Fluorescence quenching method for the determination of sodium carboxymethyl cellulose with acridine yellow or acridine orange

In near neutral to weak basic media, sodium carboxymethyl cellulose (NaCMC) will dissociate to become a macro polymeric anion, which can react with acridine yellow (AY) or acridine orange (AO) to form an ion-association complex resulting in fluorescence quenching of the acridine dyes. The maximum fluorescence quenching wavelength is 505 nm (lambda(ex)=440 nm) for AY system and 530 nm (lambda(ex)=493 nm) for AO system, respectively. The fluorescence quenching values (DeltaF) are directly proportional to the concentrations of NaCMC and the linear ranges are 20.0-4000 microg/L for AY system and 20.0-7000 microg/L for AO system, separately. This method has high sensitivity and the detection limits for NaCMC are 58.0 microg/L (AY system) and 157.2 microg/L (AO system). The effects of coexistent substance have been investigated, and the results show that this method has a relatively good selectivity. A fluorescence quenching method for the determination of NaCMC based on the ion-association reactions of CMC polymeric anion with a basic acridine dye was developed. The method is sensitive, simple and fast.     

Slow solvent relaxation dynamics of nanometer sized reverse micellar systems through tryptophan metabolite, kynurenine

Exploration of environmental dynamics using intrinsic biological probe tryptophan is very important; however, it suffers from various difficulties. An alternative probe, kynurenine (KN), has been found to be an efficient probe for the ultrafast dynamics in the biological environment ( 1 ] J. Phys. Chem. B., 114, 15236–15243). In the present study, we have investigated the efficacy of KN for the exploration of relatively slower dynamics of biologically relevant environments. A detailed investigation involving UV–Vis, steady‐state/time‐resolved fluorescence spectroscopy and Förster resonance energy transfer (FRET) studies on KN compared to a well‐known solvation probe, H33258, a DNA‐minor groove binder in a model nonionic reverse micelle reveals that ultrafast internal conversion associated with the hydrogen‐bonding dynamics masks KN to become a dynamical reporter of the immediate environments of the probe.     

Excited state characteristics of acridine dyes: acriflavine and acridine orange

The magnitude of the Stokes shift (frequency shifts in absorption and fluorescence spectra) is observed on changing the solvents and further has been used to calculate experimentally the dipole moments (ground state and excited state) of acriflavine and acridine orange dye molecules. Theoretically, dipole moments are calculated using PM 3 Model. The dipole moments of excited states, for both molecules investigated here, are higher than the corresponding values in the ground states. The increase in the dipole moment has been explained in terms of the nature of the excited state. Acriflavine dye overcomes the non-lasing behaviour of acridine orange due to quaternization of the central nitrogen atom.     

The electronic structures and spectra of proflavine,acridine orange,and their DNA complexes

The electronic structure of the proflavine cation is studied by the SCF –ASMO –CI method using the Pariser–Parr–Pople approximations. It is shown that the band at 445 mμ may be assigned to the ¹A₁ → ¹B₁, transition polarized along the long axis of the molecule. The bands in the neighbourhood of 260 mμ, which are composed of three absorption bands, are tentatively assigned to the ¹A₁ → ¹B₁, ¹A₁ → ¹B₁, and ¹A₁ → ¹A₁ transitions, respectively, in order of decreasing wavelength. The spectrum of the acridine orange cation may be understood to have the same assignment as that of the proflavine cation. The acridine dye cations are well known for their dimerization with concentration. The intermolecular distances in these dimers are estimated from the band shifts due to the formation of dimers, using the exciton theory. The main contribution to the molecular interaction is shown to be the electrostatic dipole–dipole interaction. Since the first excitation band of the dye molecule which exhibits a remarkable change due to the formation of the DNA–acridine dye complex, is suggested to be polarized along the long axis, preference of the outside stacking or the intercalation model is qualitatively discussed from the spectral shift of the acridine dye bound to the DNA, assuming simple models.     

