Presence Of Riboflavin Research Articles

Oxidative mechanisms for the biotransformation of 1-methyl-1,6-dihydropyridine-2-carbaldoxime to pralidoxime chloride

AimsDue to pralidoxime chloride's (2-PAM) positive charge, it's penetration through the blood brain barrier (BBB) and reactivation of organophosphate (OP) inhibited central nervous system (CNS) acetylcholinesterase (AChE) is poor. The results of CNS inhibited AChE are seizures. Pro-2-PAM (1-methyl-1,6-dihydropyridine-2-carbaldoxime), a pro-drug of 2-PAM, due to higher hydrophobicity, penetrates the BBB better but must be oxidized to 2-PAM, the active form of the oxime to reactivate CNS AChE in order to abrogate seizures. In this study, we characterize the in vivo mechanism of pro-2-PAM oxidation. Main methodsA high pressure liquid chromatography (HPLC) assay was developed to quantify the conversion of pro-2-PAM to 2-PAM. NADPH oxidase activity was measured by a photo-luminescence assay using lucigenin substrate. Upon analysis, the rate of NADPH induced oxidation suggested that an alternate mechanism may be involved. Therefore, various enzyme co-factors of oxidation–reduction enzyme systems were evaluated, including nicotinamide adenine dinucleotide (NAD), nicotinamide adenine dinucleotide phosphate (NADP), flavin adenine dinucleotide (FAD), riboflavin 5′-phosphate (FMN), and riboflavin. Next, a spectrophotometric assay was developed to measure the conversion of pro-2-PAM to 2-PAM in the presence of riboflavin. Key findingsIn guinea pig brain homogenate, diphenyleneiodonium (DPI), a specific NADPH oxidase inhibitor, reduced pro-2-PAM to 2-PAM conversion to less than 25%. In contrast, riboflavin, FAD, and FMN rapidly oxidized all pro-2-PAM to 2-PAM in an in vitro assay. Riboflavin oxidized pro-2-PAM reactivated diisopropylfluorophosphate (DFP) inhibited AChE. SignificanceThe present study shows that pro-2-PAM was rapidly oxidized by riboflavin to 2-PAM, which reactivated organophosphate (OP)-inhibited AChE.

Solar Disinfection of Water for Inactivation of Enteric Viruses and its Enhancement by Riboflavin

Solar disinfection (SODIS) has been described as a cheap and effective method of treating contaminated water to inactivate pathogenic microorganisms. In this study, SODIS was assessed for its efficacy in inactivating three enteric viruses (coxsackievirus B3, coxsackievirus B5 and poliovirus), either on its own or in the presence of riboflavin as a disinfection enhancer. On its own, SODIS produced a reduction of virus infectivity of 4–6 log10 in 6 h. In the presence of riboflavin, inactivation was more rapid in all viruses studied, and with coxsackievirus B5 and poliovirus an extra 1–2 log10 increase in reduction of infectivity was observed after 6 h exposure. This study provides a practical example of low technology methods which could be utilised to provide safe drinking water in various circumstances.

Photosensitized Amino Acid Degradation in the Presence of Riboflavin and Its Derivatives

The addition of photosensitizers to water can accelerate disinfection in sunlight-based systems by enhancing oxidation of target compounds through direct reaction with the excited sensitizer or through production of another oxidant, such as singlet oxygen (¹O₂). The kinetics of the oxidation of selected amino acids in the presence of the sensitizer riboflavin (Vitamin B2), its primary photoproduct lumichrome, and its derivative riboflavin tetraacetate (2',3',4',5'-tetraacetylriboflavin; RTA) were quantified and the mechanisms of reaction were determined during exposure to 365 ± 9 nm light. ¹O₂-mediated reactions contributed to the rapid photodegradation of the four amino acids, but its contribution was sensitizer-dependent and varied from 5.4-10.2% for tyrosine, 7.1-12.4% for tryptophan, 18.7-69.0% for methionine, and 64.7-100.2% for histidine. Riboflavin was subject to rapid photodegradation (t½ < 8 min), while the half-lives of lumichrome and RTA were 100 and 30 times longer, respectively. Lumichrome and RTA also were more efficient ¹O₂ sensitizers (quantum yield (Φ) = 0.63 and 0.66) compared to riboflavin (Φ = 0.48). Of the three flavin-based compounds, RTA shows the most promise as a sensitizer in sunlight-based disinfection systems because it absorbs both visible and UV light, is an efficient ¹O₂ sensitizer, is a strong oxidant in its triplet state, and exhibits greater photostability.

