Single-Molecule Catalysis in the Palladium Cross-Coupling Reaction Cycle.

Single-molecule (SM) methods are applied to study various types of catalytic processes in chemical and biochemical reactions. In this study, the Suzuki-Miyaura cross-coupling reaction forming a fluorescent product is investigated within the SM approximation. Stochastic analysis of emission intermittency in selected nanoscopic spots allows us to determine the single-molecule turnover frequency (SM-TOF) of the Pd catalyst in a specific probe reaction. We generate and analyze simulated intermittency time traces of a single catalyst surrounded by reactant molecules to assess the reliability of the method applied to real intermittency time trace data from hundreds of nanoscopic fluorescence spots. The results demonstrate that the proposed method can be used to evaluate the average SM-TOF of Pd in a cross-coupling reaction.

Acid selenites as new selenium precursor for CdSe quantum dot synthesis.

Chemical precursors for nanomaterials synthesis have become essential to tune particle size, composition, morphology, and unique properties. New inexpensive precursors investigation that precisely controls these characteristics is highly relevant. We studied new Se precursors, the acid selenites (R-O-SeOOH), to synthesize CdSe quantum dots (QDs). They were produced at room temperature by the Image 1 reaction with alcohols having different alkyl chains and were characterized by 1H NMR confirming their structures. This unprecedented precursor generates high-quality CdSe nanocrystals with narrow size distribution in the zinc-blend structure showing controlled optical properties. Advanced characterization detailed the CdSe structure showing stacking fault defects and its dependence on the used R-O-SeOOH. The QDs formation was examined using a time-dependent growth kinetics model. Differences in the nanoparticle surface structure influenced the optical properties, and they were correlated to the Se-precursor nature. Small alkyl chain acid selenites generally lead to more controlled QDs morphology, while the bigger alkyl chain leads to slightly upper quantum yields. Acid selenites can potentially replace Se-precursors at competitive costs in the metallic chalcogenide nanoparticles. Image 1 is chemically stable, and alcohols are cheap and less toxic than the reactants used today, making acid selenites a more sustainable Se precursor.

Single-Molecule Observations Provide Mechanistic Insights into Bimolecular Knoevenagel Amino Catalysis.

While single-molecule (SM) methods have provided new insights to various catalytic processes, bimolecular reactions have been particularly challenging to study. Here, the fluorogenic Knoevenagel condensation of an aromatic aldehyde with methyl cyanoacetate promoted by surface-immobilized piperazine is quantitatively characterized using super-resolution fluorescence imaging and stochastic analysis using hidden Markov modeling (HMM). Notably, the SM results suggest that the reaction follows the iminium intermediate pathway before the formation of a fluorescent product with intramolecular charge-transfer character. Moreover, the overall process is limited by the turnover rate of the catalyst, which is involved in multiple steps along the reaction coordinate.

Highly fluorescent hybrid pigments from anthocyanin- and red wine pyranoanthocyanin-analogs adsorbed on sepiolite clay.

Flavylium cations serve as models for the chemical and photochemical reactivity of anthocyanins, the natural plant pigment responsible for many of the red, blue and purple colors of fruits and flowers. Likewise, pyranoflavylium cations serve as models of the fundamental chromophoric moiety of pyranoanthocyanins, molecules that can form from reactions of grape anthocyanins in red wines during their maturation. In the present work, hybrid pigments are prepared by the adsorption of a series of five synthetic flavylium cations (FL) and five synthetic pyranoflavylium cations (PFL) on sepiolite clay (SEP). The FL are smaller in size than the PFL, but both can in principle fit into the tunnels and/or external grooves (with dimensions of 3.7 × 10.6 Å) of SEP. Measurements of the fluorescence quantum yields of the adsorbed dyes indicate that they are at least as fluorescent as in acidic acetonitrile solution, and in a few cases substantially more fluorescent. The observation of biexponential fluorescence decays is consistent with emission from dye molecules adsorbed at two distinct sites, presumably tunnels and grooves. These hybrid materials also have improved properties in terms of stability of the color in contact with pH 10 aqueous solution and resistance to thermal degradation of the dye. SEP thus appears to be a promising substrate for the development of highly fluorescent flavylium or pyranoflavylium cation-derived hybrid pigments with improved color and thermal stability.

Styryl dye formation promoted by catalytic centers of piperazine bound to a silica surface traced by single molecule fluorescence microscopy.

