Oxidation of resveratrol catalyzed by soybean lipoxygenase

In this work the oxidative degradation of resveratrol catalyzed by lipoxygenase-1 (LOX-1) has been studied. The process has been characterized by spectroscopic and polarographic measurements. The oxidation of resveratrol was dependent on the concentration of resveratrol and the enzyme. When resveratrol was incubated in the presence of lipoxygenase at pH 9.0, the reaction displayed a k(M) value of 18.6 x 10(-)(6) M and a catalytic efficiency (k(cat)/k(M)) of 4.3 x 10(4) s(-)(1) M(-)(1). These values are close to those shown by the enzyme when linoleic acid is used as the substrate. The effect of lipoxygenase inhibitors on the lipoxygenase-catalyzed resveratrol oxidation was also evaluated. The rate of resveratrol oxidation was markedly decreased by the presence of NDGA in the incubation mixture. From HPLC measurements, it can be deduced that resveratrol is oxidatively decomposed to a complex mixture of products similar to those obtained when the molecule is oxidized by hydrogen peroxide.     

Pseudoperoxidase activity of myoglobin: kinetics and mechanism of the peroxidase cycle of myoglobin with H2O2 and 2,2-azino-bis(3-ethylbenzthiazoline-6-sulfonate) as substrates

Using 2,2-azino-bis(3-ethylbenzthiazoline-6-sulfonate) (ABTS) as substrate, it has been shown that the increased peroxidase activity for decreasing pH of myoglobin activated by hydrogen peroxide is due to a protonization of ferrylmyoglobin, MbFe(IV)=O, facilitating electron transfer from the substrate and corresponding to pK(a) approximately 5.2 at 25.0 degrees C and ionic strength 0.16, rather than due to specific acid catalysis. On the basis of stopped flow absorption spectroscopy with detection of the radical cation ABTS(.+), the second-order rate constant and activation parameters for the reaction between MbFe(IV)=O and ABTS were found to have the values k = 698 +/- 32 M(-1) s(-1), DeltaH# = 66 +/- 4 kJ mol(-1), and DeltaS# = 30 +/- 15 J mol(-1) K(-1) at 25.0 degrees C and physiological pH (7.4) and ionic strength (= 0.16 M NaCl). At a lower pH (5.8) corresponding to the conditions in meat, values were found as follows: k = 3.5 +/- 0.3 x 10(4) M(-1) s(-1), DeltaH# = 31 +/- 6 kJ mol(-1), and DeltaS# = -53 +/- 19 J mol(-1) K(-1), indicative of a shift from outersphere electron transfer to an innersphere mechanism. For steady state assay conditions, this shift is paralleled by a shift from saturation kinetics at pH 7.4 to first-order kinetics for H2O2 as substrate at pH 5.8. In contrast, the activation reaction between myoglobin and hydrogen peroxide was found at 25.0 degrees C to be slow and independent of pH with values of 171 +/- 7 and 196 +/- 19 M(-1) s(-1) found at physiological and meat pH, respectively, as determined by sequential stopped flow spectroscopy, from which a lower limit of k = 6 x 10(5) M(-1) s(-1) for the reaction between perferrylmyoglobin, .MbFe(IV)=O, and ABTS could be estimated. As compared to the traditional peroxidase assay, a better characterization of pseudoperoxidase activity of heme pigments and their denatured or proteolyzed forms is thus becoming possible, and specific kinetic effects on activation, substrate oxidation, or shift in rate determining steps may be detected.     

