Surfactant-based supramolecular dye assembly: A highly selective and economically viable platform for quantification of heparin antidote.

Herein, we have employed a supramolecular assembly of a cationic dye, LDS-698 and a common surfactant sodium dodecyl sulfate (SDS) as a turn-on fluorescent sensor for protamine (Pr) detection. Addition of cationic Pr to the solution of dye-surfactant complex brings negatively charged SDS molecules together through strong electrostatic interaction, assisting aggregation of SDS way before its critical micellar concentration (CMC). These aggregates encapsulate the dye molecules within their hydrophobic region, arresting non-radiative decay channels of the excited dye. Thus, the LDS-698•SDS assembly displays substantial enhancement in fluorescence intensity that follows a nice linear trend with Pr concentration, providing limit of detection (LOD) for Pr as low as 3.84(±0.11) nM in buffer, 124.4(±6.7) nM in 1% human serum and 28.3(±0.5) nM in 100% human urine. Furthermore, high selectivity, low background signal, large stokes shift, and emission in the biologically favorable deep-red region make the studied assembly a promising platform for Pr sensing. As of our knowledge it is the first ever Pr sensory platform, using a very common surfactant (SDS), which is economically affordable and very easily available in the market. This innovative approach can replace the expensive, exotic and specialized chemicals considered for the purpose and thus showcase its potential in practical applications.

Cationic Conjugated Polymer Fluorescence Resonance Energy Transfer for DNA Methylation Assessment to Discriminate the Geographical Origins of Lonicerae japonicae flos.

The flavor and taste of Lonicerae japonicae flos (LJF) products are heavily influenced by geographical origin. Tracing the geographical origin is an important aspect of LJF quality assessment. Here, DNA methylation analysis coupled with chemometrics revealed that, in 10 CpG islands upstream of genes in the chlorogenic acid and iridoid biosynthetic pathways, DNA methylation differences appear close association with LJF geographical origin. DNA methylation status in these CpG islands was determined using the cationic conjugated polymer fluorescence resonance energy transfer method. As a result, LJFs from 39 geographical origins were classified into four groups corresponding to Northern China, Central Plain of China, Southeast China, and Western China, according to cluster analysis and principal component analysis. Our findings contribute to an understanding of the modulation of LJF taste and can assist in understanding how DNA methylation in LJF varies with geographical origin.

Accurate Description of Cation-π Interactions in Proteins with a Nonpolarizable Force Field at No Additional Cost.

Cation-π interactions play a significant role in a host of processes eminently relevant to biology. However, polarization effects arising from the interaction of cations with aromatic moieties have long been recognized to be inadequately described by pairwise additive force fields. In the present work, we address this longstanding shortcoming through the nonbonded FIX (NBFIX) feature of the CHARMM36 force field, modifying pair-specific Lennard-Jones (LJ) parameters, while circumventing the limitations of the Lorentz-Berthelot combination rules. The potentials of mean force (PMFs) characterizing prototypical cation-π interactions in aqueous solutions are first determined using a hybrid quantum mechanical/molecular mechanics (QM/MM) strategy in conjunction with an importance-sampling algorithm. The LJ parameters describing the cation-π pairs are then optimized to match the QM/MM PMFs. The standard binding free energies of nine cation-π complexes, i.e., toluene, para-cresol, and 3-methyl-indole interacting with either ammonium, guanidinium, or tetramethylammonium, determined with this new set of parameters agree well with the experimental measurements. Additional simulations were carried out on three different classes of biological objects featuring cation-π interactions, including five individual proteins, three protein-ligand complexes, and two protein-protein complexes. Our results indicate that the description of cation-π interactions is overall improved using NBFIX corrections, compared with the standard pairwise additive force field. Moreover, an accurate binding free energy calculation for a protein-ligand complex containing cation-π interactions (2BOK) shows that using the new parameters, the experimental binding affinity can be reproduced quantitatively. Put together, the present work suggests that the NBFIX parameters optimized here can be broadly utilized in the simulation of proteins in an aqueous solution to enhance the representation of cation-π interactions, at no additional computational cost.

In vitro toxicity assessment of hydrogel patches obtained by cation-induced cross-linking of rod-like cellulose nanocrystals.

