Effect of temperature, cholesterol content, and antioxidant structure on the mobility of vitamin E constituents in biomembrane models studied by laterally diffusion-controlled fluorescence quenching

Kinetic parameters relevant for the antioxidant activity of the vitamin E constituents (alpha, beta, gamma, and delta homologues of tocopherols and tocotrienols) and of an amphiphilic vitamin C derivative, l-ascorbyl 6-palmitate, were determined. Fluorescence quenching experiments of 2,3-diazabicyclo[2.2.2]oct-2-ene in homogeneous acetonitrile-water mixtures afforded reactivity trends in terms of intermolecular quenching rate constants, while the quenching of Fluorazophore-L in liposomes provided the lateral diffusion coefficients relevant for understanding their biological activity in membranes. The reactivity in homogeneous solution was not influenced by the nature of the isoprenoid tail (tocopherol versus tocotrienol), but was dependent on the methylation pattern. The resulting order (alpha > beta = gamma > delta) was found to be in line with their reactivities toward peroxyl radicals as well as the phenolic O-H bond dissociation energies. The mutual lateral diffusion coefficient in POPC liposomes was the same, within error, for different tocopherols and tocotrienols (D(L) = (1.6 +/- 0.2) x 10(-7) cm(2) s(-1)). l-Ascorbyl 6-palmitate exhibited a reactivity similar to that of delta-tocopherol in homogeneous solution, but displayed a 1 order of magnitude lower fluorescence quenching efficiency in liposomes than the vitamin E constituents. Temperature effects on the laterally diffusion-controlled fluorescence quenching were large, with activation energies of 44 +/- 6 kJ mol(-1). The addition of cholesterol (0-30%) to POPC liposomes resulted only in slightly reduced diffusion coefficients. The combined results demonstrate that Fluorazophore-L can provide important physicochemical parameters for the understanding of antioxidant activity in biological environments.     

2-Methyl Naphthalene as Fluorescence Probe in Ionic and Nonionic Micelles: Fluorescence Quenching by Halides

The fluorescence characteristics of 2-methyl naphthalene have been studied in ionic micelles of sodium dodecyl sulphate (SDS) and cetyl trimethyl ammonium bromide (CTAB) and in nonionic micellar medium of p-t-octylphenyl polyethoxyethanol (Triton X-100). The fluorescence quenching of fluorophore by halides and pseudohalide obeys the Stern-Volmer Equation up to a certain concentration of quencher. A quenching sphere of action model has been considered to explain the deviations from Stern-Volmer behaviour. The distribution of quenchers in the micellar phase has been calculated.     

On the localization of water-soluble porphyrins in micellar systems evaluated by static and time-resolved frequency-domain fluorescence techniques

Fluorescence quenching of meso-tetrakis-4-sulfonatophenyl (TPPS(4)) and meso-tetrakis-4-N-methylpyridil (TMPyP) porphyrins is studied in aqueous solution and upon addition of micelles of sodium dodecylsulfate (SDS), cetyltrimethylammonium chloride (CTAC), N-hexadecyl-N,N-dimethyl-3-ammonio-1-propanesulfonate (HPS) and t-octylphenoxypolyethoxyethanol (Triton X-100). Potassium iodide (KI) was used as quencher. Steady-state Stern-Volmer plots were best fitted by a quadratic equation, including dynamic (K(D)) and static (K(S)) quenching. K(S) was significantly smaller than K(D). Frequency-domain fluorescence lifetimes allowed estimating bimolecular quenching constants, k(q). At 25 degrees C, in aqueous solution, TMPyP shows k(q) values a factor of 2-3 higher than the diffusional limit. TPPS(4) shows collisional quenching with pH dependent k(q) values. For TMPyP quenching results are consistent with reported binding constants: a significant reduction of quenching takes place for SDS, a moderate reduction is observed for HPS and almost no change is seen for Triton X-100. Similar data were obtained at 50 degrees C. For CTAC-TPPS(4) system an enhancement of quenching was observed as compared to pure buffer. This is probably associated to accumulation of iodide at the cationic micellar interface. The attraction between CTAC headgroups and I(-), and repulsion between SDS and I(-), enhances and reduces the fluorescence quenching, respectively, of porphyrins located at the micellar interface. The small quenching of TPPS(4) in Triton X-100 is consistent with strong binding as reported in the literature.     

