Pulse Radiolysis and Inactivation of Trypsin

A study of the mechanism of aqueous trypsin inactivation by radiation was carried out by pulse radiolysis and steady irradiation in the presence of various scavengers. The initial products consist of OH radical and H atom adducts to tryptophan residues, and of electron adduct to cystine residues. Essentially, all OH react with trypophan with k(OH + trypsin) = (8.2 ± 1.2) × 10¹⁰ M^?1 sec^?1. The electron adduct to the disulphide bond accounts for about 60 per cent of the electron yield, with k(e_aq + trypsin) = (3.5 ± 0.8) × 10¹⁰ M^?1 sec^?1. Measurements of tryptophan residue loss and sulphydryl formation under different irradiation conditions show that the initial reactions are substantially reversible, except for the destruction of tryptophan under aerobic conditions. Many primary radicals induce inactivation with low independent probabilities, indicating that only a small fraction of the possible reaction sites for each species influence the integrity of the active center.     

Kinetic Study of the Photoexcited Triplet State of Free Base Porphyrins by EPR

An electron paramagnetic resonance (EPR) study of the photoexcited triplet state of four free base porphyrins is presented. The zero field splitting parameters (ZFS) |D| and |E| were calculated from the EPR spectra of the porphyrins dissolved in n-octane matrices at 80°K. |D| = 0.0359 cm^?1, |E| = 0.0079 cm^?1 for tetra phenyl porphyrin (H₂ TPP), |D| = 0.0432 cm^?1, |E| = 0.0037 cm^?1 for tetra (per-fluoro) phenyl porphyrin H₂T (per-F) PP, |D| = 0.0366 cm^?1, |E| = 0.0078 cm^?1 for tetra (para-chloro) phenyl porphyrin H₂T(P-Cl)PP, |D| = 0.0369 cm^?1, |E| = 0.0076 cm^?1 for tetra (para-methyl) phenyl porphyrin H₂T(P-Me)PP. The transient behavior of the EPR signal intensities in the last two porphyrins is discussed. The depopulation rate constants of the triplet sublevels k_p, the ratio between the population rate constants A_p (at zero field, p = x,y,z), and the spin lattice relaxation rate W within the triplet manifold, were calculated. k_x = (12 ± 2) × 10² sec^?1, k_y = (0.5 ± 0.1) × 10² sec^?1, k_z = (1.2 ± 0.4) × 10² sec^?1, A_x:A_y:A_z ? 0.63:0.01:0.33, W = (0.4 ± 0.1) × 10⁴ sec^?1 for H₂T(P-Cl)PP, k_x = (7 ± 2) × 10² sec^?1, k_y = (4 ± 1) × 10² sec^?1, k_z = (1.5 ± 0.5) × 10² sec^?1, A_x:A_y:A_z ? 0.56:0.31:0.13, W = (1.7 ± 0.4) × 10³ sec^?1 for H₂T(P-Me)PP.     

Fast Consecutive Intramolecular Processes Involving Protein and Fe(III) in Ferri-Cytochrome-c in Aqueous Solution

The reaction of H atoms (produced by nanosecond pulse radiolysis) with ferri-cytochrome-c in aqueous solution results in the second order formation of an absorption spectrum due to the addition of H atoms to the enzyme protein. This spectrum is not specific to the iron moiety. The rate constant of the addition is 1×10¹⁰M^?1 sec^?1. This is followed by two first order intramolecular processes with specific rates of ～ 1×10⁵ sec^?1 and ～ 2×10⁴ sec^?1. In these processes, specific spectra related to the α and β bands of ferro-cytochrome-c appear. The results are interpreted to show reduction equivalent transfer through the protein to the iron moiety.     