A kinetic model for the glucose-fructose-glycine browning reaction

The browning of glucose-fructose-glycine mixtures involves parallel glucose-glycine and fructose-glycine reactions, which share a common intermediate, the immediate precursor of melanoidins in the kinetic model. At pH 5.5, 55 degrees C glucose is converted into this intermediate in a two step process where k(1) = (7.8 +/- 1.1) x 10(-)(4) mol L(-)(1) h(-)(1) and k(2) = (1.84 +/- 0.31) x 10(-)(3) h(-)(1) according to established kinetics, whereas fructose is converted into this intermediate in a single step where k(4) = 5.32 x 10(-)(5)()()mol L(-)(1) h(-)(1). The intermediate is converted to melanoidins in a single rate limiting process where k(mix) = 0.0177 h(-)(1) and the molar extinction coefficient (based on the concentration of sugar converted) of the melanoidins so formed is 1073 +/- 4 mol(-)(1) L cm(-)(1). Whereas the value of k(mix) is the same when the individual sugars undergo browning, the value of the molar extinction coefficient is similar to that for melanoidins from the glucose-glycine reaction (955 +/- 45 mol(-)(1) L cm(-)(1)) but it is approximately double the value for melanoidins from the fructose-glycine reaction (478 +/- 18 mol(-)(1) L cm(-)(1)). This is the reason that the effects of glucose and fructose on the rate of browning are synergistic.     

Compost as a soil supplement increases the level of antioxidant compounds and oxygen radical absorbance capacity in strawberries

Compost as a soil supplement significantly enhanced levels of ascorbic acid (AsA) and glutathione (GSH) and ratios of AsA/dehydroascorbic acid (DHAsA) and GSH/oxidized glutathione (GSSG) in fruit of two strawberry (Fragaria x ananassa Duch.) cultivars, Allstar and Honeoye. The peroxyl radical (ROO(*)) as well as the superoxide radical (O(2)(*)(-)), hydrogen peroxide (H(2)O(2)), hydroxyl radical (OH(*)), and singlet oxygen ((1)O(2)) absorbance capacity in strawberries increased significantly with increasing fertilizer strength and compost use. The planting medium (compost) x fertilizer interaction for phenolics and flavonoids was significant. Fruit from plants grown in full-strength fertilizer with 50% soil plus 50% compost and 100% compost yielded fruit with the highest levels of phenolics, flavonol, and anthocyanin content. A positive relationship between antioxidant activities and contents of AsA and GSH and ratios of AsA/DHAsA and GSH/GSSG existed in fruit of both strawberry cultivars. Correlation coefficients for the content of antioxidant components versus antioxidant activity [against ROO(*), O(2)(*)(-), H(2)O(2), OH(*), or (1)O(2)] ranged from r( )()= 0.7706 for H(2)O(2) versus GSH/GSSH in cv. Allstar to r = 0.9832 for O(2)(*)(-) versus total flavonoids in cv. Allstar.     

Transformation of the fungicide chlorothalonil by Fenton reagent

A modified Fenton reagent (Fe(3+)/H(2)O(2)) transformed the fungicide chlorothalonil within 60 min in aqueous solution at unadjusted pH. Transformation varied with ferric salt. Transformation was greatest with ferric nitrate and least when ferric sulfate was used. UV irradiation enhanced the transformation of chlorothalonil. The transformation of chlorothalonil was enhanced, which increased with ferric ion or hydrogen peroxide concentration. Maximum transformation was achieved at 2 mM ferric ion and 100 mM hydrogen peroxide. Additionally, chlorothalonil was more dechlorinated in the UV irradiation condition. The proposed reaction pathway includes reduction of chlorothalonil to trichloroisophthalonitrile, dichloroisophthalonitrile, and monochloroisophthalonitrile; oxidation of trichloroisophthalonitrile to trichloro-3-cyanobenzoic acid and 3-carbamyltrichlorobenzoic acid; and oxidation of hydroxychlorothalonil to trichloro-3-cyanohydroxybenzoic acid and trichlorocyanophenol.     

