Marcos Gugliotti

Influence of Negatively Charged Interfaces on the Ground and Excited State Properties of Methylene Blue

Abstract Properties of the ground and excited states of methylene blue (MB) were studied in negatively charged vesicles, normal and reverse micelles and sodium chloride solutions. All these systems induce dimer formation as attested by the appearance of the dimer band in the absorption spectra (λD ∼ 600 nm). In reverse micelles the dimerization constant (KD) corrected for the aqueous pseudophase volume fraction is two–three orders of magnitude smaller than KD of MB in water, and it does not change when W0 is increased from 0.5 to 10. Differences in the fluorescence intensity as a function of dimer–monomer ratio as well as in the resonance light scattering spectra indicate that distinct types of dimers are induced in sodium dodecyl sulfate (SDS) micelles and aerosol-OT (sodium dioctyl sulfoxinate, AOT) reversed micelles. The properties of the photoinduced transient species of MB in these systems were studied by time-resolved near infrared (NIR) emission (efficiency of singlet oxygen generation), by laser flash photolysis (transient spectra, yield and decay rate of triplets) and by thermal lensing (amount of heat deposited in the medium). The competition between electron transfer (dye*–dye) and energy transfer (dye*–O2) reactions was accessed as a function of the dimer–monomer ratio. The lower yield of electron transfer observed for dimers in AOT reverse micelles and intact vesicles compared with SDS micelles and frozen vesicles at similar dimer–monomer ratios is related with the different types of aggregates induced by each interface.

Modulation of methylene blue photochemical properties based on adsorption at aqueous micelle interfaces

Methylene Blue (MB+) is a sensitizer that has been used for a variety of applications including energy conversion and photodynamic therapy (PDT). Although its photochemical properties in isotropic solution are well established, its effect in vivo and in restricted reaction environments is somewhat erratic. In order to understand its photochemical behavior when it interacts with biomolecules, in particular with membranes, MB+ properties were studied in sodium dodecyl sulfate (SDS) and cetyl trimethylammonium bromide (CTAB) solutions. Because of an electrostatic attraction, SDS and MB+ form complexes, changing the properties of both the micelles and the MB+ solutions. Surface tension measurements show that the c.m.c. of SDS decreases from ∼7 mM to ∼70 μM when the MB+ concentration increases from 0 to 45 μM. Above the c.m.c., binding of MB+ in the micelle pseudo-phase causes the formation of aggregates (mostly dimers) as attested by the increase in the absorption at 580 nm and the decrease in fluorescence emission. The extent of dimer formation is dependent on the relative concentrations of MB+ and SDS. In the presence of excess of SDS, MB+ is mainly in the monomer form and at low SDS concentration dimers are favored. Such effect, which was not observed in CTAB micelles, was modeled qualitatively by considering that MB+ molecules partition to the micelle pseudo-phase which favors or disfavors dimers as a function of its volume. MB+ transient species were characterized by laser flash photolysis and NIR emission showing the presence of triplets and subsequently singlet oxygen at high SDS concentration and semi-reduced and semi-oxidized MB+ radicals at low SDS concentration. Therefore it was shown that, depending on the ground state MB+ monomer/dimer equilibrium, induced by the micelles, the photochemical properties of MB+ can be shifted from a Type II (energy transfer to oxygen forming singlet oxygen) to a Type I mechanism (electron transfer forming the semi-reduced and the semi-oxidized radicals of MB+).

The role of the gel liquid-crystalline phase transition in the lung surfactant cycle.

Lipid polymorphism plays an important role in the lung surfactant cycle. A better understanding of the influence of phase transitions on the formation of a lipid film from dispersions of vesicles will help to describe the mechanism of action of lung surfactant. The surface pressure (or tension) of dispersions of DPPC, DMPC, and DPPE unilamellar vesicles was studied as a function of temperature. These aggregates rapidly fuse with a clean air-water interface when the system is at their phase transition temperature (Tm), showing a direct correlation between phase transition and film formation. Based on these results, an explanation on how fluid aggregates in the alveolar subphase can form a rigid monolayer at the alveolar interface is proposed.

Reduction of evaporation of natural water samples by monomolecular films

The ability of monomolecular films of fatty alcohols in reducing evaporation of natural water samples collected from two reservoirs in Sao Paulo (Guarapiranga and Billings) was studied in the laboratory. Evaporation reductions were determined directly by measuring the volume of water evaporated from the test recipients after a certain period of time. Mixed films of hexadecanol and octadecanol showed high potential of evaporation reduction (up to 57%) and persistence on the water surface of ca. 48 hours. A brief discussion on the better efficiency of these mixed films in reducing water evaporation is also presented.

Measurement of refractive-index change at a liquid–solid interface close to the critical angle

We measured the refractive-index change on a liquid sample, using the reflection of a polarized Gaussian laser beam close to the angle of total reflection. We applied this technique to a solution of nickel (ii) phthalocyanine tetrasulfonated (NiPTS) in water-ethanol (1/1 v/v), in which the nonlinearity of the refractive index is due to optically induced thermal effects. We show that close to the angle of total reflection the sensitivity of this technique is four times bigger than at normal incidence.

Fusion of vesicles with the air-water interface: the influence of polar head group, salt concentration, and vesicle size.

Fusion of vesicles with the air-water interface and consequent monolayer formation has been studied as a function of temperature. Unilamellar vesicles of DMPC, DPPC, and DODAX (X=Cl(-), Br(-)) were injected into a subphase containing NaCl, and the surface pressure (tension) was recorded on a Langmuir Balance (Tensiometer) using the Wilhelmy plate (Ring) method. For the zwitterionic vesicles, plots of the initial surface pressure increase rate (surface tension decrease rate) as a function of temperature show a peak at the phase transition temperature (T(m)) of the vesicles, whereas for ionic ones they show a sharp rise. At high concentrations of NaCl, ionic DODA(Cl) vesicles seem to behave like zwitterionic ones, and the rate of fusion is higher at the T(m). The influence of size was studied comparing large DODA(Cl) vesicles with small sonicated ones, and no significant changes were found regarding the rate of fusion with the air-water interface.

Single-beam interface thermal lensing.

A single-beam photothermal-lensing technique to study interfaces is presented. By analysis of the reflection from a quartz-solution interface with a low-power laser in a single-beam configuration, a photothermal signal is detected. The data were fitted with a conventional thermal lens model, and the results show that the optical element formed at the interface resembles an inverted thermal lens.

A simple surface tension method for demonstrating the Lβ‐Lα transition in biological membranes

The rate of vesicle formation at the air/water interface monitored by surface tension measurements as a function of temperature is presented as a simple student experiment to determine the phase transition temperature of biological membranes.

Quantitative determination of singlet oxygen by laser deflection calorimetry

ABSTRACT A sensitive method for the quantitative determination of singlet oxygen based on a low-cost laser deflection calorimeter (LDC) apparatus is presented. The heat released from the singlet oxygen generation and its nonradiative decay in an aqueous phase causes the formation of a thermally driven refractive index gradient in an adjacent organic phase. This effect, which is proportional to the amount of singlet oxygen generated, is monitored by the deflection of a laser beam. Limits of detection (LOD) obtained were in the sub-μmol level which is one order of magnitude lower than the LOD obtained by measuring the “dimol” emission at 633 run. This method should be specially designed for studying chemical and biochemical processes at interfaces by measuring low amounts of heat released.