Fluorescence quenching of naphthols by Cu2+ in micelles

Effect of the micelles of anionic, cationic and non-ionic surfactants on the fluorescence quenching of 1- and 2-naphthols has been studied in the presence of copper ion. The excited state lifetime, dynamic and static quenching constants for these systems have been determined. Fluorescence quenching in water and SDS micelle is due to the collision of the fluorophore with the quencher with a small static component. The negatively charged naphtholate ions in the excited state are quenched with significantly higher rates than the neutral naphthol molecules, which are located further inside the mesophase. CTAB micelle is less effective than the SDS micelle for fluorescence quenching. The effect of CTAB on water-assisted excited-state deprotonation has been investigated in the presence of ZnSO₄. For TX-100 micelle there is negligible quenching even at higher concentration of the quencher.     

The size of sodium dodecyl sulfate micelles in the presence ofn-alcohols as determined by fluorescence quenching measurements

The steady-state fluorescence quenching technique was used to investigate the effect of the presence of a series of alcohol homologues of mid-sized straight chain on the size of mixed micelles of sodium dodecyl sulfate (SDS). We used pyrene at concentration of ca. 10^–6M, where only its monomer exhibits any fluorescence, as fluorescent probe, and cetylpiridinium chloride at concentrations in the range (1–9)×10^–5 M as quencher. This technique allows one to determine the micellar aggregation number. The number of alcohol molecules per micelle was calculated from reported values for the micelle-water partition coefficient. On the assumption of spherical micelles, their hydrophobic radii was then calculated. The hypothesis that micelle size is determined by the available surface area per charged headgroup is discussed in the light of the results obtained.     

The aggregation of Tyr-Phe dipeptide and Val-Tyr-Val tripeptide in aqueous solution and in the presence of SDS and PEO-PPO-PEO triblock copolymer: fluorescence spectroscopic studies

The micelle formation of Tyr-Phe dipeptide and Val-Tyr-Val tripeptide has been studied for the first time. The aggregation numbers were determined for both the peptides in aqueous solution and in the presence of SDS and PEO-PPO-PEO triblock copolymer additive environments. The results obtained by steady state and time-resolved fluorescence spectroscopic techniques are in good agreement with each other. The higher value of aggregation number confirms the formation of mixed micelles. The fluorescence lifetime of tyrosine in various micellar and mixed micellar systems were also determined. The decreased lifetime values with the quencher suggested the dynamic nature of the quenching process. However, the possibility of static quenching cannot be ruled out. The accessibility of DPC quencher was found to be more in dipeptide than tripeptide.     

Daunomycin binding to detergent micelles: a model system for evaluating the hydrophobic contribution to drug-DNA interactions

The interaction of daunomycin with sodium dodecyl sulfate and Triton X-100 micelles was investigated as a model for the hydrophobic contribution to the free energy of DNA intercalation reactions. Measurements of visible absorbance, fluorescence lifetime, steady-state fluorescence emission intensity, and fluorescence anisotropy indicate that the anthraquinone ring partitions into the hydrophobic micelle interior. Fluorescence quenching experiments using both steady-state and lifetime measurements demonstrate reduced accessibility of daunomycin in sodium dodecyl sulfate micelles to the anionic quencher iodide and to the neutral quencher acrylamide. Quenching of daunomycin fluorescence by iodide in Triton X-100 micelles was similar to that seen with free daunomycin. Studies of the energetics of the interaction of daunomycin with micelles by fluorescence and absorbance titration methods and by isothermal titration calorimetry in the presence of excess micelles revealed that association with sodium dodecyl sulfate and Triton X-100 micelles is driven by a large negative enthalpy. Association of the drug with both types of micelles also has a favorable entropic contribution, which is larger in magnitude for Triton X-100 micelles than for sodium dodecyl sulfate micelles. The thermodynamic profile for the interaction of daunomycin with both types of micelles is characteristic of the "nonclassical" hydrophobic effect. The enthalpy for the interaction of daunomycin with sodium dodecyl sulfate micelles increases nonlinearly with temperature, indicating a positive (and temperature dependent) heat capacity change. The binding isotherm for daunomycin association with sodium dodecyl sulfate micelles was cooperative, with a Hill coefficient of 1.6. The cooperative behavior and the positive heat capacity change suggest that the drug alters micelle size or imposes order on the hydrocarbon interior of the micelle.     

