Laser-induced Optoacoustic Spectroscopy Research Articles

Laser-Induced Optoacoustic Spectroscopy Studies of Inorganic Functionalized Metallic Nanorods

Plasmonic nanoparticles can be used in photothermal therapy thanks to their ability to generate local heat. The light-to-heat conversion efficiency is crucial for therapeutic and diagnostic applica...

Photophysics of Xanthene Dyes at High Concentrations in Solid Environments: Charge Transfer Assisted Triplet Formation

The photophysical behavior of two xanthene dyes, Eosin Y and Phloxine B, included in microcrystalline cellulose particles is studied in a wide concentration range, with emphasis on the effect of dye concentration on fluorescence and triplet quantum yields. Absolute fluorescence quantum yields in the solid-state were determined by means of diffuse reflectance and steady-state fluorescence measurements, whereas absolute triplet quantum yields were obtained by laser-induced optoacoustic spectroscopy and their dependence on dye concentration was confirmed by diffuse reflectance laser flash photolysis and time-resolved phosphorescence measurements. When both quantum yields are corrected for reabsorption and reemission of radiation, ΦF values decrease strongly on increasing dye concentration, while a less pronounced decay is observed for ΦT . Fluorescence concentration quenching is attributed to the formation of dye aggregates or virtual traps resulting from molecular crowding. Dimeric traps are however able to generate triplet states. A mechanism based on the intermediacy of charge-transfer states is proposed and discussed. Calculation of parameters for photoinduced electron transfer between dye molecules within the traps evidences the feasibility of the proposed mechanism. Results demonstrate that photoactive energy traps, capable of yielding dye triplet states, can be formed even in highly-concentrated systems with random dye distributions.

Silver nanowires as infrared-active materials for surface-enhanced Raman scattering.

Surface-enhanced Raman scattering (SERS) is increasing in significance as a bioanalytical tool. Novel nanostructured metal substrates are required to improve performances and versatility of SERS spectroscopy. In particular, as biological tissues are relatively transparent in the infrared wavelength range, SERS-active materials suitable for infrared laser excitation are needed. Nanowires appear interesting in this respect as they show a very broad localized surface plasmon resonance band, ranging from near UV to near infrared wavelengths. The SERS activity of silver nanowires has been tested at three wavelengths and a fair enhancement at 1064 and 514 nm has been observed, whereas a very weak enhancement was present when exciting close to the nanowire extinction maximum. These experimentally measured optical properties have been contrasted with finite element method simulations. Furthermore, laser-induced optoacoustic spectroscopy measurements have shown that the extinction at 1064 nm is completely due to scattering. This result has an important implication that no heating occurs when silver nanowires are utilized as SERS-active substrates, thereby preventing possible thermal damage.

Intrinsic Photoprotective Mechanisms in Chlorophylls

Photosynthetic energy conversion competes with the formation of chlorophyll triplet states and the generation of reactive oxygen species. These may, especially under high light stress, damage the photosynthetic apparatus. Many sophisticated photoprotective mechanisms have evolved to secure a harmless flow of excitation energy through the photosynthetic complexes. Time-resolved laser-induced optoacoustic spectroscopy was used to compare the properties of the T1 states of pheophytin a and its metallocomplexes. The lowest quantum yield of the T1 state is always observed in the Mg complex, which also shows the least efficient energy transfer to O2 . Axial coordination to the central Mg further lowers the yield of both T1 and singlet oxygen. These results reveal the existence of intrinsic photoprotective mechanisms in chlorophylls, embedded in their molecular design, which substantially suppress the formation of triplet states and the efficiency of energy transfer to O2 , each by 20-25 %. Such intrinsic photoprotective effects must have created a large evolutionary advantage for the Mg complexes during their evolution as the principal photoactive cofactors of photosynthetic proteins.