Decay of the HO2 and O−2 Radicals Catalyzed by Superoxide Dismutase. A Pulse Radiolytic Investigation

The pulse radiolysis technique has been employed in the investigation of the dismutation of superoxide radicals, O^?₂ and HO₂, in the presence of superoxide dismutase in aqueous solutions. The decay of superoxide radicals in the presence of the enzyme was found to be first order in both enzyme and superoxide concentrations. An apparent second order reaction rate constant was found to be about 2 × 10⁹ M^?1 sec^?1, decreasing slightly as the pH is increased from 5 to 9.5. A mechanism which accounts for all our observations is proposed. It includes two steps: (1) formation of a product (EO^?₂ or E^?) from one enzyme (E) molecule and one O^?₂ radical ion; (2) regeneration of E by a reaction of this product with an additional O^?₂ ion radical. The reaction rate constants k = (1.4 ± 0.2) × 10⁹ and k = (1.9 ± 0.6) × 10⁹ M^?1 sec^?1 were measured at pH = 7 in an oxygenated 0.16 M sodium formate solution.     

Radiolytic Studies of the Cumyloxyl Radical in Aqueous Solutions

Formation and reactions of the cumyloxyl radical in aqueous solutions were studied by steady-state and pulse radiolytic techniques. Cumene hydroperoxide reacts with e⁻_aq (k = 4.4 × 10⁹ M⁻¹s⁻¹) to yield the cumyloxyl radical. The spectrum recorded after the pulse indicates formation of a species absorbing at 250 nm. This product was identified as acetophenone, which is formed by the fragmentation of the cumyloxyl radical. By comparison of the pseudo-first-order rates of e⁻_aq decay at 600 nm with the rate of production of acetophenone at 245 nm at increasing concentrations of cumene hydroperoxide, it was possible to derive a rate constant of 1.0 × 10⁷ s⁻¹ for the cleavage of cumyloxyl to acetophenone and methyl radical. This value is higher than that measured previously in organic solvents (1 × 10⁶ s⁻¹), as expected. HPLC analysis of the radiation products acetophenone and cumyl alcohol permitted determination of rate constants for hydrogen abstraction by the cumyloxyl radical, in competition with the fragmentation. The rate constants for H abstraction from i-PrOH, EtOH, and MeOH by CmO were found to be 9.9 × 10⁶, 3.8 × 10⁶, and 8.5 × 10⁵ M⁻¹ s⁻¹, respectively.     

Flash photolysis investigation on primary processes of the sensitized polymerization of vinyl monomers: 2. Experiments with benzoin and benzoin derivatives

Benzene solutions of benzoin (B), benzoin acetate (BA), benzoinmethyl ether (BME) and benzoinisopropyl ether (BIPE) were irradiated with 0.1 or 25 nsec flashes of 347.1 nm light at room temperature. With B and BA optical absorptions due to radical precursors were detected. The first order decay constants of these absorptions are 1.1 × 10⁸ sec^?1 (B) and 5.3 × 10⁷ sec^?1 (BA). The radical spectra decayed according to second order laws. In the case of B only one mode of decay was observed which accounts for the major part of the reaction (benzaldehyde formation by disproportionation of unlike radicals). During the experiments with BA, BME and BIPE two modes of radical-radical reactions were detected. Styrene was found to react with the rapidly decaying species with k_M = (1.3 ± 0.2) × 10⁵ M^?1 sec^?1, but did not react with the slowly decaying species. By considering the end product analyses of other authors it was concluded that the rapidly decaying mode of the absorption decay is due to the combination of benzoyl radicals forming benzil. The slowly decaying mode was assigned to the combination of alkoxybenzyl radicals (in the case of BME and BIPE) or of acetonbenzyl radicals (in the case of BA) forming pinacol derivatives. The rate constants of the reaction of naphthalene with triplet excited benzoin (7.7 ± 0.8) × 10⁹ M^?1 sec^?1 and benzoin acetate (5.0 ± 0.5) × 10⁹ M^?1 sec^?1 were determined.     