Atrazine photodegradation in aqueous solution induced by interaction of humic acids and iron: photoformation of iron(II) and hydrogen peroxide

The photochemical formation of Fe(II) and hydrogen peroxide (H 2O 2) coupled with humic acids (HA) was studied to understand the significance of iron cycling in the photodegradation of atrazine under simulated sunlight. The presence of HA significantly enhanced the formation of Fe(II) and H 2O 2, and their subsequent product, hydroxyl radical ( (*)OH), was the main oxidant responsible for the atrazine photodegradation. During 60 h of irradiation, the fraction of iron presented as Fe(II) (Fe(II)/Fe(t)) decreased from 20-32% in the presence of the Fe(III)-HA complex to 10-22% after adding atrazine. The rate of atrazine photodegradation in solutions containing Fe(III) increased with increasing HA concentration, suggesting that the complexation of Fe(III) with HA accelerated the Fe(III)/Fe(II) cycling. Using fluorescence spectrometry, the quenching constant and the percentage of fluorophores participating in the complexation of HA with Fe(III) were estimated by the modified Stern-Volmer equation. Fourier transform infrared spectroscopy (FTIR) offered the direct evidence that Fe(III)-carboxylate complex could be formed by ligand exchange of HA with Fe(III). Based on all the information, a possible reaction mechanism was proposed.     

Isomerization and degradation kinetics of hop (Humulus lupulus) acids in a model wort-boiling system

The rate of isomerization of alpha acids to iso-alpha acids (the compounds contributing bitter taste to beer) was determined across a range of temperatures (90-130 degrees C) to characterize the rate at which iso-alpha acids are formed during kettle boiling. Multiple 12 mL stainless steel vessels were utilized to heat samples (alpha acids in a pH 5.2 buffered aqueous solution) at given temperatures, for varying lengths of time. Concentrations of alpha acids and iso-alpha acids were quantified by high-pressure liquid chromatography (HPLC). The isomerization reaction was found to be first order, with reaction rate varying as a function of temperature. Rate constants were experimentally determined to be k1 = (7.9 x 10(11)) e(-11858/T) for the isomerization reaction of alpha acids to iso-alpha acids, and k2 = (4.1 x 10(12)) e(-12994/T) for the subsequent loss of iso-alpha acids to uncharacterized degradation products. Activation energy was experimentally determined to be 98.6 kJ per mole for isomerization, and 108.0 kJ per mole for degradation. Losses of iso-alpha acids to degradation products were pronounced for cases in which boiling was continued beyond two half-lives of alpha-acid concentration.     

The role of EDTA in malonaldehyde formation from DNA oxidized by Fenton reagent systems

Calf thymus DNA was oxidized by various Fenton reagent systems [Fe(II)/H(2)O(2)] with or without ethylenediamine tetraacetic acid (EDTA) under different reaction conditions. Calf DNA was also oxidized by a modified Fenton reagent (Fe(III)/H(2)O(2)/ascorbic acid) with EDTA. Malonaldehyde (MA) formed from DNA was derivatized into 1-methyl hydrazine, which was subsequently analyzed by gas chromatography with a nitrogen-phosphorus detector. MA formation increased linearly with an increase of Fe(II) concentration. MA formation reached a plateau at nearly 2 mmol/L of Fe(II) with 0.5 mmol/L of H(2)O(2). Addition of EDTA increased MA formation from DNA nearly 5 times. When DNA was oxidized with various amount of ethanol, MA formation decreased with an increase of ethanol concentration, either with or without EDTA. The rate of inhibition was greater without EDTA than with EDTA. When DNA was oxidized by a modified Fenton reagent, MA formation linearly increased with the increase of DNA. Ascorbic acid alone produced some MA upon oxidation.     

