Detoxification of chlorinated organic compound is an important and urgent issue in water remediation nowadays. Trichloroethylene (TCE), as a model compound in this study, has been proved to be degraded effectively by bimetallic nanoparticles (NPs) in solution phase. In this study, Fe/Pd bimetallic NPs were synthesized in poly (acrylic acid) (PAA) functionalized polyvinylidene fluoride (PVDF) microfiltration membranes. TCE dechlorination with these bimetallic NPs was conducted under different pH values and different metal loadings to study the role of corrosion on reaction rates. One-dimensional mathematical model with pseudo first-order reaction kinetic was introduced to discuss the TCE dechlorination profile in membrane system. Reduction reaction in pores is affected by several parameters including NP loading and size, TCE diffusivity, void volume fraction and surface-area-based reaction rates. This model result indicated that modification is needed to correct the reaction rate obtained from bulk solution in order to represent the actual efficiency of NPs on reduction reaction. In addition, TCE dechlorination mainly occurred near NPs’ surface. Second part of model indicated that reduction mechanism with TCE adsorption-desorption behavior could be used to discuss dechlorination with a high TCE concentration.

The focus of this research is on the optical measurement of uranyl in a solid matrix using fluorescence spectroscopy. Nanoporous silica-based materials were used to extract uranyl from contaminated soil and to enhance the fluorescence intensity and lifetime. The fluorescence lifetime and intensity of uranyl ions adsorbed on porous silica-based materials of varying pore size was measured as a function of pH and in the presence of fluoride. The feasibility of uranyl fluorescence detection on the top of soil by silica gel is carried out by four types of natural soil. The results show that the uranyl fluorescence intensity can be enhanced by approximately two orders of magnitude by the silica nanoporous matrix from pH 4-12 with the greatest enhancement occurring from pH 4-7. The enhanced fluorescence lifetime can be used in time-gated measurements to help minimize the influence of background environmental fluorophores. The pH and the fluoride variation causes different uranyl speciation and results in a peak shift in the fluorescence spectrum. The mechanism of the uranyl ion on the silica nanoporous matrix was studied through 15 different silica materials with different water content ratios and various concentrations of uranium on different silica structures. The result shows that the particle size, pore size, water content and uranyl concentration on silica surfaces are all important factors for optimizing the fluorescence intensity. The spacing between silica materials, either the pore inside materials or the space between particles, causes the variety of uranyl distribution on the material surface and changes the fluorescence performance. Also, X-Ray Photoelectron Spectroscopy (XPS) is used to identify the possible uranyl surface species on silica. The fluorescence emission spectra from silica materials and the XPS results are consistent with the presence of two different uranyl compounds. The specific surface area of silica materials plays an important role on uranyl adsorption mechanism. To further enhance the sensitivity, an optical ball lens was used to preferentially direct the fluorescence signal toward the excitation source in standoff measurements. The application of the ball lens was found to increase the detection distance up to 14 times.

Os fungos filamentosos acionam mecanismos eficientes para sobrevivência nas diferentes condições ambientais e modulam, por exemplo, a produção de proteínas. A abordagem proteômica tem sido utilizada com sucesso para investigar os mecanismos fisiológicos envolvidos nas adaptações metabólicas desses microrganismos, frente às mudanças de parâmetros físicos e químicos do meio extracelular. No presente trabalho foi avaliado o efeito das variações de pH, fontes de carbono e nitrogênio (aminoácidos e fontes complexas) sobre a produção de peptidases e sobre o proteoma intra- e extracelular do fungo Aspergillus niveus em bioprocessos submersos. As atividades proteolíticas foram avaliadas durante 96 h de cultivo e as melhores induções, entre os estudos comparativos, foram observadas em pH 6, xilose, cisteína e farinha de pena. O efeito dessas variações sobre o proteoma de A. niveus foi avaliado em 96 h de cultivo submerso e, para isso, foram realizados estudos proteômicos nos quais os conteúdos proteicos foram avaliados por eletroforese bidimensional (2-DE); e os spots proteicos selecionados foram identificados por espectrometria de massas (MALDI-TOF/TOF-MS/MS). As proteínas apresentadas no estudo demonstram que as variações de pH, fontes de carbono e nitrogênio influenciaram as proteínas intra- e extracelulares de A. niveus. Os perfis de proteínas mostram que os parâmetros avaliados foram capazes de influenciar a síntese ou degradação de polímeros biológicos e a resposta aos efeitos químicos e/ou disponibilidade de nutrientes do meio extracelular modularam proteínas envolvidas em diversas rotas metabólicas. Além disso, a suplementação com fontes de nitrogênio orgânicas promoveram a indução de enzimas com potencial biotecnológico para aplicação industrial, o que reforça a importância dos estudos proteômicos para a determinação do comportamento de fungos filamentosos frente às condições ambientais / The filamentous fungi activate efficient