Are "Carbon Dots" Always Carbon Dots? Evidence for their Supramolecular Nature from Structural and Dynamic Studies in Solution and in the Pure Solid.

A model for the morphology (size, shape, and crystallinity) of carbon dots (CDs) in the solid state consistent with the observed photoluminescence in solution is proposed herein. Overwhelming evidence has been collected that links the data coming from solid-state analysis (high-resolution transmission electron microscopy (HRTEM), atomic force microscopy (AFM), and matrix-assisted laser desorption/ionization (MALDI) mass spectrometry (MS)) to that of solution (pulsed-field gradient (PFG)-NMR spectroscopy, time-resolved fluorescence anisotropy (TRFA), and steady-state/time-resolved fluorescence), allowing the establishment of an overall structural model for CDs. According to this model, the so-called carbon dots, observed under HRTEM imaging, are in fact supramolecular organized structures dynamically assembled from small to medium-sized molecular species when the solvent is removed to give the solid form. In this way, the imaged nanoparticles (TEM/AFM) are not covalently bound entities formed during the synthetic process, but instead supramolecular entities formed by noncovalent interactions. These particles, if at all present in solution, have the form of loose associations of relatively small molecules. This study was conducted on CDs obtained from the hydrothermal carbonization (HTC) of a biomass waste (olive wet pomace).

Are "Carbon Dots" Always Carbon Dots? Evidence for Their Supramolecular Nature from Structural and Dynamic Studies in Solution and in the Pure Solid.

Invited for the cover of this issue are José Prata and co-workers at Instituto Superior de Engenharia de Lisboa and Instituto Superior Técnico. The image depicts a model for the supramolecular nature of carbon dots. Read the full text of the article at 10.1002/chem.202302955.

Correction: Miranda et al. Anion Binding by Fluorescent Ureido-Hexahomotrioxacalix[3]arene Receptors: An NMR, Absorption and Emission Spectroscopic Study. Molecules 2022, 27, 3247.

In the original publication [...].

Critical Analysis of Association Constants between Calixarenes and Nitroaromatic Compounds Obtained by Fluorescence. Implications for Explosives Sensing.

The binding behaviour of two ureido-hexahomotrioxacalix[3]arene derivatives bearing naphthyl (1) and pyrenyl (2) fluorogenic units at the lower rim towards selected nitroaromatic compounds (NACs) was evaluated. Their affinity, or lack of it, was determined by UV-Vis absorption, fluorescence and NMR spectroscopy. Different computational methods were also used to further investigate any possible complexation between the calixarenes and the NACs. All the results show no significant interaction between calixarenes 1 and 2 and the NACs in either dichloromethane or acetonitrile solutions. Moreover, the fluorescence quenching observed is only apparent and merely results from the absorption of the NACs at the excitation wavelength (inner filter effect). This evidence is in stark contrast with reports in the literature for similar calixarenes. A naphthyl urea dihomooxacalix[4]arene (3) is also subject to the inner filter effect and is shown to form a stable complex with trinitrophenol; however, the equilibrium association constant is greatly overestimated if no correction is applied (9400 M-1 vs 3000 M-1), again stressing the importance of taking into account the inner filter effect in these systems.

Anion Binding by Fluorescent Ureido-Hexahomotrioxacalix[3]arene Receptors: An NMR, Absorption and Emission Spectroscopic Study.

Fluorescent receptors (4a-4c) based on (thio)ureido-functionalized hexahomotrioxacalix[3]arenes were synthesised and obtained in the partial cone conformation in solution. Naphthyl or pyrenyl fluorogenic units were introduced at the lower rim of the calixarene skeleton via a butyl spacer. The binding of biologically and environmentally relevant anions was studied with NMR, UV-vis absorption, and fluorescence titrations. Fluorescence of the pyrenyl receptor 4c displays both monomer and excimer fluorescence. The thermodynamics of complexation was determined in acetonitrile and was entropy-driven. Computational studies were also performed to bring further insight into the binding process. The data showed that association constants increase with the anion basicity, and AcO-, BzO- and F- were the best bound anions for all receptors. Pyrenylurea 4c is a slightly better receptor than naphthylurea 4a, and both are more efficient than naphthyl thiourea 4b. In addition, ureas 4a and 4c were also tested as ditopic receptors in the recognition of alkylammonium salts.