Photophysics of an unsymmetrical Zn(II) phthalocyanine substituted with terminal alkynyl group

This paper examines photophysical properties of an unsymmetrical zinc phthalocyanine substituted with a terminal alkynyl group. The studies were concentrated on absorption (in the UV–vis–IR range) and fluorescence (steady-state and time resolved) in chloroform of unsymmetrical 9(10), 16(17), 23(24)-tri-tert-butyl-2-(pent-4-yloxy)phthalocyaninato zinc(II) of different concentrations. Moreover, dye photodegradation studies were also done. Besides, a laser-induced optoacoustic spectroscopy was also used to determine thermal deactivation as well as singlet oxygen generation yield. The Langmuir layers of the dye were formed and the in-situ absorption technique was applied to follow ability of the dye to aggregate formation. The experimental data were supported by the quantum chemical calculations with the use of the time-dependent density functional theory (TD-DFT) to obtain information on the distribution of electron density and electronic transitions in the molecular systems. The spectroscopic results of the unsymmetrical zinc phthalocyanine were confronted with the data of the symmetric zinc phthalocyanine.

The impact of solvents on the singlet and triplet states of selected fluorine corroles - absorption, fluorescence, and optoacoustic studies.

This paper examines the influence of aprotic solvents on the spectroscopic properties as well as the energy deactivation of two free-base corrole dyes substituted with C6F5 and/or 4-NO2C6H4 groups. Absorption, fluorescence and laser-induced optoacoustic spectroscopy have been used to follow the singlet and triplet states of fluorine corroles belonging to the A2B and A3 type in toluene (TL), chloroform (CL), dimethylformamide (DMF), dimethyl sulfoxide (DMSO) and also in solvent mixtures. Changes in the absorption and fluorescence spectra are influenced by the type of solvent mixture. The fluorescence behaviors of the two investigated corroles were extremely different - fluorescence of the nitro-corrole in TL is dramatically quenched in the presence of DMF. In contrast, fluorescence quenching of fluorine corroles in DMF-TL mixtures is substantially weakened. Absorption, fluorescence, triplet population as well as singlet oxygen generation parameters are evaluated. The spectral experimental data are supported by quantum chemical calculations - time-dependent density functional theory (TD-DFT) and cyclic voltammetry experiments. The presented results are discussed from a viewpoint of aggregation, tautomerization, and deprotonation effects occurring in the corroles.

Effect of concentration on the formation of rose bengal triplet state on microcrystalline cellulose: a combined laser-induced optoacoustic spectroscopy, diffuse reflectance flash photolysis, and luminescence study.

Laser-induced optoacoustic spectroscopy (LIOAS), diffuse reflectance laser flash photolysis (DRLFP), and laser-induced luminescence (LIL) have been applied in conjunction to the determination of triplet state quantum yields of Rose Bengal (RB) supported on microcrystalline cellulose, a strongly light-scattering solid. Among the three used methods, the only one capable of providing absolute triplet quantum yields is LIOAS, but DRLFP and LIL aid in demonstrating that the LIOAS signal arises in fact from the triplet state and confirm the trend found with RB concentration. The coherence found for the three techniques demonstrates the usefulness of the approach. Observed triplet quantum yields are nearly constant within a limited concentration range, after which they decay strongly due to the generation of inactive dye aggregates or energy trapping centers. When quantum yields are divided by the fraction of absorbed light exciting the dye, the quotient falls off steadily with concentration, following the same trend as the observed fluorescence quantum yield. The conditions that maximize triplet formation are determined as a compromise between the rising light absorption and the decrease of quantum yield with RB concentration.

Effect of Concentration on the Photophysics of Dyes in Light‐Scattering Materials.

Photoactive materials based on dye molecules incorporated into thin films or bulk solids are useful for applications as photosensitization, photocatalysis, solar cell sensitization and fluorescent labeling, among others. In most cases, high concentrations of dyes are desirable to maximize light absorption. Under these circumstances, the proximity of dye molecules leads to the formation of aggregates and statistical traps, which dissipate the excitation energy and lower the population of excited states. The search for enhancement of light collection, avoiding energy wasting requires accounting the photophysical parameters quantitatively, including the determination of quantum yields, complicated by the presence of light scattering when particulate materials are considered. In this work we summarize recent advances on the photophysics of dyes in light-scattering materials, with particular focus on the effect of dye concentration. We show how experimental reflectance, fluorescence and laser-induced optoacoustic spectroscopy data can be used together with theoretical models for the quantitative evaluation of inner filter effects, fluorescence and triplet formation quantum yields and energy transfer efficiencies.

