脂溶性铕螯合物的水溶化及其荧光信号增强

以本文合成的脂溶性的含丙烯酸配体的铕离子螯合物为聚合单体,以合成的具有聚乙二醇单甲醚结构单元的大分子为引发剂,合成了以聚乙二醇单甲醚为水溶端、铕离子螯合物的聚合单元为脂溶端的两亲性嵌段荧光共聚物,并在超声波作用下制备了该共聚物的水溶性荧光胶束纳米粒。对此纳米粒进行了TEM、粒径分布和荧光光谱表征,同时研究了荧光信号增强、时间稳定性、光漂白性能和荧光的酸碱稳定性。结果表明,该水溶性的铕荧光纳米粒的粒径分布均一(约298nm),荧光信号增强作用明显,对紫外光和酸碱的抵抗能力显著增强。本文实现了脂溶性铕螯合物的水溶化及其荧光信号增强,预示着铕荧光螯合物纳米粒可以直接用于生物标记,并具有高灵敏生物分析的应用潜能。     

Determination of DNA based on fluorescence quenching of terbium doped carbon dots

This work describes the preparation of carbon dots doped with terbium(III) (Tb-CDs) via a hydrothermal method, starting from terbium ion and ethylenediamine. The size, composition and spectral properties of the Tb-CDs were characterized by transmission electron microscopy, infrared spectra, and fluorescence spectra. The results show that doping of the CDs with Tb(III) reduces the particle size and results in more uniform particles, while fluorescence (at excitation/emission peaks of 380/475 nm) is strongly enhanced. The interaction between Tb-CDs and ct-DNA results in fluorescence quenching of Tb-CDs. The findings were exploited to design a quenchometric method for the determination of ct-DNA. The signal drops linearly in the 80 ng·mL^?1 to 50 μg·mL^?1 ct-DNA concentration range, and the detection limit is 53 ng·mL^?1. The method was applied to the determination of ct-DNA in spiked samples and gave satisfactory results. The possible fluorescence quenching mechanism (which is mainly static) was investigated using the Stern–Volmer equation and thermodynamic equations.

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Graphical abstract A kind of carbon dots doped with terbium(III) (Tb-CDs) were prepared via a hydrothermal method, using terbium ion and ethylenediamine as precursor. Doping with Tb(III) reduced the particle size of CDs and results in uniform particle size and stronger fluorescence. The interaction between the Tb-CDs and dsDNA results in quenching of the fluorescence of Tb-CDs and can be applied to determination of dsDNA.

     

Aggregation numbers of cationic oligomeric surfactants: a time-resolved fluorescence quenching study

The micelle aggregation numbers (N(agg)) of several series of cationic oligomeric surfactants were determined by time-resolved fluorescence quenching (TRFQ) experiments, using advantageously 9,10-dimethylanthracene as fluorophore. The study comprises six dimeric ("gemini"), three trimeric, and two tetrameric surfactants, which are quaternary ammonium chlorides, with medium length spacer groups (C(3)-C(6)) separating the individual surfactant fragments. Two standard cationic surfactants served as references. The number of hydrophobic chains making up a micellar core is relatively low for the oligomeric surfactants, the spacer length playing an important role. For the dimers, the number decreases from 32 to 21 with increasing spacer length. These numbers decrease further with increasing degree of oligomerization down to values of about 15. As for many conventional ionic surfactants, the micelles of all oligomers studied grow only slightly with the concentration, and they remain in the regime of small micelles up to concentrations of at least 3 wt %.     

Ionic quenching of naphthalene fluorescence in sodium dodecyl sulfate micelles

Micellar effects on luminescense of organic compounds or probes are well established, and here we show that quenching is highly favored in aqueous sodium dodecyl sulfate (SDS) micelles, which concentrate a naphthalene probe and cations of lanthanides, transition metals, and noble metals. Interactions have been studied by steady state and time-resolved fluorescence in examining the fluorescence suppression of naphthalene by metal ions in anionic SDS micelles. The quenching is collisional and correlated with the unit charge and the reduction potential of the metal ion. The rate constants, calculated in terms of local metal ion concentrations, are close to the diffusion control limit in the interior of SDS micelles, where the microscopic viscosity decreases the transfer rate, following the Stokes-Einstein relation.     

