Riboflavin In Aqueous Solution Research Articles

Evaluation of visible light and natural photosensitizers against Staphylococcus epidermidis and Staphylococcus saprophyticus planktonic cells and biofilm

Antimicrobial photoinactivation (API) has shown some promise in potentially treating different nosocomial bacterial infections, however, its application on staphylococci, especially other than Staphylococcus aureus or methicillin-resistant S. aureus (MRSA) species is still limited. Although S. aureus is a well-known and important nosocomial pathogen, several other species of the genus, particularly coagulase-negative Staphylococcus (CNS) species such as Staphylococcus epidermidis and Staphylococcus saprophyticus, can also cause healthcare-associated infections and foodborne intoxications. CNS are often involved in resilient biofilm formation on medical devices and can cause infections in patients with compromised immune systems or those undergoing invasive procedures. In this study, the effects of chlorophyllin and riboflavin-mediated API on S. epidermidis and S. saprophyticus planktonic cells and biofilm are demonstrated for the first time. Based on the residual growth determination and metabolic reduction ability changes, higher inactivating efficiency of chlorophyllin-mediated API was determined against the planktonic cells of both tested species of bacteria and against S. saprophyticus biofilm. Some insights on whether aqueous solutions of riboflavin and chlorophyllin, when illuminated with optimal exciting wavelength (440 nm and 402 nm, respectively) generate O2−•, are also provided in this work.

Photodegradation of Riboflavin under Alkaline Conditions: What Can Gas-Phase Photolysis Tell Us about What Happens in Solution?

The application of electrospray ionisation mass spectrometry (ESI-MS) as a direct method for detecting reactive intermediates is a technique of developing importance in the routine monitoring of solution-phase reaction pathways. Here, we utilise a novel on-line photolysis ESI-MS approach to detect the photoproducts of riboflavin in aqueous solution under mildly alkaline conditions. Riboflavin is a constituent of many food products, so its breakdown processes are of wide interest. Our on-line photolysis setup allows for solution-phase photolysis to occur within a syringe using UVA LEDs, immediately prior to being introduced into the mass spectrometer via ESI. Gas-phase photofragmentation studies via laser-interfaced mass spectrometry of deprotonated riboflavin, [RF − H]−, the dominant solution-phase species under the conditions of our study, are presented alongside the solution-phase photolysis. The results obtained illustrate the extent to which gas-phase photolysis methods can inform our understanding of the corresponding solution-phase photochemistry. We determine that the solution-phase photofragmentation observed for [RF − H]− closely mirrors the gas-phase photochemistry, with the dominant m/z 241 condensed-phase photoproduct also being observed in gas-phase photodissociation. Further gas-phase photoproducts are observed at m/z 255, 212, and 145. The value of exploring both the gas- and solution-phase photochemistry to characterise photochemical reactions is discussed.

Fluorescence resonance energy transfer from pyrene nanoparticles to riboflavin: Spectroscopic insights and analytical application

The aqueous suspension of fluorescent pyrene nanoparticles (PyNPs) were prepared by a reprecipitation method in the presence of sodium dodecyl sulphate (SDS) as a stabilizer. The PyNPs shows bathochromically shifted aggregation induced enhanced emission (AIEE) in the spectral region 400 nm to 600 nm peaking at 466 nm where Riboflavin (RF) absorbs strongly. The systematic FRET experiments performed by measuring fluorescence quenching of PyNPs with successive addition of RF analyte exploited the use of PyNPs as nano probe for detection of RF in aqueous solution with lower limit of detection 10.163 × 10 -5 mol.L -1 . The fluorescence of PyNPs was quenched by RF and quenching is in accordance with the Stern-Volmer relation. The distance r between the donor (PyNPs) and acceptor (RF) molecules was obtained according to the fluorescence resonance energy transfer. The evaluation of photo kinetic and thermodynamic parameters such as quenching rate constant (k q ), enthalpy change (ΔH), Gibbs free energy change (ΔG) and entropy change (ΔS) was calculated by quenching results obtained at different constant temperatures. The proposed FRET method based on fluorescence quenching of PyNPs was used further to develop an analytical relation for estimation of RF from pharmaceutical samples available commercially in the market.