Azo and anthraquinone dye decolorization in relation to its molecular structure using soluble Trichosanthes dioica peroxidase supplemented with redox mediator

In this study we investigated the decolorizing ability of Trichosanthes dioica -derived soluble peroxidase on structurally complex azo and anthraquinone dyes in the presence of redox mediators. Our results show that riboflavin acted as a better redox mediator than antraquinone-2, 6-disulfonate (AQDS). Riboflavin served as an efficient electron transferor than AQDS in the reduction of azo dyes, by contrast with anthraquinone dyes. Although the extent of decolorization (expressed as percent dye decolorization) varied from one dye to the other, maximum decolorization was achieved for the case when suspensions containing 0.45 EU (Enzyme Units)/ml and 0.2 mM riboflavin at pH 5.0 were incubated for 2 h at 40 °C. Redox mediator supplementation remarkably improved the decolorizing potential of T. dioica peroxidase. Azo dyes exhibited higher decolorization as compared to anthraquinone dyes in the presence of riboflavin. On the contrary, percent decolorization in the absence of redox mediators was inadequate and slow. Redox mediators studied herein facilitated decolorization of stable anthraquinone dyes. Therefore, the effect of redox mediators on T. dioica peroxidase-dye decolorization is affected by stability, thus the molecular structure. ► Riboflavin enhances decolorization of azo and anthraquinone dyes more than AQDS. ► Anthraquinone dyes decolorized at higher concentrations of redox mediators. ► Cleavage of chromophore for azo dyes is favorable as compared to anthraquinone. ► Redox mediated dye decolorization is related to the dye molecular structure.

Effects of Ultraviolet-A and Riboflavin on the Interaction of Collagen and Proteoglycans during Corneal Cross-linking

Corneal cross-linking using riboflavin and ultraviolet-A (RFUVA) is a clinical treatment targeting the stroma in progressive keratoconus. The stroma contains keratocan, lumican, mimecan, and decorin, core proteins of major proteoglycans (PGs) that bind collagen fibrils, playing important roles in stromal transparency. Here, a model reaction system using purified, non-glycosylated PG core proteins in solution in vitro has been compared with reactions inside an intact cornea, ex vivo, revealing effects of RFUVA on interactions between PGs and collagen cross-linking. Irradiation with UVA and riboflavin cross-links collagen α and β chains into larger polymers. In addition, RFUVA cross-links PG core proteins, forming higher molecular weight polymers. When collagen type I is mixed with individual purified, non-glycosylated PG core proteins in solution in vitro and subjected to RFUVA, both keratocan and lumican strongly inhibit collagen cross-linking. However, mimecan and decorin do not inhibit but instead form cross-links with collagen, forming new high molecular weight polymers. In contrast, corneal glycosaminoglycans, keratan sulfate and chondroitin sulfate, in isolation from their core proteins, are not cross-linked by RFUVA and do not form cross-links with collagen. Significantly, when RFUVA is conducted on intact corneas ex vivo, both keratocan and lumican, in their natively glycosylated form, do form cross-links with collagen. Thus, RFUVA causes cross-linking of collagen molecules among themselves and PG core proteins among themselves, together with limited linkages between collagen and keratocan, lumican, mimecan, and decorin. RFUVA as a diagnostic tool reveals that keratocan and lumican core proteins interact with collagen very differently than do mimecan and decorin.

