Limits of Visual Detection for Finasteride Polymorphs in Prepared Binary Mixtures: Analysis by X-ray Powder Diffraction

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Interaction of the meso-tetrakis (4-N-methylpyridyl) porphyrin with gel and liquid state phospholipid vesicles

The interaction of the cationic meso-tetrakis 4-N-methylpyridyl porphyrin (TMPyP) with large unilamellar vesicles (LUVs) was investigated in the present study. LUVs were formed by mixtures of the zwitterionic 1,2-dipalmitoyl-sn-glycero-phosphatidylcholine (DPPC) and anionic 1,2-dipalmitoyl-sn-glycero-3-phosphoglycerol (DPPG) phospholipids, at different DPPG molar percentages. All investigations were carried out above (50 degrees C) and below (25 degrees C) the main phase transition temperature of the LUVs (similar to 41 degrees C). The binding constant values, K-b, estimated from the time-resolved fluorescence study, showed a significant increase of the porphyrin affinity at higher mol% DPPG. This affinity is markedly increased when the LUVs are in the liquid crystalline state. For both situations, the increase of the K-b value was also followed by a higher porphyrin fraction bound to the LUVs. The displacement of the vesicle-bound porphyrins toward the aqueous medium, upon titration with the salt potassium chloride (KCl), was also studied. Altogether, our steady-state and frequency-domain fluorescence quenching data results indicate that the TMPyP is preferentially located at the LUVs Stern layer. This is supported by the zeta potential studies, where a partial neutralization of the LUVs surface charge, upon porphyrin titration, was observed. Dynamic light scattering (DLS) results showed that, for some phospholipid systems, this partial neutralization leads to the LUVs flocculation. (C) 2012 Elsevier Inc. All rights reserved.

Application of Electrochemical Degradation of Wastewater Composed of Mixtures of Phenol-Formaldehyde

The industrial wastewater from resin production plants contains as major components phenol and formaldehyde, which are traditionally treated by biological methods. As a possible alternative method, electrochemical treatment was tested using solutions containing a mixture of phenol and formaldehyde simulating an industrial effluent. The anode used was a dimensionally stable anode (DSAA (R)) of nominal composition Ti/Ru0.3Ti0.7O2, and the solution composition during the degradation process was analyzed by liquid chromatography and the removal of total organic carbon. From cyclic voltammetry, it is observed that for formaldehyde, a small offset of the beginning of the oxygen evolution reaction occurs, but for phenol, the reaction is inhibited and the current density decreases. From the electrochemical degradations, it was determined that 40 mA cm(-2) is the most efficient current density and the comparison of different supporting electrolytes (Na2SO4, NaNO3, and NaCl) indicated a higher removal of total organic carbon in NaCl medium.

Crystal structure of isotibolone: a major degradation product of tibolone

Isotibolone is frequently found as an impurity in tibolone, a drug used for hormone reposition of post-menopause women, due to some inadequate tibolone synthesis or as a result of degradation during drug storage. The presence of isotibolone impurities should be detected and quantified in active pharmaceutical ingredient products of tibolone before its use in the manufacturing of medicaments. The X-ray powder diffraction technique offers the possibility of quantifying isotibolone amounts at different stages of drug production and storage, from the chemical synthesis to the final formulation. In the course of a study involving the quantitative analysis of isotibolone by X-ray powder diffraction, the authors determined the structure of the title compound using a recently developed approach (A. Gomez and S. Kycia, J. Appl. Crystallogr. 2011, 44, 708-713). The structure is monoclinic, space group P2(1) (4), with unit cell parameters a = 6.80704(7) angstrom, b = 20.73858(18) angstrom, c = 6.44900(6) angstrom, beta = 76.4302(5)degrees, V = 884.980(15) angstrom(3) and two molecules per unit cell (Z = 2). The molecules are hydrogen bonded in the ab plane forming layers that are held together in the crystal by van der Waals interactions along the c-axis.

