Fine-tuning of radiative properties by "mild" substituents: searching for a perfectly soft chromophore.

Controlling spectral properties to achieve desired characteristics is an attractive goal in application-oriented research, e.g., in the design of fluorescence sensors. "Soft" chromophores, molecules with strong spectral responses to internal or external perturbations are good candidates for such studies. In this work, absorption, fluorescence, and magnetic circular dichroism (MCD) spectra were obtained for a series of porphyrins, substituted at the meso-positions with n-hexyl groups. As the number of substituents increases from 1 to 4, significant changes are observed. The intensity of the S0-S1 transition (Qx) in the 0-0 region strongly decreases in mono-substituted porphyrin, but upon additional substitutions it increases to values larger than in the parent, unsubstituted molecule. Such behavior can be explained, using the perimeter model, by changes in the energy splittings between the two highest (HOMO) and two lowest (LUMO) frontier molecular orbitals. Single substitution makes porphyrin a nearly perfect soft chromophore, but upon introduction of a larger number of n-hexyl groups it is transformed into a hard one. DFT simulations incorrectly predict a continuous transition from a soft to hard chromophore, because the calculated ordering of two HOMO orbitals is opposite to that obtained by experiment. On the other hand, for those porphyrins that can be classified as hard chromophores, the calculations nicely reproduce contributions of Franck-Condon and Herzberg-Teller terms to absorption and fluorescence spectra.

Photodegradation of free base and zinc porphyrins in the presence and absence of oxygen.

Comparison of photostability in degassed and aerated toluene solutions is reported for 5,10,15,20-tetraphenylporphyrin, 5,10,15-tri(p-tolyl)porphyrin, and their zinc analogues. After degassing, quantum yields of photodegradation are higher, but the photodecomposition rates decrease. Lower stability in deoxygenated solutions is due to much longer triplet lifetimes: 200-300 microseconds, compared to 200-360 ns in non-degassed toluene. For the zinc porphyrins, the LC-MS results show that the initial photoproduct contains two oxygen atoms. Based on electronic absorption and calculations, it is assigned to dehydrated zinc biladienone structure, relatively stable in toluene, but readily demetallated in dichloromethane. A similar species is formed also in the case of free bases, but it then undergoes hydration due to traces of water present in the solvent. Zinc derivatives were found to form biladienones even in degassed solutions. To explain this observation, we postulate formation of a complex with remaining oxygen or oxygen-containing species which is not removed by freeze-thaw procedure. This hypothesis is confirmed by MS results and by the analysis of photodegradation products obtained when zinc porphyrin is complexed with dimethylsulfoxide (DMSO). Under these circumstances, changes in absorption are the same as in the absence of DMSO when non-degassed toluene is used, but irradiation of deoxygenated solutions leads to a different photoproduct. For both degassed and non-degassed solvents, complexation with DMSO results in the enhancement of photostability.

Solvent effects on the photooxidation of indolepyrazines.

Photodestruction of 2-(pyrazin-2'-yl)-1H-indole and 2,5-di(1H-indol-2'-yl)pyrazine involves singlet oxygen generation and its rapid insertion into the indole ring with the formation of benzoxazinone derivatives: 2-(pyrazin-2-yl)-4H-3,1-benzoxazin-4-one and 2-[5-(1H-indol-2-yl)pyrazin-2-yl]-4H-3,1-benzoxazin-4-one. The quantum yield of this reaction strongly depends on the environment. It is definitely smaller in protic methanol than in aprotic acetonitrile or n-hexane. The observed effect of photostabilization is explained by formation of hydrogen bonded complexes between the chromophore and alcohol, which results in lower triplet formation efficiency and, in consequence, decrease of singlet oxygen formation quantum yield.

Anisotropic coupling of individual vibrational modes to a Cu(110) substrate.

We present a study on the excitation of individual vibrational modes with ballistic charge carriers propagating along the Cu(110) surface. By means of the molecular nanoprobe technique, where the reversible switching of a molecule-in this case tautomerization of porphycene-is utilized to detect excitation events, we reveal anisotropic coupling of two distinct vibrational modes to the substrate. The N-H bending mode, excited below |E| ≈ 376 meV, exhibits maxima perpendicular to the rows of the Cu(110) substrate and minima along the rows. In contrast, the N-H stretching mode, excited above |E| ≈ 376 meV, displays maxima along the rows and is constant otherwise. This inversion of the anisotropy reflects the orthogonality between the N-H bending and stretching mode. Additionally, we observe an energy-dependent asymmetry in the propagation direction of charge carriers injected into the Cu(110) surface state. Hereby, the anisotropic band structure results in a correlation between the group velocity and the tunneling probability into electronic states of the substrate.