The Reduction of Ferricytochrome c Studied by Pulse Radiolysis

The formation of ferrocytochrome c upon pulse radiolysis of aqueous solutions of ferricytochrome c has been studied by following spectral shifts at 550 nm and 700 nm. The reaction between ferricytochrome c and hydrated electrons was studied by following the decay of hydrated electrons as well as the ferrocytochrome c formation. The two processes proceed with the same rate which indicate diffusion-controlled processes with a second order rate constant of 1.1 × 10¹¹ M^?1 sec^?1. The reaction between ferricytochrome c and H atoms was studied in a system containing H atoms only and found to be a second order reaction with a rate constant of 1.5 × 10¹⁰ M^?1 sec^?1. The formation of ferrocytochrome c by reaction between ferricytochrome c and OH radicals proceed by a first step with a second order rate constant of 1.4 × 10¹⁰ M^?1 sec^?1 during which OH radicals react with the protein part of ferricytochrome c forming an organic radical. Some of these radicals then undergo intramolecular reduction with a rate constant of 3.1 × 10³ sec^?1.     

The Formation of CrO2+2 in the Reaction of Cr2+ + O2 in Aqueous Acid Solutions

The reduction rate of Cr³⁺ by e_aq was determined, k₈ = 1.7 × 10¹⁰ M^?1 s^?1. The reaction of Cr²⁺ with O₂ was studied, k₂ = 1.6 × 10⁸ M^?1 s^?1. The spectrum of CrO²⁺₂ was obtained both with the pulse radiolysis method and by mixing Cr²⁺ with excess of O₂. It was shown that CrO²⁺₂ decays slowly to yield HCrO^?₄. The results suggest that the reaction of Cr²⁺ with O₂ is a two electron transfer process.     

Anionic polymerization of styrene in triglyme-benzene mixtures

Living anionic polymerization of styrene was kinetically investigated in triglyme-benzene mixtures. At low concentrations of triglyme the overall propagation rate constant, k_p, was much larger than at the same concentration of monoglyme (DME) in DME-benzene mixtures. The Szwarc-Schulz plot did not have negative slopes for lithium and sodium salts at triglyme contents of 5～20vol%, and no contribution of free anions to the propagation was observed for the sodium salt. The sodium ion pair was more highly reactive than the lithium ion pair; thus at 25°C, the ion pair rate constant, k′_p, for the lithium salt was 43, 102, 135 and 165 M^?1sec^?1 at triglyme concentrations of 5, 10, 15, and 20%, respectively, while that for the sodium salt was 410, 920, and 1460 M^?1sec^?1 in 5, 10, and 15% triglyme, respectively. The dissociation constant, K, for the lithium salt was 2·4×10^?11, 1·9×10^?10 and 1·3×10^?9 M in 10, 15, and 20% triglyme, respectively and the free ion rate constant, k″_p, was 2～2·5×10⁴ M^?1sec^?1 for the lithium salt.     

Novel real-time method using the stopped-flow for evaluating bisphenol A degradation kinetics by molecular ozone and radical mechanisms

A novel real-time method was developed to evaluate the bisphenol A degradation kinetics by molecular ozone and radical pathway using the stopped-flow technique. The second-order kinetics was determined under pseudo-first-order conditions for the molecular pathway by the absolute rate constant method and for the radical pathway by the R_ct concept involving the hydroxyl radical and ozone ratio. Bisphenol A degradation by ozone was performed and evaluated at a pH ranging from 2 to 10. At pH?4?M^?1?s^?1 and for the radical pathway at pH?>?10, the constant was 3.43?×?10⁹?M^?1?s^?1. To validate the method, ciprofloxacin degradation kinetics was determined at pH 8 by radical pathway, in 4.55?×?10⁹?M^?1?s^?1. The method permits the determination of kinetic parameters for the design of chemical reactors; avoiding the generation of undesirable reactions and by-products in the degradation of emerging compounds.     