Multiproperty correlation of experimental data of the binaries propyl ethanoate+alkanes (pentane to decane). New experimental information for vapor-liquid equilibrium and mixing properties

[EN]A thermodynamic study is carried out on binary systems composed of propyl ethanoate with six alkanes, from pentane to decane. Vapor pressures of the ester and the isobaric vapor−liquid equilibria of these six mixtures were measured at 101.32 kPa in a small-capacity ebulliometer and also the mixing properties yE = vE,hE over a range of temperatures and at atmospheric pressure. Adequate correlations are drawn for the surfaces yE = yE(x,T) with an interpretation on the behavior of the mixtures and also using cp E data from literature.

Materiali Ceramici per Edilizia con Funzionalità Fotocatalitica

This thesis wad aimed at the study and application of titanium dioxide photocatalytic activity on ceramic materials. As a matter of fact, photocatalysis is a very promising method to face most of the problems connected with the increasing environmental pollution. Furthermore, titanium dioxide, in its anatase crystallographic phase, is one of the most investigated photocatalytic material and results to be perfectly compatible with silicate body mixes. That goal was pursued by two different strategies: 1. the addition to a body mix used for heavy clay products of several titania powders, with different mean crystallite size, surface area, morphology and anatase/rutile ratio and a titania nanosuspension as well. The titania addition followed two procedures: bulk and spray addition over the ceramic samples surface. Titania was added in two different percentages: 2.5 and 7.5 wt.% in both of the methods. The ceramic samples were then fired at three maximum temperatures: 900, 950 and 1000 °C. Afterwards, the photocatalytic activity of the prepared ceramic samples was evaluated by following the degradation of an organic compound in aqueous medium, under UV radiation. The influence of titania morphological characteristics on the photoactivity of the fired materials was studied by means of XRD and SEM observations. The ceramic samples, sprayed with a slip containing 7.5 wt.% of titania powder and fired at 900 °C, have the best photoactivity, with a complete photo-decomposition of the organic compound. At 1000 °C no sample acted as a photocatalyst due to the anatase-to-rutile phase transformation and to the reaction between titania and calcium and iron oxides in the raw materials. 2. The second one foresaw the synthesis of TiO2-SiO2 solid solutions, using the following stoichiometry: Ti1-xSixO2 where x = 0, 0.1, 0.3 and 0.5 atoms per formula unit (apfu). The mixtures were then fired following two thermal cycles, each with three maximum temperatures. The effect of SiO2 addition into the TiO2 crystal structure and, consequently, on its photocatalytic activity when fired at high temperature, was thoroughly investigated by means of XRD, XPS, FE-SEM, TEM and BET analysis. The photoactivity of the prepared powders was assessed both in gas and liquid phase. Subsequently, the TiO2-SiO2 solid solutions, previously fired at 900 °C, were sprayed over the ceramic samples surface in the percentage of 7.5 wt.%. The prepared ceramic samples were fired at 900 and 1000 °C. The photocatalytic activity of the ceramic samples was evaluated in liquid phase. Unfortunately, that samples did not show any appreciable photoactivity. In fact, samples fired at 900 °C showed a pretty low photoactivity, while the one fired at 1000 °C showed no photoactivity at all. This was explained by the excessive coarsening of titania particles. To summarise, titania particle size, more than its crystalline phase, seems to have a relevant role in the photocatalytic activity of the ceramic samples.