With the purpose of designing active patches for photodynamic therapy of melanoma, transparent and soft hydrogel membranes (HMs) have been fabricated by cation-induced gelation of rod-like cellulose nanocrystals (CNCs) bearing negatively charged carboxylic groups. Na+ , Ca2+ , Mg2+ have been used as cross-linkers of cellulose nanocrystal (CNC). The biosafety of this material and of its precursors has been evaluated in vitro in cell cultures. Morphological changes, cell organelles integrity, and cell survival with the tetrazolium salt reduction (MTT) assay were utilized as tests of cytotoxicity. Preliminary investigation was performed by addition of the hydrogel components to the cell culture medium and by incubations of the CNC-HM in direct and indirect contact with a confluent monolayer of A375 melanoma cells. Direct contact assays suffered from interference of physical stress. Careful evaluation of cytotoxicity was obtained considering the overall picture provided by microscopy and biochemical tests performed with the CNC-HM in indirect contact with two melanoma cell lines (A375, M14) and human fibroblasts. CNCs have been demonstrated to be a safe precursor material and CNC-HMs have a good biocompatibility provided that the excess of cations, in particular of Ca2+ is removed. These results indicate that CNC and can be safely used to fabricate biomedical devices such as transparent hydrogel patches, although attention must be paid to the fabrication procedure.

Synthesis and assessment of metallic ion migration through a novel calcium carbonate coating for biomedical implants.

Titanium (Ti) implants are commonly regarded as well accepted by the body. However, metal ion release is still a cause for concern. A small decrease in pH, which can be caused by inflammation, may produce a large increase in the corrosion rate of Ti implants. Coating the alloy with a buffer layer could have a significant protective effect. In this study, a calcium carbonate coating was developed on commercially pure Ti and a Ti-6Al-4V alloy through a hydrothermal treatment of previously NaOH-treated surfaces in calcium-citric acid chelate complexes. The results showed that a superstructured calcite coating layer formed on the Ti substrate after treatment at 170°C for 3 hr. The coating was approx. 1 µm thick and covered the substrate surface uniformly. When prolonging the hydrothermal treatment from 5 hr to 24 hr, the rhombohedral structure of calcite was observed in addition to the superstructure of calcite. Dissolution test results showed no significant differences in solution pH between the coated- and un-coated samples. However, the CaCO3 coating reduced by approx. 2-5 times the Ti and V ion release from the substrate as compared to the uncoated material, at pH 4. CaCO3 and hydroxyapatite (HA) coatings gave nonsignificant effects at neutral pH although the HA coating showed a trend for better results at the longer time points. The reduction in metal ion release from the substrate and the buffering ability of the CaCO3 coating encourage further studies on this coating for clinical applications.

Replica Ornstein-Zernike Theory Applied for Studying the Equilibrium Distribution of Electrolytes across Model Membranes.

By means of replica Ornstein-Zernike theory (supplemented in a few cases by Monte Carlo simulations) we examined the distribution of an annealed primitive model +1:-1 electrolyte in a mixture with uncharged hard spheres, or another model +1:-1 or +2:-1 electrolyte inside and outside the quenched vesicles, decorated by a model membrane, and across the membrane phase. We explored the influence of the size and charge of the annealed fluid on the partition equilibrium, as well as the effect of the vesicle size and membrane interaction parameters (repulsive barrier height, attractive depth, and membrane width). A hydrophobic cation, present in the mixture with NaCl, slightly enhanced the concentration of sodium ions inside the model vesicle, compared to pure NaCl solution. The replica theory was in good agreement with computer simulations and as such adequate for studying partitioning of small and hydrophobic ions or hydrophobic solutes across model membranes.

The comparative study of micellar TLC and RP-TLC as potential tools for lipophilicity assessment based on QSRR approach.

Lipophilicity of compound is well known as vital physicochemical property of a molecule, which determines its biological activity. Nonetheless, the assessment of a lipophilicity is still problematic and focuses attention of scientists. Although, the shake-flask method is still considered as a gold standard for experimental determination of lipophilicity, currently the chromatographic approach is mostly used. Among chromatographic methods used for lipophilicity assessment, thin layer chromatography (TLC) is still one of the most popular tools. The main goal of this study was to compare classical reversed-phase thin layer chromatography (RP-TLC) and micellar TLC as potential tools for lipophilicity assessment. Micellar liquid chromatography has significantly lover environment impact than classical reversed-phase liquid chromatography. Additionally comparison of cationic and anionic surfactants (CTAB and SDS), which have different chemical properties, as modifiers of mobile phase in micellar TLC were investigated. The Quantitative Structure-Retention Relationships (QSRR) approach was used in order to present molecular mechanisms of retention in investigated chromatographic systems. The study was based on chemically diverse model set compounds, with a proved therapeutic or toxic potential. According to obtained results the micellar TLC with CTAB as surfactant can be recommended to logP prediction. The obtained QSRR models indicated that adsorption of CTAB monomers on CN modified surface of silica gel and the silanol-quaternary ammonium interactions are possible. Consequently, the reduction of interaction between molecules and free silanol, contributes to the improvement of logP predictions. These result were confirmed by regression and classification methods.

