KINETIC AND SPECTROSCOPIC FEATURES OF SOME CAROTENOID TRIPLET STATES: SENSITIZATION BY SINGLET OXYGEN

Abstract— Triplet-triplet absorption spectra of a series of carotenoid pigments in benzene solution have been determined by pulse radiolysis experiments. The natural lifetimes in deaerated solution have also been measured. They fall in the range 2–10 µ s as found for other carotenoids under similar conditions. Pulsed laser (337 nm) excitation of benzene solutions containing oxygen, carotenoid and a photosensitized molecule (anthracene) showed the generation of absorption spectra of the triplet states. These absorptions decayed by first order kinetics in such a way as to indicate that they were formed in reactions with singlet oxygen, itself generated by interaction with the anthracene triplet state. Bimolecular rate constants for energy transfer from O*₂ (¹△_g), to carotenoid have been evaluated.     

PHOTOOXIDATION OF 8-METHOXYPSORALEN WITH SINGLET OXYGEN*

Abstract— The in vitro photooxidation of 8-methoxypsoralen (8-MOP) with singlet oxygen is studied. Irradiation of 8-MOP(295–400 or320–400 nm) in the presence of oxygen for 72 h results in the formation of a product (1.4%) which is identified as 6-formyl-7-hydroxy-8-methoxycoumarin by aid of IR, NMR, MS and co-chromatography with an authentic sample. A study of this reaction in the presence of l,4-diazobicyclo(2,2,2)octane, a singlet oxygen scavenger, indicates the involvement of ¹O₂ in the formation of this compound. In addition to this, formation of a novel dimer of 8-MOP is reported.     

PHOTOCHEMISTRY OF cis-POLYISOPRENE AND ITS SINGLET OXYGEN ADDUCT

Abstract— Reaction of singlet oxygen (¹Δ_g, ¹O₂) with cis -polyisoprene yields an allylic hydroperoxide with an olefinic double bond shifted in the polymer chain. The photochemical decomposition of the resultant hydro-peroxide and the subsequent polymer chain scission kinetics have been studied in the absence of oxygen. Quantum yields of hydroperoxide decomposition range from 3.1 to 8.4 in cyclohexane, depending on the initial amount of hydroperoxide in the polymer. On the other hand, the quantum yields for polymer chain scission are low, and vary with the frequency of the incident light. The ratio for number of polymer scissions per number of hydroperoxy groups decomposed is of the order of 10^-2. The polymer chain degradation is sensitized by the addition of ketones. Based on these data, a reaction mechanism for the overall photodegradation of the cis -polyisoprene initiated by singlet oxygen is proposed.     

PHOTOOXIDATION OF A REFINED PETROLEUM OIL: INHIBITION BY β-CAROTENE AND ROLE OF SINGLET OXYGEN

Abstract— β-Carotene, at concentrations 0.1 m M , inhibits the formation of hydroperoxides and other oxidation products in a refined petroleum oil exposed to Pyrex-filtered UV. The effect appears to be due to ¹O₂ quenching. A mechanism incorporating ¹O₂ and radical processes is proposed as a model for environmental photooxidation of petroleum.     

PHOTOOXIDATION OF EPINEPHRINE SENSITIZED BY METHYLENE BLUE—EVIDENCE FOR THE INVOLVEMENT OF SINGLET OXYGEN AND OF SUPEROXIDE

Abstract— The photooxidation of epinephrine, sensitized by methylene blue or by chlorophylls, excited with red light, involves the reduction of two molecules of oxygen to hydrogen peroxide per molecule of epinephrine oxidized to adrenochrome. The initial rates of these reactions are not affected by low concentrations of catalase. In 99 mol % D₂O, rates of methylene blue sensitized photooxidations are accelerated as much as 5.2 times over rates in ordinary water. Azide anion is a more effective inhibitor of this reaction in D₂O than in H₂O. Half maximal inhibitions are obtained by 1.3 mM azide in H₂O and by 0.1 mAf azide in D₂O. Isotope effects and azide sensitivities point to photooxidation of epinephrine in D₂O primarily by a singlet oxygen pathway; in H₂O, non-singlet oxygen pathways become more predominant. Superoxide dismutase (SOD) markedly inhibits rates of the photooxidations in H₂O and in D₂O; about 25% at 10^-9 M SOD, and 50% at 10^-6 M SOD in H₂O. In the photooxidation in H₂O, both by non-singlet and singlet oxygen mechanisms, the amount of superoxide produced is equivalent to the amount of O₂ consumed in the photooxidation of epinephrine; the superoxide thus formed participates in the oxidation of epinephrine.     