The styryl dye (E)-2-[3-[4-(diphenylamine) phenyl]-1-(p-tolyl)-allylidene]-malononitrile (DFTAM) was prepared by Knoevenagel condensation using homogeneous and surface bound amino catalysts. The catalysis by surface bound piperazine allowed the study of the condensation reaction at a single molecule (SM) level using total internal fluorescence microscopy (TIRFM). The turnover rate of the surface catalyst is dependent on the presence of the byproduct water of the condensation reaction. The addition of zeolite particles as water traps improves the catalytic activity indicated by the highest number of emissive centers mapped with super resolved fluorescence imaging and longest intensity-time trajectories showing SM bursting events. This particular condensation reaction highlights the applicability of the method to study multi-step organic reactions in the condensed phase in which the product is a fluorescent species.

Aryl-Substituted Ruthenium(II) Complexes: A Strategy for Enhanced Photocleavage and Efficient DNA Binding.

Ruthenium polypyridine complexes have shown promise as agents for photodynamic therapy (PDT) and tools for molecular biology (chromophore-assisted light inactivation). To accomplish these tasks, it is important to have at least target selectivity and great reactive oxygen species (ROS) photogeneration: two properties that are not easily found in the same molecule. To prepare such new agents, we synthesized two new ruthenium complexes that combine an efficient DNA binding moiety (dppz ligand) together with naphthyl-modified (1) and anthracenyl-modified (2) bipyridine as a strong ROS generator bound to a ruthenium complex. The compounds were fully characterized and their photophysical and photochemical properties investigated. Compound 2 showed one of the highest quantum yields for singlet oxygen production ever reported (ΦΔ= 0.96), along with very high DNA binding (log Kb = 6.78). Such photochemical behavior could be ascribed to the lower triplet state involving the anthracenyl-modified bipyridine, which is associated with easier oxygen quenching. In addition, the compounds exhibited moderate selectivity toward G-quadruplex DNA and binding to the minor groove of DNA, most likely driven by the pendant ligands. Interestingly, they also showed DNA photocleavage activity even upon exposure to a yellow light-emitting diode (LED). Regarding their biological activity, the compounds exhibited an exciting antibacterial action, particularly against Gram-positive bacteria, which was enhanced upon blue LED irradiation. Altogether, these results showed that our strategy succeeded in producing light-triggered DNA binding agents with pharmacological and biotechnological potential.

Methyltransferase-directed covalent coupling of fluorophores to DNA.

We report an assay for determining the number of fluorophores conjugated to single plasmid DNA molecules and apply this to compare the efficiency of fluorophore coupling strategies for covalent DNA labelling. We compare a copper-catalyzed azide-alkyne cycloaddition reaction, amine to N-hydroxysuccinimidyl ester coupling reaction and strain-promoted azide-alkyne cycloaddition reaction for fluorescent DNA labelling. We found increased labelling efficiency going from the amine to N-hydroxysuccinimidyl ester coupling reaction to the copper-catalyzed azide-alkyne cycloaddition and found the highest degree of DNA labelling with the strain-promoted azide-alkyne cycloaddition reaction. We also examined the effect of labelling on the DNA structure using atomic force microscopy. We observe no distortions or damage to the DNA that was labeled using the amine to N-hydroxysuccinimidyl ester and strain-promoted azide-alkyne cycloaddition coupling reactions. This was in contrast to the copper-catalyzed azide-alkyne cycloaddition reaction, which, despite the use of copper-coordinating ligands in the labelling mixture, leads to some structural DNA damage (single-stranded DNA breaks).

Imaging the photoinduced charge injection in CdS/TiO2 nanoparticles by the sequential fluorescence mapping method.

The combination of a sensitizer and TiO2 nanoparticles forming a photocatalytic material is a central issue in many fields of applied photochemistry. The charge injection of emissive sensitizers into the conduction band of the semiconductor TiO2 may form a photoactive region that becomes dark, or it has a very low emission signal due to the generation of sensitizer radicals. However, by sequential coupling of a selected photoredox dye, such as resazurin, the dark region may become fluorescent at the interfaces where the charge injection has taken place due to the concomitant formation of fluorescent resorufin by cascade electron transfer. Using this strategy and a total internal reflection fluorescence microscopy (TIRFM) image, the charge injection in TiO2/CdS and SiO2/TiO2/CdS nanoparticles is investigated The method allows the charge injection efficiency of the excited CdS into TiO2 to be evaluated qualitatively, explaining the differences observed for these photocatalytic materials in H2 generation.