Luminescent Carbon Dots from Wet Olive Pomace: Structural Insights, Photophysical Properties and Cytotoxicity.

Carbon nanomaterials endowed with significant luminescence have been synthesized for the first time from an abundant, highly localized waste, the wet pomace (WP), a semi-solid by-product of industrial olive oil production. Synthetic efforts were undertaken to outshine the photoluminescence (PL) of carbon nanoparticles through a systematic search of the best reaction conditions to convert the waste biomass, mainly consisting in holocellulose, lignin and proteins, into carbon dots (CDs) by hydrothermal carbonization processes. Blue-emitting CDs with high fluorescence quantum yields were obtained. Using a comprehensive set of spectroscopic tools (FTIR, Raman, XPS, and 1H/13C NMR) in combination with steady-state and time-resolved fluorescence spectroscopy, a rational depiction of WP-CDs structures and their PL properties was reached. WP-CDs show the up-conversion of PL capabilities and negligible cytotoxicity against two mammalian cell lines (L929 and HeLa). Both properties are excellent indicators for their prospective application in biological imaging, biosensing, and dynamic therapies driven by light.

Reversible Electronic Energy Transfer (Homo-FRET) in Cyclic Molecular and Supramolecular Systems: Fluorescence Anisotropy Decays for the Isotropic Interaction.

Reversible electronic energy transfer (homo-FRET) in cyclic multichromophoric systems is studied for sets of n identical fluorophores arranged in regular polygons (triangle, square, pentagon, etc.). A general analytic expression for the anisotropy decay is obtained for a regular polygon of any order, under the assumptions of isotropic interaction and nearest-neighbor FRET. A graphical way of connecting the decay form and polygon geometry based on the Frost circle is also presented. The consequences of the relaxation of these assumptions on the anisotropy decay are also discussed and analyzed in detail for the heptagon.

Conventional vs. Microwave- or Mechanically-Assisted Synthesis of Dihomooxacalix[4]arene Phthalimides: NMR, X-ray and Photophysical Analysis.

Direct O-alkylation of p-tert-butyldihomooxacalix[4]arene (1) with N-(bromopropyl)- or N-(bromoethyl)phthalimides and K2CO3 in acetonitrile was conducted under conventional heating (reflux) and using microwave irradiation and ball milling methodologies. The reactions afforded mono- and mainly distal di-substituted derivatives in the cone conformation, in a total of eight compounds. They were isolated by column chromatography, and their conformations and the substitution patterns were established by NMR spectroscopy (1H, 13C, COSY and NOESY experiments). The X-ray structures of four dihomooxacalix[4]arene phthalimide derivatives (2a, 3a, 3b and 5a) are reported, as well as their photophysical properties. The microwave (MW)-assisted alkylations drastically reduced the reaction times (from days to less than 45 min) and produced higher yields of both 1,3-di-substituted phthalimides (3a and 6a) with higher selectivity. Ball milling did not reveal to be a good method for this kind of reaction.

Finding Value in Wastewaters from the Cork Industry: Carbon Dots Synthesis and Fluorescence for Hemeprotein Detection.

Valorisation of industrial low-value waste residues was preconized. Hence, carbon dots (C-dots) were synthesized from wastewaters of the cork industry-an abundant and affordable, but environmentally-problematic industrial effluent. The carbon nanomaterials were structurally and morphologically characterised, and their photophysical properties were analysed by an ensemble of spectroscopy techniques. Afterwards, they were successfully applied as highly-sensitive fluorescence probes for the direct detection of haemproteins. Haemoglobin, cytochrome c and myoglobin were selected as specific targets owing to their relevant roles in living organisms, wherein their deficiencies or surpluses are associated with several medical conditions. For all of them, remarkable responses were achieved, allowing their detection at nanomolar levels. Steady-state and time-resolved fluorescence, ground-state UV-Vis absorption and electronic circular dichroism techniques were used to investigate the probable mechanisms behind the fluorescence turn-off of C-dots. Extensive experimental evidence points to a static quenching mechanism. Likewise, resonance energy transfer and collisional quenching have been discarded as excited-state deactivating mechanisms. It was additionally found that an oxidative, photoinduced electron transfer occurs for cytochrome c, the most electron-deficient protein. Besides, C-dots prepared from citric acid/ethylenediamine were comparatively assayed for protein detection and the differences between the two types of nanomaterials highlighted.