Determination of environmentally important metal ions by fluorescence quenching in anionic micellar solution

This work describes the effect of a variety of metal ions as quenchers of the fluorescence of naphthalene, in aqueous micellar solutions of sodium dodecyl sulfate (SDS). The quenching by the metal ions can be adequately described by the Stern-Volmer equation and the best signal to noise ratios are obtained with low micellized detergent concentrations. Apparent Stern-Volmer constants decrease in the order: Fe3+ > Cu2+ > Pb2+ > Cr3+ > Ni2+ and directly reflect the relative sensitivity of the method for these ions. Detection limits (defined as three times the standard deviation of the blank for n= 10) for the fluorescence quenching of naphthalene by the metal ions in aqueous micellar SDS are in the range of 1.0 x 10(-6) to 1.0 x 10(-5) mol dm(-3). The proposed fluorescence quenching method shows good repeatibility for a variety of added quencher metal ions, indicating that anionic micelle-enhanced fluorescence quenching by metal ions constitutes an analytical method of rather general application.     

Fluorescence quenching method for the determination of sodium dodecyl sulphate with near-infrared hydrophobic dye in the presence of Triton X-100

A fluorophotometric method for the determination of anionic surfactant sodium dodecyl sulphate (SDS) was proposed. The method is based on the quenching effect of SDS on the fluorescence of near-infrared (NIR) hydrophobic dye, 2-[4'chloro-7'(3'hexadecyl-2'benzothiazolinylidene)-3',5'-(1',3'-propanediyl)-1',3',5'-heptatriene-1'-yl]-3-ethylbenzothiazolium iodide (dye I) in the presence of Triton X-100. The calibration graph is linear in the concentration range from 0 to 2 x 10(-6) mol L(-1) of SDS with a detection limit (LOD) of 8.3 x 10(-8) mol L(-1). The relative standard deviation for the determination of 7 x 10(-7) mol L(-1) SDS was 4.1%. Recoveries of 95.3-110.3% were found for the addition to 1.0 x 10(-6) mol L(-1) SDS in the analysis of environmental water samples. Preliminary research shows that the fluorescence quenching is due to the formation of dye aggregate facilitated by SDS.     

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.     

Photoinduced electron transfer in a protein-surfactant complex: probing the interaction of SDS with BSA

Photoinduced fluorescence quenching electron transfer from N,N-dimethyl aniline to different 7-amino coumarin dyes has been investigated in sodium dodecyl sulfate (SDS) micelles and in bovine serum albumin (BSA)-SDS protein-surfactant complexes using steady state and picosecond time resolved fluorescence spectroscopy. The electron transfer rate has been found to be slower in BSA-SDS protein-surfactant complexes compared to that in SDS micelles. This observation has been explained with the help of the "necklace-and-bead" structure formed by the protein-surfactant complex due to coiling of protein molecules around the micelles. In the correlation of free energy change to the fluorescence quenching electron transfer rate, we have observed that coumarin 151 deviates from the normal Marcus region, showing retardation in the electron transfer rate at higher negative free energy region. We endeavored to establish that the retardation in the fluorescence quenching electron transfer rate for coumarin 151 at higher free energy region is a result of slower rotational relaxation and slower translational diffusion of coumarin 151 (C-151) compared to its analogues coumarin 152 and coumarin 481 in micelles and in protein-surfactant complexes. The slower rotational relaxation and translational diffusion of C-151 are supposed to be arising from the different location of coumarin 151 compared to coumarin 152 and coumarin 481.     

