Synthesis and Photophysical Properties of Some PEG Substitued Titanyl Phthalocyanine Derivatives

Summary: Polyethylene glycol substituted titanyl phthalocyanine was prepared in two steps starting from phthalonitrile and characterized by FT-IR, UV-vis spectrophotometry, and fluorescence spectrophotometry. The titanyl phthocyanine derivatives (TiOPcs) showed high solubility in common organic solvents, such as, CH₃Cl and DMF. These compounds decreased in absorbance intensity with increase of molecular weight of polyethylene glycol at maxima wavelength of visible range. The fluorescence spectra showed a fluorescence emission near 690 nm with a quantum yield of 0.05–0.32 (λ_ex = 625 nm).     

Chemiluminescence upon Photooxidation of 4,4"-Diazidodiphenyl with Molecular Oxygen

It was found that irradiation of the solution of 4,4"-diazidodiphenyl in ethyl alcohol with UV-light at a wavelength longer than 280 nm in the presence of dissolved oxygen resulted in appearance of post-luminescence. The luminescence intensity increases with increasing the oxygen concentration; the luminescence virtually disappears after purging the solution with argon. The observed emission of light was attributed to chemiluminescence of the products of photooxidation of 4,4"-diazidodiphenyl. The rate dependence of the afterglow shows two maxima. The first is observed immediately after cessation of the UV irradiation of the solution and its transfer to a photometric cell. The intensity of the corresponding chemiluminescence decreases by an exponential law with _e= 5 s. The second maximum appears after a certain time interval after the first one. Addition of a triplet sensitizer (Michler's ketone) to the reaction mixture results in quenching of the chemiluminescence from the photooxidation products, whereas the addition of substances that stabilize electrophilic species in the singlet state leads to an increase in the chemiluminescence intensity. It was suggested that the chemiluminescence resulted from reactions involving the singlet nitrene adduct with oxygen.     

Evidence for a pterin-derivative associated with the molybdenum cofactor of Neurospora crassa nitrate reductase

Abstract— Following the method of Johnson and Rajagopalan (1982) for obtaining Form B of molybdopterin cofactors, we observed a prominent fluorescence band at480–482 nm in purified NR of Neurospora crassa mutant albino-band after boiling the enzyme at acidic pH and readjusting the sample to alkaline pH. This fluorescence band is maximally excited at 410 nm and maximally emitting at pH 11 (“F-480_pH11”); at pH4–7 only a featureless fluorescence band of low intensity remains (“F-480_pH5”). The fluorescence ΔF-480 = F-480_pH11 - F-480_pHS is examined here. ΔF-480 is associated specifically with NADPH-dependent and MVH-dependent nitrate reduction activities and with cytochrome b-557 absorption. In a protease-digested preparation lacking NADPH-dependent NR activity, ΔF-480 is associated with MVH-dependent nitrate reduction. The ΔF-480 signal is followed during the course of purification of NR. Its size increases with increasing purity of the enzyme. In partially purified NR preparations and especially in aqueous extracts from mycelia of N. crassa, a second, strong fluorescence signal with a pH-dependent emission maximum at around 450 nm (maximally excited at350–370 nm) was found beside ΔF-480. This “unspecific” signal was lost during NR purification. A procedure is developed to demonstrate AF-480 also in presence of the unspecific (350-370/450 nm) signal as well as flavins. We deduce that the ΔF-480 component is part of the Mo cofactor of N. crassa NR and that the signal is caused by a pterin derivative. From calculations of total content of the AF-480 component in mycelia it is likely that in vivo it is shared also by other enzymes.     

The influence of annealing in a vacuum on the concentration of radicals in fullerite C<Subscript>60</Subscript>

For fullerite C₆₀ with intercalated oxygen, a sharp (by three orders of magnitude) increase in the intensity of the EPR signal with a g-factor of 2.0024 was observed at ～200°C. Studies of gases formed in heating of the sample in a vacuum showed that molecular oxygen was largely released at temperatures below 100°C, whereas the gas phase formed as the temperature increased to 200°C contained carbon oxides CO and CO₂ in addition to oxygen. The conclusion was drawn that the intensity of the EPR signal was determined by the products of oxygen interaction with fullerene rather than the concentration of oxygen in the sample.     

