good agreementwith the rate constant determined in our studies. The relative magnitude of the rate constants for the reactions of hydroxyl radical with the individual amino acids and appropriate surrogates, suggest 60-70% reactions of hydroxyl radical occur at the benzene and diene functional groups of the Adda moiety.

Rate constants for the gas phase reactions of hydroxyl radicals and chlorine atoms with a number of ethers have been determined at 300 ± 3 K and at a total pressure of 1 atmosphere. Both OH radical and chlorine atom rate constants were determined using a relative rate technique. Values for the rate constants obtained are as follows. compound kOH×1012(cm3 molecule−1 s−1) kC1×1011(cm3 molecule−1 s−1) Hexane 5.53 ± 1.55 — 2‐Chloro ethyl methyl ether 4.92 ± 1.09 14.4 ± 5.0 2,2‐Dichloro ethyl methyl ether 2.37 ± 0.50 4.4 ± 1.6 2‐Bromo ethyl methyl ether 6.94 ± 1.38 16.3 ± 5.4 2‐Chloro,1,1,1‐trifluoro ethyl ethyl ether &lt;0.3 0.30 ± 0.10 Isoflurane &lt;0.3 &lt;0.1 Enflurane &lt;0.3 &lt;0.1 Di‐i‐propyl ether 11.08 ± 2.26 16.3 ± 5.4 Diethyl ether — 25.8 ± 4.4 The above relative rate constants are based on the values of k(OH + pentane)=[3.94 ± 0.98]×10−12 and k(OH + diethyl ether)=[13.6 ± 2.26] × 10−12 cm3 molecule−1 s−1 in the case of the hydroxyl reactions. In the case of the chlorine atom reactions, the above rate constants are based on values of k(Cl + ethane)=[5.84 ± 0.88] × 10−11 and k(Cl + diethyl ether)=[25.4 ± 8.05] × 10−11 cm3 molecule−1 s−1. The quoted errors include ±2σ from a least squares analysis of our slopes plus the uncertainty associated with the reference rate constants.Atmospheric lifetimes calculated with respect to reaction with OH radicals are based on a tropospheric OH radical concentration of (7.7 ± 1.4) × 105 radicals cm−3, and lifetimes with respect to reaction with Cl atoms are based on a tropospheric Cl atom concentration of 1 × 103 atoms cm−3. Observed trends in the relative rates of reaction of hydroxyl radicals and chlorine atoms with the ethers studied is discussed. The significance of the calculated tropospheric lifetimes is also reviewed. © 1993 John Wiley &amp; Sons, Inc.

Pulse radiolysis and density functional theory (DFT) calculations at B3LYP/6-31+G(d,p) level have been carried out to probe the reaction of the water-derived hydroxyl radicals (*OH) with 5-azacytosine (5Ac) and 5-azacytidine (5Acyd) at near neutral and basic pH. A low percentage of nitrogen-centered oxidizing radicals, and a high percentage of non-oxidizing carbon-centered radicals were identified based on the reaction of transient intermediates with 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonate), ABTS2-. Theoretical calculations suggests that the N3 atom in 5Ac is the most reactive center as it is the main contributor of HOMO, whereas C5 atom is the prime donor for the HOMO of cytosine (Cyt) where the major addition site is C5. The order of stability of the adduct species were found to be C6-OH_5Ac*>C4-OH_5Ac*>N3-OH_5Ac*>N5-OH_5Ac* both in the gaseous and solution phase (using the PCM model) respectively due to the additions of *OH at C6, C4, N3, and N5 atoms. These additions occur in direct manner, without the intervention of any precursor complex formation. The possibility of a 1,2-hydrogen shift from the C6 to N5 in the nitrogen-centered C6-OH_5Ac* radical is considered in order to account for the experimental observation of the high yield of non-oxidizing radicals, and found that such a conversion requires activation energy of about 32 kcal/mol, and hence this possibility is ruled out. The hydrogen abstraction reactions were assumed to occur from precursor complexes (hydrogen bonded complexes represented as S1, S2, S3, and S4) resulted from the electrostatic interactions of the lone pairs on the N3, N5, and O8 atoms with the incoming *OH radical. It was found that the conversion of these precursor complexes to their respective transition states has ample barrier heights, and it persists even when the effect of solvent is considered. It was also found that the formation of precursor complexes itself is highly endergonic in solution phase. Hence, the abstraction reactions will not occur in the present case. Finally, the time dependent density functional theory (TDDFT) calculations predicted an absorption maximum of 292 nm for the N3-OH_5Ac* adduct, which is close to the experimentally observed spectral maxima at 290 nm. Hence, it is assumed that the addition to the most reactive center N3, which results the N3-OH_5Ac* radical, occurs via a kinetically driven process.

