Aqueous Bulk Research Articles

Ferrofluid based dispersive-solid phase extraction for spectrophotometric determination of dyes

For the first time, ferrofluid based dispersive-solid phase extraction (D-SPE) has been applied for determination of trace levels of dyes in aqueous and fish samples. The contaminant used as a model compound was crystal violet (CV), a cationic dye, and was preconcentrated without any derivatization or ion-pair formation. The method is based on rapid injection of ferrofluid into the aqueous sample by a syringe. The sample preparation time is decreased by the fact that the sorbent dispersed in the bulk solution and extraction can be achieved very fast. In this way, the separation of sorbent from the aqueous bulk was achieved by a magnet, and no centrifugation is required. These significant features which obtained with this method are of key interest for routine trace laboratory analysis. The influence of different variables on D-SPE was investigated. Under optimum conditions, the calibration graph was linear over the range of 3.3–90μgL−1, and the enrichment factor (EF) 267 was obtained. Detection limit was 1.51μgL−1 (n=7), and the relative standard deviation of 5.6% at 50ngmL−1 was obtained (n=7). The proposed method was successfully applied for the determination of crystal violet in various samples.

Estimation of LPC/water partition coefficients using molecular modeling and micellar liquid chromatography

Lysophosphatidylcholine (LPC) is a lipid based, micelle forming emulsifier and thus a potential solubilizer for hydrophobic components in pharmaceutical and food products. Since many components of interest are very hydrophobic, the determination of the partition equilibrium between the micelles and the aqueous bulk phase is challenging. In this work several methods (micellar liquid chromatography (MLC), molar solubilization ratio (MSR) and an indirect method, using an organic solvent phase) for the experimental determination of partition coefficients are compared. For the first time the emulsifier LPC was employed in the MLC. It is shown, that the partition coefficients between micelles and water can be determined with good agreement to literature data and between the different methods for solutes in a wide hydrophobicity range.Moreover, the experimental data are compared to a priori predictions based on the thermodynamic model COSMO-RS. The prediction assuming the pseudophase approach is compared to the prediction using COSMOmic, thus taking into account the anisotropic structure of the micelles, obtained from molecular dynamics all-atom simulations. Both methods agree well with the experimental data for the partition coefficients between LPC and water and between the anionic surfactant SDS and water. Using COSMOmic, beside the partition coefficient, the location of the solute within the micelle is predicted. It is shown, that the combination of experimental methods and models like COSMO-RS reveals a deeper understanding of the specific interactions, which is needed for an efficient use of emulsifiers in a variety of applications.

Coacervative extraction of trace lead from natural waters prior to its determination by electrothermal atomic absorption spectrometry

In this work, a relatively simple and sensitive method for separation/preconcentration of trace lead from natural waters prior to its determination by electrothermal atomic absorption spectrometry has been proposed. The method is based on the extraction of Pb–dithizone chelate with coacervates made up of lauric acid in the presence of potassium ions and methanol. Several important factors affecting extraction efficiency such as pH, concentration of lauric acid and dithizone, ionic strength, incubation and centrifugation time were investigated and optimized. After separation of aqueous bulk solution from surfactant-rich phase, the final extract was redissolved by using 500μl of methanol acidified with 0.2moll−1 HNO3. Under the optimized conditions (using initial sample volume of 10ml), enrichment factor of 17.0, detection limit of 0.12μgl−1, quantification limit of 0.38μgl−1, relative standard deviation of 4.2% (for 2μgl−1 of Pb; n=26), linearity of the calibration graph in the range of 0.5–4.0μgl−1 (with correlation coefficient better than 0.995) were achieved. The method was validated by the analysis of certified reference material (TMDA-61). Extraction recoveries for the CRM, spiked model solutions and spiked natural water samples were in the range of 91–96%. Finally, the method was applied to the separation/preconcentration and determination of trace lead in natural waters.

Electron at the Surface of Water: Dehydrated or Not?