Chemiluminescence arising from the decomposition of 1,4-dimethylnaphthalene endoperoxide applied to silica gel in the presence of NdIII,YbIII, and EuIII β-diketonates

Visible and near-IR chemiluminescence was observed upon the decomposition of 1,4-dimethylnaphthalene endoperoxide applied to silica gel in the presence of the β-diketonate complexes Nd(L)3·nH₂O, Yb(L)3·nH₂O, and Eu(L)3·nH₂O (L is 6,6,7,7,8,8,8-heptafluoro-2,2-dimethyloctane-3,5-dionate, 1,1,1-trifluoro-3-thenoylacetonate, and acetylacetonate). Excited lanthanide ions serve as luminescent emitters with emission maxima at λ = 870 and 1060 nm for Nd^III, 990 nm for Yb^III, and 615 nm for Eu^III. Singlet oxygen generated by decomposing endoperoxide was found to play a key role in the chemiluminescence mechanism.     

Photoinitiation by radical-cationic mechanism in the polymerization of isobutylene with lewis acids

ESR spectra of Lewis acids (VCl₄, TiCl₄, TiBr₄, SnCl₄, and AlBr₃) and their mixtures with isobutylene were investigated in a n-heptane solution in the dark and under irradiation at 400–480 nm at ?80 to ?150°C. A signal was observed only upon irradiating mixtures of VCl₄, TiCl₄, or TiBr₄ and isobutylene. The signal was identified as an isobutylene radical-cation by comparison with a simulated spectrum. A signal indicating the presence of peroxy radicals were recorded in measurements carried out in the presence of oxygen; these radicals originated from reaction of the isobutylene radical-cation with oxygen. Radical-cation initiation by visible light is indicated by the polymerization of isobutylene by VCl₄, TiCl₄, and TiBr₄ and by ESR spectra. The inhibiting effect of oxygen in photochemically initiated polymerization of isobutylene was also elucidated.     

Ultrafast fluorescence resonance energy transfer in a bile salt aggregate: Excitation wavelength dependence

Fluorescence resonance energy transfer (FRET) from Coumarin 153 (C153) to Rhodamine 6G (R6G) in a secondary aggregate of a bile salt (sodium deoxycholate, NaDC) is studied by femtosecond up-conversion. The emission spectrum of C153 in NaDC is analysed in terms of two spectra-one with emission maximum at 480 nm which corresponds to a non-polar and hydrophobic site and another with maximum at ∼530 nm which arises from a polar hydrophilic site. The time constants of FRET were obtained from the rise time of the emission of the acceptor (R6G). In the NaDC aggregate, FRET occurs in multiple time scales — 4 ps and 3700 ps. The 4 ps component is assigned to FRET from a donor (D) to an acceptor (A) held at a close distance (R _DA ∼ 17 ?) inside the bile salt aggregate. The 3700 ps component corresponds to a donor-acceptor distance ∼48 ?. The long (3700 ps) component may involve diffusion of the donor. With increase in the excitation wavelength (λ _ex) from 375 to 435 nm, the relative contribution of the ultrafast component of FRET (∼4 ps) increases from 3 to 40% with a concomitant decrease in the contribution of the ultraslow component (∼3700 ps) from 97 to 60%. The λ _ex dependence is attributed to the presence of donors at different locations. At a long λ _ex (435 nm) donors in the highly polar peripheral region are excited. A short λ _ex (375 nm) ‘selects’ donor at a hydrophobic location.     

γ-Fe2O3 Nanoparticles Covered with Glutathione-Modified Quantum Dots as a Fluorescent Nanotransporter

The present paper describes the synthesis, characterization, and utilization of multi-functional magnetic conjugates that integrate optical and magnetic properties in a single structure for use in many biomedical applications. Spontaneous interaction with eukaryotic cell membrane (HEK-239 cell culture) was determined using fluorescence microscopy, and fluorescence analyses. Both, differences in excitation, and emission wavelength were observed, caused by glutathione intake by cells, resulting in disintegration of core–shell structure of quantum dots, as well as adhesion of conjugate onto cell surface. When compared with quantum dots fluorescent properties, HEK-239 cells with incorporated nanoconjugate exhibited two excitation maxima (λ _ex = 430 and 390 nm). Simultaneously, application of ideal λ _ex for quantum dots (λ _ex = 430 nm), resulted in two emission maxima (λ = 740 and 750 nm). This nanoconjugate fulfills the requirements of term theranostics, because it can be further functionalized with biomolecules as DNA, proteins, peptides or antibodies, and thus serves as a tool for therapy in combination with simultaneous treatment.