Ionic strength effects on the photodegradation reactions of riboflavin in aqueous solution

A study of the effect of ionic strength on the photodegradation reactions (photoreduction and photoaddition) of riboflavin (RF) in phosphate buffer (pH7.0) has been carried out using a specific multicomponent spectrometric method. It has been found that the rates of photodegradation reactions of RF are dependent upon the ionic strength of the solutions at different buffer concentrations. The apparent first-order rate constants (kobs) for the photodegradation of riboflavin at ionic strengths of 0.1–0.5 (0.5M phosphate) lie in the range of 7.35–30.32×10−3min−1. Under these conditions, the rate constants for the formation of the major products, lumichrome (LC) by photoreduction pathway, and cyclodehydroriboflavin (CDRF) by photoaddition pathway, are in the range of 3.80–16.03 and 1.70–6.07×10−3min−1, respectively. A linear relationship has been observed between log kobs and √μ/1+√μ. A similar plot of log k/ko against √μ yields a straight line with a value of ~+1 for ZAZB showing the involvement of a charged species in the rate determining step. NaCl appears to promote the photodegradation reactions of RF probably by an excited state interaction. The implications of ionic strength on RF photodegradation by different pathways and flavin–protein interactions have been discussed.

Effect of Nicotinamide on the Photolysis of Riboflavin in Aqueous Solution.

The photolysis of riboflavin (RF) in aqueous solution in the presence of nicotinamide (NA) by visible light has been studied in the pH range 1.0–12.0 and the various photoproducts have been identified as known compounds. RF has been determined in degraded solutions by a specific multicomponent spectrometric method in the presence of its photoproducts and NA. The second-order rate constants (k2) for the bimolecular interaction of RF and NA range from 0.54 (pH 1.0) to 9.66 M–1 min–1 (pH 12.0). The log k2–pH profile for the photolysis reaction follows a sigmoid curve showing a gradual increase in the rate of pH due to a change in the ionization behavior of the molecule. The lower rate in the acid region is probably due to protonation of the molecule since the cationic form of RF is less susceptible to photolysis than the neutral form. Similarly, a slowing of the rate in the alkaline region is due to anion formation of the molecule. NA is involved as an electron acceptor during the sequence of reactions and thus enhances the rate of photolysis of RF. Absorption and fluorescence measurements did not provide evidence for the complex formation between the two compounds under the present conditions.

Ratio-metric sensor to detect riboflavin via fluorescence resonance energy transfer with ultrahigh sensitivity

In this paper, a novel fluorescence resonance energy transfer (FRET) ration-metric fluorescent probe based on heteroatom N, S doped carbon dots (N, S-CDs) was developed to determine riboflavin in aqueous solutions. The ratio of two emission intensities at different wavelengths is applied to determine the concentration of riboflavin (RF). This method is more effective in reducing the background interference and fluctuation of diverse conditions. Therefore, this probe obtains high sensitivity with a low limit of detection (LOD) of 1.9nM (0.7ng/ml) which is in the highest level of all riboflavin detection approaches and higher than single wavelength intensity detection (1.9μM). In addition, this sensor has a high selectivity of detecting riboflavin in deionized water (pH=7) with other biochemical like amino acids. Moreover, riboflavin in aqueous solution is very sensitive to sunlight and can be degraded to lumiflavin, which is toxic. Because the N, S doped carbon dots cannot serve as an energy donor for N, S doped carbon dots and lumiflavin system, this system makes it easy to determine whether the riboflavin is degraded or not, which is first to be reported. This platform may provide possibilities to build a new and facile fluorescence resonance energy transfer based sensor to detect analytes and metamorphous analytes in aqueous solution.

Reactions of flavosemiquinone radicals in the presence of metal ions

The rate constants of disproportionation of flavosemiquinone radicals were obtained by pulsed spectroscopy. The yield of the flavosemiquinone radical increased when Mohr’s salt was introduced in the aqueous solutions of riboflavin. The spectral kinetic characteristics of complexes of flavosemiquinone radical anions with Zn2+ and Cd2+ ions were determined.