Linear free energy study of ring-substituted aniline ozonation for developing treatment of aniline-based pesticide wastes

The relative rate constants for the reaction of ozone were determined for several substituted anilines in aqueous solutions at pH 6.5 and 1.5. At pH 6.5, with the exception of m- and p-nitroaniline, the rate constants obey Hammett's equation: log(k(X)/k(H)) = rho sigma. The departure of m- and p-nitroaniline may be explained by direct conjugation of the reaction center. The commonly used sigma(p)(-) value of 1.27, which extends the range of applicability of the Hammett equation, was insufficient to account for the conjugation effects on ozonation of p-nitroaniline; rho = -1.48 (R = 0.973). Use of amine group atomic charge determinations significantly improved correlations: (k(X)/k(H)) = 48.7 delta - 18.2 (R = 0.996). A linear plot of Hammett constants versus relative rate data at pH 1.5 showed poor correlation: rho = 0.72 (R = 0.572). Poor correlation was similarly observed for amine group atomic charge determinations, suggesting varied reaction mechanisms.     

The role of iron in peroxidation of polyunsaturated fatty acids in liposomes

This work investigated iron-catalyzed lipid oxidation in marine phospholipid liposomes. Oxygen consumption was used as a method to study lipid oxidation at pH 5.5 and 30 degrees C. The relationship between consumed oxygen and amount of peroxides (PV) and thiobarbituric reactive substances (TBARS) formed showed that both Fe2+ and Fe3+ catalyzed lipid oxidation. When Fe2+ was added to liposomes at a concentration of approximately 10 microM, an initial drop in dissolved oxygen (oxygen uptake rate >258 microM/min), followed by a slower linear oxygen uptake (oxygen uptake rate 4-6 microM/min), was observed. Addition of Fe3+ induced only the linear oxygen uptake. The initial fast drop in dissolved oxygen was due to oxidation of Fe2+ to Fe3+ by preexisting lipid peroxides (rate 79 microM Fe2+/min). Fe3+ is reduced by peroxides to Fe2+ at a slow rate (0.25 microM Fe3+/min at 30 degrees C) in a pseudo-first-order reaction. The redox cycling between Fe2+ and Fe3+ leads to an equilibrium between Fe2+ and Fe3+ resulting in a linear oxygen uptake. During the linear oxygen uptake, the interaction of Fe (3+) with lipid peroxide is the rate-limiting factor. Both alkoxy and peroxy radicals are formed by breakdown of peroxides by Fe2+ and Fe3+. These radicals react with fatty acids giving lipid radicals reacting with oxygen.     

Use of differential scanning calorimetry to study lipid oxidation. 1. Oxidative stability of lecithin and linolenic acid

The oxidation of linolenic acid (LNA) and soy lecithin was studied by differential scanning calorimetry (DSC) with linear programmed heating rates (non-isothermal mode). The interpretation of the shape of DSC curves is discussed, and it has been concluded that temperatures of the extrapolated start of heat release are the most reliable data for the rapid estimation of the oxidative stability of lipid materials. The Ozawa-Flynn-Wall method was used to calculate the kinetic parameters of the process: for LNA autoxidation the activation energy, Ea, and pre-exponential factor, Z, are 66 +/- 4 kJ/mol and 1.5 x 10(7) s(-1), respectively, and the autoxidation of lecithin is described by Ea = 98 +/- 6 kJ/mol and Z = 9.1 x 10(10) s(-1). Values of Ea and Z can be applied for calculation of the overall first-order rate constant of autoxidation at various temperatures, k(T). For the two studied lipids the comparison of k(T) values shows the inversion of their oxidative stabilities; that is, below 167 degrees C lecithin is more stable than LNA, k(T)lecithin < k(T)LNA, and above that temperature (termed the isokinetic temperature) k(T)lecithin > k(T)LNA. The calculated inversion of oxidative stabilities can be an explanation of similar observations for other pairs of lipids if the results of accelerated tests measured at temperatures above 100 degrees C are compared with the results obtained at temperatures below 100 degrees C.     