mechanisms to survive in different environment conditions and protein production modulation may be a tool of these survival adaptation. Proteomic approach has been used as methodology to investigate physiological processes related to these microorganism adaptation after chemical and physical changes in extracelullar medium. In this study, the influence of pH variation, carbon and nitrogen sources (amino acid and complex sources) was analyzed on protease production and in intra- and extracellular proteomics of filamentous fungus Aspergillus niveus during submerged bioprocess. The proteolytic activities were evaluated for 96 hours of incubation and as the best indutors among those studied, there were proteases in pH 6, xylose, cystein and feather meal conditions. The variation effects over A. niveus proteome 96 hours submerged bioprocess was analyzed, the protein contents was separated, evaluated and identified by bidimensional electrophoresys followed by mass spectometry (MALDI-TOF/TOFMS/ MS). The identified proteins showed that bioprocess condition variations could influence on intra- and extracelullar proteins of A. niveus. The protein profiles presented a modulation on synthesis and degradation of biological polymers, and on proteins from metabolic pathway in response to chemical effects and/or extracellular nutrient available. Moreover, the supplementaiton with nitrogen sources promoted the production of enzymes with biotechnological potential on industrial application, highlighting the importance of proteomics to determine the filamentous fungus behavior under environment changes.

Neste trabalho de Mestrado, foram feitos estudos do comportamento fotofísico da fisetina em diversos solventes através de medidas de absorção de luz UV-Visível, fluorescência estática e resolvida no tempo e da interação entre fisetina (3,3\',4\',7-tetrahidroxiflavona) e 7-hidroxiflavona com ciclodextrinas ( beta e gama) (CDs) através de experimentos de absorção de luz UV-Visível, sinal induzido de dicroísmo circular, fluorescência estática e resolvida no tempo, anisotropia de estado estacionário e RMN de 1H, focando a dependência destas medidas em função da temperatura e do pH. Os resultados experimentais foram comparados com estudos mecânico-quânticos baseados no modelo semi-empírico SAM1 (AMPAC), e nos funcionais B3LYP e MPW1PW91, da Teoria do Funcional de Densidade, empregando os conjuntos de funções de base 6-311G* e 3-21G**. Os estudos da fisetina em diferentes solventes próticos e apróticos mostraram que a fluorescência da sonda é fortemente dependente do solvente. Resultados experimentais indicam a formação de complexos de inclusão entre as CDs e os flavonóides acima. A fisetina com ?-CD forma complexo de estequiometria 1:1 nos meios neutro/ácido e básico, cujos valores de constante de complexação são 900 +- 100 +_ 240 +_ 90 (L/mol), respectivamente. Os dados teóricos evidenciaram que a inclusão da fisetina na beta-CD ocorre preferencialmente pelo anel fenila. O complexo com a gama-CD apresenta estequiometria de 1:1 em meio ácido/neutro e de 1:2 em meio básico, com constantes de complexação de 94 +- 30 e 130 +- 10 (L/mol), respectivamente. Os estudos com a 7-hidroxiflavona revelaram que somente ocorre a formação de complexos com a beta-CD de estequiometria 1:1 e não há dependência do pH. As constantes de complexação obtidas nos meios ácido/neutro e básico são similares, 1430 +-- 510 e 1220 +- 165 L/mol, respectivamente. / In this work, the photophysics of fisetin (3,3\',4\',7-tetrahydroxyflavone) in several solvents was studied through UV-vis absorption spectra, steady-state and time-resolved fluorescence measurements. The interaction between fisetin and 7-hydroxyflavone and cyclodextrins (b- e g-) (CDs) was also investigated by UV-vis absorption spectra, induced signal of circular dichroism, steady-state and time-resolved fluorescence, steady-state anisotropy, and 1H NMR, with dependence on pH and temperature. Some experimental data were compared with quantum-mechanics studies based on the SAM1 (AMPAC) semi-empirical model, as well as with the B3LYP and MPW1PW91 functional models from the Density Functional Theory using the 6-311G* and 3-21G* basis sets. The study of the photophysics of fisetin in protic and aprotic solvents showed a complex behavior and a strong dependence on the solvent. The occurrence of many equilibria between the possible structures of fisetin, e.g. the normal, a few deprotonated ones, and the tautomer due to the excited state intramolecular proton transfer can be responsible for the complex analyses of these experimental data. The spectroscopic measurements show that, at pH 4.0 and 6.5, the complex fisetin - b-CD is formed in a Fis:b-CD 1:1 stoichiometry and an equilibrium constant (K) of 900 ± 100 L/mol. In basic medium (pH 11.5), K decreases to 240 ± 90 L/mol. Molecular modeling points out that the inclusion complex is formed preferentially via entry of the fisetin phenyl group into b-CD. On the other hand, the fisetin - g-CD has a stoichiometry of 1:1 in acid/neutral solutions and of 1:2 in basic conditions. The K values are 94 ± 30 e 130 ± 10 (L/mol), respectively. The 7-hydroxyflavone can only form inclusion complexes with b-CD. The stoichiometry is 1:1 and there is no dependence on pH. Both equilibrium constants determined either in acid or basic medium is very similar to each other, 1430 ± 510, and 1220 ± 165 L/mol, respectively. For this reason, we suppose that the inclusion of this compound into b-CD is also through the phenyl ring of the flavonoid.