Thermal Destruction of Riboflavin in Different Aggregate States

The influence of temperature on the spectroscopic properties of powders and aqueous solutions of riboflavin was studied using NMR, linear dichroism, IR and UV absorption spectroscopy, and luminescence. It was found that exposure of riboflavin powders over a wide temperature range (290–423 K) did not change significantly its properties and spectral characteristics. The absorbance and fluorescence of aqueous solutions of riboflavin decreased under the same conditions due to riboflavin thermal destruction. The fraction of destroyed molecules in aqueous solutions of riboflavin was in the range 4–28% and depended on the drug concentration and thermal conditions.

Effect of acetate and carbonate buffers on the photolysis of riboflavin in aqueous solution: a kinetic study.

The photolysis of riboflavin (RF) in the presence of acetate buffer (pH 3.8-5.6) and carbonate buffer (pH 9.2-10.8) has been studied using a multicomponent spectrophotometric method for the simultaneous assay of RF and its photoproducts. Acetate and carbonate buffers have been found to catalyze the photolysis reaction of RF. The apparent first-order rate constants for the acetate-catalyzed reaction range from 0.20 to 2.86 × 10(-4) s(-1) and for the carbonate-catalyzed reaction from 3.33 to 15.89 × 10(-4) s(-1). The second-order rate constants for the interaction of RF with the acetate and the carbonate ions range from 2.04 to 4.33 × 10(-4) M(-1) s(-1) and from 3.71 to 11.80 × 10(-4) M(-1) s(-1), respectively. The k-pH profile for the acetate-catalyzed reaction is bell shaped and for the carbonate-catalyzed reaction a steep curve. Both HCO3(-) and CO3(2-) ions are involved in the catalysis of the photolysis reaction in alkaline solution. The rate constants for the HCO3(-) and CO3(2-) ions catalyzed reactions are 0.72 and 1.38 × 10(-3) M(-1) s(-1), respectively, indicating a major role of CO3(2-) ions in the catalysis reaction. The loss of RF fluorescence in acetate buffer suggests an interaction between RF and acetate ions to promote the photolysis reaction. The optimum stability of RF solutions is observed in the pH range 5-6, which is suitable for pharmaceutical preparations.

Stabilizing effect of citrate buffer on the photolysis of riboflavin in aqueous solution

In the present investigation the photolysis of riboflavin (RF) in the presence of citrate species at pH 4.0–7.0 has been studied. A specific multicomponent spectrophotometric method has been used to assay RF in the presence of photoproducts during the reactions. The overall first-order rate constants (kobs) for the photolysis of RF range from 0.42 to 1.08×10–2min−1 in the region. The values of kobs have been found to decrease with an increase in citrate concentration indicating an inhibitory effect of these species on the rate of reaction. The second-order rate constants for the interaction of RF with total citrate species causing inhibition range from 1.79 to 5.65×10–3M−1min−1 at pH 4.0–7.0. The logk–pH profiles for the reactions at 0.2–1.0M citrate concentration show a gradual decrease in kobs and the value at 1.0M is more than half compared to that of k0, i.e., in the absence of buffer, at pH 5.0. Divalent citrate ions cause a decrease in RF fluorescence due to the quenching of the excited singlet state resulting in a decrease in the rate of reaction and consequently leading to the stabilization of RF solutions. The greater quenching of fluorescence at pH 4.0 compared to that of 7.0 is in accordance with the greater concentration of divalent citrate ions (99.6%) at that pH. The trivalent citrate ions exert a greater inhibitory effect on the rate of RF photolysis compared to that of the divalent citrate ions probably as a result of excited triplet state quenching. The values of second-order rate constants for the interaction of divalent and trivalent citrate ions are 0.44×10–2 and 1.06×10–3M–1min–1, respectively, indicating that the trivalent ions exert a greater stabilizing effect, compared to the divalent ions, on RF solutions.