QUENCHING AND PHOTOBLEACHING OF EXCITED POLYCYCLIC AROMATIC HYDROCARBONS BY CARBON TETRACHLORIDE AND CHLOROFORM IN MICELLAR SYSTEMS

—Aromatic hydrocarbons incorporated into cetyltrimethylammonium chloride (CTACl) or sodium dodecylsulphate (SDS) micelles are efficiently quenched and photobleached by carbon tetrachloride. Similar results were obtained employing chloroform, but the efficiency of this compound as a quencher is nearly 10³ times smaller than that of carbon tetrachloride.
Bimolecular quenching constants, with the quencher concentration given in moles per liter of micellar pseudophase, have been evaluated and compared with those obtained in homogeneous solutions. For slow processes, the values obtained would indicate that the polarity sensed by the probe depends both upon the probe and the surfactant. For diffusionally controlled processes (i.e. quenching of biphenyl), bimolecular quenching constants are considerably smaller in the micellar pseudophase. For these processes the pseudo-first-order quenching rate constants are larger in SDS than in CTACl micelles. This effect is partly due to the smaller size of the SDS micelle but also is a consequence of a lower "microviscosity" of these micelles.     

Spectrochemical investigations in molecularly organized solvent media: evaluation of pyridinium chloride as a selective fluorescence quenching agent of polycyclic aromatic hydrocarbons in aqueous carboxylate-terminated poly(amido) amine dendrimers and anionic micelles

The ability of pyridinium chloride (PC) to selectively quench alternant as opposed to nonaltemant polycyclic aromatic hydrocarbons (PAHs) in organized media is examined. PC was previously shown to be a selective quenching agent of alternant PAHs in neat polar solvents. Carboxylate-terminated poly(amido) amine (PAMAM-CT) dendrimers and anionic surfactants--sodium dodecanoate (SD), sodium octanoate (SO), and sodium dodecylsulfate (SDS)--were chosen as the solubilizing media for this study. Selective quenching of alternant PAHs is observed in the presence of the SDS and SO micelles. However, the extent of PAH quenching in SO is significantly reduced compared to PAHs dissolved in either water or SDS micelles. In the case of the smaller generation 4.5 (G4.5) PAMAM-CT dendrimers, PC was prevented from quenching both alternant and nonalternant PAHs to any appreciable extent. The dendrimer is able to "protect" the PAHs from the PC quencher that resides at the dendrimer surface. Both, SD and G5.5 PAMAM-CT precipitated out of solution with the addition of PC. Differences between traditional micelles and "unimolecular micelle" dendrimers were also examined. These studies further confirm that the PAHs did not reside in the "analogous" palisade region of the dendrimers as they do in micelles. The PAHs must reside in the outermost branches of the dendrimer, but sufficiently far enough away from the charged surface groups, where PC associated, to prevent fluorescence quenching. This work further illustrates the differences between "unimolecular micelle" dendrimers and traditional micelles.     

Photophysical Behavior of a New Gemini Surfactant in Neat Solvents and in Micellar Environments

