ON THE LUMINESCENCE OF L-TRYPTOPHANE AND l-TYROSINE IN AQUEOUS SOLUTION AT 77dK INDUCED BY X-RAYS AND U.V.-LIGHT

Abstract— The lumincscence arising from L-tryptophane and L-tyrosine in aqueous solutions at 77K during irradiation with u.v.-light and with X-rays has been studied. The spectra obtained with the two types of radiation were largely similar, differing only in that the yields of phosphorescence relative to fluorescence were considerably enhanced in the case of X-irradiation. The decay times observed for the exponentially decaying phosphorescence, being 6.6 sec and 2.7 sec for tryptophane and tyrosine respectively, were the same for both kinds of irradiation. The G-value of the X-ray induced luminescence was about 10 for both tryptophane and tyrosine. Thus, about 30 per cent of the total energy absorbed from X-rays in these compounds was re-emitted as light.
It was concluded that the X-ray induced fluorescence and phosphorescence originate from the same levels as does the luminescence caused by u.v.-light, i.e. the lowest excited singlet and the lowest triplet level of the aromatic structure of these compounds. In the case of X-irradiation the enhanced ratios between the yields of phosphorescence and fluorescence indicated that some process other than excitation directly from the ground state contributed considerably to the luminescence yields. Assuming this process to be a recombination between the ionized molecule and its electron, it was calculated that the contribution to the luminescence yield from excitations directly from the ground state relative to that from ionizations, was negligible for both compounds.     

BIPHOTONIC PROCESSES IN PROTEINS: COMPARATIVE STUDIES OF DELAYED FLUORESCENCE AND ISOTHERMAL OR OPTICALLY STIMULATED RECOMBINATION LUMINESCENCE

Abstract —The properties of delayed fluorescence (DF) of proteins are studied in parallel with their properties of recombination luminescence, produced after ultraviolet excitation and photoionization of chromophoric residues at low temperature, in water-ethylene glycol glasses. It is shown that the DF emission of proteins, originating from tryptophan residues, is produced via triplet-triplet absorption processes, photoionization of tryptophan residues, and recombination of the charged species. Electron scavengers that reduce the DF yield act differently upon the DF emission and the recombination luminescence as a consequence of the protein structure. Influence of energy transfer on the protein DF yield is also emphasized.     

PHOTOPHYSICAL PROCESSES IN TROPOLONE, α-METHOXY-TROPONE AND COLCHICINE

Abstract— The spectral characteristics of the emission observed from tropolone, α-methoxy-tropone and colchicine in EPA at 77 K are reported. Luminescence polarization, lifetime, quantum yield and energy transfer experiments provide evidence that the luminescence observed is S₁→ S₀ fluorescence and that the lowest excited singlet state is π, π* The two compounds α-methoxy-tropone and colchicine have been found to exhibit dual luminescence. No phosphorescence could be detected in these three molecules. The photophysical processes for each molecule are discussed and their respective energy level diagrams are also suggested.     

DELAYED EMISSION OF CHLOROPHYLL a AGGREGATES AND RHODAMINE 6G EMBEDDED IN POLYMER MATRIX*

Delayed luminescence (in the microsecond time range) of the chlorophyll (Chl) a“dry” form as well as hydrated dimers located in a polyvinylalcohol film was measured from room temperature down to 8 K. In the same matrix the delayed luminescence of rhodamine 6G (Rhod) was investigated. The delayed emission both of Chl a and Rhod is probably due to the formation and delayed recombination of a radical pair. It seems that this process occurs without participation of triplet states, as it does not reflect their well-known sensitivity to oxygen. The temperature dependence of the delayed luminescence of vanous Chl forms is different. In the region around 678 nm (dry monomer) delayed luminescence needs a thermal activation energy of about 0.03 eV, whereas at 740 nm (wet aggregates) delayed luminescence intensity increases linearly with decreasing temperature. Its assignment as a-type delayed luminescence from the low-lying triplet state can consistently be excluded from both the weak temperature dependence of the delayed fluorescence and its large intensity as compared to the prompt fluorescence. Delayed luminescence of Rhod is almost independent of temperature between 8 K and 300 K. The dependence of delayed luminescence intensity on exciting light intensity is linear at lower intensities and tends to saturation at higher. Therefore the delayed luminescence is not related to exciton annihilation. Positions and intensities of the Chl delayed luminescence bands show that it is not phosphorescence (β-type delayed luminescence). The aggregation of both Chl and Rhod molecules strongly influences delayed luminescence since it differs in several properties if excited in the monomer or in the aggregate absorption range. Every aggregational form of dye emits its characteristic delayed luminescence band.     