Photosensitized degradation in water of the phenolic pesticides bromoxynil and dichlorophen in the presence of riboflavin, as a model of their natural photodecomposition in the environment

Within the context of environmentally friendly methods for the elimination of surface-water pollutants, the photodegradation of the phenolic pesticides bromoxynil (BXN) and dichlorophen (DCP) under simulated natural conditions has been studied. The work was done in the presence of the visible-light absorber photosensitizer riboflavin (Rf), usually present in trace quantities in natural waters. Under aerobic conditions, an efficient photooxidation of both pesticides was observed. The relatively intricate photochemical mechanism involves pesticide and oxygen consumption and, to a lesser extent, Rf degradation. The kinetic and mechanistic study supports that both H2O2 and singlet molecular oxygen, O2(1Δg), are involved in the process. Kinetic data for the O2(1Δg)-mediated oxidation indicate that BXN and DCP are photodegraded with this species faster than the parent compound phenol, very frequently employed as a model for aquatic contaminants, likely due to their lower pKa values. This observation allows the design of phenolic pesticides with different photodegradation rates under environmental conditions.

Antimicrobial Activity of Simulated Solar Disinfection against Bacterial, Fungal, and Protozoan Pathogens and Its Enhancement by Riboflavin

Riboflavin significantly enhanced the efficacy of simulated solar disinfection (SODIS) at 150 watts per square meter (W m(-2)) against a variety of microorganisms, including Escherichia coli, Fusarium solani, Candida albicans, and Acanthamoeba polyphaga trophozoites (>3 to 4 log(10) after 2 to 6 h; P < 0.001). With A. polyphaga cysts, the kill (3.5 log(10) after 6 h) was obtained only in the presence of riboflavin and 250 W m(-2) irradiance.

IMMUNOHEMATOLOGY: Understanding loss of donor white blood cell immunogenicity after pathogen reduction: mechanisms of action in ultraviolet illumination and riboflavin treatment

BACKGROUND: Donor white blood cells (WBCs) present in transfusion products can lead to immune sequelae such as production of HLA antibodies or graft‐versus‐host disease in susceptible transfusion recipients. Eliminating the immunogenicity of blood products may prove to be of clinical benefit, particularly in patients requiring multiple transfusions in whom allosensitization is common. This study examines a method of pathogen reduction based on ultraviolet light illumination in the presence of riboflavin. In addition to pathogens, WBCs treated with this system are affected and fail to stimulate proliferation of allogeneic peripheral blood mononuclear cells (PBMNCs) in vitro.STUDY DESIGN AND METHODS: This study sought to determine the mechanisms regulating this loss of immunogenicity. Treated cells were examined for surface expression of a number of molecules involved in activation and adhesion, viability, cell‐cell conjugation, and ability to stimulate immune responses in allogeneic PBMNCs.RESULTS: Compared with untreated controls, ultraviolet (UV)‐irradiated antigen‐presenting cells showed slightly reduced surface expression of HLA Class II and costimulatory molecules and had more significant reductions in surface expression of a number of adhesion molecules. Furthermore, treated cells had a severe defect in cell‐cell conjugation. The observed loss of immunogenicity was nearly complete, with UV‐irradiated cells stimulating barely measurable interferon‐γ production and no detectable STAT‐3, STAT‐5, or CD3‐ε phosphorylation in allospecific primed T cells.CONCLUSION: These results suggest that defective cell‐cell adhesion prevents UV‐irradiated cells from inducing T‐cell activation.

Degradation of alkylphenols in the presence of riboflavin under near-ultraviolet light

Degradation of alkylphenols in the presence of riboflavin under near-ultraviolet light

Visible-light promoted photoprocesses on aqueous gallic acid in the presence of riboflavin. Kinetics and mechanism

