Hydroxyl-Radical Reaction Pathways for the Fast Photochemical Oxidation of Proteins Platform As Revealed by 18O Isotopic Labeling.

Fast photochemical oxidation of protein (FPOP) has become an important mass spectrometry-based protein footprinting approach. Although the hydroxyl radical (•OH) generated by photolysis of hydrogen peroxide (H2O2) is most commonly used, the pathways for its reaction with amino-acid side chains remain unclear. Here, we report a systematic study of •OH oxidative modification of 13 amino acid residues by using 18O isotopic labeling. The results differentiate three classes of residues on the basis of their oxygen uptake preference toward different oxygen sources. Histidine, arginine, tyrosine, and phenylalanine residues preferentially take oxygen from H2O2. Methionine residues competitively take oxygen from H2O2 and dissolved oxygen (O2), whereas the remaining residues take oxygen exclusively from O2. Results reported in this work deepen the understanding of •OH labeling pathway on a FPOP platform, opening new possibilities for tailoring FPOP conditions in addressing many biological questions in a profound way.

Effect of crystalline structure on terbuthylazine degradation by H2O2-assisted TiO2 photocatalysis under visible irradiation.

Various methods for shifting the optical response of TiO2 into the visible (Vis) range have been reported. Herein, we reported the application of a TiO2/H2O2/Vis process and the effects of TiO2 crystalline structure on the degradation of terbuthylazine. The results indicated that TiO2 crystalline structure and H2O2 addition had significant effects on terbuthylazine degradation: its degradation rate could be increased from 7% to 70% with H2O2 addition after 180 min of reaction, the synergistic degradation of terbuthylazine by TiO2-Fe3+ was substantially accelerated, with the degradation rate reaching up to 100% after 20 min of reaction, and rutile TiO2 showed better photocatalytic activity and a more obvious synergistic effect than anatase TiO2. The addition of free-radical scavengers (tert-butyl alcohol or methanol) inhibited the degradation efficiency of rutile TiO2, but had a relatively minor effect on anatase TiO2. Fluorescence spectrophotometry analysis indicated that hydroxyl free radicals could be continuously produced when using rutile TiO2 as the photocatalyst. Degradation of terbuthylazine catalyzed by rutile TiO2 occurred mainly in solution, but occurred on the particle surface of the photocatalyst when catalyzed by anatase TiO2. This study provides new insight into the role of TiO2 crystalline structure on the degradation of terbuthylazine and its photocatalytic degradation mechanism.

Photolytic and photocatalytic transformation of an antipsychotic drug asenapine: Comparison of kinetics, identification of transformation products, and in silico estimation of their properties.

The photolytic and photocatalytic transformation of an antipsychotic drug asenapine with the use of H2O2 and TiO2 was studied. A method employing irradiation with a simulated full solar spectrum in the photostability chamber was applied, then the reverse-phase ultra high performance liquid chromatography with diode array detector, coupled with electrospray quadrupole time-of-flight mass spectrometer (RP-UHPLC-DAD - ESI-Q-TOF) was used for the quantitative and qualitative analysis of the processes. The developed quantitative method was fully validated, according to the International Conference on Harmonization of Technical Requirements for Registration of Pharmaceuticals for Human Use (ICH) guidelines, and the kinetic parameters of asenapine photodecomposition were compared. Nineteen phototransformation products were detected, and their probable structures - mainly hydroxylated and oxidized asenapine derivatives - were suggested. On the basis of the elucidated structures the computational prediction of their toxicity at the various endpoints, as well as bioconcentration factors and biodegradability was performed. The obtained results were then subjected to the principal component analysis (PCA). This chemometric technique facilitated comparison of the applied models, calculated properties of the TPs, and enabled visualization of relationships between them.

Mechanistic study of photo-oxidation of Bisphenol-A (BPA) with hydrogen peroxide (H2O2) and sodium persulfate (SPS).