PHOTOSENSITIZATION BY ANTIMALARIAL DRUGS

Abstract The antimalarial drugs, chloroquine, hydroxychloroquine, quinine, quinacrine, amodiaquine and primaquine and the local anaesthetic, dibucaine, were tested for in vitro photosensitizing capability by irradiation with 365 nm UV light in aqueous solutions. The ability of these compounds to photosensitize the oxidation of 2,5-dimethylfuran, histidine, tryptophan or xanthine, and to initiate the free radical polymerization of acrylamide was examined in the pH range 2-12. Chloroquine and hydroxychloroquine show maximal photooxidative behaviour when in the monocation form at pH 9, in contrast to quinine which is extremely efficient as the dication below pH 4. This pattern appears to relate to the fluorescence yield as a function of pH. Chloroquine in the monocation or neutral form was found to undergo dechlorination upon irradiation, and this correlates directly with its ability to initiate photo-polymerization of acrylamide. Quinine also gives rise to small polymerization rates, attributed to photo-ionization in the quinoline ring, yielding a cation radical. Amodiaquine, primaquine and quinacrine do not have significant photochemical activity in aqueous solution. Dibucaine exhibits a strong photosensitizing capability at low pH, similar to quinine.     

FREE RADICALS PRODUCED IN A RIGID SOLUTION OF TRYPTOPHAN BY A BIPHOTONIC PROCESS

Abstract— Free radicals were trapped by irradiation of tryptophan in an ethylene glycol/water glass with 250 nm U.V. light at 77°K. The ESR-spectrum was attributed to approximately equal amounts of trapped electrons and ethyl radicals. The electrons could be transformed into the ethyl type radicals by heating or bleaching with near u.v. or visible light. The quantum yield of trapped radicals increased with increasing intensity of the irradiation by more than one order of magnitude over the range of intensities studied. A kinetic model was formulated on the assumption that the radicals were produced by the successive absorption of two light quanta, the absorption of the first giving rise to a transient intermediate which subsequently absorbs a second quantum to produce the radicals. The experiments confirmed the applicability of this model to the present case. The decay time obtained for the intermediate was 6.8 sec which equals the phosphorescence decay time of tryptophan under similar conditions, indicating that the intermediate involved was the lowest triplet level of this molecule. The absorption cross section of this intermediate at 250 nm with regard to the formation of trapped radicals was found to be 8000 l./mol cm.     

FLUORESCENCE OF TRYPTOPHAN IN KERATIN

The excitation and emission spectra have been determined for the fluorescence from trypto-phan residues in dry keratin. The fluorescence decay was also measured and shown to be a single exponential with a rather long lifetime of 6.9 ns. It is suggested that the emission takes place from a state formed by interaction between the ¹L_a state of the tryptophan residues and neighbouring polar or polarizable groups in the protein. The fluorescence excitation spectrum displays a peak at 290 nm, and its appearance at this position rather than at 280 nm, which is the absorption maximum of tryptophan, is believed to arise from inner filtering by the tyrosine residues in keratin.     

喹哪啶酸合铕/二氧化硅发光材料的溶胶-凝胶法合成与性能

通过红外光谱、紫外可见光谱、荧光光谱和X射线粉末衍射等测试手段,对溶液中合成的喹哪啶酸合铕(Eu-qina)配合物进行了表征,且将溶胶-凝胶法合成出的Eu-qina/SiO2杂化发光材料,与溶液中合成的配合物Eu-qina进行对比.结果表明,配合物Eu-qina中由于喹哪啶酸能有效地将能量传递给Eu3+离子,使Eu3+发出较强的红色特征荧光,而杂化材料Eu-qina/SiO2因SiO2的加入使荧光强度增大.当Eu-qina配合物的掺杂量达到一定值时,Eu-qina/SiO2发较强的红光,并由晶态的Eu-qina转化为非晶态的Eu-qina/SiO2.     