Dynamic adsorption behaviors between Cu2+ ion and water‐insoluble amphoteric starch in aqueous solutions

Dynamic adsorption behavior between Cu²⁺ ion and water‐insoluble amphoteric starch was investigated. The sorption process occurs in two stages: external mass transport occurs in the early stage and intraparticle diffusion occurs in the long‐term stage. The diffusion rate of Cu²⁺ ion in both stages is concentration dependent. In the external mass‐transport process, the diffusion coefficient (D₁) increases with increasing initial concentration in the low‐ (1 × 10^?3‐4 × 10^?3M) and high‐concentration regions (6 × 10^?3‐10 × 10^?3M). The values of adsorption activation energy (k_d1) in the low‐ and high‐concentration regions are 15.46–24.67 and ?1.80 to ?11.57 kJ/mol, respectively. In the intraparticle diffusion process, the diffusion coefficient (D₂) increases with increasing initial concentration in the low‐concentration region (1 × 10^?3‐2 × 10^?3M) and decreases with increasing initial concentration in the high‐concentration region (4 × 10^?3‐10 × 10^?3M). The k_d2 values in the low‐ and high‐concentration regions are 9.96–15.30 and ?15.53 to ?10.71 kJ/mol, respectively. These results indicate that the diffusion process is endothermic in the low‐concentration region and is exothermic in the high‐concentration region for both stages. The external mass‐transport process is more concentration dependent than the intraparticle diffusion process in the high‐concentration region, and the dependence of concentration for both processes is about equal in the low‐concentration region. © 2001 John Wiley & Sons, Inc. J Appl Polym Sci 81: 2849–2855, 2001     

Laser flash photolysis investigations on primary processes of the sensitized polymerization of vinyl monomers: 1. Experiments with benzophenone

The laser flash photolysis technique was used in order to evaluate kinetic parameters concerning the efficiency of benzophenone (BP) as a photosensitizer for polymerizations. Rate constants (in M^?1 sec^?1) of the reaction of triplet BP with monomers were measured (e.g. styrene, 3.3 × 10⁹; methyl methacrylate, 6.9 × 10⁷; acrylonitrile, 3.4 × 10⁷; vinyl acetate, 5.4 × 10⁶). The rate constant of the reaction of triplet BP with tetrahydrofuran is 3 × 10⁶M^?1sec^?1. From these results it can be derived, for example, that the BP photosensitized polymerization of styrene is not feasible. Ketyl radicals of BP were found to react relatively slowly with vinyl acetate (5.5 × 10³M^?1sec^?1), acrylonitrile (3.8 × 10³M^?1sec^?1) and methyl methacrylate (9.0 × 10³M^?1sec^?1). Based on these data it was estimated that benzpinacol should not be formed as a major reaction product at relatively low incident light intensities and at monomer concentrations greater than 1 mol/l.     

Absolute reactivity in the cationic polymerization of methyl and other alkyl vinyl ethers

The cationic polymerizations of methyl-, 2-chloroethyl-, ethyl-, cyclohexyl- and t-butyl- vinyl ethers initiated by cycloheptatrienyl hexachloroantimonate in methylene chloride solutions have been studied in detail. Reaction rates were measured by an adiabatic calorimetric technique and rate constants for propagation of each of the monomers, k_p (obs), were determined by appropriate kinetic analysis of the experimental curves. The results obtained are discussed in terms of current theories regarding ion pair/free ion equilibria in non-aqueous solvents. Although ethyl-, cyclohexyl- and t-butyl- vinyl ethers behave very similarly to isobutyl vinyl ether, and their reactivities are comparable [k_p (obs) ～ 3 × 10³M^?1sec^?1 at 0°C] both methyl- and 2-chloroethyl- vinyl ethers show markedly different characteristics to the others, and in particular exhibit a reactivity approximately one order of magnitude less [k_p (obs) ～ 2 × 10²M^?1sec^?1 at 0°C]. These variations in reactivity are discussed in terms of preferred monomer conformations, and the resulting differences in activation energy which are likely to arise when such conformers are approached by an electrophile.     