Surface layer formation in polymer melts and in solutions

In this work the surface layer formation in polymer melts and in polymer solutions have been investigated with the atomic force microscope (AFM). In polymer melts, the formation of an immobile surface layer results in a steric repulsion, which can be measured by the AFM. From former work it is know, that polydimethyl siloxane (PDMS) forms a stable surface layer for molecular weights above 12 kDa. In the present thesis, polyisoprene (PI) was investigated apart from PDMS, by a)measuring the steric surface interactions and b)measuring the surface slip in hydrodynamic experiments. If a polymer flows over a surface, the flow velocity at the surface is larger then zero. If case of a surface layer formation the flow plane changes to the top of the adsorbed layer and the surface slip is reduced to zero. By measuring the surface slip in hydrodynamic experiments, it is therefore possible to determine the presence of a stable surface layer. The results show no stable repulsion for PI and only a small decrease of the surface slip. This indicates that PI does not form a stable surface layer, but is only adsorbed weakly to the surface. Furthermore for 8 kDa PDMS the timescale of the formation of a surface layer was investigated by changing themaximal force the tip applied to the surface. With a repulsive force present, applying a higher force than 15 nN resulted in a destruction of the surface layer, indicated by attractive forces. Reducing the applied force below 15 nN then resulted in an increase of the repulsion to the former state during one minute, thus indicating that a surface layer can be formed within one minute even under the influence of continuous measurements. As a next step, mixtures of two PDMS homopolymers with different chain lengths have been investigated. The aim was to verify theoretical predictions that shorter chains should predominate at the surface due to their smaller loss in conformational entropy. The measurements where done in dependence of the volume fractions of short and long chain PMDS. The results confirmed the short chain dominance for all mixtures with less then 90 vol.% long chain PDMS. Surface layer formation in solution was investigated for superplasticizers which are industrially used as an additive to cement. They change the surface interaction between the cement grains from attractive to repulsive and the freshlymixed cement paste therefore becomes liquid. The aimin this part of the thesis was, to investigate cement particle interactions in a close to real environment. Therefore calcium silicate hydrate phases have been precipitated onto an AFM tip and onto a calcite crystal and the interaction between these surfaces have beenmeasured with and without addition of superplasticizers. The measurements confirmed the change from attraction to repulsion upon addition of superplasticizers. The repulsive steric interaction increased with the length of the sidechain of the superplasticizer, and the dependence of the range of the steric interactions on the sidechain length indicated that the sidechains are in a coiled conformation.

Preparation of new crystal forms via photochemical, mechanochemical and sol-gel methods

This work of thesis involves various aspects of crystal engineering. Chapter 1 focuses on crystals containing crown ether complexes. Aspects such as the possibility of preparing these materials by non-solution methods, i.e. by direct reaction of the solid components, thermal behavior and also isomorphism and interconversion between hydrates are taken into account. In chapter 2 a study is presented aimed to understanding the relationship between hydrogen bonding capability and shape of the building blocks chosen to construct crystals. The focus is on the control exerted by shape on the organization of sandwich cations such as cobalticinium, decamethylcobalticinium and bisbenzenchromium(I) and on the aggregation of monoanions all containing carboxylic and carboxylate groups, into 0-D, 1-D, 2-D and 3-D networks. Reactions conducted in multi-component molecular assemblies or co-crystals have been recognized as a way to control reactivity in the solid state. The [2+2] photodimerization of olefins is a successful demonstration of how templated solid state synthesis can efficiently synthesize unique materials with remarkable stereoselectivity and under environment-friendly conditions. A demonstration of this synthetic strategy is given in chapter 3. The combination of various types of intermolecular linkages, leading to formation of high order aggregation and crystalline materials or to a random aggregation resulting in an amorphous precipitate, may not go to completeness. In such rare cases an aggregation process intermediate between crystalline and amorphous materials is observed, resulting in the formation of a gel, i.e. a viscoelastic solid-like or liquid-like material. In chapter 4 design of new Low Molecular Weight Gelators is presented. Aspects such as the relationships between molecular structure, crystal packing and gelation properties and the application of this kind of gels as a medium for crystal growth of organic molecules, such as APIs, are also discussed.