Neste trabalho de Mestrado, foram feitos estudos do comportamento fotofísico da fisetina em diversos solventes através de medidas de absorção de luz UV-Visível, fluorescência estática e resolvida no tempo e da interação entre fisetina (3,3\',4\',7-tetrahidroxiflavona) e 7-hidroxiflavona com ciclodextrinas ( beta e gama) (CDs) através de experimentos de absorção de luz UV-Visível, sinal induzido de dicroísmo circular, fluorescência estática e resolvida no tempo, anisotropia de estado estacionário e RMN de 1H, focando a dependência destas medidas em função da temperatura e do pH. Os resultados experimentais foram comparados com estudos mecânico-quânticos baseados no modelo semi-empírico SAM1 (AMPAC), e nos funcionais B3LYP e MPW1PW91, da Teoria do Funcional de Densidade, empregando os conjuntos de funções de base 6-311G* e 3-21G**. Os estudos da fisetina em diferentes solventes próticos e apróticos mostraram que a fluorescência da sonda é fortemente dependente do solvente. Resultados experimentais indicam a formação de complexos de inclusão entre as CDs e os flavonóides acima. A fisetina com ?-CD forma complexo de estequiometria 1:1 nos meios neutro/ácido e básico, cujos valores de constante de complexação são 900 +- 100 +_ 240 +_ 90 (L/mol), respectivamente. Os dados teóricos evidenciaram que a inclusão da fisetina na beta-CD ocorre preferencialmente pelo anel fenila. O complexo com a gama-CD apresenta estequiometria de 1:1 em meio ácido/neutro e de 1:2 em meio básico, com constantes de complexação de 94 +- 30 e 130 +- 10 (L/mol), respectivamente. Os estudos com a 7-hidroxiflavona revelaram que somente ocorre a formação de complexos com a beta-CD de estequiometria 1:1 e não há dependência do pH. As constantes de complexação obtidas nos meios ácido/neutro e básico são similares, 1430 +-- 510 e 1220 +- 165 L/mol, respectivamente. / In this work, the photophysics of fisetin (3,3\',4\',7-tetrahydroxyflavone) in several solvents was studied through UV-vis absorption spectra, steady-state and time-resolved fluorescence measurements. The interaction between fisetin and 7-hydroxyflavone and cyclodextrins (b- e g-) (CDs) was also investigated by UV-vis absorption spectra, induced signal of circular dichroism, steady-state and time-resolved fluorescence, steady-state anisotropy, and 1H NMR, with dependence on pH and temperature. Some experimental data were compared with quantum-mechanics studies based on the SAM1 (AMPAC) semi-empirical model, as well as with the B3LYP and MPW1PW91 functional models from the Density Functional Theory using the 6-311G* and 3-21G* basis sets. The study of the photophysics of fisetin in protic and aprotic solvents showed a complex behavior and a strong dependence on the solvent. The occurrence of many equilibria between the possible structures of fisetin, e.g. the normal, a few deprotonated ones, and the tautomer due to the excited state intramolecular proton transfer can be responsible for the complex analyses of these experimental data. The spectroscopic measurements show that, at pH 4.0 and 6.5, the complex fisetin - b-CD is formed in a Fis:b-CD 1:1 stoichiometry and an equilibrium constant (K) of 900 ± 100 L/mol. In basic medium (pH 11.5), K decreases to 240 ± 90 L/mol. Molecular modeling points out that the inclusion complex is formed preferentially via entry of the fisetin phenyl group into b-CD. On the other hand, the fisetin - g-CD has a stoichiometry of 1:1 in acid/neutral solutions and of 1:2 in basic conditions. The K values are 94 ± 30 e 130 ± 10 (L/mol), respectively. The 7-hydroxyflavone can only form inclusion complexes with b-CD. The stoichiometry is 1:1 and there is no dependence on pH. Both equilibrium constants determined either in acid or basic medium is very similar to each other, 1430 ± 510, and 1220 ± 165 L/mol, respectively. For this reason, we suppose that the inclusion of this compound into b-CD is also through the phenyl ring of the flavonoid.