Tautomerization of single asymmetric oxahemiporphycene molecules on Cu(111).

We have studied 22-oxahemiporphycene molecules by a combination of scanning tunneling microscopy at low temperatures and density functional theory calculations. In contrast to other molecular switches with typically two switching states, these molecules can in principle exist in three different tautomers, due to their asymmetry and three inequivalent binding positions of a hydrogen atom in their macrocycle. Different tautomers are identified from the typical appearance on the surface and tunneling electrons can be used to tautomerize single molecules in a controllable way with the highest rates if the STM tip is placed close to the hydrogen binding positions in the cavity. Characteristic switching processes are explained by the different energy pathways upon adsorption on the surface. Upon applying higher bias voltages, deprotonation occurs instead of tautomerization, which becomes evident in the molecular appearance.

SERS Signature of SARS-CoV-2 in Saliva and Nasopharyngeal Swabs: Towards Perspective COVID-19 Point-of-Care Diagnostics.

In this study, the intrinsic surface-enhanced Raman spectroscopy (SERS)-based approach coupled with chemometric analysis was adopted to establish the biochemical fingerprint of SARS-CoV-2 infected human fluids: saliva and nasopharyngeal swabs. The numerical methods, partial least squares discriminant analysis (PLS-DA) and support vector machine classification (SVMC), facilitated the spectroscopic identification of the viral-specific molecules, molecular changes, and distinct physiological signatures of pathetically altered fluids. Next, we developed the reliable classification model for fast identification and differentiation of negative CoV(-) and positive CoV(+) groups. The PLS-DA calibration model was described by a great statistical value-RMSEC and RMSECV below 0.3 and R2cal at the level of ~0.7 for both type of body fluids. The calculated diagnostic parameters for SVMC and PLS-DA at the stage of preparation of calibration model and classification of external samples simulating real diagnostic conditions evinced high accuracy, sensitivity, and specificity for saliva specimens. Here, we outlined the significant role of neopterin as the biomarker in the prediction of COVID-19 infection from nasopharyngeal swab. We also observed the increased content of nucleic acids of DNA/RNA and proteins such as ferritin as well as specific immunoglobulins. The developed SERS for SARS-CoV-2 approach allows: (i) fast, simple and non-invasive collection of analyzed specimens; (ii) fast response with the time of analysis below 15 min, and (iii) sensitive and reliable SERS-based screening of COVID-19 disease.

Spectroscopic investigation of photophysics and tautomerism of amino- and nitroporphycenes.

Parent, unsubstituted porphycene and its two derivatives: 2,7,12,17-tetra-n-propylporphycene and 2,7,12,17-tetra-t-butylporphycene were substituted at the meso position with amino and nitro groups. These two families of porphycenes were characterized in detail with respect to their spectral, photophysical, and tautomeric properties. Two trans tautomers of similar energies coexist in the ground electronic state, but only one form dominates in the lowest excited singlet state. Absorption, magnetic circular dichroism (MCD), and emission anisotropy combined with quantum-chemical calculations led to the assignment of S1 and S2 transitions in both tautomers. Compared with the parent porphycene, the S1-S2 energy gap significantly increases; for one tautomeric form, the effect is twice as large as for the other. Both amino- and nitroporphycenes emit single fluorescence; previously reported dual emission of aminoporphycenes is attributed to a degradation product. Introduction of bulky t-butyl groups leads to a huge decrease in fluorescence intensity; this effect, arising from the interaction of the meso substituent with the adjacent t-butyl moiety, is particularly strong in the nitro derivative.

Solving the Puzzle of Unusual Excited-State Proton Transfer in 2,5-Bis(6-methyl-2-benzoxazolyl)phenol.