Mixtures of colloidal rods and spheres in bulk and in confinement

When non-adsorbing polymers are added to an isotropic suspension of rod-like colloids, the colloids effectively attract each other via depletion forces. Monte Carlo simulations were performed to study the phase diagram of such rod-polymer mixtures. The colloidal rods were modelled as hard spherocylinders; the polymers were described as spheres of the same diameter as the rods. The polymers may overlap with no energy cost, while overlap of polymers and rods is forbidden. In this thesis the emphasis was on the depletion effects caused by the addition of spheres on the isotropic phase of rod-like particles. Although most of the present experimental studies consider systems close to or beyond the isotropic-nematic transition, the isotropic phase with depletion interactions turns out to be a not less interesting topic. First, the percolation problem was studied in canonical simulations of a system of hard rods and soft spheres, where the amount of depletant was kept low to prevent phase separation of the mixture. The lowering of the percolation threshold seen in experiment is confirmed to be due to the depletion interactions. The local changes in the structure of the fluid of rods, which were measured in the simulations, indicated that the depletion forces enhance local alignment and aggregation of the rods. Then, the phase diagram of isotropic-isotropic demixing of short spherocylinders was calculated using grand canonical ensemble simulations with successive umbrella sampling. Finite size scaling analysis allowed to estimate the location of the critical point. Also, estimates for the interfacial tension between the coexisting isotropic phases and analyses of its power-law behaviour on approach of the critical point are presented. The obtained phase diagram was compared to the predictions of the free volume theory. After an analysis of the bulk, the phase behaviour in confinement was studied. The critical point of gas-liquid demixing is shifted to higher concentrations of rods and smaller concentrations of spheres due to the formation of an orientationally ordered surface film. If the separation between the walls becomes very small, the critical point is shifted back to smaller concentrations of rods because the surface film breaks up. A method to calculate the contact angle of the liquid-gas interface with the wall is introduced and the wetting behaviour on the approach to the critical point is analysed.

