Nuclear Spin Singlet States in Photoactive Molecules: From Fluorescence/NMR Bimodality to a Bimolecular Switch for Spin Singlet States

Nuclear spin singlet states are silent states in nuclear magnetic resonance (NMR). However, they can be probed indirectly and offer great potential for the development of contrast agents for magnetic resonance imaging (MRI). Introduced here are two novel concepts: Firstly, the bimodal NMR/fluorescence properties of ¹³C₂‐tetraphenylethylene. It possesses a long‐lived singlet state in organic solvents, and it shortens upon the addition of water. This simultaneously increases the aggregation‐induced emission (AIE) of the molecule, resulting in a substantial enhancement of fluorescence. Secondly, introduced is a bimolecular switch for singlet states based on 3‐²H‐coumarin containing an isolated proton. Upon UV‐light exposure, a dimer forms, leading to a coupling between two previously isolated protons. A nuclear spin singlet state can now be populated. Excitation with a wavelength of 254 nm results in partial ring cleavage of the molecule back to its monomer.     

Interaction between encapsulated excited organic molecules and free nitroxides: communication across a molecular wall

Communication between two molecules, one confined and excited (triplet or singlet) and one free and paramagnetic, has been explored through quenching of fluorescence and/or phosphorescence by nitroxides as paramagnetic radical species. Quenching of excited states by nitroxides has been investigated in solution, and the mechanism is speculated to involve charge transfer and/or exchange processes, both of which require close orbital interaction between excited molecule and quencher. We show in this report that such a quenching, which involves electron-electron spin communication, can occur even when there is a molecular wall between the two. The excited state molecule is confined within an organic capsule made up of two molecules of a deep cavity cavitand, octa acid, that exists in the anionic form in basic aqueous solution. The nitroxide is kept free in aqueous solution. (1)H NMR and EPR experiments were carried out to ascertain the location of the two molecules. The distance between the excited molecule and the paramagnetic quencher was manipulated by the use of cationic, anionic, and neutral nitroxide and also by selectively including the cationic nitroxide within the cavity of cucurbituril. Results presented here highlight the role of the lifetime of the encounter complex in electron-electron spin communication when the direct orbital overlap between the two molecules is prevented by the intermediary wall.     

Photoactivatable Aggregation‐Induced Emission Fluorophores with Multiple‐Color Fluorescence and Wavelength‐Selective Activation

Photoactivatable (caged) fluorophores are widely used in chemistry, materials, and biology. However, the development of such molecules exhibiting photoactivable solid‐state fluorescence is still challenging due to the aggregation‐caused quenching (ACQ) effect of most fluorophores in their aggregate or solid states. In this work, we developed caged salicylaldehyde hydrazone derivatives, which are of aggregation‐induced emission (AIE) characteristics upon light irradiation, as efficient photoactivatable solid‐state fluorophores. These compounds displayed multiple‐color emissions and ratiometric (photochromic) fluorescence switches upon wavelength‐selective photoactivation, and were successfully applied for photopatterning and photoactivatable cell imaging in a multiple‐color and stepwise manner.     

Generating long-lasting 1H and 13C hyperpolarization in small molecules with parahydrogen-induced polarization

Recently, Levitt and co-workers demonstrated that conserving the population of long-lasting nuclear singlet states in weak magnetic fields can lead to a preservation of nuclear spin information over times substantially longer than governed by the (high-field) spin-lattice relaxation time T1. Potential benefits of the prolonged spin information for magnetic resonance imaging and spectroscopy were pointed out, particularly when combined with the parahydrogen induced polarization (PHIP) methodology. In this contribution, we demonstrate that an increase of the effective relaxation time by a factor up to three is achieved experimentally, when molecules hyperpolarized by PHIP are kept in a weak magnetic field instead of the strong field of a typical NMR magnet. This increased lifetime of spin information makes the known PHIP phenomena more compatible with the time scales of biological processes and, thus, more attractive for future investigations.     