The hydrated electron is a crucial species in radiative processes, and it has been speculated that its behavior at the water surface could lead to specific interfacial chemical properties. Here, we address fundamental questions concerning the structure and energetics of an electron at the surface of water. We use the method of ab initio molecular dynamics, which was shown to provide a faithful description of solvated electrons in large water clusters and in bulk water. The present results clearly demonstrate that the surface electron is mostly buried in the interfacial water layer, with only about 10 % of its density protruding into the vapor phase. Consequently, it has a structure that is very similar to that of an electron solvated in the aqueous bulk. This points to a general feature of charges at the surface of water, namely, that they do not behave as half-dehydrated but rather as almost fully hydrated species.

Aqueous core nanocapsules: a new solution for encapsulating doxorubicin hydrochloride

In this study, we propose a new solution for the nanoencapsulation of hydrophilic anticancer drug, doxorubicin hydrochloride (DOX). The drug molecules are solubilized in the core of aqueous nanoreservoirs, so-called aqueous core nanocapsules (ACN) recently developed by our team, and dispersed in aqueous bulk media. Since it is well acknowledged that the nanoencapsulation of DOX has many advantages, like reducing the sides effects (e.g. cardiac toxicity), we propose through the present study a novel formulation solution for this purpose. After focusing on the formulation process for optimizing the drug encapsulation yield, the DOX-release profiles were followed up and analyzed. Different physicochemical and in vitro characterization were performed, and complement activation experiments. ACN were shown efficient to encapsulate DOX reaching yields as high as 80%, followed by a sustained release governed by a diffusion-controlled mechanism. The loaded nanocarriers showed low levels of complement activation, compatible with stealth properties. To summarize, this study brings out a new tool for the nanoencapsulation of hydrophilic anticancers and could open new doors for the administration of this particular class of drugs.

Comparisons between Hygroscopic Measurements and UNIFAC Model Predictions for Dicarboxylic Organic Aerosol Mixtures

Hygroscopic behavior was measured at 12°C over aqueous bulk solutions containing dicarboxylic acids, using a Baratron pressure transducer. Our experimental measurements of water activity for malonic acid solutions (0–10 mol/kg water) and glutaric acid solutions (0–5 mol/kg water) agreed to within 0.6% and 0.8% of the predictions using Peng’s modified UNIFAC model, respectively (except for the 10 mol/kg water value, which differed by 2%). However, for solutions containing mixtures of malonic/glutaric acids, malonic/succinic acids, and glutaric/succinic acids, the disagreements between the measurements and predictions using the ZSR model or Peng’s modified UNIFAC model are higher than those for the single-component cases. Measurements of the overall water vapor pressure for 50 : 50 molar mixtures of malonic/glutaric acids closely followed that for malonic acid alone. For mixtures of malonic/succinic acids and glutaric/succinic acids, the influence of a constant concentration of succinic acid on water uptake became more significant as the concentration of malonic acid or glutaric acid was increased.

Synthesis, Growth and Characterization of Organic Nonlinear Optical L-Asparagine L-Tartrate Single Crystals

L-asparagine L-tartrate (LAT), an organic compound has been synthesized from aqueous solution and bulk single crystal has been grown by slow evaporation technique. Powder X-ray diffraction studies confirmed the monoclinic structure of the grown LAT crystal. The presence of functional groups of the grown crystal was identified by FTIR studies. Linear optical property of the grown crystal was studied by UV-Vis spectral analysis. Microhardness studies reveal that the crystal possesses relatively higher hardness compared to other organic nonlinear optical crystals. Dielectric response of the L-asparagine L-tartrate crystal was analyzed for different frequencies at various temperatures. Kurtz-Perry powder second harmonic generation test confirmed the nonlinear optical properties of the as-grown LAT crystal.