A novel fluorescent gemini surfactant, 1,4-bis-(2'-(N-dodecyl pyridinio-4"-yl)ethenyl)benzene dibromide, abbreviated BDPEBB, has been synthesized and its photophysical properties have been studied in different environments. BDPEBB has a limited solubility in alcohols where it is found in aggregate form at concentrations>/=1 mM. In other solvents, e.g., water, it is only found in aggregate form, even at much lower concentrations. Solvent polarity has a small and insignificant solvatochromic effect but alcohols give a specific interaction with BDPEBB, causing a significant hypsochromic shift in absorption maxima and a large increase in relative fluorescence efficiency. Pyrene fluorescence is effectively quenched by BDPEBB. Pyrene also forms associative complexes with BDPEBB in water. These complexes are partly dissociated in the presence of surfactant micelles. Triton X-100 micelles provide a favorable environment for BDPEBB solubilization well distinguished from the behavior of ionic surfactants. Small quantities of BDPEBB have a large influence on the behavior of aqueous sodium dodecylsulfate (SDS) and sodium decylsulfate (SDeS) micelles, inducing the formation of large aggregates, visible by the naked eye. These large aggregates are most probably microcrystals of BDPEBB(2+)/2DS(-) or BDPEBB(2+)/2DeS(-). The aggregation number of SDS and SDeS micelles in the absence and in the presence of BDPEBB has been calculated by exploitation of the static luminescence quenching kinetics of Ru(bpy)(3)(2+) by 9-methylanthracene, both solubilized in the micellar phase. It has been observed that Ru(bpy)(3)(2+) inhibits the precipitation of SDeS micelles in the presence of BDPEBB. Our results suggest that double-chain surfactant chromophores should be employed with particular care if they are to be used as probes of the micellar phase. Copyright 2000 Academic Press.     

Spectrochemical investigations in molecularly organized solvent media: evaluation of pyridinium chloride as a selective fluorescence quenching agent of polycyclic aromatic hydrocarbons in aqueous carboxylate-terminated poly(amido) amine dendrimers and anionic micelles

The ability of pyridinium chloride (PC) to selectively quench alternant as opposed to nonalternant polycyclic aromatic hydrocarbons (PAHs) in organized media is examined. PC was previously shown to be a selective quenching agent of alternant PAHs in neat polar solvents. Carboxylate-terminated poly(amido) amine (PAMAM-CT) dendrimers and anionic surfactants – sodium dodecanoate (SD), sodium octanoate (SO), and sodium dodecylsulfate (SDS) – were chosen as the solubilizing media for this study. Selective quenching of alternant PAHs is observed in the presence of the SDS and SO micelles. However, the extent of PAH quenching in SO is significantly reduced compared to PAHs dissolved in either water or SDS micelles. In the case of the smaller generation 4.5 (G4.5) PAMAM-CT dendrimers, PC was prevented from quenching both alternant and nonalternant PAHs to any appreciable extent. The dendrimer is able to “protect” the PAHs from the PC quencher that resides at the dendrimer surface. Both, SD and G5.5 PAMAM-CT precipitated out of solution with the addition of PC. Differences between traditional micelles and “unimolecular micelle” dendrimers were also examined. These studies further confirm that the PAHs did not reside in the “analogous” palisade region of the dendrimers as they do in micelles. The PAHs must reside in the outermost branches of the dendrimer, but sufficiently far enough away from the charged surface groups, where PC associated, to prevent fluorescence quenching. This work further illustrates the differences between “unimolecular micelle” dendrimers and traditional micelles. Received: 27 July 2000 / Revised: 25 September 2000 / Accepted: 27 September 2000     

稳态荧光猝灭法确定胶束聚集数的研究

选用芘作为荧光探针, 二苯酮作为猝灭剂, 以稳态荧光猝灭法测定了十二烷基硫酸钠、十二烷基磺酸钠和十六烷基三甲基演化铵的胶束聚集致, 并对其测定胶束聚集数方法的有效范围进行了讨论. 实验结果表明, 选择的探针-猝灭剂体系适用于稳态荧光猝灭法确定阴离子和阳离子表面活性剂的胶束浓度和聚集数.     