Study of some reactions of acridine orange

The nitration and formylation of acridine orange were studied, and its dinitro derivatives and mono and diformyl derivatives were obtained. By measurement of the ionization constants of the conjugated cations and PMR spectroscopy it was proved that the protons attached to C₍₄₎ and C₍₅₎ are substituted.Translated from Khimiya Geterotsiklicheskikh Soedinenii, No. 2, pp. 213–215, February, 1972.     

Fluorescence properties of nanosized sulfides

The absorption spectra of nanosized sulfides and selenides (ZnS, CdS, CuS, Cu₂S, AgS, In₂S₃, SnS, PbS, Sb₂S₃, FeS, CoS, NiS, CdSe, and Ag₂Se) showed one absorption band with a maximum at wave-lengths shorter than 300 nm. The UV fluorescence spectra of all of these nanosized sulfides in a polyvinyl alcohol film contained maxima at 380–440 nm. Nanosized sulfides are thus characterized by a very large (up to 15000 cm⁻¹) Stokes shift of fluorescence. In a polyvinyl alcohol film, a decrease in the concentration of cadmium sulfide from 0.05 to 0.002 M led to a threefold increase in the fluorescence intensity. The dependence of the degree of fluorescence buildup on the sulfide concentration is nearly linear. An increase in sulfide concentration to more than 5 × 10⁻³ M in solution led to a complete coagulation of particles.     

Effect of polyion conformation on dye binding. I. Fluorescence of acridine orange in the presence of synthetic polyacids

The interaction between polyelectrolytes and Acridine Orange (AO) has been investigated in the case of poly(acrylic acid) (PAA), poly(methacrylic acid) (PMA), and a polycondensate between 1,3-benzene disulfonyl chloride and L -lysine (PLL) by visible absorption and fluorescence spectroscopy. The influence of both polymer ionization and polymer/dye ratio (P/D) on the spectral behavior of the bound dye has been studied. The stacking tendency of AO is found lower in the presence of PLL under compact conformation (in an ionization range depending on the nature of the counterion) with correlated enhancement of the green fluorescence of the monomeric species of the bound dye. The disappearance of the green fluorescence and the dimerization of bound AO are directly related to the increased flexibility of polyion chains upon ionization. Some analogy is found between the behavior of bound AO in the presence of very compact PLL chains and that reported for AO in the presence of native DNA, which might be due to specific interactions responsible for a kind of “intercalation” of the monomeric bound dye.     

Ultrafast relaxation dynamics of a biologically relevant probe dansyl at the micellar surface

We report picosecond-resolved measurement of the fluorescence of a well-known biologically relevant probe, dansyl chromophore at the surface of a cationic micelle (cetyltrimethylammonium bromide, CTAB). The dansyl chromophore has environmentally sensitive fluorescence quantum yields and emission maxima, along with large Stokes shift. In order to study the solvation dynamics of the micellar environment, we measured the fluorescence of dansyl chromophore attached to the micellar surface. The fluorescence transients were observed to decay (with time constant approximately 350 ps) in the blue end and rise with similar timescale in the red end, indicative of solvation dynamics of the environment. The solvation correlation function is measured to decay with time constant 338 ps, which is much slower than that of ordinary bulk water. Time-resolved anisotropy of the dansyl chromophore shows a bi-exponential decay with time constants 413 ps (23%) and 1.3 ns (77%), which is considerably slower than that in free solvents revealing the rigidity of the dansyl-micelle complex. Time-resolved area-normalized emission spectroscopic (TRANES) analysis of the time dependent emission spectra of the dansyl chromophore in the micellar environment shows an isoemissive point at 21066 cm-1. This indicates the fluorescence of the chromophore contains emission from two kinds of excited states namely locally excited state (prior to charge transfer) and charge transfer state. The nature of the solvation dynamics in the micellar environments is therefore explored from the time-resolved anisotropy measurement coupled with the TRANES analysis of the fluorescence transients. The time scale of the solvation is important for the mechanism of molecular recognition.     