Pathways for fluorescence quenching in 2-aminopurine π-stacked with pyrimidine nucleobases

Fluorescent analogues of nucleobases are very useful as probes to study DNA dynamics, because natural DNA does not fluoresce significantly. In many of these analogues, such as 2-aminopurine (2AP), the fluorescence is quenched when incorporated into DNA through processes that are not well understood. This work uses theoretical studies to examine fluorescence quenching pathways in 2AP-containing dimers. The singlet excited states of π-stacked dimer systems containing 2AP and a pyrimidine base, thymine or cytosine, have been studied using ab initio computational methods. Computed relaxation pathways along the excited-state surfaces reveal novel mechanisms that can lead to fluorescence quenching in the π-stacked dimers. The placement of 2AP on the 5' or 3' terminus of the dimers has different effects on the excitation energies and the relaxation pathways on the S(1) excited state. Conical intersections between the ground and first excited states exist when 2AP is placed at the 3' side, whereas the placement of 2AP at the 5' side leads to the switching of a bright state to a dark state. Both of these processes can lead to fluorescence quenching and may contribute to the fluorescence quenching observed in 2AP when incorporated in DNA.     

Graphene as a quencher of electronic excited states of photochemical probes

Graphene sheets quench the singlet and triplet excited states of a series of six photochemical probes including pyrene, acridine orange, tris(2,2?-bipyridyl)ruthenium(II) dichloride, methylene blue, meso-tetrakis(phenylsulphonate)porphyrin, and 5,10,15,20-tetraphenyl-21H,28H-porphine zinc. It was found that Stern-Volmer fluorescence quenching can fit to one or two different quenching regimes depending on the probe. In addition, the quenching can be either static or dynamic depending on the fluorophore. The occurrence of several quenching regimes has been interpreted considering that quenching arises from the crowding of the fluorophore on both graphene faces, or site isolation on the graphene sheets. Laser flash photolysis has shown that the triplet lifetime of the probes generally decreases due to graphene quenching and that no new transients appear except in the case of methylene blue, where a new absorption spectrum characterized by a continuous absorption band is observed and attributed to graphene radical ion. This spectroscopic evidence suggests that the most general quenching mechanism is energy transfer from the singlet or triplet excited state of the dye to graphene. This raises the issue of determining the energy of the electronic excited states of graphene.     

Treating molecules in arbitrary spin states using the parametric two-electron reduced-density-matrix method

Minimizing the electronic energy with respect to a parameterized two-electron reduced density matrix (2-RDM) is known as a parametric variational 2-RDM method. The parametric 2-RDM method with the M 2-RDM parametrization [D. A. Mazziotti, Phys. Rev. Lett. 101, 253002 (2008)] is extended to treat molecules in arbitrary spin states. Like its singlet counterpart, the M parametric 2-RDM method for arbitrary spin states is derived using approximate N-representability conditions, which allow it to capture more correlation energy than coupled cluster with single and double excitations at a lower computational cost. We present energies, optimized bond lengths, potential energy curves, and occupation numbers for a set of molecules in a variety of spin states using the M and K parametric 2-RDM methods as well as several wavefunction methods. We show that the M parametric 2-RDM method can describe bond breaking of open-shell molecules like triplet B(2) and singlet and triplet OH(+) even in the presence of strong correlation. Finally, the computed 2-RDMs are shown to be nearly N-representable at both equilibrium and non-equilibrium geometries.     

About long-lived nuclear spin states involved in para-hydrogenated molecules

This study deals with a spin system constituted of three nonequivalent protons, two of them originating from para-hydrogen (p-H(2)) after a hydrogenation reaction carried out in the earth magnetic field. It is shown that three singlet states are created provided indirect (J) couplings exist between the three spins, implying hyperpolarization transfer toward the third spin. Upon insertion of the sample in the NMR (Nuclear Magnetic Resonance) high field magnet, the following events occur: (i) the longitudinal two-spin orders which are parts of the singlet states survive; (ii) the other two terms (of these singlet states) tend to be destroyed by magnetic field gradients but at the same time are partly converted into differences of longitudinal polarizations. Nuclear spin relaxation is studied by appropriate NMR measurements when evolution takes place in the high field magnet or in the earth field. In the former case, relaxation is classical although complicated by numerous relaxation rates associated with both longitudinal two-spin orders and longitudinal polarizations. In the latter case, an equilibration between the singlet states first occur, their disappearance being thereafter driven by relaxation rates which remain very small because of the absence of any dipolar contribution. Thus, even in the case of a three-spin system, long-lived states exist; this unexpected property could be very useful for many applications.     