Aggregation numbers of SDS micelles formed on EHEC. A steady state fluorescence quenching study

The present investigation proves that in the interaction between an uncharged polymer and a negatively charged amphiphilic ion (surfactant) clusters are actually formed and it provides data for the cluster concentration and the cluster size and their variation with composition. The polymer bound cluster size increases after a certain critical surfactant concentration and passes through a maximum. This maximum cluster size decreases with decreasing polymer concentration and attains a limiting value at infinite dilution. For the highest polymer concentration the cluster size is close to the size of normal surfactant micelles. The cluster concentration was determined by a fluorescence quenching technique and the amount surfactant adsorbed to the polymer by dialysis equilibrium measurements. Combining these independent sets of data permits the cluster aggregation number to be unambiguously determined. Solubilization experiments indicate the possibility to regulate the amount solubilized by varying the polymer concentration. The molecular properties of the system are sensitively monitored by the variation in two vibronic peaks in the pyrene fluorescence emission spectrum which defines a hydrophobic index. Very good agreement is found between all three experimental methods. Finally, the model suggested is analyzed in terms of coil size and cluster-cluster distance. Depending upon the degree of adsorption saturation and the density of polymer segments in solution the interaction may switch from being intramolecular to becoming intermolecular.     

Determination of flunixin by sensitized terbium fluorescence in the presence of surfactant micelles

The effect of surfactant micelles and the second ligand on the efficiency of energy transfer in the Tb³⁺–flunixin complex was studied. The fluorescence intensity of the Tb³⁺–flunixin complex was shown to 100-fold increase in the presence of Tween 80 micelles and the second trioctylphosphine oxide ligand. A procedure for the fluorimetric determination of flunixin in the Flunex drug product (NITA-FARM, Saratov, Russia) was developed. The linearity range of the calibration curve was from 1 × 10^–7 to 1 × 10^–4 M, r ² = 0.991, and the limit of detection was 4 × 10^–8 M (3σ).     

Probing solubilization sites in block copolymer micelles using fluorescence quenching

The solubilization sites provided by micelles formed by a diblock copolymer with one neutral hydrophobic block, polystyrene, and one charged hydrophilic block, poly(acrylic acid) or poly(methacrylic acid), have been studied by fluorescence quenching of pyrene by polar and nonpolar quenchers. Pyrene solubilized into these micelles is distributed between the inner corona and the micelle core. The fraction of pyrene residing in the inner corona is almost unity for star micelles, where the corona-forming blocks are larger than the core-forming blocks, and around 0.5 for crew-cut micelles where the opposite situation prevails. The kinetics of the quenching process depends on the pyrene location, i.e. is static in the micelle core, and largely dynamic in the inner corona at low quencher concentration. The rate constants for fluorescence quenching by nitromethane are shown to increase with increasing pH.     

On the intramicellar fluorescence quenching rate constant in cylindrical micelles

A theoretical model developed for the intramicellar fluorescence quenching rate constant in spherical micelles is extended to cylindrical micelles for diffusion- and reaction-controlled processes. The rate constants for prolate ellipsoidal micelles are expected to be intermediate between those of spherical and those of cylindrical micelles of the same volume. The importance of diffusion transients is dicussed as a function of micellar shape and the fluorescence decay time of the excited probe.     

Electron-transfer fluorescence quenching of aromatic hydrocarbons by europium and ytterbium ions in acetonitrile

To make the effects of molecular size on photoinduced electron-transfer (ET) reactions clear, the ET fluorescence quenching of aromatic hydrocarbons by trivalent lanthanide ions M3+ (europium ion Eu3+ and ytterbium ion Yb3+) and the following ET reactions such as the geminate and free radical recombination were studied in acetonitrile. The rate constant k(q) of fluorescence quenching, the yields of free radical (phi(R)) and fluorescer triplet (phi(T)) in fluorescence quenching, and the rate constant k(rec) of free radical recombination were measured. Upon analysis of the free energy dependence of k(q), phi(R), phi(T), and k(rec), it was found that the switchover of the fluorescence quenching mechanism occurs at deltaG(fet) = -1.4 to -1.6 eV: When deltaG(fet) < -1.6 eV, the fluorescence quenching by M3+ is induced by a long-distance ET yielding the geminate radical ion pairs. When deltaG(fet) > -1.4 eV, it is induced by an exciplex formation. The exciplex dissociates rapidly to yield either the fluorescer triplet or the geminate radical ion pairs. The large shift of switchover deltaG(fet) from -0.5 eV for aromatic quenchers to -1.4 to -1.6 eV for lanthanide ions is almost attributed to the difference in the molecular size of the quenchers. Furthermore, it was substantiated that the free energy dependence of ET rates for the geminate and free radical recombination is satisfactorily interpreted within the limits of the Marcus theory.     