Interaction of acridine orange and sodium poly-α, L-glutamate

Summary The interaction of acridine orange and sodium poly-, L-glutamate in the pH range of 3.8–6.8 was studied by a spectral method and viscosity measurement. From the binding data, the molar change in entropy, enthalpy and the number of binding sites on polymer were calculated. The values of S° were positive. S° for binding decreasing extensively in pH range of 5.0. These results are explained by the transition of helix coil of the polymer. The intrinsic viscosity reduction in the presence of the dye, in pH 3.8 is due to the formation of dye bridges between the polymer helix.

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Zusammenfassung Die Wechselwirkung zwischen Natriumsalz-, L-Glutamin-Säure wurde mit den Methoden der Spektroskopie und der Viskositätsmessung untersucht. Aus den Bindungsdaten wurde die molare Änderung von Entropie und Enthalpie und die Zahl der Brückenstellen des Polymeren berechnet. Die Werte von S° sind positiv. Für die Binding ergibt sich für S° eine starke Verminderung im Bereich pH 5.0. Diese ist durch die Änderung der Konformation der Polymerketten (Helix Knäuel-Umwandlung) zu erklären.

     

Study on the formation and depolymerization of acridine orange dimer in acridine orange-sodium dodecyl benzene sulfonate-protein system

Experiment indicates that the fluorescence of acridine orange (AO) can be greatly quenched by anionic surfactant sodium dodecyl benzene sulfonate (SDBS), but when protein is added into the AO-SDBS system, the fluorescence intensity of the latter is enhanced. It is considered that SDBS can promote the formation of AO dimer, resulting in the quenching of the fluorescence of AO. When bovine serum albumin (BSA) is added into AO-SDBS system, BSA and SDBS can interact and form negative micelle-like cluster complex with "aromatic ring stacking," which destroys the formation conditions of AO dimer and makes some AO dimers turn into monomer, resulting in the fluorescence enhancement of AO-SDBS system. Whereas the positive AO and residual AO dimer are dissolved in the negative BSA-SDBS cluster through electrostatic and hydrophobic forces and form a large association. Here, the fluorescence enhancement of AO-SDBS is considered to originate from the hydrophobic microenvironment provided by BSA and SDBS, the depolymerization of AO dimer and intermolecular energy transfer between BSA and AO.     

Fluorometric determination of DNA using nano-SiO2 particles as an effective dispersant and stabilizer for acridine orange

A sensitive fluorometric method for the determination of ctDNA (calf thymus DNA) is presented. It has good selectivity and sensitivity and uses nano-SiO₂ particles as an effective dispersant and stabilizer for acridine orange (AO). Compared to resonance light scattering (RLS) and the conventional method that uses organic dyes as fluorescence probe, the new method is more tolerant towards coexisting foreign substances and also more stable. With 20 mg nano-SiO₂ particles, 10 μmol L⁻¹ AO, at pH 8.01 and an ionic strength of 0.02 mol L⁻¹, the interaction of AO with nano-SiO₂ and ctDNA results in fluorescent signal enhancement. The extent of enhancement was in good proportion to the concentration of ctDNA at excitation/emission wavelengths of 490/523 nm, respectively. The calibration curve was linear over 0.66–55.60 μg mL⁻¹. The determination limit (3σ) was 15 μg mL⁻¹. The method was applied to the determination of ctDNA in synthetic samples with satisfactory results.     