Patterns in and kinetics of phase separation in binary mixtures

This work focused mainly on two aspects of kinetics of phase separation in binary mixtures. In the first part, we studied the interplay of hydrodynamics and the phase separation of binary mixtures. A considerably flat container (a laterally extended geometry), at an aspect ratio of 14:1 (diameter: height) was chosen, so that any hydrodynamic instabilities, if they arise, could be tracked. Two binary mixtures were studied. One was a mixture of methanol and hexane, doped with 5% ethanol, which phase separated under cooling. The second was a mixture of butoxyethanol and water, doped with 2% decane, which phase separated under heating. The dopants were added to bring down the phase transition temperature around room temperature.rnrnAlthough much work has been done already on classical hydrodynamic instabilities, not much has been done in the understanding of the coupling between phase separation and hydrodynamic instabilities. This work aimed at understanding the influence of phase separation in initiating any hydrodynamic instability, and also vice versa. Another aim was to understand the influence of the applied temperature protocol on the emergence of patterns characteristic to hydrodynamic instabilities. rnrnOn slowly cooling the system continuously, at specific cooling rates, patterns were observed in the first mixture, at the start of phase separation. They resembled the patterns observed in classical Rayleigh-Bénard instability, which arises when a liquid continuously is heated from below. To suppress this classical convection, the cooling setup was tuned such that the lower side of the sample always remained cooler by a few millikelvins, relative to the top. We found that the nature of patterns changed with different cooling rates, with stable patterns appearing for a specific cooling rate (1K/h). On the basis of the cooling protocol, we estimated a modified Rayleigh number for our system. We found that the estimated modified Rayleigh number is near the critical value for instability, for cooling rates between 0.5K/h and 1K/h. This is consistent with our experimental findings. rnrnThe origin of the patterns, in spite of the lower side being relatively colder with respect to the top, points to two possible reasons. 1) During phase separation droplets of either phases are formed, which releases a latent heat. Our microcalorimetry measurements show that the rise in temperature during the first phase separation is in the order of 10-20millikelvins, which in some cases is enough to reverse the applied temperature bias. Thus phase separation in itself initiates a hydrodynamic instability. 2) The second reason comes from the cooling protocol itself. The sample was cooled from above and below. At sufficiently high cooling rates, there are situations where the interior of the sample is relatively hotter than both top and bottom of the sample. This is sufficient to create an instability within the cell. Our experiments at higher cooling rates (5K/h and above) show complex patterns, which hints that there is enough convection even before phase separation occurs. Infact, theoretical work done by Dr.Hayase show that patterns could arise in a system without latent heat, with symmetrical cooling from top and bottom. The simulations also show that the patterns do not span the entire height of the sample cell. This is again consistent with the cell sizes measured in our experiment.rnrnThe second mixture also showed patterns at specific heating rates, when it was continuously heated inducing phase separation. In this case though, the sample was turbid for a long time until patterns appeared. A meniscus was most probably formed before the patterns emerged. We attribute the reason of patterns in this case to Marangoni convection, which is present in systems with an interface, where local differences in surface tension give rise to an instability. Our estimates for the Rayleigh number also show a significantly lower number than that's required for RB-type instability.rnrnIn the first part of the work, therefore, we identify two different kinds of hydrodynamic instabilities in two different mixtures. Both are observed during, or after the first phase separation. Our patterns compare with the classical convection patterns, but here the origins are from phase separation and the cooling protocol.rnrnIn the second part of the work, we focused on the kinetics of phase separation in a polymer solution (polystyrene and methylcyclohexane), which is cooled continuously far down into the two phase region. Oscillations in turbidity, denoting material exchange between the phases are seen. Three processes contribute to the phase separation: Nucleation of droplets, their growth and coalescence, and their subsequent sedimentation. Experiments in low molecular binary mixtures had led to models of oscillation [43] which considered sedimentation time scales much faster than the time scales of nucleation and growth. The size and shape of the sample therefore did not matter in such situations. The oscillations in turbidity were volume-dominated. The present work aimed at understanding the influence of sedimentation time scales for polymer mixtures. Three heights of the sample with same composition were studied side by side. We found that periods increased with the sample height, thus showing that sedimentation time determines the period of oscillations in the polymer solutions. We experimented with different cooling rates and different compositions of the mixture, and we found that periods are still determined by the sample height, and therefore by sedimentation time. rnrnWe also see that turbidity emerges in two ways; either from the interface, or throughout the sample. We suggest that oscillations starting from the interface are due to satellite droplets that are formed on droplet coalescence at the interface. These satellite droplets are then advected to the top of the sample, and they grow, coalesce and sediment. This type of an oscillation wouldn't require the system to pass the energy barrier required for homogenous nucleation throughout the sample. This mechanism would work best in sample where the droplets could be effectively advected throughout the sample. In our experiments, we see more interface dominated oscillations in the smaller cells and lower cooling rates, where droplet advection is favourable. In larger samples and higher cooling rates, we mostly see that the whole sample becomes turbid homogenously, which requires the system to pass the energy barrier for homogenous nucleation.rnrnOscillations, in principle, occur since the system needs to pass an energy barrier for nucleation. The height of the barrier decreases with increasing supersaturation, which in turn is from the temperature ramp applied. This gives rise to a period where the system is clear, in between the turbid periods. At certain specific cooling rates, the system can follow a path such that the start of a turbid period coincides with the vanishing of the last turbid period, thus eliminating the clear periods. This means suppressions of oscillations altogether. In fact we experimentally present a case where, at a certain cooling rate, oscillations indeed vanish. rnrnThus we find through this work that the kinetics of phase separation in polymer solution is different from that of a low molecular system; sedimentation time scales become relevant, and therefore so does the shape and size of the sample. The role of interface in initiating turbid periods also become much more prominent in this system compared to that in low molecular mixtures.rnrnIn summary, some fundamental properties in the kinetics of phase separation in binary mixtures were studied. While the first part of the work described the close interplay of the first phase separation with hydrodynamic instabilities, the second part investigated the nature and determining factors of oscillations, when the system was cooled deep into the two phase region. Both cases show how the geometry of the cell can affect the kinetics of phase separation. This study leads to further fundamental understandings of the factors contributing to the kinetics of phase separation, and to the understandings of what can be controlled and tuned in practical cases. rn

Development and validation of a liquid chromatography-tandem mass spectrometry method for the quantitation of synthetic cannabinoids of the aminoalkylindole type and methanandamide in serum and its application to forensic samples