Dihomooxacalix[4]arene-Based Fluorescent Receptors for Anion and Organic Ion Pair Recognition.

Fluorescent dihomooxacalix[4]arene-based receptors 5a-5c, bearing two naphthyl(thio)ureido groups at the lower rim via a butyl spacer, were synthesised and obtained in the cone conformation in solution. The X-ray crystal structures of 1,3- (5a) and 3,4-dinaphthylurea (5b) derivatives are reported. Their binding properties towards several anions of different geometries were assessed by 1H-NMR, UV-Vis absorption and fluorescence titrations. Structural and energetic insights of the naphthylurea 5a and 5b complexes were also obtained using quantum mechanical calculations. The data showed that all receptors follow the same trend, the association constants increase with the anion basicity, and the strongest complexes were obtained with F-, followed by the oxoanions AcO- and BzO-. Proximal urea 5b is a better anion receptor compared to distal urea 5a, and both are more efficient than thiourea 5c. Compounds 5a and 5b were also investigated as heteroditopic receptors for biologically relevant alkylammonium salts, such as the neurotransmitter γ-aminobutyric acid (GABA·HCl) and the betaine deoxycarnitine·HCl. Chiral recognition towards the guest sec-butylamine·HCl was also tested, and a 5:2 selectivity for (R)-sec-BuNH3+·Cl- towards (P) or (M) enantiomers of the inherently chiral receptor 5a was shown. Based on DFT calculations, the complex [(S)-sec-BuNH3+·Cl-/(M)-5a] was indicated as the more stable.

Calixarenes as High Temperature Matrices for Thermally Activated Delayed Fluorescence: C70 in Dihomooxacalix[4]arene.

Thermally activated delayed fluorescence (TADF) of 12C70 and 13C70 was observed up to 140 °C in a p-tert-butyldihomooxacalix[4]arene solid matrix, a temperature range significantly higher than that of previous TADF quantitative studies. An effective singlet-triplet energy gap of 29 kJ/mol and triplet formation quantum yields of 0.97 and 0.99 were measured for 12C70 and 13C70, respectively. The photophysical properties of the two fullerenes in this new matrix are comparable to those obtained in polystyrene at a lower temperature range. Calixarenes are proposed to be suitable matrices for high temperature TADF studies and applications.

Towards the Development of a Low-Cost Device for the Detection of Explosives Vapors by Fluorescence Quenching of Conjugated Polymers in Solid Matrices.

Conjugated polymers (CPs) have proved to be promising chemosensory materials for detecting nitroaromatic explosives vapors, as they quickly convert a chemical interaction into an easily-measured high-sensitivity optical output. The nitroaromatic analytes are strongly electron-deficient, whereas the conjugated polymer sensing materials are electron-rich. As a result, the photoexcitation of the CP is followed by electron transfer to the nitroaromatic analyte, resulting in a quenching of the light-emission from the conjugated polymer. The best CP in our studies was found to be poly[(9,9-dioctylfluorenyl-2,7-diyl)-co-bithiophene] (F8T2). It is photostable, has a good absorption between 400 and 450 nm, and a strong and structured fluorescence around 550 nm. Our studies indicate up to 96% quenching of light-emission, accompanied by a marked decrease in the fluorescence lifetime, upon exposure of the films of F8T2 in ethyl cellulose to nitrobenzene (NB) and 1,3-dinitrobenzene (DNB) vapors at room temperature. The effects of the polymeric matrix, plasticizer, and temperature have been studied, and the morphology of films determined by scanning electron microscopy (SEM) and confocal fluorescence microscopy. We have used ink jet printing to produce sensor films containing both sensor element and a fluorescence reference. In addition, a high dynamic range, intensity-based fluorometer, using a laser diode and a filtered photodiode was developed for use with this system.

Dinuclear Zinc(II) Macrocyclic Complex as Receptor for Selective Fluorescence Sensing of Pyrophosphate.