Parallel stacking of caffeine with riboflavin in aqueous solutions: The potential mechanism for hydrotropic solubilization of riboflavin

Hydrotropy is a phenomenon where the presence of a large quantity of one solute enhances the solubility of another solute. The mechanism of this phenomenon remains elusive and a topic of debate. This study employed molecular dynamics simulation to investigate solute interactions of a model system consisting of a hydrotropic agent, caffeine (CAF), a poorly water-soluble solute, riboflavin (RBF), and water. The study demonstrates that CAF and RBF undergo molecular parallel stacking in the aqueous solution, a result correlating closely to the self-stacking of CAF under the same conditions. The correlations are found both structurally and dynamically, suggesting that the self-stacking of CAF is the primary effect, and incorporation of RBF is the secondary effect. The solute stacking gives rise to the partitioning of solutes and water, which helps restoring the normal water structure and drives down the system energy. The interactions between the solutes are found insignificant to the solute clustering. The dynamic data confirm that the solute stacks are dynamically active. These results suggest that hydrotropic effect of CAF may be attributed to solute parallel stacking.

Effect of divalent anions on photodegradation kinetics and pathways of riboflavin in aqueous solution

The present investigation is based on a study of the effect of buffer and non-buffer divalent anions (phosphate, sulphate, tartrate, succinate, malonate) on the kinetics, product distribution and photodegradation pathways of riboflavin (RF) at pH 6.0–8.0. RF solutions (5 × 10 −5 M) were photodegraded in the presence of divalent anions (0.2–1.0 M) using a visible light source and the photoproducts, cyclodehydroriboflavin (CDRF), formylmethylflavin (FMF), lumichrome (LC) and lumiflavin (LF) were assayed by a specific multicomponent spectrophotometric method. RF degradation in the presence of divalent anions follows parallel first-order kinetics to give CDRF and LC as the final products through photoaddition and photoreduction reactions, respectively. The divalent anion-catalysed CDRF formation is affected in the order: phosphate > sulphate > tartrate > succinate > malonate, showing maximum activity of the anions around pH 7. The divalent anions cause deviation of the photoreduction pathway in favour of the photoaddition pathway to form CDRF. The first- and second-order rate constants for the reactions involved in the photodegradation of RF have been determined and the rate–pH profiles and pathway relationships discussed. The catalytic activity of the divalent anions appears to be a function of the relative strength and chemical reactivity of the RF–divalent anion complex acting as a mediator in the photoaddition reaction.

Determination of water-soluble vitamin B and vitamin C in combined feed, premixes, and biologically active supplements by reversed-phase HPLC

A procedure was developed for determining water-soluble vitamins B and vitamin C in premixes, biologically active supplements, and combined feed by reversed-phase HPLC using sodium heptanesulfonate as an ion-pair reagent. A procedure was proposed for purifying a combined food extract by solid-phase extraction on a column packed with a Sep-Pak C18 adsorbent. The stability of ascorbic acid and riboflavin in aqueous solutions with different pH values was studied.

Effect of Caffeine Complexation on the Photolysis of Riboflavin in Aqueous Solution: A Kinetic Study

The effect of caffeine complexation with riboflavin on the kinetics of riboflavin photolysis in the pH range 2.0-10.5 has been studied. The photolysis of riboflavin solutions (5x10(-5) M) was carried out in the presence of caffeine (0.5-2.5x10(-4) M) using a visible radiation source. A specific multicomponent spectrophotometric method was used for the determination of riboflavin and photoproducts in photolysed solutions. The apparent first-order rate constants (k) for the photolysis reactions range from 2.71x10(-4) to 4.26x10(-2) min(-1). The values of the rate constants decrease with increasing concentrations of caffeine indicating its inhibitory effect on the reactions. The second-order rate constants (k') for the caffeine inhibited reactions lie in the range of 0.13 to 5.10x10(-3) M(-1) min(-1). The log k-pH profiles for the photolysis reactions at various caffeine concentrations involve multiple steps indicating a gradual increase in the rate up to pH 10. The lower rates at pH 2.0 and 10.5 are due to the ionization of riboflavin as evident from fluorescence measurements. The k'-pH profile for the interaction of riboflavin with caffeine represents a bell-shaped curve in the pH range 3-6 followed by a sigmoid curve in the pH range 7-10. The inhibition of photolysis of riboflavin in the presence of caffeine is a result of the monomeric interaction and complex formation of caffeine with riboflavin. The photochemical interaction of riboflavin with caffeine suggests that a pH around 6 is most appropriate for the stabilization of the vitamin. At this pH the complex shows the highest stability constant.