封闭系统水稻土甲烷氧化的模拟

Methane oxidation by paddy soils in a closed system could be simulated by the equation x=k1xo/(k1 k2x0)exp(k2t)-k2x0 where x0 and x are the CH4 concentrations at time zero and t,respectively;k1 and k2 are constants related to the constant of first-order-kinetics.According to the equation the change of soil ability to oxidize CH4 could be estimated by the equation Ac=k2/k1(x0-x)x0k2/k1-1.The results showed that the soil ability to oxidize CH4 varied,depending on the initial CH4 concentration.High initial CH4 concentration stimulated soil ability to consume CH4,while low concentration depressed the ablility.This characteristic of paddy soil seemed to be of considerable significance to self-adjusting CH4 emission from flooded rice fields if there exist oxic microsites in the soil.     

Browning evaluation of ready-to-eat apples as affected by modified atmosphere packaging

The color and polyphenol oxidase (PPO) activity of fresh-cut Golden delicious apples were evaluated throughout cold storage under modified atmospheres. The shelf life of cut apples was extended to several weeks, especially when an initial atmosphere of 90.5% N(2) + 7% CO(2) + 2.5% O(2) and plastic pouches of 30 cm(3)/cm(2) x bar x 24 h were used. Under these conditions, a maximum 62% PPO activity depletion was observed. In all cases, the faster the initial PPO activity decays, the less the color changes. A fractional conversion first-order model was proposed for predicting color changes in minimally processed apples. Browning was better described through lightness (L) (k(L) = 0.017 - 0.07 day(-1)) and color difference (Delta E*) values (k(Delta E) = 0.015 - 0.073 day(-1)), which fitted the model with enough accuracy.     

Treatment of cork process wastewater by a successive chemical-physical method

In cork processing, the operation of boiling the raw cork generates large volumes of wastewater which are more often than not released directly into the environment untreated. Even when the wastewater is treated, this is usually by retention in evaporation ponds. This procedure, however, causes bad odors and may pollute surface water and groundwater. The present study evaluates a physicochemical method involving Fenton oxidation and coagulation/flocculation for the removal of chemical oxygen demand (COD), total polyphenols (TP), and aromatic compounds (A) from cork manufacturing process wastewater. The experimental variables studied were the dosages of iron salts (from 0.001 to 0.2 mol/L) and hydrogen peroxide (between 0.06 and 1 mol/L). The integrated Fenton-coagulation/flocculation process reduced the COD of the effluent by from 22% to 85%. The removal of total polyphenols ranged from 4% to 98%, and of aromatic compounds from 2% to 97%. A further two experiments were performed modifying the manner in which the reagents were added, splitting the reagent dose (of hydrogen peroxide and ferrous salt) into two and three fractions. Finally, an economic study was made of the chemical costs deriving from the application of this purification system. The cost of a treatment with an [H2O2](o)/COD(o) ratio of 1.8 g/g (splitting the reagent dose into three fractions) that yields a COD removal of 73% was estimated to be 11.5 euros/m(3) of wastewater.     

Development of Singlet Oxygen Absorption Capacity (SOAC) Assay Method. 3. Measurements of the SOAC Values for Phenolic Antioxidants

Measurements of the singlet oxygen ((1)O(2)) quenching rates (k(Q) (S)) and the relative singlet oxygen absortpion capacity (SOAC) values were performed for 16 phenolic antioxidants (tocopherol derivatives, ubiquinol-10, caffeic acids, and catechins) and vitamin C in ethanol/chloroform/D(2)O (50:50:1, v/v/v) solution at 35 °C. It has been clarified that the SOAC method is useful to evaluate the (1)O(2)-quenching activity of lipophilic and hydrophilic antioxidants having 5 orders of magnitude different rate constants from 1.38 × 10(10) M(-1) s(-1) for lycopene to 2.71 × 10(5) for ferulic acid. The logarithms of the k(Q) (S) and the SOAC values for phenolic antioxidants were found to correlate well with their peak oxidation potentials (E(p)); the antioxidants that have smaller E(p) values show higher reactivities. In previous works, measurements of the k(Q) (S) values for many phenolic antioxidants were performed in ethanol. Consequently, measurements of the k(Q) (S) and relative SOAC values were performed for eight carotenoids in ethanol to investigate the effect of solvent on the (1)O(2)-quenching rate. The k(Q) (S) values for phenolic antioxidants and carotenoids in ethanol were found to correlate linearly with the k(Q) (S) values in ethanol/chloroform/D(2)O solution with a gradient of 1.79, except for two catechins. As the relative rate constants (k(Q)(AO) (S)/k(Q)(α-Toc) (S)) of antioxidants (AO) are equal to the relative SOAC values, the SOAC values do not depend on the kinds of solvent used, if α-tocopherol is used as a standard compound. In fact, the SOAC values obtained for carotenoids in mixed solvent agreed well with the corresponding ones in ethanol.     