The removal of Bisphenol-A (BPA) from contaminated water using advanced oxidation methods such as UV-C assisted oxidation by hydrogen peroxide (H2O2) and sodium persulfate (SPS) has been reported by the authors earlier (Sharma et al., 2015a). In the present study, the authors report the removal of BPA from aqueous solution by the above two methods and its degradation mechanism. UV-C light (254 nm wavelength, 40 W power) was applied to BPA contaminated water at natural pH (pHN) under room temperature conditions. Experiments were carried out with the initial BPA concentration in the range of 0.04 mM-0.31 mM and the oxidant/BPA molar ratio in the range of 294:1-38:1 for UV-C/H2O2 and 31.5-4.06:1 for UV-C/SPS systems. The removal of BPA enhanced with decreasing BPA concentration. The total organic carbon also decreased with the UV-C irradiation time under optimum conditions ([H2O2]0 = 11.76 mM; [SPS]0 = 1.26 mM; temperature (29 ± 3 °C). Competition of BPA for reaction with HO or [Formula: see text] radicals at its higher concentrations results in a decrease in the removal of BPA. The intermediates with smaller and higher molecular weights than that of BPA were found in the treated water. Based on GC-MS and FTIR spectra of the reaction mixture, the formation of hydroxylated by-products testified the HO mediated oxidation pathway in the BPA degradation, while the formation of quinones and phenoxy phenols pointed to the [Formula: see text] dominating pathway through the formation of hydroxycyclohexadienyl (HCHD) and BPA phenoxyl radicals. The main route of BPA degradation is the hydroxylation followed by dehydration, coupling and ring opening reactions.

Degradation and mineralization of Bisphenol A (BPA) in aqueous solution using advanced oxidation processes: UV/H2O2 and UV/S2O8(2-) oxidation systems.

This work reports on the removal and mineralization of an endocrine disrupting chemical, Bisphenol A (BPA) at a concentration of 0.22 mM in aqueous solution using inorganic oxidants (hydrogen peroxide, H2O2 and sodium persulfate, Na2S2O8;S2O8(2-)) under UV irradiation at a wavelength of 254 nm and 40 W power (Io = 1.26 × 10(-6) E s(-1)) at its natural pH and a temperature of 29 ± 3 °C. With an optimum persulfate concentration of 1.26 mM, the UV/S2O8(2-) process resulted in ∼95% BPA removal after 240 min of irradiation. The optimum BPA removal was found to be ∼85% with a H2O2 concentration of 11.76 mM. At higher concentrations, either of the oxidants showed an adverse effect because of the quenching of the hydroxyl or sulfate radicals in the BPA solution. The sulfate-based oxidation process could be used over a wider initial pH range of 3-12, but the hydroxyl radical-based oxidation of BPA should be carried out in the acidic pH range only. The water matrix components (bicarbonate, chloride and humic acid) showed higher scavenging effect in hydroxyl radical-based oxidation than that in the sulfate radical-based oxidation of BPA. UV/S2O8(2-) oxidation system utilized less energy (307 kWh/m(3)) EE/O in comparison to UV/H2O2 system (509 kWh/m(3)) under optimum operating conditions. The cost of UV irradiation far outweighed the cost of the oxidants in the process. However, the total cost of treatment of persulfate-based system was much lower than that of H2O2-based oxidation system.

The role of iron species on the turbidity of oxidized phenol solutions in a photo-Fenton system.

This work aims at establishing the contribution of the iron species to the turbidity of phenol solutions oxidized with photo-Fenton technology. During oxidation, turbidity increases linearly with time till a maximum value, according to a formation rate that shows a dependence of second order with respect to the catalyst concentration. Next, the decrease in turbidity shows the evolution of second-order kinetics, where the kinetics constant is inversely proportional to the dosage of iron, of order 0.7. The concentration of iron species is analysed at the point of maximum turbidity, as a function of the total amount of iron. Then, it is found that using dosages FeT=0-15.0 mg/L, the majority iron species was found to be ferrous ions, indicating that its concentration increases linearly with the dosage of total iron. This result may indicate that the photo-reaction of ferric ion occurs leading to the regeneration of ferrous ion. The results, obtained by operating with initial dosages FeT=15.0 and 25.0 mg/L, suggest that ferrous ion concentration decreases while ferric ion concentration increases in a complementary manner. This fact could be explained as a regeneration cycle of the iron species. The observed turbidity is generated due to the iron being added as a catalyst and the organic matter present in the system. Later, it was found that at the point of maximum turbidity, the concentration of ferrous ions is inversely proportional to the concentration of phenol and its dihydroxylated intermediates.

Application of the electro-Fenton process for cutting fluid mineralization.