Partition and binding constants in micelles and vesicles from fluorescence quenching data

A method of analysing steady-state fluorescence quenching in compartmentalized systems is proposed which allows the evaluation of partition and binding constants where both association processes occur simultaneosly. The number of binding sites is shown to be dependent on the nature of the quencher.     

Quenching and dequenching of pyrene fluorescence by nucleotide monophosphates in cationic micelles

The fluorescence behavior of pyrene solubilized in the hexadecyltrimethylammonium bromide aqueous micellar solution in the presence of adenosine 5'-monophosphate (AMP) and uridine 5'-monophosphate (UMP) was investigated. AMP and UMP were found to influence oppositely the fluorescence of micellized pyrene. UMP acts as quencher, while AMP acts as dequencher. Both effects saturate at high nucleotide concentration (about 40 mM). Dequenching of micellized pyrene fluorescence is induced also by addition of disodium hydrogen orthophosphate (Na 2HPO 4), while loading with sodium bromide (NaBr) quenches the fluorescence. Furthermore, in absence of micelles, pyrene fluorescence depends on the UMP, according to the Stern-Volmer relation, but is unaffected by AMP. Dynamic light scattering experiments showed that the size and shape of aggregates is not affected by different types of nucleotide loaded into the solution; thus, we conclude that the opposite photophysical effect exploited by AMP and UMP are uncorrelated to any change in micellar microstructure. The whole fluorescence data set was successfully accounted for by assuming that the anionic nucleotides compete with the surfactant counterion (bromide) for the surface of the micelle. Accordingly, substitution of bromide with the more effective quencher UMP results in a strong decrease of the pyrene fluorescence, while the substitution of bromide with the nonquencher AMP results in an increase in the pyrene fluorescence.     

Nonlinear two-phase equilibrium model for the binding of arsenic anions to cationic micelles

The binding of arsenic ions to cationic cetylpyridinium chloride (CPC) micelles has been investigated using the semiequilibrium dialysis (SED) technique. In SED experiments, it has been shown that CPC micelles are very effective in binding arsenic ions in the retentate. At the studied pH (pH 8), the unbound and bound arsenic exists primarily as divalent anions (HAsO₄²⁻) while CPC molecules exist as monovalent cations. Therefore, arsenic ions are bound electrostatically to the cationic micelle. The resultant colloid is large enough not to pass through the dialysis membrane, producing a rejection greater than 99.59%. The concentration of the unbound arsenic anions passing through the dialysis membrane is practically the same as the permeate concentration of these species in the analogous micellar-enhanced ultrafiltration (MEUF) experiments. Therefore, a nonlinear equilibrium model which combines thermodynamic relations, charge balance equations, and material balance equations with the Oosawa two-phase polyelectrolyte theory has been developed to correlate the binding of arsenic to CPC micelles in SED and MEUF. It was shown that the equilibrium model successfully accounts for the experimental data in both SED and MEUF in the absence and presence of monovalent (HCO₃¹⁻) and divalent co-ions (HPO₄²⁻). Because of their small sizes (less than 10 nm), micelles should retain their equilibrium shapes in the presence of hydrodynamic shear typically attained in most dynamic processes. Therefore, the equilibrium model can be used to predict separation efficiencies in other ultrafiltration units such as in crossflow ultrafiltration processes.     

Effects of cationic micelles on fluorescence of indole and indolecarboxylic acids. Analytical determinations

The effect of cationic micelles of benzyldimethylhexadecylammonium chloride and hexadecyltrimethylammonium bromide on the fluorescence spectra of indole and its carboxylic derivatives were determined. The larger solute-micelle binding constants were provided by benzyldimethylhexadecylammonium chloride, ranging from 0.10·10⁵ and 2.15·10⁵ M ^–1 for indole-5-carboxylic and indole-2-carboxylic acid, respectively. The determination of indole-4-carboxylic acid with a detection limit of 1.0 ng ml^–1, in the presence of the 3-and 5-substituted derivatives, shows the analytical potential of the synchronous spectrofluorimetry combined with the enhancement effects caused by micellar media.     

Observations on photochemical fluorescence enhancement of the terbium(III)-sparfloxacin system.