Hydroxyl radical generation in electro-Fenton process with in situ electro-chemical production of Fenton reagents by gas-diffusion-electrode cathode and sacrificial iron anode

Hydroxyl radical reactions and the radical scavenging activity of β-carboline alkaloids

Reactions of hydroxyl radical with bergenin, a natural poly phenol studied by pulse radiolysis

This paper describes how the homogeneous decomposition of hydrogen peroxide vapour has been successfully used as a source of hydroxyl radicals in a kinetic study of the relative rates of reaction of hydroxyl radicals with methane, carbon monoxide, formaldehyde and hydrogen peroxide. It has been found that the method is of general applicability provided subsequent reactions of radicals formed can be controlled. With hydroxyl radicals so produced from the decomposition of hydrogen peroxide, surprisingly consistent values were obtained for the relative rates of reaction of methane and carbon monoxide with hydroxyl radicals. Thus, within an accuracy of 10%, the ratios of rate constants for reaction of hydroxyl radicals with methane and carbon monoxide were found to be 3.6 at 650 °C, 2.1 at 525 °C and 0.85 at 400 °C. The rates of reaction of hydroxyl radicals with formaldehyde and hydrogen peroxide were less accurately determined because of expected additional reactions. Within an accuracy of about 50% it was estimated that hydroxyl radicals reacted ten times as fast with hydrogen peroxide as with carbon monoxide at 525 °C. At the same temperature hydroxyl radicals reacted 33 ± 6 times as fast with formaldehyde as with methane. Data were obtainable at temperatures as low as 400 °C even though heterogeneous decomposition of hydrogen peroxide was relatively more important at these temperatures. This was because heterogeneous decomposition of hydrogen peroxide did not cause oxida­tion of the other gases which were also present. A preliminary account (Hoare 1962) of this work has previously been published.

Rate coefficients for the reactions of hydroxyl (OH) radicals with the dimethylbenzaldehydes have been determined at 295 ± 2K and atmospheric pressure using the relative rate technique. Experiments were performed in an atmospheric simulation chamber using gas chromatography for chemical analysis. The rate coefficients (in units of cm3 molecule−1 s−1) are: 2,3‐dimethylbenzaldehyde, (25.9 ± 2.8) × 10−12; 2,4‐dimethylbenzaldehyde, (27.5 ± 4.4) × 10−12; 2,5‐dimethylbenzaldehyde, (27.6 ± 5.1) × 10−12; 2,6‐dimethylbenzaldehyde, (30.7 ± 3.0) × 10−12; 3,4‐dimethylbenzaldehyde, (24.6 ± 4.0) × 10−12; and 3,5‐dimethylbenzaldehyde, (28.2 ± 2.5) × 10−12. The reactivity of the dimethylbenzaldehydes is compared with other aromatic compounds and it is shown that the magnitude of the OH rate coefficients does not depend significantly on the position of the CH3 substituent on the aromatic ring. The rate coefficient data are explained in terms of known mechanistic features of the reactions and the atmospheric implications are also discussed. © 2006 Wiley Periodicals, Inc. Int J Chem Kinet 38: 563–569, 2006

A kinetic model has been developed, taking into account both decomposition of ozone molecules and interactions between ozone and hydrogen peroxide for formation of hydroxyl radical and subsequent reactions. Experiments were carried out at 25°C in the pH range of 3 to 13, indicating that the depletion rate of ozone increases with the concentrations of ozone, hydrogen peroxide and hydroxyl ion, as predicted by the kinetic model. Adverse scavenging reactions, however, also play significant roles at sufficient concentration ratios of hydrogen peroxide to ozone and high concentrations of hydroxyl ion in reducing the depletion rate. Results of this research suggest, that it is most desirable to conduct the peroxone oxidation for pollutant destruction by the hydroxyl radical reaction in alkaline solutions of pH below 11, while maintaining about the same concentration of ozone and hydrogen peroxide.

Photoinduced Carbene for Effective Photodynamic Therapy Against Hypoxic Cancer Cells