Within the frame of possible precursory photoreactions in the generation of humic substances, the visible-light promoted interaction between riboflavin (Rf), a native photosensitizer in aqueous systems, and gallic acid (GA), a polyphenol naturally formed after lignin degradation, was investigated. A systematic kinetic and mechanistic study was conducted under aerobic conditions in aqueous media, through visible-light continuous photolysis, polarographic detection of oxygen uptake, stationary and time resolved fluorescence spectroscopy, time resolved near-IR phosphorescence detection and laser flash photolysis techniques. GA is degraded relatively fast in pH 7 aqueous solutions, where singlet molecular oxygen (O 2( 1Δ g)), superoxide radical anion (O 2 −) and hydrogen peroxide (H 2O 2) – all three species photogenerated from triplet excited Rf – participate in the photoprocess. The general conclusion is that in natural waters GA can undergo spontaneous phototodegradation under environmental conditions. Radical species generated in the presence of Rf can participate in condensation or polymerization reactions promoting the natural synthesis of humic products.

5-Methyltetrahydrofolate can be photodegraded by endogenous photosensitizers

Folate deficiency is linked to serious birth defects, pregnancy complications, male infertility, cardiovascular diseases, and even the evolution of human skin color. Conflicting data exist on whether exposure to sun or artificial UV sources may deplete the levels of blood folate in humans. Blood contains several photosensitizers and proteins, as well as antioxidants, which when exposed to UV radiation and visible light may be involved in the degradation of folate. In this study the photodegradation of 5-methyltetrahydrofolate (5MTHF) in aqueous and deuterious solutions exposed to UVB, UVA, or visible light in the absence or presence of riboflavin, uroporphyrin, and conjugated bilirubin was investigated by absorption spectroscopy. 5MTHF is stable under exposure to visible light and UVA radiation, whereas it is slowly photooxidized under UVB exposure. However, it is rapidly oxidized by UVA or visible radiation in the presence of riboflavin or uroporphyrin, but not in the presence of conjugated bilirubin, which acts in a protective manner. Reactive oxygen species produced in type I and/or type II reactions were involved. This study suggests that 5MTHF in blood can be photodegraded in the presence of the flavins and porphyrins, but protected by bilirubins. This may have health and evolutionary implications.

Dermal Carbon Monoxide Excretion in Neonatal Rats During Light Exposure

Total body, head, and trunk carbon monoxide (CO) excretion rates were measured separately by gas chromatography in 1- to 7-d-old Wistar rat pups exposed to the dark and to mixed blue (one Special Blue-F20T12/BB) and white (two Cool White-F20T12/CW) fluorescent light or blue light emitting diode (LED) sources. During 48-min cycled exposures to the dark and to either light source, total body CO excretion rapidly increased 1.9- and 1.4-fold, respectively, over dark control levels. When CO excretion rates from the head and trunk were measured separately during exposure to either light source, CO excretion from the head did not change significantly; however, a large mean 4.4-fold increase in CO excretion from the trunk was observed. When light intensity delivered by the blue LED source was varied, we found that trunk CO excretion increased with increasing light intensities. In the presence of riboflavin (10 micromol/kg), total body CO excretion increased 2.8- and 2.1-fold during exposure to the mixed fluorescent light and blue LED sources, respectively. We conclude that light-induced elevations in total body CO excretion may be caused by transdermally excreted CO, which is most likely produced through endogenous photosensitizer-mediated photooxidation of dermal biomolecules.

Effects of Trolox and ascorbic acid on the riboflavin photosensitised oxidation of aromatic amino acids

Abstract The effects of 0, 100, 250, 500, 750 or 1000 ppm Trolox and ascorbic acid on the singlet oxygen oxidation of aromatic amino acids viz. phenylalanine, tryptophan, tyrosine in the presence of 25 ppm riboflavin were determined by measuring depleted headspace oxygen by gas chromatography and aromatic amino acid content by high performance liquid chromatography. The coefficients of variations (CVs) for headspace oxygen analysis and HPLC analysis of aromatic amino acids were 1.4% and 4.7%, respectively. Samples were stored under light (1000 lux) at 30 °C for 10 h. Both Trolox and ascorbic acid acted as antioxidants. As the concentration of Trolox and ascorbic increased from 0 to 1000 ppm, the head space oxygen depletion increased. This was due to the oxidation of Trolox and ascorbic acid along with amino acids in the presence of riboflavin. High performance liquid chromatography analysis of the samples clearly indicated that both Trolox and ascorbic acid minimised the degradation of phenylalanine, tryptophan, tyrosine significantly ( p