THE ROLE OF SINGLET OXYGEN IN THE PHOTOOXIDATION OF POLYMERS

Abstract— This paper is a critical review of the singlet oxygen oxidation of polymers in solid state and in solution, referring in particular to polydienes, polystyrene and polyvinyl chloride). The singlet oxygenation of polydienes resulted in formation of allylic hydroperoxide groups with shifted double bonds, according to the "ene"-type process. The singlet oxygenation of polystyrene and polyvinyl chloride) occurs only when the new double bonds are formed in these polymers. During dye-photosensitized singlet oxygenation of polydienes in methanol-benzene solution, a very rapid decrease in the molecular weight was observed. For the chain-scission which occurs, not only singlet oxygen but several intermediates such as radicals, bi-radicals and cation-radicals which are formed during light fading of dyes are responsible. At the end of this paper a short review appears which has been focused on the quenching behavior of stabilizers, particularly interactions with singlet oxygen.     

SINGLET OXYGEN FORMATION BY SENSITIZATION OF FUROCOUMARINS COMPLEXED WITH, OR BOUND COVALENTLY TO DNA

Abstract— The formation of singlet molecular oxygen (¹O₂) by sensitization of the furocoumarins 5-methoxypsoralen (5-MOP), 8-methoxypsoralen (8-MOP) and psoralen complexed with DNA was investigated. From the results it is concluded that 5-MOP complexed with native DNA is able to generate ¹O₂, even in a larger extent than 5-MOP free in solution. Also, with 8-MOP and especially with psoralen, ¹O₂ formation by the complexed compound could be observed. The ¹O₂ formation sensitized by covalently bound furocoumarin was demonstrated with psoralen as a model compound. 4',5'-Dihydropsoralen, a model compound for the UVA light absorbing 4',5'monoadducts of furocoumarins to DNA, is also able to generate ¹O₂.     

SINGLET MOLECULAR OXYGEN QUENCHING BY SATURATED AND UNSATURATED FATTY-ACIDS AND BY CHOLESTEROL

The rate constants of molecular singlet oxygen quenching by saturated and unsaturated fatty-acids and by cholesterol-membrane critical components - membrane critical components - have been measured by time resolved detection of the 1270 nm phosphorescence of singlet molecular oxygen [O2(1deltag)]. We have determined (i) an increment of 5.7 x 10(2)M(-1)s(-1) per -CH2- in C6D6 and CD3OD for saturated fatty acids between C4 and C20, (ii) an increment of 3 x 10(4)M(-1)s(-1) per non-conjugated cis-double bond for C18 unsaturated fatty acids, identical in C6D6 and DC3OD, (iii) a lower quenching rate constant by a factor of 2.7 for the trans-C16 and trans-C18 as compared to the corresponding cis-monounsaturated fatty acids, (iv) a rate constant of O2x(1deltag) quenching by cholesterol of 5.7 x 10(4)M(-1)s(-1) in benzene. These rate constants are compared to those obtained for other membrane cellular components.     