THE EFFECTS OF PORPHYRIN STRUCTURE AND AGGREGATION STATE ON PHOTOSENSITIZED PROCESSES IN AQUEOUS AND MICELLAR MEDIA

The efficiency of several porphyrins at 10 μM and 83 μM as sensitizers of the photooxidation of 0.1 mM tryptophan and histidine via a singlet oxygen-mechanism was studied in pH 7.4-buffered aqueous solutions and in aqueous dispersions of Triton X-100 micelles. The porphyrins were either solubilized in the bulk aqueous medium or associated with the micellar phase, whereas the amino acids were always located in the aqueous phase. With those porphyrins, such as uroporphyrin I, meso-tetra (4-sulfonatophenyl)porphine, meso-tetra(4-carboxyphenyl)porphine and meso-tetra)N,N,N-trimethylanilinium)porphine, which are > 98% monomeric in both media, the efficiency of histidine photooxidation was independent of the site of O₂(¹Δ_g) generation, as shown by the closely similar values for the photooxidation rate constant and oxygen-consumption quantum yield in the presence and absence of Triton micelles; the same indications were provided by photokinetic experiments with tryptophan. Actually, laser flash photolysis studies showed that the micelle-incorporation of the above mentioned porphyrins brought about only minor changes in their photophysical properties, including the relative yield of O₂(¹Δ_g) generation. On the other hand, hematoporphyrin IX, its Zn²⁺-complex, and coproporphyrin III are largely aggregated in homogeneous aqueous solution; their incorporation into Triton micelles caused an increase of the triplet quantum yield and an enhancement of the oxygen-consumption quantum yield and photooxidation rate constant for both histidine and tryptophan. The lower photosensitizing efficiency of aggregated porphyrin species in comparison with the corresponding monomeric porphyrin was confirmed by measuring the initial rate and quantum yield of oxygen consumption upon irradiation of 1 mM histidine and tryptophan in the presence of different hematoporphyrin concentrations within the 0.3-100μM range.     

PHOTOCHEMICAL REACTIONS IN RIGID MEDIA-I. THE EFFECT OF GELATIN ON THE AEROBIC PHOTOCHEMISTRY OF RIBOFLAVIN IN AQUEOUS SOLUTION

Abstract— The addition of potassium iodide to dilute aqueous solutions of riboflavin reduced both the rate of the aerobic photolysis and the fluorescence quantum yield of riboflavin in the same proportions. This indicated that under these conditions the photolysis proceeded from the singlet excited state. The addition of gelatin to aqueous solutions of riboflavin also reduced the rate of the aerobic photolysis but increased slightly the quantum yield of fluorescence. The rates and the fluorescence of solutions of riboflavin to which gelatin had been added were also reduced by the addition of potassium iodide but in this case the effect on the rate was proportionately greater than the effect on the fluorescence. The data suggests that in the presence of gelatin the mechanism of the reaction is changed and that the triplet state becomes more important.     

FEMTOSECOND STUDIES OF HEMATOPORPHYRIN DERIVATIVE IN SOLUTION: EFFECT OF pH AND SOLVENT ON THE EXCITED STATE DYNAMICS

Abstract We report direct femtosecond measurements of the excited state dynamics of hematoporphyrin derivative (HpD) in solution. The dynamics are found to be very sensitive to the solvent and pH of aqueous solutions. The decay of the excited singlet states is much faster in acidic and pH 7 buffer aqueous solutions (<230 ps) than in basic aqueous solutions or organic solvents (> 10 ns). The dynamical results show strong correlation with static fluorescence measurements: weaker fluorescence in acidic and pH 7 buffer solutions corresponding to shorter-lived excited states. A new fast decay component with a time constant around 5 ps is identified both in acidic aqueous solutions and in organic solvents such as acetone and attributed to internal conversion from the second to the first excited singlet state of aggregates or certain oligomers in HpD, in accord with the observation that the fast decay component is larger at a higher concentration. Oxygen is found to have no effect on the dynamics on the time scale investigated, 1 ns, indicating that oxygen quenching of the singlet excited states is insignificant on this time scale. The sensitive solvent and pH dependence of the excited state dynamics has important clinical implications in the use of HpD as a photosensitizing agent.     

PHOTOREDUCTION OF CRYSTAL VIOLET IN SOLUTION AND IN POLY(VINYL ALCOHOL) FILMS

Abstract— The photolysis of crystal violet oxalate has been studied in aqueous and non-aqueous solutions and in films of poly(viny1 alcohol) at room temperature. Evidence has been obtained from electron-spin-resonance, absorption and fluorescence spectroscopy for the formation of the semireduced dye radical, tri-( p -dimethylaminophenyl)methyl, both in solution and in the solid state. Electron spin resonance and luminescence spectra of the radical produced by electrolytic reduction of the dye are also reported.     