Photochemistry on surfaces: Fluorescence emission of monomers and dimers and triplet state absorption of acridine orange adsorbed on microcrystalline cellulose

Prompt fluorescence as well as delayed fluorescence emission of acridine orange was detected at room temperature from samples where this dye is adsorbed on microcrystalline cellulose. Ground state absorption studies provided evidence for dimer formation of the dye when adsorbed on cellulose, and the equilibrium constant for dimerisation was determined as 1.6±0.1 × 10⁶mol⁻¹g. At low loadings of acridine orange on cellulose (<1 μmol g⁻¹) the fluorescence emission is mainly due to the monomer and is similar to that observed in ethanolic solutions where little aggregation occurs, and peaks at 530 nm. A linear dependence of the fluorescence intensity on the amount of light absorbed by the dye was established for these “diluted” samples. However, at higher loadings (>20 μmol g⁻¹), the fluorescence intensity decreases, and the emission is broad with its maximum at 620 nm, and is mainly due to the dimer. By assuming that the excited monomer and dimer of acridine orange are the only emitting species, it was possible to determine the fluorescence quantum yields for these two species when adsorbed on microcrystalline cellulose as 0.95±0.05 and 0.40±0.10, respectively. Pulsed emission studies at room temperature in the millisecond time-range also revealed monomer and dimer emissions on this longer time-scale. These are shown to be due to thermally activated delayed fluorescence arising from the triplet states of monomer and dimer acridine orange as confirmed by diffuse reflectance transient absorption studies.     

Fluorescence quenching mechanisms of aromatic hydrocarbon-dicyanobenzene and-n.n-dimethylaniline systems in aqueous micellar solutions

Fluorescence quenching of pyrene and several other aromatic hydrocarbons by dicyanobenzenes as well as N,N-dimethylaniline has been observed in aqueous micellar solutions. The experimental results have been explained by means of equations for fluorescence intensity and decay curve derived on the basis of a simplified model.     

Different sources of 4-aminophthalimide solvation dynamics retardation inside micellar systems

The solvation dynamics of 4-aminophthalimide (4-AP) in two micellar systems (cetyltrimethylammonium bromide (M-CTAB) and Triton X-100 (M-TX-100)) has been studied. The results presented are in agreement with our earlier findings in sodium dodecyl sulfate (M-SDS) micelle (J. Phys. Chem. B 107 (2003) 13,986, J. Phys. Chem. B 109 (2005) 9422). They have confirmed that the main reason for the observed shape and position of the time changes in 4-AP time-resolved emission spectra (TRES) is the process of establishing a new equilibrium between two emissive species present in micellar systems, the excited 4-AP in the intramolecular charge transfer state (S(1)-ICT) and the exciplex formed between 4-AP in the S1-ICT state and water molecules dissolved inside micelles. In M-TX-100 and in M-CTAB this process has been found to be slower than in the earlier studied M-SDS. The presence of two emitting species has been concluded on the basis of observation of the isoemissive point in the time-resolved area-normalized emission spectra (TRANES) of 4-AP in micellar systems studied. It has been shown that the distance between the 4-AP molecule and the water molecules present inside the micelles can be one of the parameters responsible for the long-time duration of the exciplex formation and solvation process in the micelles.     

Investigation of the interaction between acridine orange and bovine serum albumin

The results from the measurement of the fluorescence spectrum showing the binding characteristics of acridine orange (AO) and bovine serum albumin (BSA) are reported. It was found that the equilibrium constant k=4848.64 l mol⁻¹, and the number of binding sites n=0.82. Based on the mechanism of the Forster energy transference, the transfer efficiency of energy and distance between the acceptor AO and BSA were found. The interaction between AO and BSA have been verified as consistent with the static quenching procedure and the quenching mechanism is related to the energy transfer.     

Reactions in micellar systems

The notion of "green chemistry" has encouraged even synthetic organic chemists to include water as a solvent. Incredible selectivities and activities can be achieved through the addition of amphiphiles with a defined structure. The morphology of supramolecular assemblies or associates formed by surfactants vary according to the temperature and concentration. As a rule, reactions are typically conducted using simple spherical aggregates, that is, micelles in the nanometer range. The strong polarity gradient present between the hydrophilic surface and the hydrophobic core of the micelle means that both nonpolar and polar reagents can be solubilized. This solubilization results in reactants becoming more concentrated within the micelle than in the surrounding water phase and leads to an acceleration of the reaction and causes selective effects. The kinetic treatment of reactions in micellar systems can be accomplished by considering them as microheterogeneous two-phase systems.