Photodegradation of the acaricide abamectin: a kinetic study

The acaricide abamectin is a mixture of two colorless homologues in a molar ratio of at least 4:1 with the same structure of macrocyclic lactone. The kinetics of its degradation under direct (254 nm) and dye-sensitized (>400 nm) photoirradiation in methanol solution has been studied by UV-vis spectrophotometry, potentiometric detection of dissolved oxygen, stationary fluorescence, laser flash photolysis, and time-resolved detection of singlet molecular oxygen (O2((1)Delta(g))) phosphorescence. The results indicate that the degradation is very efficient under direct irradiation with UV light (254 nm), with a quantum yield of 0.23. On the contrary, under visible-light irradiation, using the natural pigment riboflavin or the synthetic dye rose bengal as sensitizers, the degradation is very inefficient and proceeds through a O2((1)Delta(g))-mediated mechanism, with a bimolecular rate constant for the overall O2((1)Delta(g)) quenching (the sum of physical and chemical quenching) of 5.5 x 10(5) M(-1) s(-1). This value is similar to those reported for the rate constants of the reactions of O2((1)Delta(g)) with isolated double bonds or conjugated dienes and points to similar processes in the case of abamectin.     

Antiphoto-oxidative activity of sesamol in methylene blue- and chlorophyll-sensitized photo-oxidation of oil

The effects and mechanism of sesamol on the methylene blue- or chlorophyll-sensitized photo-oxidations of soybean oil have been studied. Sesamol showed strong antiphoto-oxidative activity in both methylene blue-and chlorophyll-sensitized photo-oxidations of soybean oil in a dose-dependent manner. The 1.0 x 10(-3) M sesamol treatments showed 84.7 and 43.4% inhibitions of methylene blue- and chlorophyll-sensitized photo-oxidations of soybean oil in methylene chloride. The antiphoto-oxidative activity of sesamol was comparable to that of delta-tocopherol in both methylene blue- and chlorophyll-sensitized photo-oxidations, at the same molar basis. Sesamol effectively inhibited rubrene oxidation with a chemical source of singlet oxygen in microemulsion, showing its strong singlet oxygen quenching ability. The results suggested that the antiphoto-oxidative activity of sesamol in the photo-oxidation of oil was, at least in part, due to its singlet oxygen scavenging activity. The singlet oxygen quenching rate constant (k(ox-Q) + k(q)) of sesamol was determined to be 1.9 +/- 0.3 x 10(7) M(-1) s(-1). This represents the first report on the antiphoto-oxidative activity of sesamol in the sensitized photo-oxidation of oil, and its bimolecular singlet oxygen quenching ability.     