The Suggested Physiologic Aryl Hydrocarbon Receptor Activator and Cytochrome P4501 Substrate 6-Formylindolo[3,2-b]carbazole Is Present in Humans

Dioxins and other polycyclic aromatic compounds formed during the combustion of waste and fossil fuels represent a risk to human health, as well as to the well being of our environment. Compounds of this nature exert carcinogenic and endocrine-disrupting effects in experimental animals by binding to the orphan aryl hydrocarbon receptor (AhR). Understanding the mechanism of action of these pollutants, as well as the physiological role(s) of the AhR, requires identification of the endogenous ligand(s) of this receptor. We reported earlier that activation of AhR by ultraviolet radiation is mediated by the chromophoric amino acid tryptophan (Trp), and we suggested that a new class of compounds derived from Trp, in particular 6-formylindolo[3,2-b]carbazole (FICZ), acts as natural high affinity ligands for this receptor. Here we describe seven new FICZ-derived indolo[3,2-b]carbazole-6-carboxylic acid metabolites and two sulfoconjugates, and we demonstrate the following. (i) FICZ is formed efficiently by photolysis of Trp upon exposure to visible light. (ii) FICZ is an exceptionally good substrate for cytochromes P450 (CYP) 1A1, 1A2, and 1B1, and its hydroxylated metabolites are remarkably good substrates for the sulfotransferases (SULTs) 1A1, 1A2, 1B1, and 1E1. Finally, (iii) sulfoconjugates of phenolic metabolites of FICZ are present in human urine. Our findings indicate that formylindolo[3,2-b]carbazols are the most potent naturally occurring activators of the AhR signaling pathway and may be the key substrates of the CYP1 and SULT1 families of enzymes. These conclusions contradict the widespread view that xenobiotic compounds are the major AhR ligands and CYP1 substrates.

Riboflavin Is an Active Redox Cofactor in the Na+-pumping NADH:Quinone Oxidoreductase (Na+-NQR) from Vibrio cholerae

Here we present new evidence that riboflavin is present as one of four flavins in Na+-NQR. In particular, we present conclusive evidence that the source of the neutral radical is not one of the FMNs and that riboflavin is the center that gives rise to the neutral flavosemiquinone. The riboflavin is a bona fide redox cofactor and is likely to be the last redox carrier of the enzyme, from which electrons are donated to quinone. We have constructed a double mutant that lacks both covalently bound FMN cofactors (NqrB-T236Y/NqrC-T225Y) and have studied this mutant together with the two single mutants (NqrB-T236Y and NqrC-T225Y) and a mutant that lacks the noncovalently bound FAD in NqrF (NqrF-S246A). The double mutant contains riboflavin and FAD in a 0.6:1 ratio, as the only flavins in the enzyme; noncovalently bound flavins were detected. In the oxidized form, the double mutant exhibits an EPR signal consistent with a neutral flavosemiquinone radical, which is abolished on reduction of the enzyme. The same radical can be observed in the FAD deletion mutant. Furthermore, when the oxidized enzyme reacts with ubiquinol (the reduced form of the usual electron acceptor) in a process that reverses the physiological direction of the electron flow, a single kinetic phase is observed. The kinetic difference spectrum of this process is consistent with one-electron reduction of a neutral flavosemiquinone. The presence of riboflavin in the role of a redox cofactor is thus far unique to Na+-NQR.