Organic compound is the main pollutant in industrial effluent. Conventional wastewater treatment processes are inefficient for the removal of toxic or non-biodegradable organic pollutants. Advanced electrochemical depollution is a very efficient and economic method, suitable when the wastewater contains toxic and recalcitrant organic pollutants. The aim of the present study was to investigate the application of the electro-Fenton (EF) process for the degradation and mineralization of a stable oil-in-water emulsion (0.01% in v/v). The effects of operating parameters such as cathode material (graphite, Ti/Pt and steel), nature (Na2SO4, NaNO3 and NaCl) and dose of electrolyte (25-75 mM), initial ferrous ions concentration (1-75 mM), current intensity (0.1-0.2 A) and operating time, on chemical oxygen demand (COD) removal efficiency, were studied. Results showed that the EF method can be used efficiently for the degradation of stable cutting oil emulsion. For considered initial conditions (bubbling compressed air at 1 L/min, 0.15 A, pH 3, [Na2SO4]=0.05 M, [FeSO4]=0.015 M, COD0=400 mg O2/L), the best removal efficiencies were obtained under the following conditions: graphite as cathode material, 180 min for treatment duration and 0.05 M [Na2SO4]. For these conditions, treatment of 250 mL of emulsion led to 93.6% of cutting fluid mineralization, which correspond to 25 mg O2/L of final COD, 19 kWh/m3 of treated wastewater and 24.039 kWh/kg of COD removal.

Mineralization of hetero bi-functional reactive dye in aqueous solution by Fenton and photo-Fenton reactions.

This study focused on the advanced oxidation of the hetero bi-functional reactive dye Sumifix Supra Yellow 3RF (CI Reactive Yellow 145) using dark Fenton and photo-Fenton conditions in a lab-scale experiment. A 2(3) factorial design was used to evaluate the effects of the three key factors: temperature, Fe(II) and H2O2 concentrations, for a dye concentration of 250 mg L(-1) with chemical oxygen demand (COD) of 172 mg L(-1) O2 at pH=3. The response function was the COD reduction. This methodology lets us find the effects and interactions of the studied variables and their roles in the efficiency of the treatment process. In the optimization, the correlation coefficients for the model (R2) were 0.948 and 0.965 for Fenton and photo-Fenton treatments, respectively. Under optimized reaction conditions: pH=3, temperature=298 K, [H2O2]=11.765 mM and [Fe(II)]=1.075 mM; 60 min of treatment resulted in a 79% and 92.2% decrease in COD, for the dye taken as the model organic compound, after Fenton and photo-Fenton treatments, respectively.

Mineralization of wastewater from the pharmaceutical industry containing chloride ions by UV photolysis of H2O2/Fe(II) and ultrasonic irradiation.

The mineralization of pharmaceutical wastewater containing chloride ions using a UV/H2O2/Fe(II) process was studied. The addition of Fe(II) to the UV/H2O2 system did not improve the degradation efficiency due to inhibition of the photo-Fenton reaction, at acid pH, in the presence of chloride ions in these wastewaters. The increase of pH from 2 to 7 increased the degree of mineralization under UV photolysis of H2O2 because more HO radicals are available by HOCl dissociation reaction. Under the selected operation conditions ([H2O2]o = 11,500 ppm, [Fe(II)] = 0 ppm, [TOC]o = 125 ppm and pH = 7), 100% of TOC removal was attained in 120 min. A significant synergistic effect of combining photolysis (UV/H2O2) and sonolysis was observed. Sonophotolysis (UV/H2O2/ultrasound) technique significantly increased the degree of mineralization (100% TOC removal in 90 min using 6500 ppm H2O2) when compared with each individual process. Sonochemical reaction was favored by the presence of chloride ions since the concentration of contaminants at the gas-liquid interface increased. Free radicals reaction was the controlling mechanism in the UV/H2O2/ultrasound system. HO radicals were the main oxidative intermediate species in the process, although hydroperoxyl radicals (HO2) also played a role. The contribution of thermal-pyrolytic reaction (in gas-phase) to sonophotolysis process was negligible.

Advanced photochemical oxidation of emergent micropollutants: carbamazepine.

The combination of UV radiation with hydrogen peroxide has been widely used for the photodegradation of pollutants in aqueous solutions. Statistical design of experiments is a powerful tool to optimize this kind of process. Initial hydrogen peroxide concentration, pH and temperature were considered as the variables for the process optimization. The interactions existing between these three variables were analyzed. Initial concentration of hydrogen peroxide proved to be the most important variable conditioning the removal efficiency, followed by temperature, and pH shows a non-significant positive influence along the whole operation interval. The ANOVA test reported significance for five of the nine involved variables. The Response Surface Methodology technique was used to optimize carbamazepine degradation. Under optimal conditions (hydrogen peroxide concentration = 0.38·10(-3) mol L(-1), pH = 1 and temperature = 35.6°C) total carbamazepine degradation was achieved.