Molecular symmetry determines the mechanism of a very efficient ultrafast excitation-to-heat conversion in Ni-substituted chlorophylls

In the Ni-substituted chlorophylls, an ultrafast (<60fs) deactivation channel is created, which is not present in Ni-porphyrins. This observation prompted us to investigate in detail the mechanism of excitation-to-heat conversion in Ni-substituted chlorophylls, experimentally, using time-resolved laser-induced optoacoustic spectroscopy, and theoretically, using group theory approach. The Ni-substituted chlorophylls show exceptional photostability and the optoacoustic measurements confirm the prompt and very efficient (100%) excitation-into-heat conversion in these complexes. Considering their excellent spectral properties and the loss-free excitation-into-heat conversion they are likely to become a new class of versatile photocalorimetric references. The curious features of the Ni-substituted chlorophylls originate from the symmetry of a ligand field created in the central cavity. The central NNi2+ bonds, formed via the donation of two electrons from each of the sp2 orbitals of two central nitrogens to an empty s−dx2−y2 hybrid centered on Ni2+, have a considerable covalent character. The extreme rate of excited state relaxation is then not due to a ladder of the metal centered d-states, often invoked in metalloporphyrins, but seems to result from a peculiar topology of the potential energy surface (a saddle-shaped crossing) due to the covalent character of the NNi2+ bonds. This is confirmed by a strong 0→0 character of electronic transitions in these complexes indicating a similarity of their equilibrium geometries in the ground (S0) and the excited states (both QX and QY). The excitation energy is very efficiently converted into molecular vibrations and dissipated as heat, involving the central Ni2+. These Ni-substituted pigments pose a fine exemplification of symmetry control over properties of excited states of transition metal complexes.

Triplet quantum yields in light-scattering powder samples measured by laser-induced optoacoustic spectroscopy (LIOAS)

In recent years, different methods and techniques have been applied to study the primary photophysical processes occurring in dye-loaded light-scattering powdered samples. In spite of this, there are still no reliable methods for the determination of triplet quantum yields for this kind of systems. Laser-induced optoacoustic spectroscopy (LIOAS) has been extensively used for the determination of triplet quantum yields of dyes in solution. In a previous work, LIOAS was applied to the measurement of absolute emission quantum yields of highly fluorescent powdered samples. Excellent agreement was found with values obtained from reflectance data. In this work, we apply the same technique for the determination of triplet quantum yields of Rose Bengal and Erythrosine B adsorbed on microcrystalline cellulose. In contrast to water and other solvents, internal conversion cannot be neglected in the cellulose environment. The triplet quantum yield for both dyes is around 0.55 and does not change with dye concentration.

Structural volume changes upon triplet formation of water-soluble porphyrins depend on the resonant effect of the substituents

Laser-induced optoacoustic spectroscopy was applied to evaluate the photoinduced structural volume changes upon triplet state formation, Δ(T)V, of an aqueous solution of 5,10,15,20-tetrakis-(4-carboxyphenyl)-porphin. Two molecular forms differing in the ionization state of the carboxylic groups at the para-position of the phenyl ring were studied. The contractions, Δ(T)V, for all water-soluble 5,10,15,20-tetra-aryl-porphyrin compounds studied to date show a linear correlation with the Hammett resonant σ(R) constant. This indicates that the resonance electronic communication spreading over the π-orbitals between the meso-aryl rings and the porphyrin macrocycle determines the molecular contraction of the aqueous sphere around the macrocycle in the triplet state.

π-Cation interactions as the origin of the weak absorption at 532 nm observed in tryptophan-containing polypeptides

We have previously reported that bovine serum albumin (BSA) and other proteins that do not contain prosthetic groups exhibited a weak light absorption in the visible, only detectable by pulsed laser-induced optoacoustic spectroscopy (LIOAS). Human serum albumin (HSA) exhibited signals 25% higher than those observed with BSA. Signals comparable to those obtained with BSA were observed with poly(L-Trp, L-Lys), poly(L-Trp, L-Arg) or poly(L-Trp, L-Orn) at pH 7.0. No signals were obtained when tryptophan was replaced by other amino acids or when free tryptophan or the tripeptide Lys-Trp-Lys was assayed (pH 7.0). Tryptophan in HCl 5 N produced LIOAS signals similar to those produced by tryptophan-containing copolymers. Moreover, the absorption peak could be observed in a UV-VIS spectrophotometer. Therefore, the LIOAS signals obtained with BSA, HSA, and tryptophan-containing random copolymers may be attributed to a new transition of the indole moiety of their tryptophan residues when "protonated". Tryptophan residues of proteins are known to participate in π-cation interactions, which are important in protein stability and function. As a matter of fact, HSA and BSA contain an internal tryptophan in close proximity to lysine and arginine residues and therefore suitable for π-cation interactions. The strength of this type of interaction strongly depends on distances and relative orientations of both amino acid residues. Accordingly, these interactions should be highly sensitive to conformational changes. Based on preliminary results that have shown that LIOAS signal at 532 nm depended on the aggregation state of BSA and/or on the oxidation state of its Cys-34, we postulate that the LIOAS signal observed with proteins and tryptophan-containing polypeptides are related to Trp-Lys or Trp-Arg interactions and that the intensity of the signal depends on the strength of such interactions.