     

Chemiluminescence produced by oxidation of fullerene hydride C60H36 with oxygen in solution

It was found that the oxidation of fullerene hydride C₆₀H₃₆ with oxygen in solution is accompanied by chemiluminescence (CL). The CL spectrum maxima are assigned to the emission by the oxidation products of C₆₀H₃₆, namely, the excited hydride C₆₀H₁₈* (495, 535 nm) and singlet-excited fullerene ¹C₆₀* (720—750 nm).     

γ-Fe2O3 Nanoparticles Covered with Glutathione-Modified Quantum Dots as a Fluorescent Nanotransporter

The present paper describes the synthesis, characterization, and utilization of multi-functional magnetic conjugates that integrate optical and magnetic properties in a single structure for use in many biomedical applications. Spontaneous interaction with eukaryotic cell membrane (HEK-239 cell culture) was determined using fluorescence microscopy, and fluorescence analyses. Both, differences in excitation, and emission wavelength were observed, caused by glutathione intake by cells, resulting in disintegration of core–shell structure of quantum dots, as well as adhesion of conjugate onto cell surface. When compared with quantum dots fluorescent properties, HEK-239 cells with incorporated nanoconjugate exhibited two excitation maxima (λ _ex = 430 and 390 nm). Simultaneously, application of ideal λ _ex for quantum dots (λ _ex = 430 nm), resulted in two emission maxima (λ = 740 and 750 nm). This nanoconjugate fulfills the requirements of term theranostics, because it can be further functionalized with biomolecules as DNA, proteins, peptides or antibodies, and thus serves as a tool for therapy in combination with simultaneous treatment.     

STUDIES ON AMINODIOXETANES AS A MODEL OF BIOLUMINESCENCE INTERMEDIATES. 1-(1-METHYL-3-INDOLYL)-6-PHENYL-2,5,7,8-TETRAOXABICYCLO[4,2,0]OCTANE,AN AMINODIOXETANE RESULTING IN EFFICIENT ULTRAVIOLET AND EXCIPLEX CHEMILUMINESCENCE*

Abstract— The title aminodioxetane prepared by photooxygenation of 2–(l-methyl-3-indolyl-3-phenyl-1,4-dioxene is fairly stable at ?46°C and gives on warming to room temperature an ultraviolet light at Λ_max 320 nm, which corresponds to 377kJ/mol; the highest energy ever observed among the efficient chemiluminescent compounds. The efficiency of chemiluminescence and excited singlet molecule formation in n-hexane were 3.6% and at least 50%, respectively. Substitution and solvent effects suggest polar nature of the transition state as expected from the ‘CIEEL’ mechanism. In polar solvents, the dioxetane gives visible light (Λ_max 400 nm in dichloromethane) as well as the UV light; the former being quenched by methanol. This is interpreted in terms of an intramolecular exciplex formation between the indole and phenyl groups. This is the first example of an intramolecular exciplex produced by the dioxetane decomposition.     

Horseradish Peroxidase-Catalyzed Generation of Acetophenone and Benzophenone in the Triplet State*

Horseradish peroxidase catalyzes the aerobic oxidation of 2-phenylpropanal and diphenylacetaldehyde at physiological pH to yield acetophenone and benzophenone partly in the triplet state, respectively. These excited products plus formic acid are expected from the thermolysis of dioxetane intermediates. The presence of acetophenone was demonstrated spectrophotometr-ically and the chemiluminescence spectrum (δ_max - 430 nm) was consistent with its triplet state. Energy transfer to 9,10-dibromoanthracene-2-sulfonate ion, a triplet carbonyl counter, but not to anthracene-2-sulfonate ion, a singlet carbonyl acceptor, occurred, confirming the triplet nature of the main emitter. In the case of the diphenylacetaldehydelperoxidase system, benzophenone could also be detected spectrophotometrically but the corresponding chemiluminescence spectrum showed only red emission (δ_max - 630 nm), which was tentatively attributed to singlet oxygen formed by triplet-triplet energy transfer to ground state oxygen. Horseradish peroxidase can be replaced by other he-meproteins such as myoglobin, hemoglobin and micro-peroxidase as catalyst of the chemiluminescent reaction. The distinct emission spectra achieved with different hemeproteins suggest a role of the microenvi-ronment in totally or partly protecting the excited species from oxygen collisions, resulting in emission maxima around 430 nm, 630 nm or both.     

