Else Lemp

Singlet Oxygen Reactions with Flavonoids. A Theoretical – Experimental Study

Detection of singlet oxygen emission, λmax = 1270 nm, following laser excitation and steady-state methods were employed to measure the total reaction rate constant, kT, and the reactive reaction rate constant, kr, for the reaction between singlet oxygen and several flavonoids. Values of kT determined in deuterated water, ranging from 2.4×107 M−1s−1 to 13.4×107 M−1s−1, for rutin and morin, respectively, and the values measured for kr, ranging from 2.8×105 M−1s−1 to 65.7×105 M−1s−1 for kaempferol and morin, respectively, being epicatechin and catechin chemically unreactive. These results indicate that all the studied flavonoids are good quenchers of singlet oxygen and could be valuable antioxidants in systems under oxidative stress, in particular if a flavonoid-rich diet was previously consumed. Analysis of the dependence of rate constant values with molecular structure in terms of global descriptors and condensed Fukui functions, resulting from electronic structure calculations, supports the formation of a charge transfer exciplex in all studied reactions. The fraction of exciplex giving reaction products evolves through a hydroperoxide and/or an endoperoxide intermediate produced by singlet oxygen attack on the double bond of the ring C of the flavonoid.

Naphthoxazole-Based Singlet Oxygen Fluorescent Probes†

In this study, we report the synthesis and photochemical behavior of a new family of photoactive compounds to assess its potential as singlet oxygen (1O2) probes. The candidate dyads are composed by a 1O2 trap plus a naphthoxazole moiety linked directly or through an unsaturated bond to the oxazole ring. In the native state, the inherent great fluorescence of the naphthoxazole moiety is quenched; but in the presence of 1O2, generated by the addition and appropriate irradiation of an external photosensitizer, a photooxidation reaction occurs leading to the formation of a new chemical entity whose fluorescence is two orders of magnitude higher than that of the initial compound, at the optimal selected wavelength. The presented dyads outperform the commonly used indirect fluorescent 1O2 probes in terms of fluorescence enhancement maintaining the required specificity for 1O2 detection in solution.

Protective Effect of Boldo and Tea Infusions on the Visible Light–mediated Pro-oxidant Effects of Vitamin B2, Riboflavin¶

Abstract The effect of Boldo and black tea infusions on the pro-oxidant effects of vitamin B2, riboflavin (RF), when exposed to the action of visible light was studied. The amounts of antioxidants present in Boldo and tea infusions were evaluated by a procedure based on the bleaching of preformed 2,2′-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) radical cations and were expressed as 6-hydroxy-2,5,7,8-tetramethyl-chroman-2-carboxylic acid equivalent concentrations. The quenching rate constants of singlet oxygen (1O2; [kq]Boldo = 6.0 × 107 M−1 s−1 and [kq]Tea = 3.2 × 107 M−1 s−1) and triplet RF (3RF; [3RFkq]Boldo = 10 × 108 M−1 s−1 and [3RFkq]TEA = 3.2 × 108 M−1 s−1) with Boldo and tea were determined by flash photolysis. These data allow a quantitative interpretation of the results obtained. Our data suggest that most of the oxygen consumption observed in the photolysis of RF in the presence of tea and Boldo infusions is caused by 1O2 reactions. The oxygen consumption quantum yield is considerably smaller than the fraction of RF triplets trapped by the additives (AH) present in the infusion, indicating that their interaction with 3RF does not lead to chemical reactions or that the AH·+ radical ions initially formed participate in secondary processes that do not consume oxygen. Boldo and tea infusions have a significant protective effect when a system containing RF and tryptophan (Trp) is exposed to visible light, not only by quenching the 1O2 and interfering with the Type-I mechanism but also by repairing the damage to Trp molecules associated with the latter mechanism.

Interaction of singlet oxygen [O2(1Δ)] with aliphatic amines and hydroxylamines

Singlet oxygen quenching rates for O2(1Δ) by triethylamine, diethylamine, and N,N-diethylhydroxylamine have been measured in several solvents. The data obtained for the amines are compatible with a charge-transfer quenching mechanism; quenching is faster in solvents of high capacity to stabilize charges and low capacity to donate a proton. Quenching by N,N-diethylhydroxylamine shows a different solvent dependence: the rate constant depends upon the availability of the hydroxylic hydrogen. This fact, together with the high yields of hydrogen peroxide produced in the singlet oxygen–diethylhydroxylamine reaction, implies a different quenching mechanism.

Vesicular effect on the reactivity of anthracene derivatives towards singlet molecular oxygen

Abstract The effect of dioctadecyldimethylammonium chloride (DODAC) vesicles upon the reactivity of several anthracene derivatives towards O 2 ( 1 Δ g ) has been measured. Incorporation of the substrates into the vesicles decreases their consumption rate. The effect depends on both the substrate characteristics and the vesicle size. The most pronounced decreases are observed with those substrates for which deep incorporation of the anthryl group into the vesicles could be expected, i.e. , 9,10-dimethylanthracene and 3-(9-anthryl)propionic acid. For these substrates, the reactivity is nearly two times smaller in the large (injected) vesicles than in the small (sonicated) vesicles. The dependence of the bleaching rates on the surfactant concentration allows the evaluation of the substrate distribution between the vesicles and the aqueous solution. For 9-methylanthracene and 9-anthracenemethanol, the partition constants are nearly ten times smaller in the large vesicles than in the smaller vesicles.

