Slow solvent relaxation dynamics of nanometer sized reverse micellar systems through tryptophan metabolite, kynurenine

Exploration of environmental dynamics using intrinsic biological probe tryptophan is very important; however, it suffers from various difficulties. An alternative probe, kynurenine (KN), has been found to be an efficient probe for the ultrafast dynamics in the biological environment ( 1 ] J. Phys. Chem. B., 114, 15236–15243). In the present study, we have investigated the efficacy of KN for the exploration of relatively slower dynamics of biologically relevant environments. A detailed investigation involving UV–Vis, steady‐state/time‐resolved fluorescence spectroscopy and Förster resonance energy transfer (FRET) studies on KN compared to a well‐known solvation probe, H33258, a DNA‐minor groove binder in a model nonionic reverse micelle reveals that ultrafast internal conversion associated with the hydrogen‐bonding dynamics masks KN to become a dynamical reporter of the immediate environments of the probe.     

Photochemistry of diketones: observation of a triplet state-oxygen adduct

The presence of triplet state-oxygen adducts (a species previously proposed but never observed in a direct manner) is readily observed following laser flash photolysis studies of 2,2'-thenil. We report on the kinetic and spectroscopic parameters characteristic of this transient adduct.     

Factors affecting the ultramicro-determination of tryptophan,kynurenine and their derivatives

The factors affecting the ultramicrodetermination of tryptophan and kynurenine were investigated by paper electrophoresis, paper chromatography and spectrophotometry. The electrolyte used in electrophoresis influences the mobility, the separation and the ultraviolet spectra of tryptophan and kynurenine. The pre-treatment of filter paper for chromatography did not influence the R_F of tryptophan and kynurenine, but interfered with their quantitative determination. The effect of ultraviolet irradiation of d-, l-, and dl-tryptophan as well as d-, l-, and dl-kynurenine was studied.     

Antioxidative properties of nitroxyl radicals and hydroxyamines in reactions with triplet and deaminated kynurenine

The reactions of triplet kynurenine and 4-(2-aminophenyl)-4-oxocrotonic acid, formed upon the thermal decomposition of kynurenine, with nitroxyl radicals and cyclic N-hydroxylamines were studied. Nitroxyl radicals were found to quench efficiently the triplet state of kynurenine (rate constant 3–6·.10⁸ L mol^-1 s^-1). The quenching proceeds via the spin-exchange mechanism and affords no new products. Neither nitroxyl radicals, nor hydroxyl-lamines react with 4-(2-aminophenyl)-4-oxocrotonic acid under conditions similar to physiological.     

Characterization, reactivity and photosensitizing properties of the triplet excited state of alpha-lapachone

alpha-Lapachone is a natural 1,4-naphthoquinone with promising biological activity. The fused dihydropyran ring present in its structure, acting as formal 2-alkoxy and 3-alkyl substituents to the quinone moiety, endows this compound with milder redox properties and lower toxicity, when compared with other bioactive 1,4-quinones. Its photochemistry, here reported, seems to originate from the triplet state, which shows pipi* character. Triplet quenching in acetonitrile solution with added hydrogen-atom donors such as 1,4-cyclohexadiene or 2-propanol is inefficient, independent of solvent polarity, and leads to formation of the semiquinone radical. With phenol and indole, quenching rate constants are two orders of magnitude higher, but smaller than the value for triethylamine. In the first two cases the semiquinone radical can be detected by laser flash photolysis and in the last case, the anion radical derived from alpha-lapachone is readily detected. The semiquinone radical can also be observed in the quenching of triplet alpha-lapachone by 2'-deoxyguanosine and by the methyl esters of L-tryptophan and L-tyrosine, whereas for L-cysteine methyl ester the quenching rate constant is very slow. Triplet alpha-lapachone is not quenched by thymine, thymidine, 2'-deoxycytosine or 2'-deoxyadenosine; this is probably due to its pipi* character and low energy, which prevents oxetane formation and triplet-triplet energy transfer, respectively. Steady-state photolysis of aerated solutions of these compounds in the presence of alpha-lapachone does not show evidence of decomposition, whereas similar experiments with 2'-deoxyguanosine result in efficient consumption of the nucleoside. Singlet oxygen is formed from triplet alpha-lapachone, and a quantum yield of 0.68 is measured.     