The spatial distribution of the photostability of thionine in zeolite L nanochannels investigated by Photobleaching Lifetime Imaging Microscopy.

Dye photobleaching is a photochemical reaction that can be investigated locally using fluorescence microscopy techniques. In this study, a user-friendly computational tool to assist photobleaching experiments called Photobleaching Lifetime Imaging Microscopy (PbLIM) is presented. With this tool it is possible to recover the photobleaching kinetics spatially, where a photobleaching lifetime is generated for each pixel of the image. Our model was applied to the photobleaching process of thionine encapsulated into the one-dimensional nano-channels of Zeolite L (ZL), from where we gained insight into the molecular oxygen distribution inside the ZL channels, as well as the detailed photobleaching of the confined thionine.

Fluorescence spectroscopy and confocal microscopy of the mycotoxin citrinin in condensed phase and hydrogel films.

The emission spectra, quantum yields and fluorescence lifetimes of citrinin in organic solvents and hydrogel films have been determined. Citrinin shows complex fluorescence decays due to the presence of two tautomers in solution and interconversion from excited-state double proton transfer (ESDPT) process. The fluorescence decay times associated with the two tautomers have values near 1 and 5 ns depending on the medium. In hydrogel films of agarose and alginate, fluorescence imaging showed that citrinin is not homogeneously dispersed and highly emissive micrometer spots may be formed. Fluorescence spectrum and decay analysis are used to recognize the presence of citrinin in hydrogel films using confocal fluorescence microscopy and spectroscopy.

Intrachain energy migration to weak charge-transfer state in polyfluorene end-capped with naphthalimide derivative.

Polyfluorene end-capped with N-(2-benzothiazole)-1,8-naphthalimide (PF-BNI) is a highly fluorescent material with fluorescence emission modulated by solvent polarity. Its low energy excited state is assigned as a mixed configuration state between the singlet S(1) of the fluorene backbone (F) with the charge transfer (CT) of the end group BNI. The triexponential fluorescence decays of PF-BNI were associated with fast energy migration to form an intrachain charge-transfer (ICCT) state, polyfluorene backbone decay, and ICCT deactivation. Time-resolved fluorescence anisotropy exhibited biexponential relaxation with a fast component of 12-16 ps in addition to a slow one in the range 0.8-1.4 ns depending on the solvent, showing that depolarization occurs from two different processes: energy migration to form the ICCT state and slow rotational diffusion motion of end segments at a longer time. Results from femtosecond transient absorption measurements agreed with anisotropy decay and showed a decay component of about 16 ps at 605 nm in PF-BNI ascribed to the conversion of S(1) to the ICCT excited state. From the ratio of asymptotic and initial amplitudes of the transient absorption measurement, the efficiency of intrachain ICCT formation is estimated in 0.5, which means that, on average, half of the excited state formed in a BNI-(F)(n)-BNI chain with n = 32 is converted to its low energy intrachain charge-transfer (ICCT) state.

Excited-state dynamics of polyfluorene derivatives in solution.

The excited-state dynamics of two polyfluorene copolymers, one fully conjugated containing phenylene vinylene units alternated with 9,9'-dihexylfluorenyl groups and the other segmented by -(CH2)8- spacer, were studied in dilute solution of different solvents using a picosecond single-photon timing technique. The excited-state dynamics of the segmented copolymer follows the Förster resonant energy-transfer model which describes intrachain energy-transfer kinetics among random oriented chromophores. Energy transfer is confirmed by analysis of fluorescence anisotropy relaxation with the measurement of a short decay component of about 60 ps. The fluorescence decay surface of the fully conjugated copolymer is biexponential with decay times of about 470 and 900 ps, ascribed to deactivation of chain moieties containing trans and cis isomers already in a photostationary condition. Thus, energy transfer is very fast due to the conjugated nature and rigid-rod-like structure of this copolymer chain.

Static and dynamic bimolecular fluorescence quenching of porphyrin dendrimers in solution.

The fluorescence quenching kinetics of two porphyrin dendrimer series (GnTPPH(2) and GnPZn) by different type of quenchers is reported. The microenvironment surrounding the core in GnPZn was probing by core-quencher interactions using benzimidazole. The dependence of quencher binding constant (K ( a )) on generation indicates the presence of a weak interaction between branches and the core of the porphyrin dendrimer. The similar free volume in dendrimers of third and fourth generation suggests that structural collapse in high generations occurs by packing of the dendrimer peripheral layer. Dynamic fluorescence quenching of the porphyrin core by 1,3-dicyanomethylene-2-methyl-2-pentyl-indan (PDCMI) in GnTPPH(2) is a distance dependent electron transfer process with an exponential attenuation factor beta = 0.33 A(-1). The quenching by 1,2-dibromobenzene occurs by diffusion process of the quencher toward to the porphyrin core, and its rate constant is practically independent of dendrimer generation.