PHOTOCHEMISTRY OF URACIL. INTERSYSTEM CROSSING AND DIMERIZATION IN AQUEOUS SOLUTION

Abstract— Ultraviolet irradiation of ¹⁴C-uracil in aqueous solution results in the formation of hydrate and dimer photoproducts. The rate of dimerization increases with increasing uracil concentration, and decreases with increasing concentration of oxygen in solution. The kinetics are in agreement with a model previously proposed to account for the reactions, in which dimerization occurs by a reaction involving the triplet state of uracil, but hydration occurs from an excited singlet state. Oxygen reduces dimer formation by quenching the triplet. The quantum yield for intersystem crossing (ISC) to the triplet depends on the irradiation wavelength, increasing from 0.0014 at 280 nm to 0.016 at 230 nm. The ratio of rate constants for reaction of the triplet with oxygen and for dimerization is 1.1; the ratio of rate constants for triplet decay and for dimerization is 5.9 × 10^-5 M. The increase in ISC with photon energy suggests that ISC is favoured from excited vibrational levels. The quantum yield for hydration is about 0.002 at pH 4.5 for all wavelengths, but increases as the pH is decreased.     

EXCITATION-ENERGY TRANSFER BETWEEN TYROSINE AND TRYPTOPHAN IN PROTEINS EVALUATED BY THE SIMULTANEOUS MEASUREMENT OF FLUORESCENCE AND ABSORBANCE

Abstract— The efficiencies of the excitation–energy transfer from tyrosine to tryptophan residues in eight globular proteins in the native and denatured states are obtained by studying the wavelength dependence of the fluorescence quantum yield. The measurements are made over a wide wavelength range using a computer-controlled spectrophotometer which can measure the fluorescence and absorbance simultaneously in one sample solution (Wada et al. , 1980). The values of the energy transfer efficiencies ranged from 0.17 ± 0.12 to 0.69 ± 0.06 in the native state and from -0.04 ± 0.09 to 0.12 ± 0.06 in the denatured state. These values are considerably lower than the values reported by Kronman and Holmes (1971); in particular, an almost complete absence of energy transfer for the denatured state is shown.     

ON THE MECHANISM OF FLUORESCENCE QUENCHING IN PYRENE- AND 3,4-BENZOPYRENE-1,3,7,9-TETRAMETHYLURIC ACID SYSTEMS

T he interaction between aromatic hydrocarbons and purine or its analogues has been widely noted in connection with the possibility of complex formation between carcinogens and DNA or its precursors. Weil-Malherbe[1], Boyland and Green[2] reported that 1,3,7,9-tetramethyluric acid (TMUA) solubilizes aromatic hydrocarbons in aqueous solution and exerts a strong quenching action on fluorescence of the hydrocarbons. The solubilization has been attributed to formation of a molecular complex [2]. Complex formation is now thought to be mainly due to van der Waal's force (polarization and dispersion force) rather than charge transfer ( CT ) one[2,3–5]. Van Duuren [3,5] examined fluorescence spectra of TMUA-hydrocarbon complexes both in solution and in potassium bromide pellets. In solution, the fluorescence spectrum of the complex showed no change in the position of the maximum and in intensity, when compared with that of the free hydrocarbon. In the solid state, however, a blue shift was observed for the fluorescence of the complex. From these results, it was concluded that the complex is entirely dissociated in solution and the fluorescence shift observed in the solid state is the result of the interaction in the excited state. The present note describes a new observation which is useful for understanding the mechanism of fluorescence quenching in the hydrocarbons-TMUA systems.     

A NEW FLUORESCENT PHOTOPRODUCT OF TRYPTOPHAN EVIDENCED BY LONG WAVELENGTH EXCITATION OF FLUORESCENCE

Abstract— Besides the normal tryptophan (Trp) fluorescence in aqueous solution (emission maximum at 350 nm), a new emission, peaking around 380 nm, appears by long wavelength excitation. Its fluorescence yield (φ_s 0.24) is higher than that of tryptophan (φ_Trp= 0.13). The growth of this emission is observed under different experimental conditions, mainly under UV anaerobic irradiation. To explain this observation, the formation of a C₃-hydroxylated derivative is tentatively suggested.     