After the discovery of synthetic cannabimimetic substances in 'Spice'-like herbal mixtures marketed as 'incense' or 'plant fertilizer' the active compounds have been declared as controlled substances in several European countries. As expected, a monitoring of new herbal mixtures which continue to appear on the market revealed that shortly after control measures have been taken by legal authorities, other compounds were added to existing mixtures and to new products. Several compounds of the aminoalkylindole type have been detected so far in herbal mixtures but still their consumption cannot be detected by commonly used drug-screening procedures, encouraging drug users to substitute cannabis with those products. There is a increasing demand on the part of police authorities, hospitals and psychiatrists for detection and quantification of synthetic cannabinoids in biological samples originating from psychiatric inpatients, emergency units or assessment of fitness to drive. Therefore, a liquid chromatography-tandem mass spectrometry method after liquid-liquid extraction for the quantitation of JWH-015, JWH-018, JWH-073, JWH-081, JWH 200, JWH-250, WIN 55,212-2 and methanandamide and the detection of JWH-019 and JWH-020 in human serum has been developed and fully validated according to guidelines for forensic toxicological analyses. The method was successfully applied to 101 serum samples from 80 subjects provided by hospitals, detoxification and therapy centers, forensic psychiatric centers and police authorities. Fifty-seven samples or 56.4% were found positive for at least one aminoalkylindole. JWH-019, JWH-020, JWH-200, WIN 55,212-2 and methanandamide were not detected in any of the analyzed samples.

Redox-Switching in a Viologen-type Adlayer: An Electrochemical Shell-Isolated Nanoparticle Enhanced Raman Spectroscopy Study on Au(111)-(1 x 1) Single Crystal Electrodes

We reported the first application of in situ shell-isolated nanoparticle enhanced Raman spectroscopy (SHINERS) to an interfacial redox reaction under electrochemical conditions. We construct gap-mode sandwich structures composed of a thiol-terminated HS-6V6H viologen adlayer immobilized on a single crystal Au(111)-(1x1) electrode and covered by Au(60 nm)@SlO(2) core shell nanoparticles acting as plasmonic antennas. We observed high-quality, potential-dependent Raman spectra of the three viologen species V(2+),V(+center dot) and V(0) on a well-defined Au(111) substrate surface and could map their potential-dependent evolution. Comparison with experiments on powder samples revealed an enhancement factor of the nonresonant Raman modes of similar to 3 x 10(5), and up to 9 x 10(7) for the resonance modes. The study illustrates the unique capability of SHINERS and its potential in the entire field of electrochemical surface science to explore structures and reaction pathways on well-defined substrate surfaces, such as single crystals, for molecular, (electro-)- catalytic, bioelectrochemical systems up to fundamental double layer studies at electrified solid/liquid interfaces.

Influence of Mechanical and Thermal Boundary Conditions on Stabilizing/Destabilizing Mechanisms in Evaporating Liquid Films

Liquid films, evaporating or non-evaporating, are ubiquitous in nature and technology. The dynamics of evaporating liquid films is a study applicable in several industries such as water recovery, heat exchangers, crystal growth, drug design etc. The theory describing the dynamics of liquid films crosses several fields such as engineering, mathematics, material science, biophysics and volcanology to name a few. Interfacial instabilities typically manifest by the undulation of an interface from a presumed flat state or by the onset of a secondary flow state from a primary quiescent state or both. To study the instabilities affecting liquid films, an evaporating/non-evaporating Newtonian liquid film is subject to a perturbation. Numerical analysis is conducted on configurations of such liquid films being heated on solid surfaces in order to examine the various stabilizing and destabilizing mechanisms that can cause the formation of different convective structures. These convective structures have implications towards heat transfer that occurs via this process. Certain aspects of this research topic have not received attention, as will be obvious from the literature review. Static, horizontal liquid films on solid surfaces are examined for their resistance to long wave type instabilities via linear stability analysis, method of normal modes and finite difference methods. The spatiotemporal evolution equation, available in literature, describing the time evolution of a liquid film heated on a solid surface, is utilized to analyze various stabilizing/destabilizing mechanisms affecting evaporating and non-evaporating liquid films. The impact of these mechanisms on the film stability and structure for both buoyant and non-buoyant films will be examined by the variation of mechanical and thermal boundary conditions. Films evaporating in zero gravity are studied using the evolution equation. It is found that films that are stable to long wave type instabilities in terrestrial gravity are prone to destabilization via long wave instabilities in zero gravity.