A new diethylenetriamine-derived macrocycle known as L, bearing 2-methylquinoline arms and containing m-xylyl spacers, was prepared in good yield by a one-pot [2 + 2] Schiff base condensation procedure, followed by reduction with sodium borohydride. Up to now this is the first hexaazamacrocycle with appended fluorophore units. Single-crystal X-ray diffraction determination of the dinuclear zinc(II) complex of L showed that metal centers are located at about 7.20(2) Å from one another. This complex exhibits only weak fluorescence in aqueous solution, but the addition of 1 equiv of pyrophosphate (PPi) caused a 21-fold enhancement of the fluorescence intensity. The sensor response is linear up to a value of 10 µM HPPi(3-) and has a detection limit of 300 nM. The receptor behaves as a highly selective sensor for pyrophosphate as other anions, including phosphate, phenylphosphate (PhP), adenosine monophosphate (AMP), adenosine diphosphate (ADP), and adenosine triphosphate (ATP), failed to induce any fluorescence change and practically do not affect the fluorescence intensity of the sensor in the presence of HPPi(3-). Competition titrations carried out in aqueous solution at pH 7.4 [in 20 mM 3-(N-morpholino)propanesulfonic acid (MOPS) buffer] by spectrofluorometry revealed a high association constant value of 6.22 log units for binding of PPi by the dinuclear zinc(II) receptor, one of the highest reported values for colorimetric/fluorometric sensors able to work under real aqueous physiological conditions, while association constant values for binding of the other phosphorylated substrates are in the 5.51-4.03 log unit range.

Influence of excited-state absorption on time-resolved luminescence: general formalism and application to the phosphorescence of polycyclic aromatic hydrocarbons.

The luminescence decay of a species in an absorbing medium whose optical thickness changes with time, as occurs with triplet-triplet absorption following excitation cut-off, is studied theoretically and experimentally. A general luminescence decay function based on a distribution of optical thicknesses is presented. A simple decay function previously used empirically is shown to result from an exponential distribution of optical thicknesses. The general approach introduced allows the adequate description of the phosphorescence decays of two polycyclic aromatic hydrocarbons, coronene and triphenylene (normal and perdeuterated forms for both molecules), in polymer films in the presence of excited-state absorption.

Sensing and imaging of oxygen with parts per billion limits of detection and based on the quenching of the delayed fluorescence of (13)C70 fullerene in polymer hosts.

We report on a new method for sensing trace oxygen in the gas phase. It is based on the extreme efficiency of the quenching of the thermally activated delayed fluorescence of isotopically enriched carbon-13 fullerene C(70) ((13)C(70)). This fullerene was dissolved in polymer matrixes of varying oxygen permeability, viz., polystyrene (PS), ethyl cellulose (EC) and an organically modified silica gel ("ormosil"; OS). The sensor films (5-10 µm thick), on photoexcitation at 470 nm, display a strong delayed photoluminescence with peaks between 670 and 700 nm. Its quenching by molecular oxygen was studied at 25 and 60 °C and at concentrations from zero up to 150 ppmv of oxygen in nitrogen. The rapid lifetime determination (RLD) method was applied to determine oxygen-dependent lifetimes and for fluorescence lifetime imaging of oxygen. The lower limits of detection (at 1% quenching) vary with the polymer used (EC ∼250 ppbv, OS ∼320 ppbv, PS ∼530 ppbv at 25 °C) and with temperature. The oxygen sensors reported here are the most sensitive ones described so far.

Fluorescent homooxacalixarenes: recent applications in supramolecular systems.

This review covers recent advances (from 2006 to date) in supramolecular systems based on fluorescent homooxacalixarenes, namely hexahomotrioxacalix[3]arenes, dihomooxacalix[4]arenes and tetrahomodioxacalix[4]arenes, focusing on fluorescence sensing using their intrinsic fluorescence (built-in mesitol-like groups) or the extrinsic fluorescence of organic fluorophores, either covalently linked to the calixarenes or forming supramolecular complexes with them. Sensing applications of ions, ion pairs and neutral molecules are discussed, as well as the potential measurement of temperature based on thermally activated delayed fluorescence.

Characterization of the Aeration and Hydrodynamics in Vertical-Wheel™ Bioreactors.