The use of treated waste water for irrigation of vegetable crops is on the increase in Botswana especially in the Glen Valley farms, a peri-urban settlement of Gaborone city. However, the effects of this practice on heavy metals uptake by vegetable crops are uninvestigated. Chromium and nickel have been reported to be accumulating in Gaborone crop soils and cultivating vegetables in these soils with treated waste water could potentially lead to an increased bio-availability of the heavy metals in the vegetable crops. The main aim of this study was therefore to compare the uptake of chromium and nickel in tomato plants, a vegetable grown in sludge amended Glen Valley soils, to those grown in sludge absent Glen Valley soils using treated waste water at different pH values and tap water for irrigation. The high water uptake and high water consumption rate of tomato plants made it suitable for this study. Twenty five pots each containing 2.5 kg sludge amended Glen Valley soils and 5 pots each containing 2.5 kg sludge absent soils were utilized. Fresh treated waste water in a 50 L plastic container on a need by need basis was used. For the control experiments 5 pots each containing 2.5 kg standard commercial soils and fresh tap water were used. The potted tomato plants were cultivated from early May to middle of October 2009. One leaf and one fruit from each tomato plant was harvested and tested in this study. The highest uptakes of chromium (0.819 mg/L) and nickel (0.327 mg/L) were experienced in the leaves where the tomato plant were cultivated in standard commercial soil and irrigated with tap water at pH 7.0. The least uptake of chromium (0.052 mg/L) and that of nickel (-0.030 mg/L) was found in the fruits, where the tomatoes were grown in sludge amended Glen Valley soil and irrigated with normal Glen Valley treated waste water at pH 8.5. Increasing the pH of the treated waste water from 5.0 to 6.0 caused increased bio-accumulation of chromium and nickel in the leaves and the fruits of the tomato plants. Normal treated waste water (pH 8.5) and treated waste water at pH 9.0, however, reduced the chromium and the nickel uptake by the tomato plants. Treated waste water at pH 10.0 bio-accumulate more chromium and more nickel in the leaves and fruits of tomato plants. The pH variation experiments suggested that the fruit tissues accumulated more chromium and the leaf tissues accumulated more nickel. The mean chromium uptake in the tomato plants exceeded the Food and Agriculture Organization permissible limits but the Botswana Bureau of Standards effluent limit was not exceeded. The mean nickel concentrations were below the threshold limits for both local and international standards. Statistical analysis showed no significant difference between the mean chromium and the mean nickel concentration in the leaves and the fruits of the tomatoes at the 5% significant level. It can be concluded from this study that cultivating tomatoes with sludge amended Glen Valley soil combined with normal treated waste water at pH 8.5 could reduce the uptake of chromium and nickel uptake in tomato plants. However, an increase in the uptake of chromium and nickel in the leaves and fruits of the tomato plants could be triggered at slightly low pH (pH 5.0 and pH 6.0) and high pH (pH 10.0) of the treated waste water. It is recommended that the current practices of using treated waste water combined with sludge amended Glen Valley soil to cultivate tomatoes at the Glen Valley farm is good practice and should be continued. Nonetheless, further studies need to be carried out at the farm to establish possible phytotoxicity effects of these heavy metals on tomatoes when using treated waste water combined with sludge amended and sludge absent soils. / Dissertation (MSc)--University of Pretoria, 2011. / Chemical Engineering / unrestricted