2,5-Bis(6-methyl-2-benzoxazolyl)phenol (BMP) exhibits an ultrafast excited-state intramolecular proton transfer (ESIPT) when isolated in supersonic jets, whereas in condensed phases the phototautomerization is orders of magnitude slower. This unusual situation leads to nontypical photophysical characteristics: dual fluorescence is observed for BMP in solution, whereas only a single emission, originating from the phototautomer, is detected for the ultracold isolated molecules. In order to understand the completely different behavior in the two regimes, detailed photophysical studies have been carried out. Kinetic and thermodynamic parameters of ESIPT were determined from stationary and transient picosecond absorption and emission for BMP in different solvents in a broad temperature range. These studies were combined with time-dependent- density functional theory quantum-chemical modeling. The excited-state double-well potential for BMP and its methyl-free analogue were calculated by applying different hybrid functionals and compared with the results obtained for another proton-transferring molecule, 2,5-bis(5-ethyl-2-benzoxazolyl)hydroquinone (DE-BBHQ). The results lead to the model that explains the difference in proton-transfer properties of BMP in vacuum and in the condensed phase by inversion of the two lowest singlet states occurring along the PT coordinate.

Influence of bulky substituents on single-molecule SERS sensitivity.

The surface-enhanced Raman spectroscopy (SERS) detection limit strongly depends on the molecular structure, which we demonstrate for a family of tert-butyl-substituted porphycenes. Even though the investigated species present very similar photophysical properties, the ratio between the SERS signal and fluorescence background depends on the number of bulky tert-butyl groups. Moreover, the probability of single molecule detection systematically drops with the number of the moieties attached to the pyrrole ring. As steric hindrance is the only significantly changing feature among the studied chromophores, we attribute the observed phenomena to the spatial structure. We also show that the sensitivity of the SERS technique can be improved by lowering the temperature. We managed to observe single-molecule spectra for derivatives for which this was unattainable at room temperature.

Controlling Emissive Properties by Intramolecular Hydrogen Bonds: Alkyl and Aryl meso-Substituted Porphycenes.

Porphycene, a porphyrin isomer, is an efficient fluorophore. However, four-fold meso substitution with alkyl groups decreases the fluorescence quantum yield by orders of magnitude. For aryl substituents, this effect is small. To explain this difference, we have synthesized and studied a mixed aryl-alkyl-substituted compound, 9,20-diphenyl-10,19-dimethylporphycene, as well as the 9,20-diphenyl and 9,20-dimethyl derivatives. Analysis of the structural, spectroscopic, and photophysical data of the six porphycenes, combined with quantum chemical calculations, shows a clear correlation between the strength of the intramolecular NH⋅⋅⋅N hydrogen bonds and the efficiency of the radiationless depopulation of the lowest-excited singlet state. This result led us to propose a model in which the delocalization of the inner protons in the cavity of the macrocycle is responsible for the nonradiative deactivation channel. The applicability of the model is confirmed by the literature data for other alkyl- or aryl-substituted porphycenes. The finding of a correlation between structural and emissive characteristics enables a rational design of porphycenes with desired photophysical properties.

Scouting for strong light-matter coupling signatures in Raman spectra.

Strong coupling between vibrational transitions and a vacuum field of a cavity mode leads to the formation of vibrational polaritons. These hybrid light-matter states have been widely explored because of their potential to control chemical reactivity. However, the possibility of altering Raman scattering through the formation of vibrational polaritons has been rarely reported. Here, we present the Raman scattering properties of different molecules under vibrational strong coupling conditions. The polariton states are clearly observed in the IR transmission spectra of the coupled system for benzonitrile and methyl salicylate in liquid phase and for polyvinyl acetate in a solid polymer film. However, none of the studied systems exhibits a signature of the polariton states in the Raman spectra. For the solid polymer film, we have used cavities with different layer structures to investigate the influence of vibrational strong coupling on the Raman spectra. The only scenario where alterations of the Raman spectra are observed is for a thin Ag layer being in direct contact with the polymer film. This shows that, even though the system is in the vibrational strong coupling regime, changes in the Raman spectra do not necessarily result from the strong coupling, but are caused by the surface enhancement effects.

Towards More Photostable, Brighter, and Less Phototoxic Chromophores: Synthesis and Properties of Porphyrins Functionalized with Cyclooctatetraene.

Free base and zinc porphyrins functionalized with cyclooctatetraene (COT), a molecule known as a good triplet-state quencher, have been obtained and characterized in detail by structural, spectral, and photophysical techniques. Substitution with COT leads to a dramatic decrease of the intrinsic lifetime of the porphyrin triplet. As a result, photostability in oxygen-free solution increases by two to three orders of magnitude. In non-degassed solutions, improvement of photostability is about tenfold for zinc porphyrins, but the free bases become less photostable. Similar quantum yields of photodegradation in free base and zinc porphyrins containing the COT moiety indicate a common mechanism of photochemical decomposition. The new porphyrins are expected to be much less phototoxic, since the quantum yield of singlet oxygen formation strongly decreases because of the shorter triplet lifetime. The reduction of triplet lifetime should also enhance the brightness and reduce blinking in porphyrin chromophores emitting in single-molecule regime, since the duration of dark OFF states will be shorter.