Vitamin B2‐sensitized Photo‐oxidation of Dopamine

Kinetics and mechanism of the photo-oxidation of the natural catecholamine-type neurotransmitter dopamine (DA) has been studied in aqueous solution, under aerobic conditions, in the presence of riboflavin (Rf, vitamin B(2)) as a photosensitizer. Results indicate the formation of a weak dark complex Rf-DA, with a mean apparent association constant K(ass) = 30 m(-1), only detectable at DA concentrations much higher than those employed in photochemical experiments. An intricate mechanism of competitive reactions operates upon photoirradiation. DA quenches excited singlet and triplet states of Rf, with rate constants of 4.2 x 10(9) and 2.2 x 10(9) m(-1) s(-1), respectively. With the catecholamine in a concentration similar to that of dissolved molecular oxygen in air-saturated water, DA and oxygen competitively quench the triplet excited state of Rf, generating superoxide radical anion (O(2)) and singlet molecular oxygen (O(2)((1)Delta(g))) by processes initiated by electron and energy-transfer mechanisms, respectively. Rate constants values of 1.9 x 10(8) and 6.6 x 10(6) m(-1) s(-1) have been obtained for the overall and reactive (chemical) interaction of DA with O(2)((1)Delta(g)). The presence of superoxide dismutase increases both the observed rates of aerobic DA photo-oxidation and oxygen uptake, due to its known catalytic scavenging of O(2), a species that could revert the overall photo-oxidation effect, according to the proposed reaction mechanism. As in most of the catecholamine oxidative processes described in the literature, aminochrome is the DA oxidation product upon visible light irradiation in the presence of Rf. It is generated with a quantum yield of 0.05.

Photodegradation of bisphenol A and related compounds under natural-like conditions in the presence of riboflavin: Kinetics, mechanism and photoproducts

The aerobic riboflavin (Rf)-sensitized photodegradation of the endocrine disruptor 4,4′-isopropylidenebisphenol (bisphenol A, BPA), and of the related compounds 4,4′-isopropylidenebis(2,6-dibromophenol) and 4,4′-isopropylidenebis(2,6-dimethylphenol) has been studied in water and water–methanol mixtures through visible-light continuous photolysis, polarographic detection of oxygen uptake, stationary and time-resolved fluorescence spectroscopy, time-resolved near-IR phosphorescence detection and laser flash photolysis techniques. Bisphenols (BPs) quench excited singlet and triplet states of Rf, with rate constants close to the diffusion limit. BPs and dissolved molecular oxygen, employed in similar concentrations, competitively quench triplet excited Rf. As a consequence, superoxide radical anion and singlet molecular oxygen (O 2( 1Δ g )) are produced by electron- and energy-transfer processes, respectively, as demonstrated by auxiliary experiments employing selective quenchers of both oxidative species and the exclusive O 2( 1Δ g ) generator Rose Bengal. As a global result, the photodegradation of Rf is retarded, whereas BPs are degraded, mainly by an O 2( 1Δ g )-mediated mechanism, which constitutes a relatively efficient process in the case of BPA. Oxidation, dimerization and fragmentation products have been identified in the photooxidation of BPA. Results indicate that BPs in natural waters can undergo spontaneous photodegradation under environmental conditions in the presence of adequate photosensitizers.

5-Methyltetrahydrofolate is photosensitive in the presence of riboflavin

5-Methyltetrahydrofolate (5MTHF) is the main form of folate in human plasma, and an important vitamin for human health. Photodegradation of folates may have played a role in the development of different human skin colours. 5MTHF can be degraded directly by exposure to ultraviolet radiation or by exposure to visible light in the presence of endogenous sensitizers like riboflavin (RF). These photochemical reactions were studied by absorption spectroscopy. While 5MTHF is stable under UV and visible light exposure in pure aqueous media, it is quickly degraded in the presence of RF during UVA and blue light exposure. The degradation of 5MTHF is dependent on the concentration of RF, but not on the concentration of 5MTHF itself. UVA and blue light gave similar reactions. Further investigations are necessary to evaluate the consequences of large light exposures in vivo in humans. Our findings should be taken into the ongoing discussion about the development of human skin colours. Due to the presence of RF in human blood, folate can be significantly degraded during prolonged or intense blue light exposure. Thus, a dark skin colour may be favourable for prevention of folate degradation under high solar fluence rates, such as in equatorial areas.