Ao ser excitado do estado fundamental para o estado singlete excitado, o Pka do equilíbrio ácido-base das antocianinas apresenta um deslocamento de ca. 4 para ca. -1. Esta característica de superfotoácido, aliado à técnica de fotólise por pulso de laser, permite a perturbação da posição do equilíbrio ácido-base no estado fundamental. O acompanhamento da cinética de reestabelecimento do equilíbrio inicial, existente antes do pulso do laser, permite a determinação das constantes de desprotonação kd e protonação kp no estado fundamental. Este método foi aplicado para a determinação das constantes de protonação e desprotonação das seguintes antocianinas sintéticas e naturais: 4-metil-7-hidroxiflavílio (HMF), 4\', 7-dihidroxiflavílio (DHF), malvidina-3-glucosídeo (Oenina), malvidina-3,5-diglucosídeo (Malvina), pelargonidina-3,5-diglucosídeo (Pelargonina) e cianidina-3 ,5-diglucosídeo (Cianina). Assim, as constantes de desprotonação do estado fundamental são: kd = 1,4 x 106 s-1 (HMF), 3,1 x 106 s-1 (DHF), 5, 1 x 106 s-1 (Oenina), 3,8 x 106 s-1 (Malvina), 1,3 x 106 s-1 (Pelargonina) e 1,8 x 106 s-1 (Cianina). As constantes de protonação do estado fundamental são:kp = 3,56 x 1010 L mol-1 s-1 (HMF), 3,06 x 1010 L mol-1s-1 (DHF), 2,51 x 1010 L mol-1s-1 (Oenina), 2,9 x 1010 L mol-1s-1 (Malvina), 3,6 x 1010 L mol-1s-1 (Pelargonina) e 2,2 x 1010 L mol-1s-1 (Cianina). Nosso método é a única técnica que permite a determinação direta destas constantes. Este método também permite a determinação de kd e kp em meios micro-heterogêneos. Através de medidas de kd , foi possível provar que ocorre a estabilização do cátion flavílio de HMF em micelas aniônicas de SOS (dodecilsulfato de sódio). Em micelas não iônicas e catiônicas, por outro lado, ocorre a desestabilização do cátion flavílio, que se manifesta através do aparecimento de reatividade ausente em solução aquosa. Os resultados da aplicação deste método revelam ainda que o principal processo de dissipação da energia da luz, absorvida pelas antocianinas, é a transferência de próton no estado excitado e não a fluorescência. / Upon excitation from the ground state to the first excited singlet state, the pKa of the acid-base equilibrium of anthocyanins exhibits a large shift, from ca. 4 to ca. -1. This characteristic of super-photoacidity, coupled with the technique of laser flash photolysis, can be employed to perturb the position of the acid-base equilibrium in the ground state. Monitoring the kinetics of relaxation back to the initial equilibrium position that existed prior to the laser pulse permits determination of the rate constants for protonation (kp) and deprotonation (kd) in the ground state. This method was applied to the following natural and synthetic anthocyanins: 4-methyl-7-hydroxyflavylium (HMF); 4\', 7-dihydroxyflavylium (DHF); malvidine-3-glucoside (Oenin); malvidine-3,5-diglucoside (Malvin); Pelargonidine-3,5-diglucoside (Pelargonin); and. cyanidine-3,5-diglucoside (Cyanin). Thus, the ground state deprotonation rate constants (kd) for the acid form were found to be: 1.4 x 106 s-1 (HMF); 3.1 x 106 s-1 (DHF); 5.1 x 106 s-1 (Oenin); 3.8 x 106 s-1 (Malvin); 1.3 x 106 s-1 (Pelargonin); 1.8 x 106 s-1 (Cyanin). The corresponding rate constants for protonation of the ground state of the base (kp) were: 3.6 x 1010 M-1s-1 (HMF); 3.1 x 1010 M-1s-1 (DHF); 2.5 x 1010 M-1s-1 (Oenin); 2.9 x 1010M-1s-1 (Malvin); 3.6 x 1010 M-1s-1 (Pelargonin); 2.2 x 1010 s-1 (Cyanin). Our method is currently the only one that permits direct determination of these rate constants. This method also allows the determination of kpand kd in microheterogeneous media. Thus, employing measurements of kd, it was possible to demonstrate that anionic SDS (sodium dodecyl sulfate) micelles stabilize the flavylium cation form of HMF. In cationic and nonionic micelles, however, the flavylium cation is destabilized, which manifests itself via the appearance of reactivity that is absent in aqueous solution in the absence of these surfactants. Finally, the results of the application of this method show that the primary energy wasting process for radiant energy absorbed by anthocyanins is excited state proton transfer rather than fluorescence.