Influence of local microenvironment on the double hydrogen transfer in porphycene.

We performed time-resolved transient absorption and fluorescence anisotropy measurements in order to study tautomerization of porphycene in rigid polymer matrices at cryogenic temperatures. Studies were carried out in poly(methyl methacrylate) (PMMA), poly(vinyl butyral) (PVB), and poly(vinyl alcohol) (PVA). The results prove that in all studied media hydrogen tunnelling plays a significant role in the double hydrogen transfer which becomes very sensitive to properties of the environment below approx. 150 K. We also demonstrate that there exist two populations of porphycene molecules in rigid media: "hydrogen-transferring" molecules, in which tautomerization occurs on time scales below 1 ns and "frozen" molecules in which double hydrogen transfer is too slow to be monitored with nanosecond techniques. The number of "frozen" molecules increases when the sample is cooled. We explain this effect by interactions of guest molecules with a rigid host matrix which disturbs symmetry of porphycene and hinders tunnelling. Temperature dependence of the number of hydrogen-transferring molecules suggests that the factor which restores the symmetry of the double-minimum potential well in porphycene are intermolecular vibrations localized in separated regions of the amorphous polymer.

2 + 2 Can Make Nearly a Thousand! Comparison of Di- and Tetra-Meso-Alkyl-Substituted Porphycenes.

Two porphycenes, substituted at the meso positions with two and four methyl groups, respectively, reveal similar absorption spectra, but their photophysical properties are completely different. 9,20-dimethylporphycene emits fluorescence with about 20% quantum yield, independent of the solvent. In contrast, fluorescence of 9,10,19,20-tetramethylporphycene is extremely weak in nonviscous solvents, but it can be recovered by placing the chromophore in a rigid environment. We propose a model that explains these differences, based on calculations and structural analogies with other extremely weakly emitting derivatives, dibenzo[cde,mno]porphycenes. The efficient S1 deactivation involves delocalization of two inner cavity protons coupled with proton translocation toward a high-energy cis tautomer. The latter process leads to distortion from planarity. The probability of deactivation increases with the strength of the intramolecular NH···N hydrogen bonds. The model also explains the observation of biexponential fluorescence decay in weakly emitting porphycenes. It can be extended to other derivatives, in particular, the asymmetrically substituted ones. We also point to the possibility of using specific porphycenes as viscosity sensors, in particular, when working in single molecule regime.

Goodbye to Quinine in Sulfuric Acid Solutions as a Fluorescence Quantum Yield Standard.

Quinine is by far the most popular fluorescence quantum yield standard used nowadays. In this work, we exploit a modified version of a recently developed SAFE (simultaneous absorption and emission measurement) method for investigating the temperature dependence of fluorescence quantum yields. The accuracy of the method was verified using rhodamine 6G and rhodamine B solutions, which have well characterized quantum yield temperature dependences. Subsequently, we investigated the quantum yield temperature dependence of quinine solutions in 0.05 M sulfuric acid and 0.1 M perchloric acid. The results show a large temperature dependence of fluorescence quantum yield for quinine in 0.05 M sulfuric acid. This temperature dependence is particularly pronounced near room temperature (from 20 to 25 °C), where the quantum yield changes by -0.45% per Celsius degree. In contrast, the fluorescence quantum yield of quinine in 0.1 M perchloric acid shows no temperature dependence between 20 and 45 °C, where, in the entire range, the luminescence quantum yield value remains constant and equal to 0.60 ± 0.007. These results clearly indicate that it should be recommended to use quinine in 0.1 M perchloric acid rather than quinine in 0.05 M sulfuric acid as a standard solution for the determination of luminescence quantum yields.

Photoinduced oxidation of an indole derivative: 2-(1'H-indol-2'-yl)-[1,5]naphthyridine.