THE SENSORY PHOTORECEPTORS OF Halobacterium halobium REVISITED: ACTION SPECTRA and INFLUENCE OF BACKGROUND LIGHT*

Abstract– Action spectra of the light-dependent behavior of Halobacterium and the effect of background light have been measured with regard to the current hypothesis of Spudich and Bogomolni [Nature 312 ,509–513 (1984)], which proposes sensory rhodopsin I (sRI₅₈₇) to be the receptor for long-wavelength light, and its photoproduct S₃₇₃ to be the receptor for UV light. The action spectrum shows three maxima for attractant responses (prolonged swimming intervals) at 565, 590, and 610 nm, and two maxima for repellent responses (shortened intervals) at 370 and 480 nm. The latter is assigned to sensory rhodopsin II (P-480). All peaks are red-shifted after substitution of the endogeneous retinal by 3, 4-dehydroretinal. The peaks at 590 and 610 nm are suppressed by long-wavelength background light. Ultraviolet background light converts all attractant peaks into repellent peaks. The response at 370 nm is strongly activated by visible background light, the maximal effect occurring with 510 nm. The activated state declines with a half-life of about 1.2 s. In a growing culture, full sensitivity to UV and blue light is restored about 10 h earlier than sensitivity to long-wavelength light. Some of the results cannot easily be explained by the sRI₅₈₇/S₃₇₃ hypothesis. Explanations for the three maxima in the long-wavelength range and for the maximal activation of the UV response by 510 nm light are discussed.     

A Femtosecond Study of Solvation Dynamics and Anisotropy Decay in a Catanionic Vesicle: Excitation‐Wavelength Dependence

The structure and dynamics of a catanionic vesicle are studied by means of femtosecond up‐conversion and dynamic light scattering (DLS). The catanionic vesicle is composed of dodecyl‐trimethyl‐ammonium bromide (DTAB) and sodium dodecyl sulphate (SDS). The DLS data suggest that 90 % of the vesicles have a diameter of about 400 nm, whereas the diameter of the other 10 % is about 50 nm. The dynamics in the catanionic vesicle are compared with those in pure SDS and DTAB micelles. We also study the dynamics in different regions of the micelle/vesicle by varying the excitation wavelength (λ_ex) from 375 to 435 nm. The catanionic vesicle is found to be more heterogeneous than the SDS or DTAB micelles, and hence, the λ_ex‐dependent variation of the solvation dynamics is more prominent in the first case. The solvation dynamics in the vesicle and the micelles display an ultraslow component (2 and 300 ps, respectively), which arises from the quasibound, confined water inside the micelle, and an ultrafast component (<0.3 ps), which is due to quasifree water at the surface/exposed region. With an increase in λ_ex, the solvation dynamics become faster. This is manifested in a decrease in the total dynamic solvent shift and an increase in the contribution of the ultrafast component (<0.3 ps). At a long λ_ex (435 nm), the surface (exposed region) of a micelle/vesicle is probed, where the solvation dynamics of the water molecules are faster than those in a buried location of the vesicle and the micelles. The time constant of anisotropy decay becomes longer with increasing λ_ex, in both the catanionic vesicle and the ordinary micelles (SDS and DTAB). The slow rotational dynamics (anisotropy decay) in the polar region (at long λ_ex) may be due to the presence of ionic head groups and counter ions.     

Spectrofluorimetric determination of titanium,zirconium and hafnium and their binary mixtures with biacetylmonoxime nicotinylhydrazone

The synthesis, characteristics and analytical applications of biacetylmonoxime nicotinylhydrazone are described. This compound forms fluorescent complexes with titanium(IV) (λ_ex = 430, λ_em = 540 nm), zirconium (λ_ex = 415 nm, λ_em = 505 nm) and hafnium (λ_ex = 400, λ_em = 500 nm) in an acidic medium. Titanium forms a 1:2 metal:ligand complex, whereas zirconium and hafnium form 1:2:1 metal:ligand:sulphate ternary complexes. Highly selective spectrofluorimetric methods for titanium (20–100 ng ml^?1), zirconium and hafnium (5–100 ng ml^?1) are proposed, and procedures for the analysis of binary mixtures of these ions are described.     