Development of singlet oxygen absorption capacity (SOAC) assay method. 2. Measurements of the SOAC values for carotenoids and food extracts

Recently a new assay method that can quantify the singlet oxygen absorption capacity (SOAC) of antioxidants was proposed. In the present work, kinetic study of the reaction of singlet oxygen ((1)O(2)) with carotenoids and vegetable extracts has been performed in ethanol/chloroform/D(2)O (50:50:1, v/v/v) solution at 35 °C. Measurements of the second-order rate constants (k(Q)(S)) and the SOAC values were performed for eight kinds of carotenoids and three kinds of vegetable extracts (red paprika, carrot, and tomato). Furthermore, measurements of the concentrations of the carotenoids included in vegetable extracts were performed, using a HPLC technique. From the results, it has been clarified that the total (1)O(2)-quenching activity (that is, the SOAC value) for vegetable extracts may be explained as the sum of the product {Σ k(Q)(Car-i)(S) [Car-i](i)} of the rate constant (k(Q)(Car-i)(S)) and the concentration ([Car (i)]) of carotenoids included in vegetable extracts.     

Kinetic solvent effects on phenolic antioxidants determined by spectrophotometric measurements

The effects of polar (acetonitrile and tert-butyl alcohol) and apolar (cyclohexane) solvents on the peroxyl-radical-trapping antioxidant activity of some flavonoids, catechol derivatives, hydroquinone, and monophenols have been studied. The inhibition rate constants k(inh) of the antioxidants have been determined by following the increase in absorbance at 234 nm of a dilute solution of linoleic acid at 50 degrees C containing small amounts of antioxidant and radical initiator. Despite the low concentration of linoleic acid, the peroxidation process has been confirmed to be a free radical chain reaction described by the classical kinetic laws for this process. However, in the evaluation of k(inh), a careful analysis of the peroxidation curve, absorbance versus time, must be done because the final oxidation products of phenols may absorb at 234 nm. Phenols with two ortho-hydroxyls are the most active antioxidants, with inhibition rate constants in the range of (3-15) x 10(5) M(-1) x s(-1) (in cyclohexane). Nevertheless, it has been observed that in tert-butyl alcohol (a strong hydrogen bond acceptor) the rate constants dramatically decline to values not detectable by the present kinetic method. In acetonitrile (a weaker hydrogen bond acceptor) instead, the phenols with two ortho-hydroxyls scavenge the peroxyl radicals with rate constants close to those in cyclohexane. From the kinetic solvent effect, the equilibrium constant of the first solvation step of hydroquinone with tert-butyl alcohol has been determined at 50 degrees C, K(1) = 2.5 +/- 0.5 M(-1).     

Reactivity of bovine whey proteins, peptides, and amino acids toward triplet riboflavin as studied by laser flash photolysis

The reaction between the triplet excited state of riboflavin and amino acids, peptides, and bovine whey proteins was investigated in aqueous solution in the pH range from 4 to 9 at 24 degrees C using nanosecond laser flash photolysis. Only tyrosine and tryptophan (and their peptides) were found to compete with oxygen in quenching the triplet state of riboflavin in aqueous solution, with second-order rate constants close to the diffusion limit, 1.75 x 10(9) and 1.40 x 10(9) L mol(-1) s(-1) for tyrosine and tryptophan, respectively, with beta-lactoglobulin and bovine serum albumin having comparable rate constants of 3.62 x 10(8) and 2.25 x 10(8) L mol(-1) s(-1), respectively. Tyrosine, tryptophan, and their peptides react with the photoexcited triplet state of riboflavin by electron transfer from the tyrosine and tryptophan moieties followed by a fast protonation of the resulting riboflavin anion rather than by direct H-atom abstraction, which could be monitored by time-resolved transient absorption spectroscopy as a decay of triplet riboflavin followed by a rise in riboflavin anion radical absorption. For cysteine- and thiol-containing peptides, second-order rate constants depend strongly on pH, for cysteine corresponding to pKaRSH = 8.35. H-atom abstraction seems to operate at low pH, which with rising pH gradually is replaced by electron transfer from the thiol anion. From the pH dependence of the second-order rate constant, the respective values for the H-atom abstraction (k = 1.64 x 10(6) L mol(-1) s(-1)) and for the electron transfer (k = 1.20 x 10(9) L mol(-1) s(-1)) were determined.