Study of enhanced low-quality coal oxidative desulphurization and deashing by using HNO3 and microwave pretreatment.

In this paper, the effect of urea-hydrogen peroxide (UHP) solution on desulphurization and demineralization of coal with high sulphur and high ash by using HNO3 and microwave pretreatment was investigated. The oxidation process is strongly dependent on irradiation power and time for microwave pretreatment, UHP concentration, leaching time and temperature of the UHP solution. X-ray diffraction and Fourier transform infrared technique have been performed for the raw and treated coals. Compared with the UHP alone, successive treatments with HNO3 and microwave pretreatment resulted in the significant removal of total sulphur and mineral matter from the coal. The proposed experimental method has the meaning of practical guide to the desulphurization and deashing of coal by microwave.

Photo-Fenton degradation of phenol, 2,4-dichlorophenoxyacetic acid and 2,4-dichlorophenol mixture in saline solution using a falling-film solar reactor.

In this work, a saline aqueous solution of phenol, 2,4-dichlorophenoxyacetic acid (2,4-D) and 2,4-dichlorophenol (2,4-DCP) was treated by the photo-Fenton process in a falling-film solar reactor. The influence of the parameters such as initial pH (5-7), initial concentration of Fe2+ (1-2.5mM) and rate of H202 addition (1.87-3.74mmol min-1) was investigated. The efficiency of photodegradation was determined from the removal of dissolved organic carbon (DOC), described by the species degradation of phenol, 2,4-D and 2,4-DCP. Response surface methodology was employed to assess the effects of the variables investigated, i.e. [Fe2+], [H202] and pH, in the photo-Fenton process with solar irradiation. The results reveal that the variables' initial concentration of Fe2+ and H202 presents predominant effect on pollutants' degradation in terms of DOC removal, while pH showed no influence. Under the most adequate experimental conditions, about 85% DOC removal was obtained in 180 min by using a reaction system employed here, and total removal of phenol, 2,4- and 2,4-DCP mixture in about 30min.

Light-activated bleaching: effects on surface mineral change on enamel.

AIM: This study evaluated the in vitro effect of 35% hydrogen peroxide (HP) on surface enamel change when activated with different light curing units (LCUs). MATERIALS AND METHODS: Enamel blocks (4 × 4 × 2 mm) were obtained from bovine incisors. The initial microhardness of the enamel was determined for each specimen. After this enamel blocks were randomly divided into four groups (n = 10) and treated as follows: Control, no bleaching procedure performed; HP - LCU, application of 35% HP gel without light activation; HP + QTH, application of 35% HP gel and light activation with a Quartz Tungsten-Halogen (QTH); and HP + Light Emitting Diode, application of 35% HP gel and light-activation with a LED. New microhardness measurements were obtained, immediately, 7 and 14 days after treatment. The percentage of surface mineral change was calculated according to the baseline and post-treatment microhardness values. Additionally, six samples from each group were randomly selected and prepared for scanning electron microscopy (SEM) characterization. The data were analyzed using an analysis of variance (ANOVA) to detect differences between the three time periods, and an ANOVA and Tukey's test with a confidence level of 95%. RESULTS: There was no significant difference between the initial hardness values and hardness values after treatment in any of the groups or time periods (p > 0.05). No major surface alterations were detected with SEM when comparing control groups to those undergoing bleaching treatments. CONCLUSION: The use of 35% HP in combination to QTH or LED light curing units LCU does not have detrimental effect on the enamel surface topography or in the mineral content, when compared with unbleached enamel or enamel submitted to 35% HP treatment alone.

Reactive oxygen species activation of MAPK pathway results in VEGF upregulation as an undesired irradiation response.

BACKGROUND: Radioresistance limits the effectiveness of radiotherapy in head and neck squamous cell carcinoma. We previously demonstrated post-radiogenic mitogen-activated protein kinase (MAPK) pathway activation and vascular endothelial growth factor (VEGF) release resulting in reduced tumor cell response. Here, we examined the association of this mechanism with the induction of reactive oxygen species (ROS) under irradiation (IR). METHODS: Intracellular ROS after IR were measured. We modeled radiation-induced ROS by exposure of two SCC lines to H2 O2 and evaluated the impact of irradiation and ROS on ERK phosphorylation by Western blot, immunohistochemistry, and ELISA. RESULTS: We found elevated pERK levels after treatment with IR and H2 O2 , which could be distinctly suppressed by U0126. Immunohistochemistry and ELISA revealed increased intracellular VEGF levels after H2 O2 application. CONCLUSIONS: Our data show that irradiation-induced ROS activate the MAPK pathway and release of VEGF. As VEGF is known to be released after cellular distress resulting in cytoprotection, the described mechanism is potentially of importance for therapy success.