Facile and green fabrication of cation exchange membrane adsorber with unprecedented adsorption capacity for protein purification.

Fabricating membrane adsorbers with high adsorption capacity and appreciable throughput for the separation and purification of protein products is challenging in biomedical and pharmaceutical industries. Herein, we report the synthesis of a novel membrane adsorber by functionalizing a nylon microfiltration membrane with alginate dialdehyde (ADA) followed by sulphonic addition, without any solvent usage, and its successful application in the purification of lysozyme. Taking advantage of abundant dual cation exchange (CEX) groups on sulphonic-ADA (S-ADA) ligands, this novel S-ADA-nylon membrane adsorber showed an unprecedented static binding capicity of 286mg/mL for lysozyme adsorption. Meanwhile, the prepared membrane adsorber could be easily regenerated (complete protein elution) under mild conditions and be reused at least for five times. Featured with a unique selectivity, the S-ADA-nylon membrane also captured lysozyme from chicken egg white solution with a high purity (100%) and a high recovery of 98%. The purified lysozyme showed similar specific activity as commercial product. The present work provides a facile, green and low-cost approach for the preparation of high-performance membrane adsorbers, which has a great potential in protein production.

Sorption of uranium(VI) onto hydrous ferric oxide-modified zeolite: Assessment of the effect of pH, contact time, temperature, selected cations and anions on sorbent interactions.

Zeolites are commonly used as adsorbents for metal removal in most applications e.g. in wastewater. However, the ubiquity of iron in such systems may, in the long-term, distort the true interactions and mechanisms of contaminant removal as a result of modification of the zeolite surface. In this study, this potential phenomenon was assessed for the removal of uranium(VI) from aqueous solution by hydrous ferric oxide-modified zeolite (HFOMZ). This was prepared by precipitating iron hydroxide (the common precipitate of iron in aqueous systems) onto zeolite. The prepared HFOMZ was characterised by the scanning electron microscope (SEM), and Fourier transform infrared spectroscopy (FTIR). Batch adsorption experiments were performed to assess the effect of: pH, initial uranium(VI) concentration, adsorbent dosage, contact time, temperature, presence of cations (Pb2+, Cr3+, Cu2+, Mn2+ and Co2+) and anions ( [Formula: see text] , [Formula: see text] and [Formula: see text] ) on the adsorption of uranium(VI). Kinetic studies under these conditions indicated that the pseudo second-order kinetic model (R2 > 0.99) best described the adsorption behaviour, implying that this could be proceeding through a chemisorption process. The experimental data was best described by the Freundlich isotherm model (R2 > 0.93), an implication that the adsorption surface was heterogeneous. The thermodynamic parameters calculated from the experimental data suggested that the adsorption of U(VI) onto HFOMZ was spontaneous and exothermic in nature. The adsorption of U(VI) onto HFOMZ was dominated by complexation with strong ionizable hydroxyl sites on the hydrous ferric oxide surfaces and the edge sites of the zeolite. At pH values from 2 to 6, increased adsorption was observed and this decreased at higher pH values (above 6). This corresponded with the changes in speciation as determined by the PHREEQC modelling code. The presence of the cations (Pb2+, Cr3+, Cu2+, Mn2+ and Co2+) and anions ( [Formula: see text] and [Formula: see text] ) resulted in a significant decrease in the adsorption capacity of U(VI) by HFOMZ, implying that in a system where these anions and cations are present in high concentrations over time, U(VI) will adsorb less onto the material.

First-Principles Interaction Analysis Assessment of the Manganese Cation in the Catalytic Activity of Glycosyltransferases.