Synthesis, Structure and Photoluminescence of Two Zinc Carboxyl－ate Polymers with Different Coordination Architectures

The hydrothermal reaction of ZnO with benzene‐1,4‐dicarboxylic add gave Zn·BDC·2H₂O (1) and Zn‐BDC·H₂O (2) (BDC = benzene‐1, 4‐dicarboxylate), respectively. Polymer 1 (C₄H₄O₃ZH_0.5) shows a one‐dimensional zigzag chain structure built up from the alternate connection of tetrahedral ZnO₄ and BDC units. Polymer 2 (C₄H₃O_2.5Zn_0.5) possesses a three‐dimensional framework containing infinite zigzag Zn·Zn·Zn pseudochains generated by five‐coordinate zinc centers and a rectangular channel system including three groups of different straight channels along the [001], [010] and [60–1] directions. The two metal‐organic polymeric compounds exhibit strong photoluminescent emission bands at 402 nm (λ_ex = 260 nm) (for 1) and at 344 nm and 385 nm (λ_ex = 279 nm) (for 2) in the solid state at room temperature.     

Study of the chromatographic behavior and inclusion constants of new fluorescence agents by use of cyclodextrin additives under conditions of steady- and dynamic-state equilibrium and with fluorescence detection

Summary We report four new derivatization agents, acridone-N-acetic acid (ARC), carbazole-9-ylacetic acid (CRA), carbazole-9-ylpropionic acid (CRP), and 2-methyl-2-carbazole-9-ylacetic acid (MCRA), with strong fluorescence emission which has low dependence on solvent polarity. The emission maxima for ARC, CRA, CRP, and MCRA were 430 nm (λ_ex 404 nm), 368 nm (λ_ex 335 nm), 356 nm (λ_ex 340 nm) and 360 nm (λ_ex 330 nm), respectively. The effects of mobile-phase composition, pH, and temperature on the liquid chromatographic retention behavior of the four fluorescence agents were investigated. An experimental model was established for calculating the inclusion constants of cyclodextrin (CD) complexes in the dynamic state, using β-cyclodextrin (β-CD) and hydroxypropyl-β-cyclodextrin (HP-β-CD) as examples, and different mobilephase compositions. On the basis of the model, the inclusion constants of the solutes in pure water (K _fw) were determined by extrapolation. The thermodynamic parameters (ΔH ^o and ΔS ^o) and dissociation constantsK _am for the solutes in this chromatographic system were obtained by means of capacity factor (k) values using a corresponding model formulation.     

Application of unfold principal component analysis and parallel factor analysis to the exploratory analysis of olive oils by means of excitation-emission matrix fluorescence spectroscopy

Discrimination between virgin olive oils and pure olive oils is of primary importance for controlling adulterations. Here, we show the potential usefulness of two multiway methods, unfold principal component analysis (U-PCA) and parallel factor analysis (PARAFAC), for the exploratory analysis of the two types of oils. We applied both methods to the excitation-emission fluorescence matrices (EEM) of olive oils and then compared the results with the ones obtained by multivariate principal component analysis (PCA) based on a fluorescence spectrum recorded at only one excitation wavelength. For U-PCA and PARAFAC, the ranges studied were λ_ex=300-400 nm, λ_em=400-695 nm and λ_ex=300-400 nm, λ_em=400-600 nm. The first range contained chlorophylls, whose peak was much more intense than those of the rest of species. The second range did not contain the chlorophylls peak but only the fluorescence spectra of the remaining compounds (oxidation products and Vitamin E). The three-component PARAFAC model on the second range was found to be the most interpretable. With this model, we could distinguish well between the two groups of oils and we could find the underlying fluorescent spectra of three families of compounds.     