Photochemical stability of 4′-azido-2′-deoxy-2′-methylcytidine hydrochloride: structural elucidation of major degradation products by LC–MS and NMR analysis

The photochemical stability of (1'R,2'S,3'S,4'R)-4'-azido-2'-deoxy-2'-methylcytidine hydrochloride, a new anti-HCV agent, was investigated. Aqueous solutions and bulk drug powder of the drug candidate were exposed to UV-visible light, complying with ICH requirements. The nucleoside analog decomposed via loss of nitrogen to yield products derived from a highly reactive azide intermediate. Major photolysis products were identified by LC-MS and NMR analysis, revealing three main photodegradation pathways. The first one led to the formation of a ring-expanded imidate ester. The other degradation pathways involved exocyclic or endocyclic bond cleavage with imine or imino lactone formation. The latter were prone to rapid hydrolysis, eventually resulting in the release of cytosine, 2-methyl malonaldehyde and (E)-cytosyl-2-methylpropenal.

Spatial Scanning Spectroelectrochemistry. Study of the Electrodeposition of Pd Nanoparticles at the Liquid/Liquid Interface

Spatial scanning spectroelectrochemistry is a new analytical technique that provides spectral information at different distances from an electrified liquid/liquid interface where an electrochemical process takes place. As a proof of concept, we have studied two different electrochemical processes at the electrified liquid/liquid interface: (1) Ru(bpy)(3)(2+) transfer through the water/1,2-dichloroethane interface and (2) electrodeposition of Pd nanoparticles at the water/1,2-dichloroethane interface. The instrumental setup developed consists of a movable slit for the light beam to sample at well-defined positions on both sides of the interface, providing important information about the chemical process occurring. If the slit is scanned at different distances from the interface during an electrochemical experiment, a complete picture of the reactions and equilibria in the diffusion layer can be obtained. For example, in the case of the Ru(bpy)(3)(2+), the experiments show clearly how the complex is transferred from one phase to the other. In the case of electrosynthesis of Pd nanoparticles, it is demonstrated that nanoparticles are not only deposited at the interface but diffuse to the aqueous bulk solution. These in situ observations were confirmed by ex situ experiments using transmission electron microscopy.

Ionic Strength and pH as Control Parameters for Spontaneous Surface Oscillations

A system far from equilibrium, where the surfactant transfer from a small drop located in the aqueous bulk to the air-water interface results in spontaneous nonlinear oscillations of surface tension, is theoretically and experimentally considered. The oscillations in this system are the result of periodically arising and terminating Marangoni instability. The surfactant under consideration is octanoic acid, the dissociated form of which is much less surface-active than the protonated form. Numerical simulations show how the system behavior can be controlled by changes in pH and ionic strength of the aqueous phase. The results of numerical simulations are in good agreement with experimental data.

Net Proton Uptake Is Preceded by Multiple Proton Transfer Steps upon Electron Injection into Cytochrome c Oxidase

Cytochrome c oxidase (COX), the last enzyme of the respiratory chain of aerobic organisms, catalyzes the reduction of molecular oxygen to water. It is a redox-linked proton pump, whose mechanism of proton pumping has been controversially discussed, and the coupling of proton and electron transfer is still not understood. Here, we investigated the kinetics of proton transfer reactions following the injection of a single electron into the fully oxidized enzyme and its transfer to the hemes using time-resolved absorption spectroscopy and pH indicator dyes. By comparison of proton uptake and release kinetics observed for solubilized COX and COX-containing liposomes, we conclude that the 1-μs electron injection into Cu(A), close to the positive membrane side (P-side) of the enzyme, already results in proton uptake from both the P-side and the N (negative)-side (1.5 H(+)/COX and 1 H(+)/COX, respectively). The subsequent 10-μs transfer of the electron to heme a is accompanied by the release of 1 proton from the P-side to the aqueous bulk phase, leaving ∼0.5 H(+)/COX at this side to electrostatically compensate the charge of the electron. With ∼200 μs, all but 0.4 H(+) at the N-side are released to the bulk phase, and the remaining proton is transferred toward the hemes to a so-called "pump site." Thus, this proton may already be taken up by the enzyme as early as during the first electron transfer to Cu(A). These results support the idea of a proton-collecting antenna, switched on by electron injection.