QUENCHING OF SINGLET MOLECULAR OXYGEN BY PHTHALOCYANINES AND NAPHTHALOCYANINES

Using the direct measurement of the photosensitized luminescence of singlet molecular oxygen (1O2) the rate constants (kq) have been determined for 1O2 quenching by the monomeric molecules of the following phthalocyanines and naphthalocyanines in chloroform: tetra-(4-tert-butyl) phthalocyanine (I); octa-(3,6-butoxy) phthalocyanine (II), tetra-(6-tert-butyl)-2,3 naphthalocyanine (III), aluminium tetra-(1-tert-phenyl)-2,3 naphthalocyanine (IV), tri-(n-hexyl-siloxy) derivatives of silicon- (V), tin- (VI), aluminium- (VII) and gallium- (VIII) 2,3 naphthalocyanine. The following kq values were obtained (kq x 10(-8) M-1 s-1): 2.9 (I), 59 (II), 100 (III), 20 (IV), 3.9 (V), 53 (VI), 33 (VII), 110 (VIII). As most of the quenchers have the low-lying triplet levels, a contribution of the quenching mechanism based on the energy transfer from 1O2 to these levels has been analysed. A formula is proposed describing the relation between kq values caused by this mechanism, and photophysical constants of the quencher triplet state. This formula was applied to phthalocyanines, naphthalocyanines, beta-carotene and bacterochlorophyll a. The data suggest that the energy transfer can fully explain the activity of V and strongly contributes into the activities of II, III and VI-VIII. A charge transfer interaction might be an additional mechanism involved in 1O2 quenching by compounds studied. As some phthalocyanines and naphthalocyanines are strong physical quenchers of singlet oxygen they can be used as efficient inhibitors for photodestructive processes in photochemical systems.     

THE ROLE OF SINGLET OXYGEN IN PHOTOOXIDATION OF EXCITABLE CELL MEMBRANES

Abstract— Giant nerve axons from lobsters were photooxidized by visible light from a Xe arc using eosin Y, rose bengal and methylene blue as sensitizers. The degree of modification was assayed as the rate constant for block of sodium channels during illumination. Protection or enhancement of modification was tested for several reagents known to interact with singlet oxygen. Deuterium oxide enhanced the modification up to 57% while sodium azide protected up to 60%. β -Carotene, triethylamine and DABCO exerted no effect. From a consideration of the probable low polarity environment of the sensitization sites and diffusional restrictions on reagents it is concluded that intramembranously created singlet oxygen plays a significant role in membrane modification.     

PHOTOOXIDATION OF CHLOROPHYLL AND PHEOPHYTIN. QUENCHING OF SINGLET OXYGEN AND INFLUENCE OF THE MICELLAR STRUCTURE

Abstract— When chlorophyll(Chl) and pheophytin(Phn) are irradiated in Triton X-100 water binary solvents, singlet oxygen is formed in the medium in a higher yield for Phn than for Chi. Chlorophyll shows an irreversible photooxidation reaction and a chemical oxidation reaction when 1,3-diphenyliso-benzofuran (DPBF) is added to the solution. During the chemical oxidation, Chi is destroyed by an oxidizing agent that is a reaction product of the endoperoxide formed in the medium by the addition of singlet oxygen to DPBF. This reaction depends on the structure of the medium and has some characteristics of an oxidation by hydroxyl radicals. The highest yield is obtained with the micellar structure. Chlorophyll and Phn are readily oxidized by hydroxyl radicals generated using the Fenton reagent. This suggests that in the presence of Triton X-100, the Mg²⁺ ion of a Chi molecule plays a key role in the irreversible oxidation of the pigment.     

THE QUENCHING OF SINGLET OXYGEN BY AMINO ACIDS AND PROTEINS

Abstract— The physical quenching of singlet molecular oxygen (¹Δ_g) by amino acids and proteins in D₂O solution has been measured by their inhibition of the rate of singlet oxygen oxidation of the bilirubin anion. Steady-state singlet oxygen concentrations are produced by irradiating the oxygenated solution with the 1–06 μm output of a Nd-YAG laser, which absorbs directly in the electronic transition ¹Δ_g+ 1 v →³Σ_g^-. The rate of quenching by most of the proteins studied is approximated by the sum of the quenching rates of their amino acids histidine, tryptophan and methionine, which implies that these amino acids in the protein structure are all about equally accessible to the singlet oxygen. The quenching constants differ from those obtained by the ruby-laser methylene-blue-photosensitized method of generating singlet oxygen, or from the results of steady-state methylene-blue-photosensitized oxidation, where singlet oxygen is assumed to be the main reactive species. The singlet oxygen quenching rates in D₂O, pD 8, are (10⁷ℒ mol^-1 s^-1): alanine 0–2, methionine 3, tryptophan 9, histidine 17, carbonic anhydrase 85, lysozyme 150, superoxide dismutase 260, aposuperoxide dismutase 250.     