The UV-induced oxidation of 2-(1'H-indol-2'-yl)-[1,5]naphthyridine acetonitrile solution in the presence of air leads to the formation of 2-(1,5-naphthyridin-2-yl)-4H-3,1-benzoxazin-4-one as a major product and N-(2-formylphenyl)-1,5-naphthyridine-2-carboxamide as a minor one. The probable reaction mechanisms are different for the two photoproducts and may involve both the reaction with singlet oxygen generated by the excited substrate or the reaction of the excited substrate with the ground state oxygen molecule. Electronic absorption and IR spectra indicate that both photoproducts are formed as mixtures of syn and anti-rotameric forms. The obtained results indicate an efficient and easy method for the synthesis of molecules with a benzoxazinone structure.

Spectroscopy and Tautomerization Studies of Porphycenes.

Tautomerization in porphycenes, constitutional isomers of porphyrins, is strongly entangled with spectral and photophysical parameters. The intramolecular double hydrogen transfer occurring in the ground and electronically excited states leads to uncommon spectroscopic characteristics, such as depolarized emission, viscosity-dependent radiationless depopulation, and vibrational-mode-specific tunneling splittings. This review starts with documentation of the electronic spectra of porphycenes: Absorption and magnetic circular dichroism are discussed, together with their analysis based on the perimeter model. Next, photophysical characteristics are presented, setting the stage for the final part, which discusses the developments in research on tautomerism. Porphycenes have been studied in different experimental regimes: molecules in condensed phases, isolated in supersonic jets and helium nanodroplets, and, recently also on the level of single molecules investigated by optical and scanning probe microscopies. Because of the rich and detailed information obtained from these diverse investigations, porphycenes emerge as very good models for studying the complex, multidimensional phenomena involved in the process of intramolecular double hydrogen transfer.

Antiaromatic or Nonaromatic? 21 H,61 H-2,6(2,5)-Dipyrrola-1,5(2,6)-dipyridinacyclooctaphane-3,7-diene: a Porphycene Derivative with 4 N π Electrons.

A porphycene-derived compound with a 20 π-electron skeleton has been obtained by replacing two pyrrolene units of porphycene by pyridine rings. NMR, electronic absorption and MCD spectra, and the lack of fluorescence are typical for 4 N cyclic π electron systems. The electronic structure and the differences with respect to porphycene can be rationalized by treating these compounds as perturbed, doubly positively charged [22]annulene and [20]annulene perimeters, respectively. Even though the spectroscopic and photophysical criteria proposed for antiaromatic systems are fulfilled, the molecule is very stable. We argue that the compound should be characterized as nonaromatic rather than antiaromatic. The perimeter model is recommended as a powerful tool for predicting the electronic structure and spectra and as a useful addition to other methods that probe the aromaticity.

Near-Field Enhanced Photochemistry of Single Molecules in a Scanning Tunneling Microscope Junction.

Optical near-field excitation of metallic nanostructures can be used to enhance photochemical reactions. The enhancement under visible light illumination is of particular interest because it can facilitate the use of sunlight to promote photocatalytic chemical and energy conversion. However, few studies have yet addressed optical near-field induced chemistry, in particular at the single-molecule level. In this Letter, we report the near-field enhanced tautomerization of porphycene on a Cu(111) surface in a scanning tunneling microscope (STM) junction. The light-induced tautomerization is mediated by photogenerated carriers in the Cu substrate. It is revealed that the reaction cross section is significantly enhanced in the presence of a Au tip compared to the far-field induced process. The strong enhancement occurs in the red and near-infrared spectral range for Au tips, whereas a W tip shows a much weaker enhancement, suggesting that excitation of the localized plasmon resonance contributes to the process. Additionally, using the precise tip-surface distance control of the STM, the near-field enhanced tautomerization is examined in and out of the tunneling regime. Our results suggest that the enhancement is attributed to the increased carrier generation rate via decay of the excited near-field in the STM junction. Additionally, optically excited tunneling electrons also contribute to the process in the tunneling regime.

Versatile Approach for Reliable Determination of Both High and Low Values of Luminescence Quantum Yields.

The determination of luminescence quantum yields by means of relative methods requires setting identical experimental conditions for both sample and reference compounds. This requirement has a critical impact on the applicability of these protocols, as it does not allow for the precise determination of low quantum-yield values using well-characterized high-quantum-yield standards. We show that using the simultaneous absorption and fluorescence-emission measurement (SAFE) approach [ Nawara and Waluk. Anal. Chem. 2017 , 89 , 8650 ], the sample and reference compounds can be effectively measured with different excitation-slit spectral bandpass or integration times, separately optimized for each chromophore. This unique feature simplifies the determination of luminescence quantum yields, allowing measurements of low quantum-yield values using well-characterized high-quantum-yield standards.