Solvent-based de-emulsification dispersive liquid–liquid microextraction of palladium in environmental samples and determination by electrothermal atomic absorption spectrometry

A new simple, rapid and efficient sample pretreatment technique for simultaneous extraction and preconcentration was developed. In this study, solvent-based de-emulsification dispersive liquid–liquid microextraction (SD-DLLME) was developed for preconcentration and highly sensitive determination of Pd(II) using the electrothermal atomic absorption spectrometry. The extraction of Pd(II) was performed in the presence of Thio-Michler's ketone (TMK) as the complexing agent. After dispersing, when an aliquot of acetonitrile was introduced as a chemical demulsifier into the aqueous bulk, the obtained emulsion cleared into two phases quickly. Therefore, an extra centrifugation step is not needed for phase separation. Several factors that influence the microextraction efficiency and ETAAS signal, such as pH, TMK concentration, counter ion, extraction time, disperser, de-emulsifier solvents and extraction solvents, stirring rate, pyrolysis and atomization temperature were investigated. Under the optimized conditions, the calibration graph was linear over the range of 0.025–0.500μgL−1 and relative standard deviation of 3.68% at 0.1μgL−1 was obtained (n=7). The limit of detection and the enrichment factor (EF) were obtained 0.007μgL−1 and 231, respectively. The obtained results indicated that the developed method is an excellent alternative for the routine analysis in the environmental field.

In-syringe demulsified dispersive liquid–liquid microextraction and high performance liquid chromatography–mass spectrometry for the determination of trace fungicides in environmental water samples

An in-syringe demulsified dispersive liquid–liquid microextraction (ISD–DLLME) technique was developed using low-density extraction solvents for the highly sensitive determination of the three trace fungicides (azoxystrobin, diethofencarb and pyrimethanil) in water samples by high performance liquid chromatography–mass spectrometry chromatography–diode array detector/electrospray ionisation mass spectrometry. In the proposed technique, a 5-mL syringe was used as an extraction, separation and preconcentration container. The emulsion was obtained after the mixture of toluene (extraction solvent) and methanol (dispersive solvent) was injected into the aqueous bulk of the syringe. The obtained emulsion cleared into two phases without centrifugation, when an aliquot of methanol was introduced as a demulsifier. The separated floating organic extraction solvent was impelled and collected into a pipette tip fitted to the tip of the syringe. Under the optimal conditions, the enrichment factors for azoxystrobin, diethofencarb and pyrimethanil were 239, 200, 195, respectively. The limits of detection, calculated as three times the signal-to-noise ratio (SN−1), were 0.026μgL−1 for azoxystrobin, 0.071μgL−1 for diethofencarb and 0.040μgL−1 for pyrimethanil. The repeatability study was carried out by extracting the spiked water samples at concentration levels of 0.02μgmL−1 for all the three fungicides. The relative standard deviations varied between 4.9 and 8.2% (n=5). The recoveries of all the three fungicides from tap, lake and rain water samples at spiking levels of 0.2, 1, 5μgL−1 were in the range of 90.0–105.0%, 86.0–114.0% and 88.6–110.0%, respectively. The proposed ISD–DLLME technique was demonstrated to be simple, practical and efficient for the determination of different kinds of fungicide residues in real water samples.

Experimental Methods and Prediction with COSMO-RS to Determine Partition Coefficients in Complex Surfactant Systems

Surfactant-based separation processes are a promising alternative to conventional organic solvent processes. A crucial parameter to describe the efficiency of such processes is the partition coefficient between the surfactant aggregates (micelles) and the aqueous bulk phase. In this work, several experimental methods to determine these partition coefficients (micellar liquid chromatography, micellar enhanced ultrafiltration, and cloud point extraction) are evaluated and compared. In addition, these results are compared to predictions with the thermodynamic model COSMO-RS. In particular, systems with the nonionic surfactant TritonX-100 are studied. The partition equilibria of various solutes (pyrene, naphthalene, phenanthrene, phenol, 3-methoxyphenol, and vanillin) and the influence of different additives (alcohols) are investigated. All experimental methods show very good reproducibility. Moreover, the results from different methods are in good agreement, supplementing one another concerning the temperature ranges. Notably, the COSMO-RS model is capable of predicting partition coefficients between micelles and water in the investigated temperature range and at different alcohol concentrations. The results demonstrate the potential of the model COSMO-RS to facilitate the selection of optimized process parameters for a given separation problem. By predicting partition equilibria in multicomponent systems, the selection of surfactant, temperature, and appropriate additives can be facilitated.