Photovoltaic and spectroscopic studies of selected halogenated porphyrins for their application in organic solar cells

Three porphyrin dyes were investigated: basic tetraphenylporphyrin (TPP), fluorine tetraphenylporphyrin symmetrically substituted with twelve fluorine atoms (TPPF 12) and chlorine tetraphenylporphyrin symmetrically substituted with twelve chlorine atoms (TPP Cl 12). TPP without external atoms was used as a standard sample. Photoelectrochemical examinations (short-circuit photocurrent in the second time scale) combined with the spectroscopic methods (absorption, fluorescence, photoacoustics supported by the laser-induced optoacoustic spectroscopy) were applied. On the basis of absorption and fluorescence spectra some spectral parameters were estimated, whereas with photoacoustic methods the thermal parameters and kinetics of thermal relaxation in the microsecond time scale were evaluated. The studies of the dyes in the range from about 10 −6 M up to 10 −3 M show the absence of aggregated structures although dye fluorescence behavior could indicate the presence of weakly interacting dye molecules when the dyes are in their excited states. The results show that the changes in fluorescence behavior indicate redistribution of the electron density in the molecular macroring upon linkage of the fluorine and chlorine atoms to the main molecular core. The results of the photocurrent and spectral examinations were interpreted in terms of occurrence of mesomeric and inductive effects; photocurrent enhancement in the chlorinated porphyrin (TPPCl 12) versus the free-based (TPP) and fluorinated (TPPF 12) porphyrins support the occurrence of the mesomeric effect which dominates over the inductive one in TPPCl 12, whereas in TPPF 12 the inductive effect is essential and can lead to photocurrent declining.

Validation of Fluorescence Quantum Yields for Light-Scattering Powdered Samples by Laser-Induced Optoacoustic Spectroscopy

Determination of quantum yields for various processes (such as fluorescence and triplet formation) in dye-loaded light-scattering powdered samples is an open issue. Here, we report the testing of laser-induced optoacoustic spectroscopy (LIOAS) for the determination of fluorescence quantum yields of Rhodamine 101 and Rhodamine 6G adsorbed at various loadings on microgranular cellulose powder. The results of the LIOAS experiments are consistent with those from the method developed in one of our laboratories based on the measurement of apparent reflectance using an integrating sphere [Mirenda, M.; Lagorio, M. G.; San Roman, E. Langmuir 2004, 20, 3690-3697], which allows the simultaneous calculation of fluorescence quantum yield and reflectance devoid of fluorescence artifacts. Criteria to quantify overall errors and detect outlying values are developed. The theory underlying the application of LIOAS to light-scattering samples and experimental details are presented.

Entropic Changes Control the Charge Separation Process in Triads Mimicking Photosynthetic Charge Separation

Laser-induced optoacoustic spectroscopy (LIOAS) measurements with carotene-porphyrin-acceptor "supermolecular" triads (C-P-A, with A = C60, a naphthoquinone NQ, and a naphthoquinone derivative, Q) were carried out with the purpose of analyzing the thermodynamic parameters for the formation and decay of the respective long-lived charge separated state C*+-P-A*-. The novel procedure of inclusion of the benzonitrile solutions of the triads in Triton X-100 micelle nanoreactors suspended in water permitted the separation of the enthalpic and structural volume change contributions to the LIOAS signals, by performing the measurements in the range 4-20 degrees C. Contractions of 4.2, 5.7, and 4.2 mL mol-1 are concomitant with the formation of C*+-P-A*- for A = C60, Q and NQ, respectively. These contractions are mostly attributed to solvent movements and possible conformational changes upon photoinduced electron transfer, due to the attraction of the oppositely charged ends, as a consequence of the giant dipole moment developed in these compounds upon charge separation ( approximately 110 D). The estimations combining the calculated free energies and the LIOAS-derived enthalpy changes indicate that entropy changes, attributed to solvent movements, control the process of electron transfer for the three triads, especially for C-P-C60 and C-P-Q. The heat released during the decay of 1 mol of charge separated state (CS) is much smaller than the respective enthalpy content obtained from the LIOAS measurements for the CS formation. This is attributed to the production of long-lived energy storing species upon CS decay.