Fluorescence of terbium(III) was sensitized when excited in the presence of sparfloxacin (SPFX) in the aqueous solution because a Tb(III)-SPFX complex was formed. The sensitized fluorescence was further enhanced when this system was exposed to 365 nm ultraviolet light. By the spectral properties and contrast experiments, it is proved that irradiation makes this system undergo photochemical reactions and a new terbium complex which is more favorable to the intramolecular energy transfer is formed. The mechanism of photochemical fluorescence enhancement of the Tb(III)-SPFX system is discussed and a new sensitive and selective photochemical fluorimetry for the determination of SPFX is established. Under the optimum conditions, the linear range is 1.0-50 x 10(-7) M for SPFX, the detection limit is 3.0 x 10(-9) M and the R.S.D. for 5.0 x 10(-7) M SPFX is 1.3% (n = 9). Without any pretreatment the recovery of SPFX in human urine was determined with satisfaction.     

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.     

Anion effects on the fluorescence spectra of europium and terbium complexes with a family of ether-amide type multipodal ligands

Three multipodal ligands: 3,3,7,7-tetra[N-methyl-N-phenyl(acetamide)-2-oxymethyl]-5-oxanonane (L(a)), 3,3,7,7-tetra[N-ethyl-N-phenyl(acetamide)-2-oxymethyl]-5-oxanonane (L(b)), 3,3,7,7-tetra[N-benzyl-N-phenyl(acetamide)-2-oxymethyl]-5-oxanonane (L(c)) and their europium and terbium nitrate and perchlorate complexes were synthesized. The 12 complexes were characterized by elemental analysis, IR, fluorescence spectroscopy and conductivity. The lanthanide atoms are coordinated by O atoms from CO, C-O-C. With the difference of the anion, the fluorescence properties of anion effect for the complexes were investigated. Some factors that influencing the fluorescent intensity were discussed.     

Multiple labelling of immunoglobulin G, albumin and testosterone with a fluorescent terbium chelate for fluorescence immunoassays

Human immunoglobulin G, human serum albumin and testosterone were labelled with the 4-aminosalicylic acid derivative of diethylenetriaminepentaacetic acid complexed with terbium ions. An exceptionally large amount of label, of the order of a few hundred moles of complex per mole of analyte, could be conjugated to the compounds tested by the use of poly-L-lysine. Self-quenching appears to be minimal, even with this high local concentration of fluorophores. The tracers were stable at 4 degrees C, and gave competitive calibration graphs at physiological concentrations.     

Conformation effects on the fluorescence spectra of europium and terbium complexes with a family of ether-amide type multipodal ligands

Three multipodal ligands: 3,3,7,7-tetra[N-methyl-N-phenyl(acetamide)-2-oxymethyl]-5-oxanonane (La), 3,3,7,7-tetra[N-ethyl-N-phenyl(acetamide)-2-oxymethyl]-5-oxanonane (Lb), 3,3,7,7-tetra[N-benzyl-N-phenyl(acetamide)-2-oxymethyl]-5-oxanonane (Lc) and their europium and terbium nitrate and perchlorate complexes were synthesized. The 12 complexes were characterized by elemental analysis, IR, fluorescence spectroscopy and conductivity. The novel cup-like crystal structure of the Eu(III) complex with Lb and fluorescence emission properties of the complexes have also been studied. It has been showed that the nature of the anion has a great effect upon the composition of the complexes. The difference in the composition of the complexes results in the difference in the emission properties of them. The [Ln(NO3)5] group is negatively effective to the fluorescence intensity of the nitrate complexes. The luminescence intensity of Ln(III) perchlorate complex is stronger than that of nitrate complex.     

Spectrophotofluorometric determination of terbium and europium in potassium carbonate solution

The conditions for the spectrophotofluorometric determination of terbium and europium, in solutions of potassium carbonate, have been established. The apparent excitation and fluorescence wavelengths used, respectively, are 245 mmu and 550 mmu for terbium and 400 mmu and 620 mmu for europium. The fluorescence varies linearly with the concentration of terbium and europium in the range 0.3-70 mug, of terbium/ml and 4-800 mug of europium/ml. Large amounts of gadolinium, lutetium and yttrium do not interfere. Cerium(IV) interferes seriously.     

Synthesis and spectroscopic studies of europium chelate complexes

The synthesis of the mixed complexes of the europium ion with 2-pyridylcarbinol-N-oxide, and 2,2′-dipyridyl and 1,10-phenanthroline were prepared. The compounds, were characterized by means of chemical analyses, vibrational spectra, molar conductivities, and electronic spectra. The emission of the [EuL₃, phen] complex at 10 K is very intense and an assignment of the point symmetry of the Eu(III) ion has been made.