CAROTENOID TRIPLET STATE AND CHLOROPHYLL FLUORESCENCE QUENCHING IN CHLOROPLASTS AND SUBCHLOROPLAST PARTICLES

Abstract— Absorption changes attributed to the triplet state of carotenoids and to primary electron donors (P-700. P-680)^: and fluorescence quenching at several wavelengths have been measured with a single apparatus. following flash excitation with a dye or a ruby laser. Spinach chloroplasts as well as subchloroplast particles enriched in Photosystem-1 (F₁), Photosystem-2 (F₁) or the light-harvesting Chl a/h (F_III) have been examined at temperatures varying between 5 and 294 K.
The triplet state of carotenoids has been identified on the basis of its difference spectrum (having a peak at 515 nm) and decay kinetics (⋍ 7 µs at low temperature; accelerated by O₂ at 294 K). It is formed in all of the materials studied. The quantum yield of carotenoid triplet formation in chloroplasts increases at low temperature, but less than the fluorescence yield.
In most cases the fluorescence quenching recovers approximately with the same kinetics as the decay of the carotenoid triplets. The fluorescence recovery is, however, significantly faster for chloroplasts at 730 nm. Fluorescence quenching occurs in all types of materials. The ratio of fluorescence quenching to the concentration of carotenoid triplets varies with the material, being maximum in chloroplasts and minimum in F_m particles.
We conclude that the formation of the carotenoid triplet state is not limited to a few sites in the chloroplast and that a carotenoid triplet is a quencher of chlorophyll fluorescence. A detailed comparison of carotenoid triplets and fluorescence quenching gives some information concerning the organization of the pigments in the photosynthetic apparatus.     

Influence of energy transfer on the photoionization of tryptophan and tyrosine in basic media

Abstract— The luminescence studies of the delayed isothermal and light stimulated recombinations of photoejected electrons with photoionized chromophores at 77°K show that in the peptide tryptophyltyrosine, where energy transfer occurs at the singlet level from tryptophan to tyrosine and at the triplet level from tyrosine to tryptophan, tryptophan photoionization is greatly enhanced. This increase in photoionization efficiency is attributed to triplet-triplet transfer.     

Photomechanical Luminescence from Through‐Space Conjugated AIEgens

Transforming molecular motions into the macroscopic scale is a topic of great interest to nanoscience. The photomechanical effect is a promising strategy to achieve this goal. Herein, we report an intriguing photomechanical luminescence driven by the photodimerization of 2‐phenylbenzo[b]thiophene 1,1‐dioxide (P‐BTO) in molecular crystals and elucidate the working mechanism and substituent effect through crystallographic analysis and theoretical calculations. Striking splitting, hopping, and bending mechanical behaviors accompanied by a significant blue fluorescence enhancement are observed for P‐BTO crystals under UV light, which is attributed to the formation of photodimer 2P‐BTO. Although 2P‐BTO is poorly π‐conjugated because of the central cyclobutane ring, it exhibits prominent through‐space conjugation and aggregation‐induced emission (AIE), affording strong solid‐state blue fluorescence at 415 nm with an excellent quantum yield of up to 96.2 %.     

FORMATION OF OXONIACHLORINS ON PHOTOOXIDATION OF 20-TRIFLUOROACETOXY- AND 20-CHLORO-CHLOROPHYLL DERIVATIVES*

Zinc complexes of both 20-trifiuoroacetoxy- and 20-chloro-pheophorbide-a methyl ester yield the same isolable 20-oxonia-pheophorbide (2b) on irradiation with visible light in methylene chloride for 90 and 10 min, respectively. In the case of the 20-chloro derivative, the yield amounts to40–50% along with 30% of starting material. Zinc complexes of 20-trifluoroacetyl- and 20-chloro-pyropheophorbide-a methyl ester behave likewise. In aqueous solution, the expected dihydro-biliverdin derivatives 2 are obtained among other products, some of them display a strong red fluorescence. In the presence of nucleophiles the oxoniachlorin macrocycle is readily cleaved, yielding the corresponding linear tetrapyrroles. Particularly, reaction with cyanide ions leads to biline derivatives, which could serve as intermediates for the synthesis of natural occurring luciferins from bioluminescent plankton. The possible role of some of the above mentioned reactions in connection with chlorophyll catabolism is discussed.