Effect of borate buffer on the photolysis of riboflavin in aqueous solution

The photolysis of riboflavin (RF) in the presence of borate buffer (0.1–0.5 M) at pH 8.0–10.5 has been studied using a specific multicomponent spectrophotometric method for the determination of RF and photoproducts, formylmethylflavin (FMF), lumichrome (LC) and lumiflavin (LF). The overall first-order rate constants for the photolysis of RF (1.55–4.36 × 10 −2 min −1) and the rate constants for the formation of FMF (1.16–3.52 × 10 −2 min −1) and LC (0.24–0.84 × 10 −2 min −1) have been determined. The values of all these rate constants decrease with an increase in buffer concentration suggesting the inhibition of photolysis reaction by borate species. The kinetic data support the formation of a RF-borate complex involving the ribityl side chain to cause the inhibition of photolysis. The second-order rate constants for the borate inhibited reaction range from 1.17–3.94 × 10 −2 M −1 min −1. The log k-pH profiles for the reaction at various buffer concentrations indicate a gradual increase in rate, with pH, up to 10 followed by a decrease in rate at pH 10.5 probably due to ionization of RF and quenching of fluorescence by borate species. A graph of second-order rate constants against pH is a sigmoid curve showing that the rate of photolysis increases with an increase in pH. The results suggest the involvement of excited singlet state, in addition to excited triplet state, in the formation of LC.

Effect of light intensity and wavelengths on photodegradation reactions of riboflavin in aqueous solution

A study of the effect of light intensity and wavelengths on photodegradation reactions of riboflavin (RF) solutions in the presence of phosphate buffer using three UV and visible radiation sources has been made. The rates and magnitude of the two major photodegradation reactions of riboflavin in phosphate buffer (i.e., photoaddition and photoreduction) depend on light intensity as well as the wavelengths of irradiation. Photoaddition is facilitated by UV radiation and yields cyclodehydroriboflavin (CDRF) whereas photoreduction results from normal photolysis yielding lumichrome (LC) and lumiflavin (LF). The ratios of the photoproducts of the two reactions at 2.0 M phosphate concentration, CDRF/RF (0.09–0.22) and CDRF/LC (0.54–1.75), vary with the radiation source and are higher with UV radiation than those of the visible radiation. On the contrary, the ratios of LF/LC (0.15–0.25) increase on changing the radiation source from UV to visible. The rate is much faster with UV radiation causing 25% degradation of a 10 −5 M riboflavin solution in 7.5 min compared to that of visible radiations in 150–330 min.

Deactivation of riboflavin triplet-excited state by phenolic antioxidants: mechanism behind protective effects in photooxidation of milk-based beverages

The water-soluble plant polyphenols rutin, (+)-catechin, and (−)-epigallocatechin gallate were each found to be efficient quenchers for the triplet-excited state of riboflavin in aqueous solution using nano-second laser flash photolysis. The deactivation followed second-order kinetics with rates close to the diffusion control: (−)-epigallocatechin gallate (1.70±0.2×109 l mol−1 s−1), (+)-catechin (1.44±0.04×109 l mol−1 s−1), and rutin (9.7±0.2×108 l mol−1 s−1) at 25 °C and pH = 6.8. No synergetic effects for mixtures of the phenolic compounds were found. Hydrogen atom transfer is calculated to be more exergonic than electron transfer, suggesting the hydrogen atom transfer mechanism to be operative. The efficient deactivation by the plant polyphenols makes this type of compounds of importance as protectors against light-induced oxidation in flavored milk-based beverages.