Comments on the reactivity of methoxy pentadecyl phenyl isocyanate isomers

The reactions of 2-methoxy-4-pentadecyl phenyl isocyanate and 4-methoxy-2-pentadecyl phenyl isocyanate with excess 2-ethyl hexanol originally reported by Ghatge and co-workers to follow zero order kinetics have been re-examined on the basis of their data and shown to follow more realistically the product catalyzed pseudo first order kinetics. The new rate constant, k_s (sec^?1) for the spontaneous reaction and k_p (li. mole^?1 sec^?1) for the product catalyzed reaction are found to be: k_s = 0.57 × 10^?6 and k_p = 34 × 10^?6 for 2-methoxy-4-pentadecyl phenyl isocyanate and k_s = 1.2 × 10^?6 and k_p = 82 × 10^?6 for 4-methoxy-2-pentadecyl phenyl isocyanate.     

Kinetics of Estrone Ozone/Hydrogen Peroxide Advanced Oxidation Treatment

Ozone/hydrogen peroxide batch treatment was utilized to study the degradation of the steroidal hormone estrone (E1). The competition kinetics method was used to determine the rate constants of reaction for direct ozone and E1, and for hydroxyl radicals and E1 at three pH levels (4, 7, and 8.5), three different molar O₃/H₂O₂ ratios (1:2, 2:1, and 4:1) and a temperature about 20°C. The average second-order rate constants for direct ozone-E1 reaction were determined as 6.2?×?10³?±?3.2?×?10³ M^?1s^?1, 9.4?×?10⁵?±?2.7?×?10⁵ M^?1s^?1, and 2.1?×?10⁷?±?3.1?×?10⁶ M^?1s^?1 at pH 4, 7, and 8.5, respectively. It was found that pH had the greatest influence on the reaction rate, whereas O₃/H₂O₂ ratio was found to be slightly statistically significant. For the hydroxyl radical-E1 reaction, apparent rate constants ranged from 1.1?×?10¹⁰ M^?1s^?1 to 7.0?×?10¹⁰ M^?1s^?1 with an average value of 2.6?×?10¹⁰ M^?1s^?1. Overall, O₃/H₂O₂ is shown to be an effective treatment for E1.     

In-chain functionalization by alternating anionic copolymerization of styrene and 1-(4-dimethylaminophenyl)-1-phenylethylene

Anionic copolymerizations of styrene (M₁) with excess 1-(4-dimethyl-aminophenyl)-1-phenylethylene (M₂) were conducted in benzene at 25°C for 24h, using sec-butyllithium as initiator. Narrow molecular weight distribution copolymers with M?;_n = 16.1 × 10³ g/mol (M?_w/M?_n = 1.04) and 38.2 × 10³g/mol (M?_w/M?_n = 1.05), and 24 and 38 moles of M₂ per macromolecule, respectively, were characterized by size exclusion chromatography, ¹H NMR spectroscopy and DSC. The monomer reactivity ratio, r₁ = 5.6, was obtained from the copolymer composition at complete consumption of M₁, assuming that the rate constant k₂₂ =0,i.e. r₂ =0. The polymers exhibited T_g values of 128 and 119°C, respectively, which correspond to an estimated T_g = 217°C for the hypothetical homopolymer of M₂.     

The Kinetics of Complexed and Free Ho2 Radical in the Reaction of Ce+4 and H2O2

The intermediates formed in the reaction of Ce⁺⁴ with H₂O₂ in 0.5 M perchloric acid were studied, spectrophotometrically and through the quenching method, using a stopped-flow system. The spectrum in the UV range and the kinetics of the generated radicals and their dependence on Ce⁺³ concentration were investigated. On adding Ce⁺³ to the mixture, a change in the spectrum as well as in the recombination rate constant was observed. This behaviour was attributed to the formation of a complex between the cerous ion and the HO₂ radical. The values of the recombination rate constant of the complexed radical is 4.0 ± 0.4·10⁶ F^?1 sec^?1 while that of the free HO₂ radical is 0.9 ± 0.1·10⁶ F^?1 sec^?1. The stabilization constant of the complex was found to be 60 ± 18 F^?1.     