THE EFFECTS OF AGGREGATION ON THE FLUORESCENCE and THE TRIPLET STATE YIELD OF HEMATOPORPHYRIN

Abstract— The fluorescence and triplet state yields of hematoporphyrin have been measured in wuter/methanol mixtures. There is a closely linked response of these yields to aggregation of the hematoporphyrin molecule. The monomer, which is the principal species present at high methanol content, has a triplet state yield of 0.91 and a fluorescence yield of 0.09. By contrast, hematoporphyrin solutions with a high water content containing aggregates have lower fluorescence and triplet state yields, e.g. 0.018 and 0.56, respectively, in water. Static, singlet state quenching in some of the aggregates is responsible for the reduced fluorescence yield. The results also show that in addition to these aggregates there are other types of aggregates where there is an increased singlet to ground state radiative transition probability, resulting from the interaction between transition dipoles in adjacent molecules.     

Electrogenerated Chemiluminescence of Donor–Acceptor Molecules with Thermally Activated Delayed Fluorescence

The electrochemistry and electrogenerated chemiluminescence (ECL) of four kinds of electron donor–acceptor molecules exhibiting thermally activated delayed fluorescence (TADF) is presented. TADF molecules can harvest light energy from the lowest triplet state by spin up‐conversion to the lowest singlet state because of small energy gap between these states. Intense green to red ECL is emitted from the TADF molecules by applying a square‐wave voltage. Remarkably, it is shown that the efficiency of ECL from one of the TADF molecule could reach about 50 %, which is comparable to its photoluminescence quantum yield.     

Intramolecular photoexcitation dynamics and magnetic field effects in an intermediate-case molecule

Molecular vibration and rotation play a significant role in the intramolecular photoexcitation dynamics of the so-called intermediate-case molecule, and the fluorescence intensity, decay and polarization of s-triazine vapor are shown to depend on the excited rovibronic level of the S₁ state. Fluorescence characteristics are interpreted by assuming three zero-order states: (1) a zero-order singlet state that carries the absorption intensity and emits fluorescence with sharp structure; (2) zero-order singlet states that do not carry the absorption intensity but emit broad fluorescence; and (3) zero-order triplet states. The interaction among these states depends not only on the vibrational level but also on the rotational level excited. It is suggested that the number of triplet states coupled to the singlet state increases with increasing excess vibrational energy. It is also suggested that K-scrambling occurs both in the triplet manifold following intersystem crossing (ISC) and in the singlet manifold following intramolecular vibrational energy redistribution (IVR). The fluorescence intensity and decay of s-triazine vapor are significantly influenced by a magnetic field, and the field effects are interpreted in terms of the spin decoupling in the triplet manifold following ISC; the role of external magnetic fields is to mix the spin sublevels of different rovibronic levels coupled to the excited singlet state. Magnetic depolarization of fluorescence also occurs because of the efficient interaction between the excited singlet state and the triplet state.     

Method for accurate experimental determination of singlet and triplet exciton diffusion between thermally activated delayed fluorescence molecules

Understanding triplet exciton diffusion between organic thermally activated delayed fluorescence (TADF) molecules is a challenge due to the unique cycling between singlet and triplet states in these molecules. Although prompt emission quenching allows the singlet exciton diffusion properties to be determined, analogous analysis of the delayed emission quenching does not yield accurate estimations of the triplet diffusion length (because the diffusion of singlet excitons regenerated after reverse-intersystem crossing needs to be accounted for). Herein, we demonstrate how singlet and triplet diffusion lengths can be accurately determined from accessible experimental data, namely the integral prompt and delayed fluorescence. In the benchmark materials 4CzIPN and 4TCzBN, we show that the singlet diffusion lengths are (9.1 ± 0.2) and (12.8 ± 0.3) nm, whereas the triplet diffusion lengths are negligible, and certainly less than 1.0 and 1.2 nm, respectively. Theory confirms that the lack of overlap between the shielded lowest unoccupied molecular orbitals (LUMOs) hinders triplet motion between TADF chromophores in such molecular architectures. Although this cause for the suppression of triplet motion does not occur in molecular architectures that rely on electron resonance effects (e.g. DiKTa), we find that triplet diffusion is still negligible when such molecules are dispersed in a matrix material at a concentration sufficiently low to suppress aggregation. The novel and accurate method of understanding triplet diffusion in TADF molecules will allow accurate physical modeling of OLED emitter layers (especially those based on TADF donors and fluorescent acceptors).

A method for measuring triplet diffusion between TADF molecules is presented, and implications of limited triplet diffusion for OLEDs discussed.     