Anomalous association and fluorophore influence on the position of dimethylaniline in micelles: fluorescence quenching of 1,8-acridinedione

Fluorescence quenching of 9,10-dimethyl-3, 4,6,7,9,10-hexahydro-1,8(2H,5H) acridinedione (ADD) dye by N,N-dimethylaniline (DMA) in SDS and CTAB were studied by steady state fluorescence and time resolved techniques. The Stern-Volmer plots for the quenching of ADD by DMA is found to be linear and the Stern-Volmer constant K(SV) depends on the micellar concentration. The fluorescence quenching analysis reveals the binding of DMA with the micelles. The perturbation of the probe on the position of DMA molecule in micelle is inferred in the present investigation. The ADD fluorophore drives the DMA molecule into the non-polar region (core) of the micelle whereas other fluorophores like pyrene and rhodamine6G do not affect the position of DMA. In this report, the importance of the nature of fluorophores in determining the position and association of the quencher molecules in the aggregated systems is being discussed.     

DEACTIVATION OF PYRENE BY INDOLE IN MICELLAR SOLUTIONS

Abstract— The deactivation rate of excited pyrene by indole strongly depends on the polarity of the media. In micellar systems (Triton X-100, cetyltrimcthylammonium chloride (CTAC) and sodium dodecylsulfate (SDS) the deactivation efficiency is enhanced due to the high local concentration of indole in the micellar pseudophase. Quantitative interpretation of the data in CTAC and SDS micelles requires to take into account indole exchange between the micelles and the aqueous phase. In SDS micelles, where due to their smaller size the exchange process is more relevant, the exit and entrance rates are (3.0 ± 0.6) x 10⁶ and (1.2 ± 0.3) x 10¹⁰ M ⁻¹s⁻¹ respectively. Intramicellar bimolecular quenching constants are (1.1 ± 0.2) x 10⁸ M⁻¹ s⁻¹ (1.4 ± 0.2) x 10⁸ M ⁻¹ s⁻¹ and (1.5 ± 0.2) x 10⁸ M ⁻¹ s⁻¹ in Triton X-100, SDS and CTAC respectively. These rates are similar to those measured in ethanol rich ethanol-water homogeneous solutions. This is in agreement with the average polarity sensed by both pyrene and indole in the micellar pseudophases.     

Distribution of fluorocarbon quencher among micelles via pyrene fluorescence probe method

 Fluorescence-quenching of pyrene in micellar system has been investigated using 1,1,2,2-tetrahydroheptadecafluorodecylpyridinium chloride (HFDePC). The new fluorocarbon quencher has a similar quenching ability as hexadecyl-pyridinium chloride (CPC) towards pyrene in hydrocarbon micelles if only a quencher molecule is solubilized in a micelle. The fluorocarbon quencher randomly distributed among micelles if the average occupancy number of probes per a micelle was small enough. The fluorescence behavior of pyrene was examined for hexadecyl-trimethylammonium chloride (CTAC) and HFDePC mixtures. The variation of fluorescence intensity gave second cmc, reflecting the micellar immiscibility of fluorocarbon and hydrocarbon surfactants. The second cmc can be simulated by material balances of both surfactants supposing the coexistence of two kinds of mixed micelles. The fluorescence-quenching behavior suggested the enhanced micellar immiscibility probably due to nonrandom distribution of fluorocarbon quenchers among micelles. Received: 13 March 1997 Accepted: 24 May 1997     

A general chemiluminescence method for the determination of surfactants based on its quenching effect on the luminol-NaIO4-cyclodextrin reaction

Here we report that all types of surfactant could be simply and sensitively determined, by directly quenching the chemiluminescence (CL) between luminol and NaIO4 in a basic solution containing one polyhydroxyl compound such as cyclodextrin (CD), glucose or glycerol. This specific quenching effect was attributed to the change of the microenvironment of the CL reaction, caused by the addition of various surfactants. Based on this fact, the potential use of this CL reaction was exemplified by the cationic surfactant CTMAB, anionic surfactant SDS and non-ionic surfactant Triton X-100. It was found that the measurable range of CTMAB, SDS and Triton X-100 were 4.0 x 10(-6)-4.0 x 10(-4) M by using a basic CD-luminol-NaIO4 CL reaction. With our simple setup, CTMAB, SDS and Triton X-100 were detectable at a concentration as low as 2 microM. Overall, this new CL reaction is quite promising for the post-column determination of surfactant mixtures.