Kinetic study of the reaction between tocopheroxyl radical and unsaturated fatty acid esters in benzene

A kinetic study of the reaction between a tocopheroxyl radical and unsaturated fatty acid esters has been undertaken. The rates of allylic hydrogen abstraction from various unsaturated fatty acid esters (ethyl oleate2, ethyl linoleate3, ethyl linolenate4, and ethyl arachidonate5) by the tocopheroxyl radical (5,7-diisopropyltocopheroxyl6) in benzene have been determined spectrophotometrically. The second-order rate constants, k₃, obtained are 1.04×10⁻⁵ M⁻¹s⁻¹ for2, 1.82×10⁻² M⁻¹s⁻¹ for3, 3.84×10⁻² M⁻¹s⁻¹ for4, and 4.83×10⁻² M⁻¹s⁻¹ for5 at 25.0°C. Thus, the rate constants, k_abstr/H, given on an available hydrogen basis are k₃/4=2.60×10⁻⁶ M⁻¹s⁻¹ for2, k₃/2=9.10×10⁻³ M⁻¹s⁻¹ for3, k₃/4=9.60×10⁻³ M⁻¹s⁻¹ for4, and k₃/6=8.05×10⁻³ M⁻¹s⁻¹ for5. The k_abstr/H values obtained for the polyunsaturated fatty acid esters3,4, and5 containing H-atoms activated by two π-electron systems are similar to each other, and are about three orders of magnitude higher than that for the ethyl oleate2 containing H-atoms activated by a single π-system. From these results, it is suggested that the prooxidant effect of α-tocopherol in edible oils and fats may be induced by the above hydrogen abstraction reaction.     

Effect of amino acids containing sulfur on the corrosion of mild steel in phosphoric acid solutions containing Cl<Superscript>−</Superscript>, F<Superscript>−</Superscript> and Fe<Superscript>3+</Superscript> ions: Behavior under polarization conditions

The effect of cysteine (RSH), methionine (CH₃SR), cystine (RSSR) and N-acetylcysteine (ACC) on the corrosion behavior of mild steel in 40% H₃PO₄ solution without and with Cl⁻, F⁻, Fe^3‰+ and their ternary mixture was studied using both potentiostatic and electrochemical impedance (EIS) techniques under anodic and cathodic polarization conditions. The inorganic additives stimulate the overall corrosion reaction while the amino acids inhibit it with a predominant effect on the dissolution of iron. Both RSH and ACC are adsorbed according to Temkin’s isotherm while adsorption of RSSR and CH₃SR follows Frumkin and Langmuir isotherms respectively. The standard free energy of adsorption (ΔG ) was found to be in the order: RSSR > RSH ≅ ACC > CH₃SR. The binary mixtures of Cl⁻ or F⁻ with RSH or CH₃SR are the best inhibitors (IE > 90%) while those containing ferric ions or blend I and amino acids are not good corrosion inhibitors. EIS measurements showed that the cathodic reaction, hydrogen evolution, is charge transfer controlled while the anodic one, iron dissolution, is a complex process.     

The Reaction of Ozone with Bisulfide (HS−) in Aqueous Solution – Mechanistic Aspects

The ozone demand to oxidize HS^?/H₂S [pK_a(H₂S) = 6.9, k(HS^? + O₃) = 3 × 10⁹ M^?1 s^?1, k(H₂S + O₃) = 3 × 10⁴ M^?1 s^?1] to SO₄ ^2? is only 2.4 mol ozone per mol SO₄ ^2? formed, much lower than stoichiometric 4.0 mol/mol if a series of O-transfer reactions would occur. As primary step, the formation of an ozone adduct to HS^?, HSOOO^–, is suggested that decomposes into HSO^– and singlet oxygen (16%) or rearranges into peroxysulfinate ion, HS(O)OO^– (84%). Potential reactions of the above intermediates are discussed. Some of these can account for the low ozone demand.