Efficacy of tooth bleaching with and without light activation and its effect on the pulp temperature: an in vitro study.

The aim of this in vitro study was to evaluate the colour stability of bleaching after light activation with halogen unit, laser, LED unit or chemical activation up to 3 months after treatment. Four groups of teeth (n = 20) were bleached with Opalescence Xtra Boost (38% hydrogen peroxide) using four different methods: activation with halogen, LED, laser or chemical activation only. All teeth were bleached in one session for four times (4 × 15 min) and the colour was evaluated using a spectrophotometer at the following time points: before bleaching, immediately after bleaching, 1 day, and 1 and 3 months after the end of bleaching. Between the tested time points, the teeth were stored in 0.9% NaCl solution. Additionally, the temperature increase in the pulp chamber was measured using a measuring sensor connected to a computer. Bleaching with the halogen unit showed the highest colour change. Halogen unit, laser and chemical activation resulted in whiter teeth after 1 and 3 months compared to the colour after the end of the bleaching procedure (p ≤ 0.05). Three months after the end of bleaching, the shade changes observed were-halogen: 7.1 > chemical activation: 6.2 > LED: 5.4 > laser: 5.2. Halogen showed the highest temperature increase (17.39°C ± 1.96) followed by laser (14.06°C ± 2.55) and LED (0.41°C ± 0.66) (p < 0.0001). Chemical activation did not affect the temperature in the pulp chamber. The use of light activation did not show any advantages compared to chemical bleaching. Although halogen unit showed the higher shade's change, its use resulted also in the higher pulp temperature. According to the present findings, light activation of the bleaching agent seems not to be beneficial compared to bleaching without light activation, concerning the colour stability up to 3 months after bleaching and the pulp temperature caused during the bleaching procedure.

Toxicity assessment of tannery effluent treated by an optimized photo-Fenton process.

In this work, an optimized photo-Fenton process was applied to remove pollutants from tannery industrial effluent (TIE) with its final toxicity level being assessed by a lettuce-seed-based bioassay test. A full 33 factorial design was applied for the optimization of long-term photo-Fenton experiments. The oPtimum conditions of the photo-Fenton process were attained at concentration values of 0.3 g Fe(2+) L(-1) and 20 g H2O2 L(-1) and pH3, for 120 min UV irradiation time. Reactor operating parameter (ROP) effects on the removal of chemical oxygen demand, colour, turbidity, total suspended solids and total volatile solids were evaluated, suggesting that a broad range of ROP values are also suitable to give results very near to those of the photo-Fenton experiments under optimal conditions. Based on the low calculated median lethal dose (LD50) values from a lettuce-seed-based bioassay test, we suggest that recalcitrant substances are present in treated TIE samples. A possible cause of the high toxicity level could partly be attributed to the nitrate concentration, which was not completely abated by the photo-Fenton process. Apart from this, the photo-Fenton process can be used as a part of an industrial effluent treatment system in order to abate high organic pollutant loads.

Degradation kinetics and mechanism of trace nitrobenzene by granular activated carbon enhanced microwave/hydrogen peroxide system.

The kinetics of the degradation of trace nitrobenzene (NB) by a granular activated carbon (GAC) enhanced microwave (MW)/hydrogen peroxide (H202) system was studied. Effects of pH, NB initial concentration and tert-butyl alcohol on the removal efficiency were examined. It was found that the reaction rate fits well to first-order reaction kinetics in the MW/GAC/H202 process. Moreover, GAC greatly enhanced the degradation rate of NB in water. Under a given condition (MW power 300 W, H202 dosage 10 mg/L, pH 6.85 and temperature (60 +/- 5)degrees C), the degradation rate of NB was 0.05214 min-1when 4 g/L GAC was added. In general, alkaline pH was better for NB degradation; however, the optimum pH was 8.0 in the tested pH value range of 4.0-12.0. At H202 dosage of 10 mg/L and GAC dosage of 4 g/L, the removal of NB was decreased with increasing initial concentrations of NB, indicating that a low initial concentration was beneficial for the degradation of NB. These results indicated that the MW/GAC/H202 process was effective for trace NB degradation in water. Gas chromatography-mass spectrometry analysis indicated that a hydroxyl radical addition reaction and dehydrogenation reaction enhanced NB degradation.