The energetic effect of water substitution reactions in hexacoordinated [Mn(H2O)6-nLzn]2+nz complexes with L = methanol, formic acid, formamide, formate, imidazole, and diphosphate is quantitatively analyzed at the MP2/triple-ζ level of theory. Subsequently, the state-of-the-art open shell symmetry-adapted perturbation theory (SAPT) analysis of the interaction energies of Mn2+···ligand dimers with selected O-, S-, and N-binding ligands is presented and compared to similar interactions of Mg2+ and Zn2+ ions. We find that the induction energies in the dimers with manganese are almost twice as large as in dimers with magnesium. The total interaction energies rise in the order Mn2+ < Mg2+ < Zn2+. The calculations of the Mn2+ → Mg2+ replacement reaction suggest that metal-dependent glycosyltransferases influence the binding preference of Mn2+ over Mg2+ by inserting amino acids that coordinate the metal via nitrogen or sulfur into their active site.

Assessment of ionic liquids' toxicity through the inhibition of acylase I activity on a microflow system.

Acylase I (ACY I) plays a role in the detoxication and bioactivation of xenobiotics as well in other physiological functions. In this context, an automated ACY I assay for the evaluation of ionic liquids' (ILs) toxicity was developed. The assay was implemented in a sequential injection analysis (SIA) system and was applied to eight commercially available ILs. The SIA methodology was based on the deacetylation of N-acetyl-l-methionine with production of l-methionine, which was determined using fluorescamine. ACY I inhibition in the presence of ILs was monitored by the decrease of fluorescence intensity. The obtained results confirmed the influence of ILs' structural elements on its toxicity and revealed that pyridinium and phosphonium cations, longer alkyl side chains and tetrafluoroborate anion displayed higher toxic effect on enzyme activity. The developed methodology proved to be robust and exhibited good repeatability (RSD < 1.3%, n = 10), leading also to a reduction of reagents consumption and effluents production. Thus, it is expected that the proposed assay can be used as a novel tool for ILs' toxicity screening.

Microhydration of Protonated Nα-Acetylhistidine: A Theoretical Approach.

Extensive exploration of the potential energy surfaces of protonated Nα-acetylhistidine hydrated by 0-3 molecules of water was performed. The methodology combined hierarchical and genealogical (Darwin family tree) approaches using polarizable AMOEBA force field and M06 functional. It is demonstrated that this mixed approach allows recovering a larger number of conformers than the number recovered by using any one of the two methods alone. Hydration enthalpies of protonated Nα-acetylhistidine and of model compounds have been computed using higher theoretical methods, up to the G4MP2 procedure. Excellent agreement with experiment is observed for successive hydration of methylamonium and imidazolium cations using MP2/6-311++G(2d,2p)//M06/6-311++G(d,p) and G4MP2 methods, thereby validating the theory levels used for hydrated protonated Nα-acetylhistidine. It is found that the first hydration enthalpy of protonated Nα-acetylhistidine is ca. 10 kJ mol(-1) lower than that of imidazolium, a result explained by the local environment of the positively charged imidazolium moiety.

Spin-inversion and spin-selection in the reactions FeO(+) + H2 and Fe(+) + N2O.

The reactions of FeO(+) with H2 and of Fe(+) with N2O were studied with respect to the production and reactivity of electronically excited (4)Fe(+) cations. The reaction of electronic ground state (6)FeO(+) with H2 was found to predominantly produce electronically excited (4)Fe(+) as opposed to electronic ground state (6)Fe(+) corresponding to a spin-allowed reaction. (4)Fe(+) was observed to react with N2O with a rate constant of 2.3 (+0.3/-0.8) × 10(-11) cm(3) molecule(-1) s(-1), smaller than the ground state (6)Fe(+) rate constant of 3.2 (±0.5) × 10(-11) cm(3) molecule(-1) s(-1) (at room temperature). While the overall reaction of (6)FeO(+) with H2 within the Two-State-Reactivity concept is governed by efficient sextet-quartet spin-inversion in the initial reaction complex, the observation of predominant (4)Fe(+) production in the reaction is attributed to a much less efficient quartet-sextet back-inversion in the final reaction complex. Average spin-inversion probabilities are estimated by statistical modeling of spin-inversion processes and related to the properties of spin-orbit coupling along the reaction coordinate. The reaction of FeO(+) with H2 served as a source for (4)Fe(+), subsequently reacting with N2O. The measured rate constant has allowed for a more detailed understanding of the ground state (6)Fe(+) reaction with N2O, leading to a significantly improved statistical modeling of the previously measured temperature dependence of the reaction. In particular, evidence for the participation of electronically excited states of the reaction complex was found. Deexcitation of (4)Fe(+) by He was found to be slow, with a rate constant <3 × 10(-14) cm(3) molecule(-1) s(-1).