Photophysical properties of benzil in solution: triplet state deactivation pathways

The fluorescence and phosphorescence from benzil in dilute benzene and cyclohexane solutions (2 × 10⁻⁴ M) were studied by both conventional luminescence and time-correlated single-photon techniques in the temperature range 8 – 69 °C. The fluorescence (λ = 502 nm) did not show a substantial temperature dependence and was free from thermal and triplet-triplet annihilation delayed contributions at the low concentration used. The phosphorescence (λ = 562 nm) was temperature dependent and its decay was controlled by an activation energy (E_a = 7.4 ± 0.5 kcal mol⁻¹) which was slightly larger than the spectroscopic single-triplet splitting (6.1 kcal mol⁻¹). The photophysical parameters derived from the lifetimes of the two emissions was not consistent with the model of thermal equilibration between S₁ and T₁.     

A theoretical study on the quenching mechanisms of triplet state riboflavin by tryptophan and tyrosine

The reactions of tryptophan (Trp) and tyrosine (Tyr) with endogenous photosensitizer riboflavin (RF) have gained much interest for their crucial roles in various photobiological processes. In this paper, the quenching mechanisms of triplet state RF by Trp and Tyr have been explored employing density functional theory calculations. It is revealed that the H-atom transfer reaction from Trp and Tyr to triplet state RF is more favorable on thermodynamic grounds compared with direct energy transfer or direct electron transfer pathways. During the photosensitization, RF can photogenerate various reactive oxygen species (ROS) as intermediates, while the present study provides some deeper insights into the photosensitizing behaviors of triplet state RF by reacting directly with Trp and Tyr.     

High-voltage electrophoretic separation of tryptophan metabolites of the kynurenine pathway

An analytical separation method for tryptophan and its seven metabolites of the kynurenine pathway by high-voltage paper electrophoresis is presented. Anthranilic acid, 3-hydroxyanthranilic acid, kynurenic acid, kynurenine, nicotinic acid, quinolinic acid, xanthurenic acid and unmetabolized tryptophan are measured in urine. Using radioactive labelling and scintillation counting as a quantification method, the relative standard deviation varied from 3.5% to 14.4%, corresponding to kynurenine and nicotinic acid, respectively. The recovery of labelled tryptophan added to urine was 95%. An advantage of the electrophoretic method is the minor tailing of spots and, hence, a good resolution of the components. For the monovalent anions of the kynurenine pathway metabolites, a linear correlation (r = 0.9996) was found between the experimental relative electrophoretic mobility and the quantity M-2/3, where M is the molecular mass of the anion.     

Photochemistry of furyl- and thienyldiazomethanes: spectroscopic characterization of triplet 3-thienylcarbene

Photolysis (λ > 543 nm) of 3-thienyldiazomethane (1), matrix isolated in Ar or N(2) at 10 K, yields triplet 3-thienylcarbene (13) and α-thial-methylenecyclopropene (9). Carbene 13 was characterized by IR, UV/vis, and EPR spectroscopy. The conformational isomers of 3-thienylcarbene (s-E and s-Z) exhibit an unusually large difference in zero-field splitting parameters in the triplet EPR spectrum (|D/hc| = 0.508 cm(-1), |E/hc| = 0.0554 cm(-1); |D/hc| = 0.579 cm(-1), |E/hc| = 0.0315 cm(-1)). Natural Bond Orbital (NBO) calculations reveal substantially differing spin densities in the 3-thienyl ring at the positions adjacent to the carbene center, which is one factor contributing to the large difference in D values. NBO calculations also reveal a stabilizing interaction between the sp orbital of the carbene carbon in the s-Z rotamer of 13 and the antibonding σ orbital between sulfur and the neighboring carbon-an interaction that is not observed in the s-E rotamer of 13. In contrast to the EPR spectra, the electronic absorption spectra of the rotamers of triplet 3-thienylcarbene (13) are indistinguishable under our experimental conditions. The carbene exhibits a weak electronic absorption in the visible spectrum (λ(max) = 467 nm) that is characteristic of triplet arylcarbenes. Although studies of 2-thienyldiazomethane (2), 3-furyldiazomethane (3), or 2-furyldiazomethane (4) provided further insight into the photochemical interconversions among C(5)H(4)S or C(5)H(4)O isomers, these studies did not lead to the spectroscopic detection of the corresponding triplet carbenes (2-thienylcarbene (11), 3-furylcarbene (23), or 2-furylcarbene (22), respectively).     