A dose-controlled system for air-liquid interface cell exposure and application to zinc oxide nanoparticles

BACKGROUND: Engineered nanoparticles are becoming increasingly ubiquitous and their toxicological effects on human health, as well as on the ecosystem, have become a concern. Since initial contact with nanoparticles occurs at the epithelium in the lungs (or skin, or eyes), in vitro cell studies with nanoparticles require dose-controlled systems for delivery of nanoparticles to epithelial cells cultured at the air-liquid interface. RESULTS: A novel air-liquid interface cell exposure system (ALICE) for nanoparticles in liquids is presented and validated. The ALICE generates a dense cloud of droplets with a vibrating membrane nebulizer and utilizes combined cloud settling and single particle sedimentation for fast (~10 min; entire exposure), repeatable (<12%), low-stress and efficient delivery of nanoparticles, or dissolved substances, to cells cultured at the air-liquid interface. Validation with various types of nanoparticles (Au, ZnO and carbon black nanoparticles) and solutes (such as NaCl) showed that the ALICE provided spatially uniform deposition (<1.6% variability) and had no adverse effect on the viability of a widely used alveolar human epithelial-like cell line (A549). The cell deposited dose can be controlled with a quartz crystal microbalance (QCM) over a dynamic range of at least 0.02-200 mug/cm(2). The cell-specific deposition efficiency is currently limited to 0.072 (7.2% for two commercially available 6-er transwell plates), but a deposition efficiency of up to 0.57 (57%) is possible for better cell coverage of the exposure chamber. Dose-response measurements with ZnO nanoparticles (0.3-8.5 mug/cm(2)) showed significant differences in mRNA expression of pro-inflammatory (IL-8) and oxidative stress (HO-1) markers when comparing submerged and air-liquid interface exposures. Both exposure methods showed no cellular response below 1 mug/cm(2 )ZnO, which indicates that ZnO nanoparticles are not toxic at occupationally allowed exposure levels. CONCLUSION: The ALICE is a useful tool for dose-controlled nanoparticle (or solute) exposure of cells at the air-liquid interface. Significant differences between cellular response after ZnO nanoparticle exposure under submerged and air-liquid interface conditions suggest that pharmaceutical and toxicological studies with inhaled (nano-)particles should be performed under the more realistic air-liquid interface, rather than submerged cell conditions.

In Situ SHINERS at Electrochemical Single-Crystal Electrode/Electrolyte Interfaces: Tuning Preparation Strategies and Selected Applications

We have studied Au(55 nm)@SiO2 nanoparticles (NPs) on two low-index phases of gold and platinum single crystal electrodes in ClO4– and SO42– ion-containing electrolytes by both electrochemical methods and in-situ shell-isolated nanoparticle enhanced Raman spectroscopy (SHINERS). We showed the blocking of the electrode with surfactants originating from the synthesis of as-prepared SHINERS NPs. We introduce an efficient procedure to overcome this problem, which provides a fundamental platform for the application of SHINERS in surface electrochemistry and beyond. Our method is based on a hydrogen evolution treatment of the SHINERS-NP-modified single-crystal surfaces. The reliability of our preparation strategy is demonstrated in electrochemical SHINERS experiments on the potential-controlled adsorption and phase formation of pyridine on Au(hkl) and Pt(hkl). We obtained high-quality Raman spectra on these well-defined and structurally carefully characterized single-crystal surfaces. The analysis of the characteristic A1 vibrational modes revealed perfect agreement with the interpretation of single-crystal voltammetric and chronoamperometric experiments. Our study demonstrates that the SHINERS protocol developed in this work qualifies this Raman method as a pioneering approach with unique opportunities for in situ structure and reactivity studies at well-defined electrochemical solid/liquid interfaces.