In this work, the oxygen transport and hydrodynamic flow of the PBS Vertical-Wheel MINI™ 0.1 bioreactor were characterized using experimental data and computational fluid dynamics simulations. Data acquired from spectroscopy-based oxygenation measurements was compared with data obtained from 3D simulations with a rigid-lid approximation and LES-WALE turbulence modeling, using the open-source software OpenFOAM-8. The mass transfer coefficients were determined for a range of stirring speeds between 10 and 100 rpm and for working volumes between 60 and 100 mL. Additionally, boundary condition, mesh refinement, and temperature variation studies were performed. Lastly, cell size, energy dissipation rate, and shear stress fields were calculated to determine optimal hydrodynamic conditions for culture. The experimental results demonstrate that the kL can be predicted using Sh=1.68Re0.551Sc13G1.18, with a mean absolute error of 2.08%. Using the simulations and a correction factor of 0.473, the expression can be correlated to provide equally valid results. To directly obtain them from simulations, a partial slip boundary condition can be tuned, ensuring better near-surface velocity profiles or, alternatively, by deeply refining the mesh. Temperature variation studies support the use of this correlation for temperatures up to 37 °C by using a Schmidt exponent of 1/3. Finally, the flow was characterized as transitional with diverse mixing mechanisms that ensure homogeneity and suspension quality, and the results obtained are in agreement with previous studies that employed RANS models. Overall, this work provides new data regarding oxygen mass transfer and hydrodynamics in the Vertical-Wheel bioreactor, as well as new insights for air-water mass transfer modeling in systems with low interface deformation, and a computational model that can be used for further studies.

External heavy-atom effect on the prompt and delayed fluorescence of [70]fullerenes.

The influence of the external heavy-atom effect (HAE) on the fluorescence properties of C(70) and a C(70) methano monoadduct is determined. For this purpose the photophysics of these [70]fullerenes is studied in polystyrene (PS) and in a related heavy-atom polymer, poly(4-bromostyrene) (PBS). In the absence of HAE (PS matrix) both fullerenes display a strong thermally activated delayed fluorescence (DF) that is more pronounced in the case of C(70). In the presence of HAE (PBS matrix) both prompt (PF) and DF intensities decrease significantly, the same happening to the delayed fluorescence lifetimes. The relative fluorescence intensities (DF intensity/PF intensity) for each fullerene are on the other hand surprisingly similar to the respective ones in PS for the full experimental temperature range. The HAE is responsible for a significant increase of the S(1)→T(1) and S(1)←T(1) intersystem crossing (ISC) rates, and of the T(1)→S(0) radiative rate. In particular, the HAE on a S(1)←T(1) ISC rate is reported here for the first time. The overall substantial insensitivity of relative fluorescence intensities to HAE is explained by a compensation effect: As the S(1)→T(1) and S(1)←T(1) ISC rates on the one hand, and the T(1)→S(0) radiative rate on the other hand work in opposition with respect to DF, a near cancellation of effects occurs.

Silica nanoparticles with thermally activated delayed fluorescence for live cell imaging.

Thermally activated delayed fluorescence (TADF) has revolutionized the field of organic light emitting diodes owing to the possibility of harvesting non-emissive triplet states and converting them in emissive singlet states. This mechanism generates a long-lived delayed fluorescence component which can also be used in sensing oxygen concentration, measuring local temperature, or on imaging. Despite this strong potential, only recently TADF has emerged as a powerful tool to develop metal-free long-lived luminescent probes for imaging and sensing. The application of TADF molecules in aqueous and/or biological media requires specific structural features that allow complexation with biomolecules or enable emission in the aggregated state, in order to retain the delayed fluorescence that is characteristic of these compounds. Herein we demonstrate a facile method that maintains the optical properties of solvated dyes by dispersing TADF molecules in nanoparticles. TADF dye-doped silica nanoparticles are prepared using a modified fluorescein fluorophore. However, the strategy can be used with many other TADF dyes. The covalent grafting of the TADF emitter into the inorganic matrix effectively preserves and transfers the optical properties of the free dye into the luminescent nanomaterials. Importantly, the silica matrix is efficient in shielding the dye from solvent polarity effects and increases delayed fluorescence lifetime. The prepared nanoparticles are effectively internalized by human cells, even at low incubation concentrations, localizing primarily in the cytosol, enabling fluorescence microscopy imaging at low dye concentrations.

How fast is a fast equilibrium? A new view of reversible reactions.

Reversible reactions are described in terms of kinetic cycles. From this description, two useful parameters arise: 1) average cycle duration and 2) average number of cycles. The latter applies to cycles with at least one unstable species. These parameters allow answering the question "How fast is a fast equilibrium?" in absolute and in relative terms, respectively. The general interest of the approach is demonstrated by application to reactions drawn from several areas: Acid-base equilibria, enzyme kinetics, excited-state proton transfer, thermally activated delayed fluorescence, monomer-excimer kinetics and homo-FRET.