Photochemistry on surfaces: Fluorescence emission of monomers and dimers and triplet state absorption of acridine orange adsorbed on microcrystalline cellulose

Prompt fluorescence as well as delayed fluorescence emission of acridine orange was detected at room temperature from samples where this dye is adsorbed on microcrystalline cellulose. Ground state absorption studies provided evidence for dimer formation of the dye when adsorbed on cellulose, and the equilibrium constant for dimerisation was determined as 1.6±0.1 × 10⁶mol⁻¹g. At low loadings of acridine orange on cellulose (<1 μmol g⁻¹) the fluorescence emission is mainly due to the monomer and is similar to that observed in ethanolic solutions where little aggregation occurs, and peaks at 530 nm. A linear dependence of the fluorescence intensity on the amount of light absorbed by the dye was established for these “diluted” samples. However, at higher loadings (>20 μmol g⁻¹), the fluorescence intensity decreases, and the emission is broad with its maximum at 620 nm, and is mainly due to the dimer. By assuming that the excited monomer and dimer of acridine orange are the only emitting species, it was possible to determine the fluorescence quantum yields for these two species when adsorbed on microcrystalline cellulose as 0.95±0.05 and 0.40±0.10, respectively. Pulsed emission studies at room temperature in the millisecond time-range also revealed monomer and dimer emissions on this longer time-scale. These are shown to be due to thermally activated delayed fluorescence arising from the triplet states of monomer and dimer acridine orange as confirmed by diffuse reflectance transient absorption studies.     

Photochemistry of sulfilimine-based nitrene precursors: generation of both singlet and triplet benzoylnitrene

Photolysis of N-benzoyl-S,S-diphenylsulfilimine or N-benzoyl dibenzothiophene sulfilimine produces PhNCO and also benzoylnitrene. Direct observation of the triplet nitrene, energetic differences between the singlet and triplet state of the nitrene, and oxygen quenching experiments suggest that the triplet nitrene derives from the triplet excited state of the sulfilimine precursors, rather than through equilibration of nearby singlet and triplet states of the nitrene itself. In acetonitrile, the formation of an ylide, followed by cyclization to the corresponding oxadiazole, is the predominant nitrene chemistry, occurring on the time scale of a few microseconds and few tens of microseconds, respectively. Trapping experiments with substrates such as cis-4-octene suggest that reactivity of the nitrene is mainly through the singlet channel, despite a fairly small energy gap between the singlet ground state and the triplet.     

Multiple spin-echoes in a photo-excited triplet state

A Carr-Purcell multiple spin-echo, as modified by Meiboom and Gill, is described for the 3598 MHz zero-field transition of phosphorescent quinoline C₉ND₇ in a durene host. The results show that the dephasing of the electron spins through spectral diffusion may be largely eliminated by the multiple echo technique.     

Is tetramethyleneethane a ground state triplet?