Laser induced optoacoustic spectroscopy of benzil: Evaluation of structural volume change upon photoisomerization

The cis-skew to trans-planar photoisomerization of benzil in the photoexcited state was studied by laser-induced optoacoustic spectroscopy (LIOAS) in a series of normal alkane solvents at room temperature. The structural volume change due to the photoisomerization process has been estimated and compared with the same calculated from the optimized skew and trans structures. The magnitude of the structural volume change is estimated to be 22.9 ± 1.2 mL/mol. This study reveals that an expansion in volume occurs during the photoinduced isomerization process. In non-polar alkane solvents, the estimated volume change has been argued to be free from electrostrictional contribution.

Photoinduced Electron Transfer from Tetrasulfonated Porphyrin to Benzoquinone Revisited. The Structural Volume-Normalized Entropy Change Correlates with Marcus Reorganization Energy

Time-resolved laser-induced optoacoustic spectroscopy was used for the determination of the enthalpy, DeltaTH, and structural volume changes, DeltaTV, concomitant with triplet state formation upon excitation of meso-tetrakis(4-sulfonatophenyl)porphyrin, TSPP(4-), as well as with the triplet state electron-transfer (ET) quenching by benzoquinone, BQ (DeltaRH and DeltaRV). The values of DeltaTH and DeltaTV for (3)TSPP(4-) formation in the presence of different cations (Li+, Na+, K+, NH4+, and Cs+) correlated with each other and afforded a value of DeltaTG = 140 +/- 20 kJ mol(-1), equal to the value of E+ at 77 K, but much larger than the DeltaTH values in solution at room temperature, due to the large entropic factor in solution. The influence of the cations on DeltaTH and DeltaTV (a contraction ranging from 5.4 to 3.8 cm3 mol(-1)) is attributed to changing chromophore-water interactions in the ground and triplet states. Upon quenching of 3TSPP(4-) by BQ, the quantum yield of free radical formation, PhiR = 0.66 +/- 0.04, is the same in the solutions of the five cations. The values of DeltaRH and DeltaRV are small and have a large error. The energy level of the free radicals formed is thus very similar to that of 3TSPP(4-). TDeltaRS and X = TDeltaRS/DeltaRV, i.e., the structural volume change-normalized entropy change for free radicals formation, were derived using average values of DeltaRH and DeltaRV together with the calculated DeltaRG degrees . The measured Marcus reorganization energy, lambda, and X fall into the lambda vs X linear dependence we previously found for the radical formation upon ET quenching of triplet flavins (3FMN and 3FAD) by amines and amino acids. Thus, X = TDeltaRS/DeltaRV in aqueous solutions is a property of the particular donor-acceptor pair linearly correlated to the corresponding Marcus reorganization energy. The value of X is much larger than the predicted value applying the electrostriction concepts in view of the noncontinuum nature of the aqueous solutions.

Photoinduced Electron Transfer to Triplet Flavins. Correlation between the Volume Change-Normalized Entropic Term and the Marcus Reorganization Energy