QUENCHING OF SINGLET OXYGEN BY HUMAN PLASMA

Direct measurements of the decay of singlet oxygen phosphorescence at 1270 nm were made in human plasma diluted with various amounts of deuterium oxide. The Stern-Volmer plot of the singlet oxygen lifetimes was linear up to 15% plasma concentration (vol/vol). Extrapolation of these measurements to 100% plasma content gave a singlet oxygen lifetime of 1.04 +/- 0.03 microseconds in human plasma. Biological molecules accounted for 77% of the total singlet oxygen quenching while water accounted for 23% of the quenching. The contributions of various types of biological molecules to the total singlet oxygen quenching were calculated from their plasma concentrations and their quenching constants. Plasma proteins quenched most of the singlet oxygen. Uric acid also quenched a significant amount of singlet oxygen (12%). Tocopherols, carotenoids, ascorbic acid and bilirubin made only small contributions to the total singlet oxygen quenching (less than or equal to 4%).     

SINGLET OXYGEN GENERATION BY FUROCOUMARINS: EFFECT OF DNA AND LIPOSOMES

The quantum yield of singlet oxygen generation by aqueous furocoumarins was measured at 365 nm using the photosensitized inactivation of subtilisin Carlsberg as the probe with the following results: psoralen (0.18), 5-methoxypsoralen (0.013), and 8-methoxypsoralen (0.035). Singlet oxygen formation was significant for dark complexes of 8-MOP with calf thymus DNA and the covalent DNA photoadducts. Incorporation of 8-MOP in sonicated egg phosphatidylcholine liposomes did not inhibit photosensitization of subtilisin Carlsberg and also led to lipid peroxidation, with positive tests for the involvement of singlet oxygen. Peroxidation of the liposomes was inhibited by the presence of α-tocopherol and promoted by the presence of cholesterol in the membranes.     

DIRECT DETECTION OF SINGLET OXYGEN SENSITIZED BY HAEMATOPORPHYRIN AND RELATED COMPOUNDS

Abstract— Direct time-resolved detection of the luminescence at 1270 nm from 'singlet oxygen' was used to estimate the quantum yield of singlet oxygen production (Φ_Δ) from a series of related porphyrins in benzene and in D₂O. From this and available data the fraction of oxygen quenching interactions leading to singlet oxygen production (S_Δ) was derived in most cases. A marked increase in Φ_Δ value was observed for di-haematoporphyrin ester (DHE) in cetyltrimethyl ammonium bromide/D₂O solution in comparison to D₂O alone, this increase is attributed to a major structural alteration of DHE on introduction of the detergent.     

PRODUCTS AND RELATIVE REACTION RATES OF THE OXIDATION OF TOCOPHEROLS WITH SINGLET MOLECULAR OXYGEN

Abstract— The relative reactivity of singlet molecular oxygen, 0₂(¹Δ_g), α-,β-,Γ-and δ with -tocopherol (vitamin E) was investigated using microwave discharge generation as a uniquely clean source of singlet oxygen and using a hydrocarbon solvent to approximate the membrane environment. The relative efficiencies of the tocopherols for O₂(¹Δg) were found to decrease in the order: D-α-tocopherol > D-β-tocopheroI > D–Γ-tocopherol > D-δ-tocopherol. The reaction products in all cases were found to be mixtures of quinone and quinone epoxides apparently resulting from decomposition of the primary product, the hydroperoxydienone.     