We have examined a number of possible ways by which tetramethyleneethane (TME) can be a ground state triplet, as claimed by experimental studies, in violation of Ovchinnikov’s theorem for alternant hydrocarbons of equal bond lengths. Model exact π calculations of the low-lying states of TME, 3,4-dimethylenefuran and 3,4-dimethylenepyrrole were carried out using a diagrammatic valence bond approach. The calculations failed to yield a triplet ground state even after (a) tuning of electron correlation, (b) breaking alternancy symmetry, and (c) allowing for geometric distortions. In contrast to earlier studies of fine structure constants in other conjugated systems, the computedD andE values of all the low-lying triplet states of TME for various geometries are at least an order of magnitude different from the experimentally reported values. Incorporation of σ-π mixing by means of UHF MNDO calculations is found to favour a singlet ground state even further. A reinterpretation of the experimental results of TME is therefore suggested to resolve the conflict.     

On the influence of silver perchlorate on the lowest triplet state properties of N-vinylcarbazole

N-vinylcarbazole is non-phosphorescent due to effective radiationless deactivation of the triplet state but forms a ground state complex with silver ions which is phosphorescent. Two different complexes are observed. From their phosphorescence spectra the T₁ state energy of N-vinylcarbazole is estimated to be 24000 cm⁻¹.     

The triplet state of 4-nitro-N,N-dimethynaphthylamine

Nanosecond laser photolytic studies of 4-nitro-N,N-dimethylnaphthylamine (4-NDMNA) in nonpolar and polar solvents at room temperature show a transient species with an absorption maximum in the 500-510-nm range. This species is assigned to the lowest triplet excited state of 4-NDMNA. The absorption maximum of this state is independent of solvent polarity, and its lifetime is a function of the hydrogen donor efficiency of the solvent. In n-hexane the lifetime 1/k of the triplet state is 9.1 × 10^?6 sec, while in acetonitrile 1/k is 2.0 × 10^?7 sec. The hydrogen abstraction rate constant k_H of the triplet state with tributyl tin hydride (Bu₃SnH) in n-hexane is 1.7 × 10⁷M^?1·sec^?1, while in the case of isopropyl alcohol as hydrogen donor, k_H is 4.0 × 10⁷M^?1·sec^?1. The activation energy for the hydrogen abstraction by the triplet state from Bu₃SnH in deaerated n-hexane is 0.6 kcal/mol. The lack of spectral shift with increasing solvent polarity, and the appreciable hydrogen abstraction reactivity of the triplet state, also independent of solvent polarity, seem to indicate that this excited state is an n-π* state which retains its n-π* character even in polar media.     

Photochemistry of 1,2-dihydronaphthalene oxide: concurrent triplet and singlet processes via singlet excitation

The photochemistry of 1,2-dihydronaphthalene oxide (254 nm) was reexamined and indan was found to be a primary photoproduct, as well as the traditionally assumed secondary photoproduct. Quenching studies demonstrated that indan, as a primary photoproduct, is derived from a triplet pathway, competing with a singlet route, back to the ground state surface. CASSCF calculations strongly suggest that the triplet pathway consists of a dissociation of the oxirane moiety to give a triplet carbene and aldehyde, which via hydrogen abstraction-decarbonylation-ISC recloses to give indan. Conical intersections corresponding to the presumed 1,2-hydrogen shift and 1,2-alkyl shift to give 2-tetralone and 1-indancarbaldehyde, respectively, were located computationally.     

Determination of neopterin, kynurenine, tryptophan and creatinine in human serum by high throuput HPLC

A new HPLC method for simultaneous determination of neopterin, creatinine, kynurenine and tryptophan in human serum was developed and validated. Monolithic stationary phase's technology (two monolithic columns RP-18e were connected with guard monolithic cartridge 4.6 mm × 50 mm + 3.0 mm × 100 mm and 4.6 × 10 mm) and special auto sampler for micro titration plates (samples are storage in dark cooled place protected against evaporation) were combined with easy sample preparation step. As mobile phase 15 mmol/L phosphate buffer at pH 4.50 was used. Neopterin and tryptophan were detected using fluorescent detection and kynurenine and creatinine were detected by diode-array detection. This method may be suitable for large sequences of samples in clinical research and routine practice.