The data obtained through the application of nanosecond laser-induced optoacoustic spectroscopy (LIOAS) to several electron donor-acceptor pairs in aqueous solution were analyzed together with the respective experimentally determined Marcus reorganization energy. Acceptors were the flavin mononucleotide and flavin-adenine dinucleotide triplet states (3FMN and 3FAD) and donors were tryptophan, tyrosine, histidine, triethanolamine, and ethylenediaminetetraacetic acid. The respective calculated Gibbs energy for electron transfer, Delta(ET)G degrees , was used together with the enthalpy change for the formation of free radicals, Delta(FR)H, obtained from the LIOAS data, to derive the entropy change for the formation of the radicals, Delta(FR)S. In all cases, variation of the monovalent cations, i.e., [CH3(CH2)3]4N+, Li+, NH4+, K+, and Cs+, resulted in variation of the enthalpy change, Delta(FR)H, and in the structural volume change, Delta(FR)V, for the free-radical production, both derived from LIOAS. Delta(FR)H and Delta(FR)V linearly correlated with each other within the cation series. From this correlation the respective entropic term TDelta(FR)S was derived as well as the ratio X = TDelta(FR)S/Delta(FR)V for each of the pairs. X linearly correlated with the respective total Marcus reorganization energy, lambda, for all systems analyzed. This observation underlines the concept that both lambda and Delta(FR)V respond to the same phenomena. The correlation also offers an experimental approach for the understanding at a molecular level of the origin of the lambda values as well as for their evaluation.

The influence of flavo nucleotide (FMN) dimerization on the efficiency of the FMN triplet states generation

Abstract The efficiency of triplet state generation in flavin mononucleotide (FMN) water solutions at various pigment concentrations illuminated at two wavelengths (380 nm and 430 nm) was established by the Laser Induced Optoacoustic Spectroscopy (LIOAS) method. It was found that the efficiency of triplet generation by the singlet–triplet intersystem crossing after illumination at 430 nm was usually lower than that after illumination at 380 nm. The yield of triplet generation in FMN at low concentrations in solutions at 380 nm illumination is rather high, about 0.6 but its value is diminished with the increase in the sample temperature and the dye concentration and depends also on the wavelength of excitation. The increase in FMN dimers content gives an increase in fast internal nonradiative conversion of excitation and a decrease in the yield of the fluorescence. LIOAS measurements of the sample and the reference dye in buffered water solutions at 3 °C suggest that only a small part of the excitation energy is used for some structural changes of FMN or possibly for the changes in FMN interactions with its surrounding solvent molecules. The changes in both types should ensue some modification in the volume of the absorbing species giving nonthermal contributions to LIOAS signal. The LIOAS results were corrected for FMN volume changes. The different yields of triplet states generation obtained by two wavelengths of illumination are not only due predominantly to various contributions to absorption from dimeric FMN but also can be partially related to different grade of the superposition of n–π ∗ and π–π ∗ transitions in a region of 380 nm and 430 nm absorption. The absorption at 430 nm band is due predominantly to π–π ∗ transition, whereas in 380 nm region the n–π ∗ transition could have higher contribution which can exhibit different efficiency of triplet generation than π–π ∗ transition. For the samples with low FMN concentrations the yields of fluorescence of FMN excited at these two regions are similar, but at higher dye concentrations they are different.

Acid−Base Equilibria in 5,10,15,20-Tetrakis(4-sulfonatophenyl)chlorin: Role of Conformational Flexibility

The acid-base equilibria in 5,10,15,20-tetrakis(4-sulfonatophenyl)chlorin were studied in aqueous solution and compared with the respective data for the corresponding porphyrin. The reduction of the pyrrole ring in the tetrapyrrolic macrocycle noticeably influences both free base/monoprotonated and mono-/diprotonated species equilibria. In strong acidic solutions protonation of 4-sulfonatophenyl groups takes place in addition to protonation of the macrocycle core. The photophysical properties of all ionic forms are influenced by an enhanced rate of internal S1 --> S0 conversion, leading to about 50% and 90% deactivation through this channel for the free base and diprotonated species, respectively. The enhancement of the rate of the radiationless transitions is explained by an increased conformational flexibility of the chlorin macrocycle with respect to that of a porphyrin. Structural volume change measurements with laser-induced optoacoustic spectroscopy support this explanation. The contraction upon triplet state formation of the free base is about one-half of that measured for the corresponding porphyrin. This contraction should be due to intramolecular structural rearrangements of the macrocycle to adopt a minimum energy conformation in case of the chlorin. On the contrary, for the more rigid porphyrin macrocycle the interactions of the molecule with the solvent environment play a more important role. The diprotonated forms of both porphyrin and chlorin show a high radiationless S1 --> S0 conversion rate and seem to have a similar conformational flexibility. In agreement with previous calculations, the conformational flexibility of the diprotonated forms appears to be higher than that of the free base molecule.