SPECTROSCOPIC STUDIES OF CUTANEOUS PHOTOSENSITIZING AGENTS. XVIII. INDOMETHACIN

Abstract— The photochemistry, photophysics, and photosensitization (Type I and II) of indomethacin (IN) (N-[p-chlorobenzoyl]-5-methoxy-2-methylindole-3-acetic acid) has been studied in a variety of solvents using NMR, high performance liquid chromatography-mass spectroscopy, transient spectroscopy, electron paramagnetic resonance in conjunction with the spin trapping technique, and the direct detection of singlet molecular oxygen (^l O₂) luminescence. Photodecomposition of IN (λ_ex > 330 nm) in degassed or air-saturated benzene proceeds rapidly to yield a major (2; N-[p-chlorobenzoyl]-5-methoxy-2-methyl-3-methylene-indoline) and a minor (3; N-[p-chlorobenzoyl]-5-methoxy-2, 3-dimethyl-indole) decarboxylated product and a minor indoline (5; 1-en-5-methoxy-2-methyl-3-methylene-in-doline), which is formed by loss of the p-chlorobenzoyl moiety. In air-saturated solvents two minor oxidized products 4 (N-[p-chlorobenzoyl]-5-methoxy-2-methylindol-3-aldehyde) and 6 (5-methoxy-2-methyl-indole-3-aldehyde) are also formed. When photolysis was carried out in ¹⁸O₂-saturated benzene, the oxidized products 4 and 6 contained ¹⁸O, indicating that oxidation was mediated by dissolved oxygen in the solvent. In more polar solvents such as acetonitrile or ethanol, photodecomposition is extremely slow and inefficient. Phosphorescence of IN at 77 K shows strong solvent dependence and its emission is greatly reduced as polarity of solvent is increased. Flash excitation of IN in degassed ethanol or acetonitrile produces no transients. A weak transient is observed at 375 nm in degassed benzene, which is not quenched by oxygen. Irradiation of IN (λ_ex > 325 nm) in N₂-gassed C₆H₆ in the presence of 5, 5-dimethyl-1-pyrroline-N-oxide (DMPO) results in the trapping of two carbon-centered radicals by DMPO. One adduct was identified as DMPO/^.COC₆H₄-p-CI, while the other was probably derived from a radical formed during IN decarboxylation. In air-saturated benzene, (hydro) peroxyl and alkoxyl radical adducts of DMPO are observed. A very weak luminescence signal from ¹O₂ at 1268 nm is observed initially upon irradiation (λ_ex= 325 nm) of IN in air-saturated benzene or chloroform. The intensity of this ¹O₂ signal increases as irradiation is continued suggesting that the enhancement in ¹O₂ yield is due to photoproduct(s). Accordingly, when 2 and 3 were tested directly, 2 was found to be a much better sensitizer of ¹O₂ than IN. In air-saturated ethanol or acetonitrile no IN ¹O₂ luminescence is detected even on continuous irradiation. The inability of IN to cause phototoxicity may be related to its photo stability in polar solvents, coupled with the low yield of active oxygen species (¹O₂, O₂^?_-) upon UV irradiation.     

CHEMILUMINESCENCE and EPR STUDIES ON THE EXCITATION SITE OF FERRIC-HEME-OXO COMPLEXES OF NATURAL and MODEL HEME SYSTEMS

Chemiluminescence was detected both in the reaction system of H₂O₂ plus heme proteins such as methemo- and metmyoglobin and ferric-protoheme complexes used as a model system. The intensity of chemiluminescence was found to be mediated by ligand binding to the sixth coordination site of the ferric-protoheme compounds, e.g. chemiluminescence was not observed with the bisimidazole ferric-protoheme complex. On the other hand the pentacoordinated histidine ferric-protoheme complex exhibited strong light emission. Comparative studies with various ligand-heme compounds elucidated that light emission was inversely correlated with the binding strength of the respective ligand at the sixth coordination site. The basic reaction mechanism causing the establishment of an excited state was studied by monitoring chemiluminescence and EPR signal formation of ligand-modified heme proteins in the presence of different electron donors. External electron donors such as Trolox C, TMPD and ascorbic acid affected a strong reduction in the development of chemiluminescence suggesting the essential involvement of an inner-molecular electron transfer process. Our results allow the conclusion that chemiluminescence is generated from the decay of an excited state of oxo-heme compounds established as a result of a one electron transfer step from a ligand group to heme iron.