Scavenging of riboflavin-photogenerated oxidative species by uric acid, xanthine or hypoxanthine: A kinetic study

Air-equilibrated aqueous solutions of the purine bases (PBs) uric acid (URA), xanthine (XAN) and hypoxanthine (HXA) have been irradiated with visible light in the presence of riboflavin (Rf) as the only light-absorbing compound, to study to what extent the reactive oxygenated species (ROSs) singlet molecular oxygen, O 2( 1Δ g), and superoxide radical anion, O 2 −, are involved in the observed photodegradation processes. Both reactive species are generated from triplet excited riboflavin ( 3Rf*) by well-known processes: energy transfer from 3Rf* to ground state oxygen to yield O 2( 1Δ g), and electron transfer from each PB to 3Rf* to produce the radical anion Rf − that, after another electron transfer step to ground state oxygen, yields O 2 − and Rf. The kinetics of the involved processes have been studied by polarographic detection of oxygen uptake and time-resolved phosphorescence detection of O 2( 1Δ g), at concentrations in the range 0.05–0.5 mM in aqueous solutions at pH values 5, 7, 9, and 12, to compare the reactivity of the neutral and ionic species of each purine base. By means of Rose Bengal photosensitized reactions, it could be shown that O 2( 1Δ g) is quenched by the three PBs with an overall rate constant decreasing in the order URA > XAN > HXA, with values ranging from 1.16 × 10 9 M −1 s −1 (URA at pH 12) to 5 × 10 6 M −1 s −1 (HXA at pH 7). When Rf is used instead, the quenching process takes place mostly by a chemical mechanism that prevents the oxidation of Rf by O 2( 1Δ g). Experiments with Rf and superoxide dismutase indicate that the PBs also react with O 2 − by a mechanism which is not simple. With URA, largely the most efficient O 2( 1Δ g)-quencher, the major ROS generated is O 2 −, and the process is characterized by the largest rate of oxygen uptake, indicating efficient overall-scavenging and URA oxidation. With HXA, the less efficient O 2( 1Δ g)-quencher, the process shows a relatively high rate of oxygen consumption, especially in alkaline solution, likely as a result of the efficient generation of O 2 − from Rf − and its concomitant reaction with HXA. Finally, with XAN the quenching efficiency takes intermediate values. At physiological pH, URA is the most efficient antioxidant, showing the highest overall rate constant for O 2( 1Δ g) quenching, the lowest efficiency for the reaction with O 2( 1Δ g), and the highest relative rate of oxygen uptake.

Biogenic sulphide plays a major role on the riboflavin-mediated decolourisation of azo dyes under sulphate-reducing conditions

The effect of high concentrations of sulphate on the reductive decolourisation of different azo dyes by anaerobic sludge was studied in batch cultures. Sludge cultures were pre-incubated under sulphate-reducing conditions prior addition of dyes. Little or no effects of sulphate (5-10 g sulphate l(-1)) on the rate of decolourisation of Reactive Orange 14 (RO14), Direct Blue 53 (DB53) and Direct Blue 71 (DB71) were observed when no external redox mediator was provided. However, an increase in sulphate concentration, in the presence of riboflavin (20 microM), enhanced the decolourisation of all dyes. The first-rate constant of decolourisation (k) was increased up to 2-, 3.6- and 2-fold for RO14, DB53 and DB71, respectively, by supplying high sulphate concentrations, compared to the controls lacking sulphate, in the presence of the redox mediator. Sulphate reduction did not take place during the course of azo reductions, but was only evident before dye addition and after complete decolourisation, suggesting azo dyes reduction out-competed sulphate reduction for the available reducing equivalents. The experimental data suggest that reduction of azo dyes by riboflavin, which had been reduced by biogenic sulphide, was the major mechanism implicated during decolourisations, which was corroborated by abiotic incubations. Riboflavin greatly accelerated the abiotic reduction of RO14, so that the k value was increased up to 44-fold compared to the control lacking riboflavin.