Ao ser excitado do estado fundamental para o estado singlete excitado, o Pka do equilíbrio ácido-base das antocianinas apresenta um deslocamento de ca. 4 para ca. -1. Esta característica de superfotoácido, aliado à técnica de fotólise por pulso de laser, permite a perturbação da posição do equilíbrio ácido-base no estado fundamental. O acompanhamento da cinética de reestabelecimento do equilíbrio inicial, existente antes do pulso do laser, permite a determinação das constantes de desprotonação kd e protonação kp no estado fundamental. Este método foi aplicado para a determinação das constantes de protonação e desprotonação das seguintes antocianinas sintéticas e naturais: 4-metil-7-hidroxiflavílio (HMF), 4\', 7-dihidroxiflavílio (DHF), malvidina-3-glucosídeo (Oenina), malvidina-3,5-diglucosídeo (Malvina), pelargonidina-3,5-diglucosídeo (Pelargonina) e cianidina-3 ,5-diglucosídeo (Cianina). Assim, as constantes de desprotonação do estado fundamental são: kd = 1,4 x 106 s-1 (HMF), 3,1 x 106 s-1 (DHF), 5, 1 x 106 s-1 (Oenina), 3,8 x 106 s-1 (Malvina), 1,3 x 106 s-1 (Pelargonina) e 1,8 x 106 s-1 (Cianina). As constantes de protonação do estado fundamental são:kp = 3,56 x 1010 L mol-1 s-1 (HMF), 3,06 x 1010 L mol-1s-1 (DHF), 2,51 x 1010 L mol-1s-1 (Oenina), 2,9 x 1010 L mol-1s-1 (Malvina), 3,6 x 1010 L mol-1s-1 (Pelargonina) e 2,2 x 1010 L mol-1s-1 (Cianina). Nosso método é a única técnica que permite a determinação direta destas constantes. Este método também permite a determinação de kd e kp em meios micro-heterogêneos. Através de medidas de kd , foi possível provar que ocorre a estabilização do cátion flavílio de HMF em micelas aniônicas de SOS (dodecilsulfato de sódio). Em micelas não iônicas e catiônicas, por outro lado, ocorre a desestabilização do cátion flavílio, que se manifesta através do aparecimento de reatividade ausente em solução aquosa. Os resultados da aplicação deste método revelam ainda que o principal processo de dissipação da energia da luz, absorvida pelas antocianinas, é a transferência de próton no estado excitado e não a fluorescência. / Upon excitation from the ground state to the first excited singlet state, the pKa of the acid-base equilibrium of anthocyanins exhibits a large shift, from ca. 4 to ca. -1. This characteristic of super-photoacidity, coupled with the technique of laser flash photolysis, can be employed to perturb the position of the acid-base equilibrium in the ground state. Monitoring the kinetics of relaxation back to the initial equilibrium position that existed prior to the laser pulse permits determination of the rate constants for protonation (kp) and deprotonation (kd) in the ground state. This method was applied to the following natural and synthetic anthocyanins: 4-methyl-7-hydroxyflavylium (HMF); 4\', 7-dihydroxyflavylium (DHF); malvidine-3-glucoside (Oenin); malvidine-3,5-diglucoside (Malvin); Pelargonidine-3,5-diglucoside (Pelargonin); and. cyanidine-3,5-diglucoside (Cyanin). Thus, the ground state deprotonation rate constants (kd) for the acid form were found to be: 1.4 x 106 s-1 (HMF); 3.1 x 106 s-1 (DHF); 5.1 x 106 s-1 (Oenin); 3.8 x 106 s-1 (Malvin); 1.3 x 106 s-1 (Pelargonin); 1.8 x 106 s-1 (Cyanin). The corresponding rate constants for protonation of the ground state of the base (kp) were: 3.6 x 1010 M-1s-1 (HMF); 3.1 x 1010 M-1s-1 (DHF); 2.5 x 1010 M-1s-1 (Oenin); 2.9 x 1010M-1s-1 (Malvin); 3.6 x 1010 M-1s-1 (Pelargonin); 2.2 x 1010 s-1 (Cyanin). Our method is currently the only one that permits direct determination of these rate constants. This method also allows the determination of kpand kd in microheterogeneous media. Thus, employing measurements of kd, it was possible to demonstrate that anionic SDS (sodium dodecyl sulfate) micelles stabilize the flavylium cation form of HMF. In cationic and nonionic micelles, however, the flavylium cation is destabilized, which manifests itself via the appearance of reactivity that is absent in aqueous solution in the absence of these surfactants. Finally, the results of the application of this method show that the primary energy wasting process for radiant energy absorbed by anthocyanins is excited state proton transfer rather than fluorescence.