Solubilization of lipid bilayers by myristyl sucrose ester: effect of cholesterol and phospholipid head group size

The solubilization of biological membranes by detergents has been used as a major method for the isolation and purification of membrane proteins and other constituents. Considerable interest in this field has resulted from the finding that different components can be solubilized selectively. Certain membrane constituents are incorporated into small micelles, whereas others remain in the so-called detergent-resistant membrane domains that are large enough to be separated by centrifugation. The detergent-resistant fractions contain an elevated percentage of cholesterol, and thus its interaction with specific lipids and proteins may be key for membrane organization and regulation of cellular signaling events. This report focuses on the solubilization process induced by the sucrose monoester of myristic acid, beta-D-fructofuranosyl-6-O-myristyl-alpha-D-glucopyranoside (MMS), a nonionic detergent. We studied the effect of the head group and the cholesterol content on the process. 1-Palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) and dioctadecyl-dimethyl-ammonium chloride (DODAC) vesicles were used, and the solubilization process was followed using Laurdan (6-dodecanoyl-2-dimethylaminonaphthalene) generalized polarization (GP) measurements, carried out in the cuvette and in the 2-photon microscope. Our results indicate that: (i) localization of the MMS moieties in the lipid bilayer depends on the characteristics of the lipid polar head group and influences the solubilization process. (ii) Insertion of cholesterol molecules into the lipid bilayer protects it from solubilizaton and (iii) the microscopic mechanism of solubilization by MMS implies the decrease in size of the individual liposomes.

Sucrose Monoester Micelles Size Determined by Fluorescence Correlation Spectroscopy (FCS)

One of the several uses of sucrose detergents, as well as other micelle forming detergents, is the solubilization of different membrane proteins. Accurate knowledge of the micelle properties, including size and shape, are needed to optimize the surfactant conditions for protein purification and membrane characterization. We synthesized sucrose esters having different numbers of methylene subunits on the substituent to correlate the number of methylene groups with the size of the corresponding micelles. We used Fluorescence Correlation Spectroscopy (FCS) and two photon excitation to determine the translational D of the micelles and calculate their corresponding hydrodynamic radius, Rh. As a fluorescent probe we used LAURDAN (6-dodecanoyl-2-dimethylaminonaphthalene), a dye highly fluorescent when integrated in the micelle and non-fluorescent in aqueous media. We found a linear correlation between the size of the tail and the hydrodynamic radius of the micelle for the series of detergents measured.

Photophysics and photochemistry of naphthoxazinone derivatives

The photophysics and photochemistry of a series of naphthoxazinones have been studied using a combination of methods ranging from steady-state and time-resolved spectroscopic techniques to product analysis. The photophysics of naphthoxazinone derivatives is very dependent on the structure: phenanthrene-like compounds exhibit higher fluorescence quantum yield than the less aromatic anthracene-like homologous. The latter, exhibit a substantial degree of charge transfer in the excited singlet state. These compounds are fairly photostable in the absence of additives, yielding a single photoproduct arising from the triplet state. The presence of electron donors such as amines increases the photoconsumption quantum yield and changes the product distribution, the primary photoproduct being a dihydronaphthoxazinone that photoreacts further yielding ultimately an oxazoline derivative.

Sensitized photoxygenation of piroxicam in neat solvents and solvent mixtures

Detection of O(2)(1Delta(g)) phosphorescence emission, lambda(max)=1270 nm, following laser excitation and steady state methods were employed to determine the total rate constant, k(T), for the reaction between the non-steroidal anti-inflammatory drug piroxicam (PRX) and singlet oxygen in several solvents. Values of k(T) ranged from 0.048+/-0.003 x 10(6) M(-1) s(-1) in chloroform to 71.2+/-2.2 x 10(6) M(-1) s(-1) in N,N-dimethylformamide. The chemical reaction rate constant, k(R), was determined by using thermal decomposition of 1,4-dimethylnaphthalene endoperoxide as the singlet oxygen source. In acetonitrile, the k(R) value is equal to 5.0+/-0.4 x 10(6) M(-1) s(-1), very close to the k(T) value. This result indicates that, in this solvent, the chemical reaction corresponds to the main reaction path. Dependence of total rate constant on the solvent parameters pi* and beta can be explained in terms of a reaction mechanism that involves the formation of a perepoxide intermediate. Rearrangement of the perepoxide to dioxetane followed by ring cleavage and transacylation accounts for the formation of N-methylsaccharine and N-(2-pyridyl)oxamic acid, the main reaction products. Data obtained in dioxane-water (pH 4) mixtures with neutral enolic and zwitterionic tautomers of piroxicam in equilibrium show that the zwitterionic tautomer reacts with singlet oxygen faster than the enolic tautomer.

Singlet Oxygen‐mediated Photobleaching of the Prosthetic Group in Hemoglobins and C‐Phycocyanin

Proteins bearing colored prosthetic groups, such as the heme group in hemoglobin or the bilin group in c‐phy‐cocyanin, quench singlet oxygen by interactions at the apoprotein and the prosthetic group levels. In both proteins, chemical modification of the chromophore constitutes only a minor reaction pathway. While total deactivation of singlet oxygen takes place with rate constants of 4.0 times 109and 4.2 times 108M‐ls‐1for hemoglobin and phycocyanin, respectively, the bleaching of the chromophore takes place with rate constants of 3.2 times 106and ˜1 times 107M‐1s‐1. Irradiation of phycocyanin with red light bleaches the chromophore with low yields (˜0.8 times 10‐4). Part of this bleaching is mediated by singlet oxygen produced by the irradiation of the bilin group. The low relevance of the singlet oxygen pathway is compatible with a low quantum yield (˜10‐3) of free singlet oxygen production after irradiation of the protein.