Fluorescence modulation of acridine and coumarin dyes by silver nanoparticles.

Silver nanoparticles were synthesized by chemical reduction of silver ions by sodium borohydride in the presence of poly-(N)-vinyl-2-pyrrolidone in solution of short chain alcohols. The nanoparticles are stable in 2-propanol, and the average diameter of the Ag colloid obtained in this solvent is about 6 nm. The photophysical properties of acridinium and coumarin dyes in 2-propanol are affected by the presence of silver nanoparticles. The interaction of silver nanoparticles with acridinium derivative leads to a spectral change of its intramolecular charge transfer (ICT) absorption band. The dye emission increases suddenly with the initial addition of the Ag metal nanoparticles, but at a high concentration of the colloid, static fluorescence quenching occurs with a progressive decrease of the fluorescence efficiency. Amino coumarin fluorescence is only quenched by the silver nanoparticles in solution.

Surfactant--polymer aggregates formed by sodium dodecyl sulfate, poly(N-vinyl-2-pyrrolidone), and poly(ethylene glycol).

The interaction of sodium dodecyl sulfate (SDS) in aqueous solution with poly(N-vinyl-2-pyrrolidone) (M(w) = 55,000 g/mol) in the presence of poly(ethylene glycol) (M(w) = 8000 g/mol) is investigated by electrical conductivity, zeta potential measurements, viscosity measurements, fluorescence spectroscopy, and small-angle X-ray scattering (SAXS). The results indicate that SDS-polymer interaction occurs at low surfactant concentration, and its critical aggregation concentration is fairly dependent on polymer composition. The polymer-supported micelles have average aggregation numbers dependent on surfactant concentration, are highly dissociated when compared with aqueous SDS micelles, and have zeta potentials that increase linearly with the fraction of PVP at constant SDS concentration. The analysis of the SAXS measurements indicated that the PVP/PEG/SDS system forms surface-charged aggregates of a cylindrical shape with an anisometry (length to cross-section dimension ratio) of about 3.0.

Charge transfer complexes of 9-vinyl-carbazole with pi acceptors in homogeneous and in micellar solutions.

The molecular association of 9-vinyl-carbazole (CBZ) with three electron acceptors, p-chloranil (CHL), 2,7-dinitro-9-fluorenone (FL), and tetracyano-p-quinodimethane (TCNQ), is studied in acetonitrile and in micellar aqueous solution of sodium dodecyl sulfate (SDS). In both media, stable charge transfer (CT) complexes are formed with association constants in the range of 8-500 M(-1). CBZ and FL form a 1:2 complex in acetonitrile, but in SDS micelles the association is 1:1 due to size restriction and occupancy statistics in the host aggregates. The combination of absorption and fluorescence emission spectroscopy data indicates that the bimolecular CT complex of CBZ with TCNQ is stabilized in two distinct environments of the SDS micelles providing then two separated CT absorption bands.

Synthesis, characterization, and photophysical studies of new bichromophoric ruthenium(II) complexes.

The photophysical properties of a series of prepared ruthenium tris(bipyridine) complexes, covalently linked to aromatic species, of type [Ru(bpy)(2)-(4-methyl-4'-(arylaminocarbonyl)-2,2'-bipyridine)](2+) ([Ru(bpy)(2)(mbpy-L)](2+), where bpy = 2,2'-bipyridine; mbpy = 4-methyl-4'-carbonyl-2,2'-bipyridine; and L = 2-aminonaphthyl (naph), 9-aminoanthryl (anth), 1-aminopyrenyl (pyr), or 9-aminoacridinyl (acrd)) were studied by electronic absorption spectroscopy and steady state and time resolved luminescence spectroscopies. The absorption spectra of the MLCT electronic transition of the complexes are similar, which is in agreement with a practically constant redox potential of Ru(III/II) close to 1.28 V versus Ag/AgCl. However, the luminescence spectra of the new complexes are red shifted compared to Ru(bpy)(3)(2+), and this effect is ascribed to solvation and inductive effects of the amide group which enhance the symmetry breakdown among the three bipyridyl ligands. The energy stabilization of the (3)MLCT state is in the range 2.1-8.4 kJ/mol. The triplet-triplet energy transfer between the Ru complex and the aromatic species linked by an amide spacer is a slow process with rate constants of 2.6 x 10(4), 3.6 x 10(4), and 4.9 x 10(4) s(-)(1) for anthracene, acridine, and pyrene as acceptors in methanol, respectively. The energy transfer rate constant increases with decreasing polarity of the solvent. In dichloromethane, the rate constants for anthracene, acridine, and pyrene acceptors are 2.6 x 10(5), 1.5 x 10(5), and 2.9 x 10(5) s(-)(1), respectively. The low efficiency of energy transfer is due to the small difference in triplet energy between donor and acceptor species, weak electronic coupling, and unfavorable Franck-Condon factors, despite the short separation distance between donor and acceptor species in an amide bridge.