The collision complex in the exchange reaction Na + Na2. I. An ortho-para pumping experiment on Na2

The exchange reaction Na + Na₂ → Na₂ + Na is studied in the gas phase. The influence of this reaction on the nuclear spin states of the molecules is investigated by two different experiments. In an ortho-para pumping experiment the ortho-para conversion rate of Na₂ is determined with aid of laser induced molecular fluorescence. This rate is found to be twice the nuclear spin relaxation of the molecules, measured in a NMR experiment with nuclear spin polarized Na₂. Furthermore the influence of dissociation- and intramolecular relaxation processes on nuclear spin relaxation and ortho-para conversion is investigated.     

Long-lived 1H singlet spin states originating from para-hydrogen in Cs-symmetric molecules stored for minutes in high magnetic fields

Nuclear magnetic resonance (NMR) is a very powerful tool in physics, chemistry, and life sciences, although limited by low sensitivity. This problem can be overcome by hyperpolarization techniques dramatically enhancing the NMR signal. However, this approach is restricted to relatively short time scales depending on the nuclear spin-lattice relaxation time T(1) in the range of seconds. This makes long-lived singlet states very useful as a way to extend the hyperpolarization lifetimes. Para-hydrogen induced polarization (PHIP) is particularly suitable, because para-H(2) possesses singlet symmetry. Most PHIP experiments, however, are performed on asymmetric molecules, and the initial singlet state is directly converted to a NMR observable triplet state decaying with T(1), in the order of seconds. We demonstrate that in symmetric molecules, a long-lived singlet state created by PHIP can be stored for several minutes on protons in high magnetic fields. Subsequently, it is converted into observable high nonthermal magnetization by controlled singlet-triplet conversion via level anticrossing.     

Quenching of pyrene fluorescence by amphiphilic nitroxide radicals embedded in cationic micelles

Steady-state and nanosecond time-resolved studies have been carried out on the fluorescence quenching of excited pyrene by n-doxylstearic methyl esters (n-DSE, N = 5, 10, 12) in an aqueous solution of cationic micelles of hexadecyltrimethylammonium chloride (HTAC, 0.1 M). The aggregation number (N = 114 ± 6) and the rate constants of intramicellar quenching (1.4 × 10⁷, 1.3 × 10⁷ and 1.05 × 10⁷ s⁻¹ for 5-, 10- and 12-DSE respectively) have been determined. The results are discussed in terms of the average location of the luminophore and the quencher molecules in the aggregates, considering previous findings of electron spin echo modulation studies of n-doxylstearic acid spin probes in micellar systems.     

Theory of long-lived nuclear spin states in solution nuclear magnetic resonance. I. Singlet states in low magnetic field

We have recently demonstrated the existence of exceptionally long-lived nuclear spin states in solution-state nuclear magnetic resonance. The lifetime of nuclear spin singlet states in systems containing coupled pairs of spins-12 may exceed the conventional relaxation time constant T1 by more than an order of magnitude. These long lifetimes may be observed if the long-lived singlet states are prevented from mixing with rapidly relaxing triplet states. In this paper we provide the detailed theory of an experiment which uses magnetic field cycling to observe slow singlet relaxation. An approximate expression is given for the magnetic field dependence of the singlet relaxation rate constant, using a model of intramolecular dipole-dipole couplings and fluctuating external random fields.     