Modeling rhamnolipids production by Pseudomonas aeruginosa from immiscible carbon source in a batch system

A mathematical model to predict the rhamnolipids production by Pseudomonas aeruginosa from oleic acid in a two phase liquid-liquid batch reaction system, was developed in this study. The model was based on two theoretical assumptions: 1) the convective oleic acid mass transfer is coupled to a bioreaction in the aqueous liquid bulk, and 2) the volume of the immiscible oleic acid drops and the saturation concentration at the interface are a function of rhamnolipids production. The model was able to accurately predict the experimental growth of the Pseudomonas aeruginosa strain, and the rhamnolipids production data with oleic acid as carbon source. This mathematical approach indicated a high correspondence between the saturation dimensionless profiles of oleic acid at the interface and the experimental profiles of surface tension difference. This modeling approach may constitute a useful tool in the design and scaling-up of bioreactors applied to the production of biosurfactants with immiscible carbon sources.

Successful preferential formation of a novel macromolecular assembly—Trilayered polymeric micelle—That can incorporate hydrophilic compounds: The optimization of factors affecting the micelle formation from amphiphilic block copolymers

We have developed a novel macromolecular assembly, trilayered polymeric micelle, which can incorporate hydrophilic compounds. The micelle can be prepared from the amphiphilic block copolymers without regard to their properties such as the copolymer's charges and the homogeneity of the copolymers forming the micelle's inner and outer parts. In this study, we investigated the optimal condition for the preferential formation of the trilayered polymeric micelle. GPC results clarified that the composition of the block copolymer, the concentration of PVA in the aqueous bulk phase, and the temperature during the preparation were the important preparation factors affecting preferential formation of the trilayered polymeric micelles. We successfully achieved the preferential formation of the trilayered polymeric micelles under optimal conditions. Furthermore, we confirmed that the model hydrophilic compound, FITC-dextran, was successfully encapsulated into the hydrophilic core of the trilayered polymeric micelles. The novel micelle that can incorporate hydrophilic compounds can have a variety of future medical applications such as a protein delivery-based therapy.

Affinity of Four Polar Neurotransmitters for Lipid Bilayer Membranes

Weak interactions of neurotransmitters and the lipid matrix in the synaptic membrane have been hypothesized to play a role in synaptic transmission of nerve signals, particularly with respect to receptor desensitization (Cantor, R. S. Biochemistry 2003, 42, 11891). The strength of such interactions, however, was not measured, and this is an obvious impediment for further evaluation and understanding of a possible role for desensitization. We have used dialysis equilibrium to directly measure the net affinity of selected neurotransmitters for lipid membranes and analyzed this affinity data with respect to calorimetric measurements and molecular dynamics simulations. We studied an anionic (glutamate), a cationic (acetylcholine), and two zwitterionic (γ-aminobutyric acid and glycine) neurotransmitters, and membranes of pure dimyristoyl phosphatidylcholine (DMPC), DMPC doped with 10% anionic lipid (dimyristoyl phosphatidylglycerol, DMPG, or dimyristoyl phosphatidylserine, DMPS), or 1:1 mixtures of dipalmitoyl phosphatidylcholine (DPPC) and dilauroyl phosphatidylcholine (DLPC). The results showed a remarkable variability among the investigated systems. For example, the chloride salt of acetylcholine interacts unfavorably with DMPC and is thus preferentially excluded from the membrane's hydration layer. Conversely, the zwitterionic neurotransmitters are attracted to membranes with 10% anionic lipid and their local concentration at the interface is 5-10 times larger than in the aqueous bulk. The simulations suggest that this attraction mainly relies on electrostatic interactions of the amino group of the neurotransmitter and the lipid phosphate. We conclude that moderate attraction to lipid membranes occurs for some polar neurotransmitters and hence that one premise for a theory of bilayer-mediated modulation of nerve transmission seems to be fulfilled. However, the strong variability in interaction strengths also shows that this attraction is not an inherent property of all neurotransmitters.