BIOLOGICAL ACTIVITIES OF PHTHALOCYANINES—VI. PHOTOOXIDATION OF L-TRYPTOPHAN BY SELECTIVELY SULFONATED GALLIUM PHTHALOCYANINES: SINGLET OXYGEN YIELDS AND EFFECT OF AGGREGATION

Abstract— The photosensitized oxidation of L-tryptophan by gallium phthalocyanines sulfonated to different degrees is studied as a function of both substrate and sensitizer concentrations in water and 50% MeOH-H₂O solutions. The maximum quantum yield of singlet oxygen was found to be nearly 0.5 for all sulfonated gallium complexes. The effect of adding sulfonate groups in the phthalocyanine backbone is to change the tendency of dye molecules to dimerize or aggregate in a particular solvent. A shift in the chemical equilibrium away from the monomeric state, which occurs at high dye concentrations and at lower degrees of dye sulfonation, results in a reduced photochemical yield. The variation of quantum yields in different solvent systems and at several wavelengths is similarly accounted for by the fraction of light absorbed by the productive monomer state.     

ACTIVATED OXYGEN: SINGLET MOLECULAR OXYGEN AND SUPEROXIDE ANION

Abstract— Elusive processes associated with molecular oxygen in chemical and biological systems are interpreted in terms of two activated oxygen species, singlet molecular oxygen (¹Σ⁺_g/¹Δ_g) and superoxide anion (X²π_g). The generation and deactivation of singlet oxygen by interaction with organic triplet states are discussed within a comprehensive theoretical framework. Experimental results indicate the anomalous molecular oxygen enhanced luminescence from organic chromophores in polymer matrices results from the deactivation of singlet (¹Δ_g) oxygen by energy transfer to electronically excited states of the chromophore, and three types of oxygen enhanced luminescence have been identified in these systems. Properties of the superoxide anion relevant to its solution chemistry are briefly discussed. Electron transfer theory is used to theoretically examine the generation of singlet oxygen in disproportionation reactions of the superoxide anion, predicting that, depending on the number of water molecules present, the disproportionation reaction is a proficient source of singlet oxygen. A competing quenching process imposes a limit to the steady state concentration of singlet oxygen in most chemical systems. Available experimental results on the quenching of singlet oxygen by superoxide anion are in good agreement with theoretical results obtained via application of electron transfer theory.     

EPR STUDIES ON SINGLET OXYGEN PRODUCTION BY PORPHYRINS

Abstract— EPR studies of the porphyrin-sensitized photooxidation of 2, 2, 6,6-tetramethyl-piperidine to the nitroxide demonstrate that all the porphyrins examined are able to generate ¹O₂, although the efficiency of the photoprocess is dependent on the nature of the side chains. Incorporation of metal ions into the porphyrin molecule depresses or even inhibits the formation of ¹O₂. Comparison of these results with previously obtained kinetic data points out that the efficiency of porphyrins as photosensitizes is controlled by the lifetime of their lowest triplet state.     

COMPETITION BETWEEN THE SINGLET OXYGEN AND ELECTRON TRANSFER MECHANISMS IN THE PORPHYRIN-SENSITIZED PHOTOOXIDATION OF l-TRYPTOPHAN AND TRYPTAMINE IN AQUEOUS MICELLAR DISPERSIONS

Abstract— Kinetic studies of the hematoporphyrin–sensitized photooxidation of l -tryptophan and tryptamine at pH 10 in either homogeneous aqueous solutions or in aqueous dispersions of Triton X–100 and cetyltrimethylammonium bromide micelles indicate that the indole substrates are attacked via a mixed type I (electron transfer from triplet dye)/type II (¹O₂-involving) mechanism. Both reactive intermediates, generated by micelle-solubilized hematoporphyrin, can diffuse to attack substrate molecules located in either the bulk aqueous phase or a different micelle. In particular, incorporation of the substrate into a micelle has only minor effects on its reactivity toward¹O₂, although the ¹O₂—indole interaction appears to be more efficient in cationic micelles owing to a favourable orientation of the target with respect to the attacking species. On the other hand, the electron transfer from triplet porphyrin to a micellized substrate is virtually non-operative when the latter is located in an anionic micelle, whereas in neutral or cationic micelles, the efficiency of the process is again controlled by the substrate orientation. Studies of tryptamine photooxidation sensitized by meso-tetra-(4-sulfonato-phenyl) porphine in the presence of sodium dodecylsulphate micelles lend further support to the abovementioned conclusions.