Time resolved fluorescence spectroscopy of quercetin and morin complexes with Al3+.

The association process of Al3+ with quercetin and morin in methanol was studied by electronic absorption and emission spectroscopies. The number of species in solution with different absorption spectra were determined by the method of Rank analysis of the absorbance matrix, and the stoichiometries of the complexes were evaluated using the Job method. The number of fluorescent species in solution were calculated from the Rank analysis method of the time resolved emission spectra (TRES), and compared with a global analysis of the decay surface using a proper multi-exponential decay model. The association of Al3+ with morin gives rise to two complexes with 1:1 and 2:1 (morin: Al3+) stoichiometries, but in both species the association of the cation involves the carbonyl and 3-hydroxyl groups of the pyrone ring. The fluorescence decay surface of this system is biexponential and the lifetimes of the 1:1 and 2:1 complexes are 4.3 and 2.0 ns, respectively. The association of Al3+ with quercetin forms preferentially two complexes with 1:1 and 1:2 (quercetin: Al3+) stoichiometries where the first cation binds to the site of the pyrone ring but the second one is bound to the cathecol group of the molecule. However, the multichelation character of the quercetin ligand allows larger aggregates to be formed, thereby the species Al2Q3 is also detected in methanol. The lifetime of the 1:1 complex is about 2.7 ns, while for 1:2 and 3:2 complexes the lifetimes measured are 3.5 and 1.8 ns, respectively.

The role of the carboxylate head group in the interaction of sodium dodecanoate with poly(ethylene oxide) investigated by electrical conductivity, viscosity, and aggregation number measurements.

The binding of sodium dodecanoate (SDoD) to poly(ethylene oxide) (PEO) in aqueous solution was investigated and compared with the well-known polymer-surfactant complexes formed between PEO and sodium dodecyl sulfate (SDS). Electrical conductivity measurements indicated that the concentration ratio of bound SDoD to PEO (on monomer basis) was greater than that for the system PEO-SDS. However, the aggregation numbers of the micelles supported on the polymer chain are practically constant and similar for both surfactants at concentrations lower than the polymer saturation point. The difference in binding capability is explained in terms of a larger PEO coil expansion upon complexation of SDoD than in the case of SDS. An increase in the polymer surface favors the binding of SDoD to PEO in aqueous solution. This conclusion is supported by the results of the viscometric studies of PEO-surfactant solution.

The Change in the Properties of Sodium Dodecyl Sulfate Micelles upon Addition of Isomeric and Unsaturated Short-Chain Alcohols Probed by Photophysical Methods.

The properties of sodium dodecyl sulfate micelles (SDS) in the presence of 1-propanol, 2-propanol, and 2-propen-1-ol (allyl alcohol) were investigated using electrical conductivity and fluorescence quenching methods. The constants of association of the alcohols to SDS micelles, obtained experimentally and calculated from linear solvation energy relationship analysis, are in the range 1-2 M(-1), indicating that these additives behave not only as cosolvents but also as micellar surface active solutes. Larger molecular volume and lower basicity of the alcohol favor partitioning to the micelle. The formation of smaller micelles upon addition of the alcohols is accompanied by an enhancement of solute intermicellar migration and intramicellar quenching rate constants. In particular, the micelle fragmentation-coagulation process of the SDS (0.04 M)/allyl alcohol (1.5 M) system occurs in the time range of 1.5 &mgr;s, as indicated by the intermicellar exchange rate constant of the N-dodecylpyridinium ion. Copyright 2001 Academic Press.