Azobenzene-linked porphyrin-fullerene dyads

A new group of porphyrin-fullerene dyads with an azobenzene linker was synthesized, and the photochemical and photophysical properties of these materials were investigated using steady-state and time-resolved spectroscopic methods. The electrochemical properties of these compounds were also studied in detail. The synthesis involved oxidative heterocoupling of free base tris-aryl-p-aminophenyl porphyrins with a p-aminophenylacetal, followed by deprotection to give the aldehyde, and finally Prato 1,3-dipolar azomethineylide cycloaddition to C60. The corresponding Zn(II)-porphyrin (ZnP) dyads were made by treating the free base dyads with zinc acetate. The final dyads were characterized by their 1H NMR, mass, and UV-vis spectra. 3He NMR was used to determine if the products are a mixture of cis and trans stereoisomers, or a single isomer. The data are most consistent with the isolation of only a single configurational isomer, assigned to the trans (E) configuration. The ground-state UV-vis spectra are virtually a superimposition of the spectral features of the individual components, indicating there is no interaction of the fullerene (F) and porphyrin (H2P/ZnP) moieties in the ground state. This conclusion is supported by the electrochemical data. The steady-state and time-resolved fluorescence spectra indicate that the porphyrin fluorescence in the dyads is very strongly quenched at room temperature in the three solvents studied: toluene, tetrahydrofuran (THF), and benzonitrile (BzCN). The fluorescence lifetimes of the dyads in all solvents are sharply reduced compared to those of H2P and ZnP standards. In toluene, the lifetimes of the free base dyads are 600-790 ps compared to 10.1 ns for the standard, while in THF and BzCN the dyad lifetimes are less than 100 ps. For the ZnP dyads, the fluorescence lifetimes were 10-170 ps vs 2.1-2.2 ns for the ZnP references. The mechanism of the fluorescence quenching was established using time-resolved transient absorption spectroscopy. In toluene, the quenching process is singlet-singlet energy transfer (k approximately 10(11) s-1) to give C60 singlet excited states which decay with a lifetime of 1.2 ns to give very long-lived C60 triplet states. In THF and BzCN, quenching of porphyrin singlet states occurs at a similar rate, but now by electron transfer, to give charge-separated radical pair (CSRP) states, which show transient absorption spectra very similar to those reported for other H2P-C60 and ZnP-C60 dyad systems. The lifetimes of the CSRP states are in the range 145-435 ns in THF, much shorter than for related systems with amide, alkyne, silyl, and hydrogen-bonded linkers. Thus, both forward and back electron transfer is facilitated by the azobenzene linker. Nonetheless, the charge recombination is 3-4 orders of magnitude slower than charge separation, demonstrating that for these types of donor-acceptor systems back electron transfer is occurring in the Marcus inverted region.     

The electronic relaxation of methylglyoxal in the vapor phase

The time-resolved fluorescence of methylglyoxal was studied as a function of excitation energy and pressure. In all circumstances a dual fluorescence was observed. From these measurements was obtained the parameters describing the singlet-triplet coupling, the disspative leak rates from singlet and triplet states and the fluorescence quenching constants. Also, the pressure induction of the thermalized phosphorescence was studied.As found previously in biacetyl, the excited manifold should be divided into a black note region and an overlap region, the former being more extensive in methylglyoxal. The decal characteristics of methylglyoxal are intermediate between those of glyoxal and biacetyl. The differences in behavior between the three molecules can be readily understood on the basis of the differences in their level densities, due to the different number of atoms in these molecules.     

A Hyperpolarizable 1H Magnetic Resonance Probe for Signal Detection 15 Minutes after Spin Polarization Storage

Nuclear magnetic resonance (NMR) and magnetic resonance imaging (MRI) are two extremely important techniques with applications ranging from molecular structure determination to human imaging. However, in many cases the applicability of NMR and MRI are limited by inherently poor sensitivity and insufficient nuclear spin lifetime. Here we demonstrate a cost‐efficient and fast technique that tackles both issues simultaneously. We use the signal amplification by reversible exchange (SABRE) technique to hyperpolarize the target ¹H nuclei and store this polarization in long‐lived singlet (LLS) form after suitable radiofrequency (rf) pulses. Compared to the normal scenario, we achieve three orders of signal enhancement and one order of lifetime extension, leading to ¹H NMR signal detection 15 minutes after the creation of the detected states. The creation of such hyperpolarized long‐lived polarization reflects an important step forward in the pipeline to see such agents used as clinical probes of disease.     

Highly Efficient Singlet–Singlet Energy Transfer in Light‐Harvesting [60,70]Fullerene–4‐Amino‐1,8‐naphthalimide Dyads

New C₆₀ and C₇₀ fullerene dyads formed with 4‐amino‐1,8‐naphthalimide chromophores have been prepared by the Bingel cyclopropanation reaction. The resulting monoadducts were investigated with respect to their fluorescence properties (quantum yields and lifetimes) to unravel the role of the charge‐transfer naphthalimide chromophore as a light‐absorbing antenna and excited‐singlet‐state sensitizer of fullerenes. The underlying intramolecular singlet–singlet energy transfer (EnT) process was fully characterized and found to proceed quantitatively (Φ_EnT≈1) for all dyads. Thus, these conjugates are of considerable interest for applications in which fullerene excited states have to be created and photonic energy loss should be minimized. In polar solvents (tetrahydrofuran and benzonitrile), fluorescence quenching of the fullerene by electron transfer from the ground‐state aminonaphthalimide was postulated as an additional path.