The Many Faces of Gold Nanorods

Gold nanorods exhibit optical properties that are tunable with their shape, leading to sensing, imaging, and biomedical therapeutic applications. Colloidal preparations of gold nanorods impart surfactants or other species on the nanorod surfaces; a popular preparation leads to a surfactant bilayer on the surface. The specific chemistry at three distinct interfaces has roles to play in the growth and subsequent usage of these nanomaterials; these interfaces are the gold−surfactant interface, the hydrophobic surfactant bilayer, and, finally, the surfactant interface with the aqueous bulk. Each one of these interfaces provides strategies for altering nanorod properties such as stability against aggregation, toxicity, and ease of assembly. It is the solvent-accessible interface that dictates nanorod interactions with other particles, macromolecules, and living cells.

Sensitive determination of 2,4,6-trichloroanisole in water samples by ultrasound assisted emulsification microextraction prior to gas chromatography–tandem mass spectrometry analysis

A novel application of an ultrasound assisted emulsification microextraction (USAEME) technique is proposed for the extraction and preconcentration of 2,4,6-trichloroanisole (2,4,6-TCA) from water samples prior to its determination by gas chromatography-tandem mass spectrometry (GC-MS/MS). USAEME employs a non-polar high-density solvent (extractant solvent), which forms an oil-in-water emulsion (O/W) in the aqueous sample bulk assisted by ultrasonic radiation. Several factors including, solvent type and volume, extraction time, extraction temperature, shaking mode and matrix modifiers were studied and optimized over the relative recovery of the target analyte. An aliquot of 5mL water sample was conditioned by adding 150microL 6.15molL(-1) sodium chloride and 300microL 0.05molL(-1) phosphate buffer (pH 6), and finally extracted with 40microL chloroform by using USAEME technique. Under the optimal experimental conditions 2,4,6-TCA was quantitatively extracted achieving an enrichment factor (EF) of 555. The detection limit (LOD), calculated as three times the signal-to-noise ratio (S/N), was 0.2ngL(-1) and the RSD was 6.3% (n=5) when 1ngL(-1) 2,4,6-TCA standard mixture was analyzed. The coefficients of estimation of the calibration curves obtained following the proposed methodology was >or=0.997 and the linear working range was 1-5000ngL(-1). Finally, the proposed technique was successfully applied for extraction and determination of the 2,4,6-TCA in water samples. Recovery studies lead values >or=94%, which showed a successfully robustness of the analytical methodology for determination of nanogram per liter of 2,4,6-TCA in water samples.

Functional interaction structures of the photochromic retinal protein rhodopsin

We studied functional interaction structures of the vertebrate membrane photoreceptor rhodopsin containing retinal as a chromophore. Using time-resolved fluorescence depolarization we analyzed real-time dynamics and conformational changes of the cytoplasmic helix 8 (H8) preceding the long C-terminal tail of rhodopsin. H8 runs parallel to the membrane surface and extends from transmembrane helix 7 whose highly conserved NPxxY(x)F motif connects that region of rhodopsin with the retinal binding pocket. Our measurements indicate that photo-induced retinal isomerization from 11-cis to all-trans provokes conformational changes of H8, including slower motion and reduced flexibility, that are specific for the active metarhodopsin-II photo-intermediate. These conformational changes are absent in the retinal-devoid state opsin and in the phosphorylated metarhodopsin-II state upon receptor deactivation. Furthermore we show that membrane rim effects can influence interfacial reactions at the cytoplasmic rhodopsin surface such as proton transfer reactions between surface and aqueous bulk phase or binding of the signaling protein transducin visualized with single-molecule widefield microscopy. These findings are important for an understanding of the effects of membrane structure on the photo-transduction mechanism.