Heterogeneous fenton degradation of azo dyes catalyzed by modified polyacrylonitrile fiber fe complexes: QSPR (quantitative structure peorperty relationship) study.

The amidoximated polyacrylonitrile (PAN) fiber Fe complexes were prepared and used as the heterogeneous Fenton catalysts for the degradation of 28 anionic water soluble azo dyes in water under visible irradiation. The multiple linear regression (MLR) method was employed to develop the quantitative structure property relationship (QSPR) model equations for the decoloration and mineralization of azo dyes. Moreover, the predictive ability of the QSPR model equations was assessed using Leave-one-out (LOO) and cross-validation (CV) methods. Additionally, the effect of Fe content of catalyst and the sodium chloride in water on QSPR model equations were also investigated. The results indicated that the heterogeneous photo-Fenton degradation of the azo dyes with different structures was conducted in the presence of the amidoximated PAN fiber Fe complex. The QSPR model equations for the dye decoloration and mineralization were successfully developed using MLR technique. MW/S (molecular weight divided by the number of sulphonate groups) and NN=N (the number of azo linkage) are considered as the most important determining factor for the dye degradation and mineralization, and there is a significant negative correlation between MW/S or NN=N and degradation percentage or total organic carbon (TOC) removal. Moreover, LOO and CV analysis suggested that the obtained QSPR model equations have the better prediction ability. The variation in Fe content of catalyst and the addition of sodium chloride did not alter the nature of the QSPR model equations.

Microbial resistance in relation to catalase activity to oxidative stress induced by photolysis of hydrogen peroxide.

The purpose of the present study was to evaluate the mechanism of microbial resistance to oxidative stress induced by photolysis of hydrogen peroxide (H(2)O(2)) in relation to microbial catalase activity. In microbicidal tests, Staphylococcus aureus and Candida albicans were killed and this was accompanied by production of hydroxyl radicals. C. albicans was more resistant to hydroxyl radicals generated by photolysis of H(2)O(2) than was S. aureus. A catalase activity assay demonstrated that C. albicans had stronger catalase activity; accordingly, catalase activity could be one of the reasons for the resistance of the fungus to photolysis of H(2)O(2). Indeed, it was demonstrated that C. albicans with strong catalase activity was more resistant to photolysis of H(2)O(2) than that with weak catalase activity. Kinetic analysis using a modified Lineweaver-Burk plot also demonstrated that the microorganisms reacted directly with hydroxyl radicals and that this was accompanied by decomposition of H(2)O(2). The results of the present study suggest that the microbicidal effects of hydroxyl radicals generated by photolysis of H(2)O(2) can be alleviated by decomposition of H(2)O(2) by catalase in microorganisms.

The effectiveness of low-intensity red laser for activating a bleaching gel and its effect in temperature of the bleaching gel and the dental pulp.

STATEMENT OF THE PROBLEM: The effectiveness of low-intensity red laser for activating a bleaching gel and its effect in pulp temperature was not investigated in dental literature. PURPOSE: The objective of this study was to assess the effectiveness of low-intensity red laser for activating a bleaching gel, as well as its effect in temperature of the bleaching gel and the dental pulp. MATERIALS AND METHODS: Forty extracted bovine teeth were immersed in a solution of coffee 14 days for darkening. The initial colors were recorded by spectrophotometric analysis. The specimens were randomly distributed into two groups (N = 20): the control, which did not receive light and the experimental group that received light from an appliance fitted with three red light-emitting laser diodes (λ = 660 nm). A green-colored, 35% H(2) O(2) -based bleaching gel was applied for 30 minutes, and changed three times. After bleaching, the colors were again measured to obtain the L*a*b* values. Color variation was calculated (ΔE) and the data submitted to the non-paired t-test (5%). To assess temperature, 10 human incisors were prepared, in which one thermocouple was placed on the bleaching gel applied on the surface of the teeth and another inside the pulp chamber. RESULTS: There was a significant difference between the groups (p = 0.016), and the experimental group presented a significantly higher mean variation (7.21 ± 2.76) in comparison with the control group (5.37 ± 1.76). There was an increase in pulp temperature, but it was not sufficient to cause damage to the pulp. CONCLUSION: Bleaching gel activation with low-intensity red laser was capable of increasing the effectiveness of bleaching treatment and did not increase pulp temperature to levels deleterious to the pulp.