Deactivation of Triplet-Excited Riboflavin by Purine Derivatives: Important Role of Uric Acid in Light-Induced Oxidation of Milk Sensitized by Riboflavin

The reactivity of purine derivatives (uric acid, xanthine, hypoxanthine, and purine) toward triplet-excited riboflavin in aqueous solution at pH 6.4 is described on the basis of kinetic (laser flash photolysis), electrochemical (square-wave voltammetry), and theoretical data (density functional theory, DFT). Direct deactivation of triplet-excited riboflavin in aqueous solution, pH 6.4 at 24 degrees C, in the presence of uric acid, xanthine, and hypoxanthine strongly suggests a direct electron transfer from the purine to the triplet-excited riboflavin with k = 2.9 x 10(9) M(-1) s(-1) (DeltaH(++) = 14.7 kJ mol(-1), DeltaS(++) = -15.6 J mol(-1) K(-1)), 1.2 x 10(9) M(-1) s(-1) (DeltaH(++) = 34.3 kJ mol(-1), DeltaS(++) = +45.3 J mol(-1) K(-1)), and 1.7 x10(8) M(-1) s(-1) (DeltaH(++) = 122 kJ mol(-1), DeltaS(++) = +319 J mol(-1) K(-1)), respectively. From the respective one-electron oxidation potentials collected in aqueous solution at pH 6.4 for uric acid (E = +0.686 vs normal hydrogen electrode, NHE), xanthine (E = +1.106 vs NHE), and hypoxanthine (E = +1.654 vs NHE), the overall free energy changes for electron transfer from the quencher to the triplet-excited riboflavin are as follows: uric acid (DeltaG(o) = -114 kJ mol(-1)), xanthine (DeltaG(o) = -73.5 kJ mol(-1)), hypoxanthine (DeltaG(o) = -20.6 kJ mol(-1)), and purine (DeltaG(o) > 0). The inertness observed for purine toward triplet-excited riboflavin corroborates with its electrochemical inactivity in the potential range from 0 up to 2 V vs NHE. These data are in agreement with the DFT results, which show that the energy of the purine highest occupied molecular orbital (HOMO) (-0.2685 arbitrary unit) is lower than the energy of the semioccupied molecular orbital (SOMO) (-0.2557 a.u.) of triplet-excited riboflavin, indicating an endergonic process for the electron-transfer process. The rate-determining step for deactivation by purine derivatives can be assigned to an electron transfer from the purine derivative to the SOMO orbital of the triplet-excited riboflavin. The results show that uric acid may compete with oxygen and other antioxidants to deactivate triplet-excited riboflavin in milk serum and other biological fluids leading to a free radical process.

Effect of phosphate buffer on photodegradation reactions of riboflavin in aqueous solution

The effect of phosphate buffer on aerobic photodegradation reactions of riboflavin (RF) at pH 7.0 has been studied. The photoproducts of the two major reactions, viz., intramolecular photoreduction and intramolecular photoaddition, have been determined by a specific multicomponent spectrophotometric method. The overall photodegradation of riboflavin in the presence of phosphate buffer involves the participation of both H 2 PO 4 - and HPO 4 2 - species. The second-order rate constants for the H 2 PO 4 - -catalysed photodegradation of riboflavin (normal photolysis) to lumichrome (LC) and HPO 4 2 - -catalysed photodegradation of riboflavin (photoaddition) to cyclodehydroriboflavin (CDRF) are 0.93 × 10 −4 and 4.0 × 10 −4 M −1 s −1, respectively. The addition of 0.25–2.00 M phosphate to RF solutions at pH 7.0 gives rise to RF – HPO 4 2 - complex and hence the quenching of 4–36% fluorescence, respectively. This results in the suppression of normal photolysis leading to the formation of LC in favour of photoaddition to yield CDRF. The present study shows the involvement of H 2 PO 4 - anions in the base-catalysed degradation of riboflavin by normal photolysis vis-a-vis the involvement of HPO 4 2 - anions in photoaddition reactions of riboflavin suggested earlier [M. Schuman Jorns, G. Schollnhammer, P. Hemmerich, Intramolecular addition of the riboflavin side chain. Anion-catalysed neutral photochemistry, Eur. J. Biochem. 57 (1975) 35–48].

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.