Increasing RO efficiency by chemical-free ion-exchange and Donnan dialysis: Principles and practical implications.

Ion-exchange (IEX) and Donnan dialysis (DD) are techniques which can selectively remove cations, limiting scaling in reverse osmosis (RO). If the RO concentrate could be recycled for regeneration of these pre-treatment techniques, RO recovery could be largely increased without the need for chemical addition or additional technologies. In this study, two different RO feed streams (treated industrial waste water and simple tap water) were tested in the envisioned IEX-RO and DD-RO hybrids including RO concentrate recycling. The efficiency of multivalent cation removal depends mainly on the ratio of monovalent to multivalent cations in the feed stream, influencing the ion-exchange efficiency in both IEX and DD. Since the mono-to-multivalent ratio was very high in the waste water, the RO recovery could potentially be increased to 92%. For the tap water, these high RO recoveries could only be reached by adding additional NaCl, because of the low initial monovalent to multivalent ratio in the feed. In both cases, the IEX-RO hybrid proved to be most cost-efficient, due to the high current cost of the membranes used in DD. The membrane cost would have to decrease from ±300 €/m² to 10-30 €/m² - comparable to current reverse osmosis membranes - to achieve a comparable cost. In conclusion, the recycling of RO concentrate to regenerate ion exchange pre-treatment techniques for RO is an interesting option to increase RO recovery without addition of chemicals, but only at high monovalent/multivalent cation-ratios in the feed stream.

Assessment of bromide-based ionic liquid toxicity toward aquatic organisms and QSAR analysis.

The toxicities of 24 bromide-based ionic liquids (Br-ILs) towards Vibrio fischeri (V. fischeri) and Daphnia magna (D. magna) were determined. These Br-ILs are composed of a bromide ion and a generic cation (i.e., pyrrolidinium, piperidinium, pyridinium or imidazolium) with different alkyl side chains. QSAR models with relatively high correlation coefficients, R(2), of 0.954 and 0.895 were developed for V. fischeri and D. magna. The model for V. fischeri indicated that the Br-IL toxicity towards V. fischeri was negatively correlated with the energy of the lowest unoccupied molecular orbitals (ELUMO) which reflects the electron affinities (EAs) and positively correlated with the volumes of Br-IL cations. For the D. magna model, the Br-IL toxicity was positively correlated with the dipole moment (µ) and negatively correlated with the total energy (TE) that is highly correlated with the molecular volume (V). For Br-ILs with the same cation ring, the toxicity increased as the length of the alkyl chains increased. For the same alkyl chain length, the toxicity order for V. fischeri was pyridinium>imidazolium>piperidinium>pyrrolidinium, except for those containing octyl side chains, while the toxicity ranking for D. magna was imidazolium~pyridinium>piperidinium>pyrrolidinium.

Neutron scattering studies of the hydration structure of Li+.

New results derived from the experimental method of neutron diffraction and isotopic substitution (NDIS) are presented for the hydration structure of the lithium cation (Li(+)) in aqueous solutions of lithium chloride in heavy water (D2O) at concentrations of 6, 3, and 1 m and at 1.5 m lithium sulfate. By introducing new and more-accurate data reduction procedures than in our earlier studies (I. Howell and G. W. Neilson, J. Phys: Condens. Matter, 1996, 8, 4455-4463), we find, in the first hydration shell of Li(+), ∼4.3(2) water molecules at 6 m, 4.9(3) at 3 m, 4.8(3) at 1 m in the LiCl solutions, and 5.0(3) water molecules in the case of Li2SO4 solution. The general form of the first hydration shell is similar in all four solutions, with the correlations for Li-O and Li-D sited at 1.96 (0.02) Å and 2.58 (0.02) Å, respectively. The results resemble those presented in 1996, in terms of ion-water distances and local coordination, but the hydration number is significantly lower for the case at 1 m than the 6.5 (1.0) given at that time. Thus, experimental and theoretical results now agree that lithium is hydrated by a small number of water molecules (4-5) in the nearest coordination shell.

Charge transfer dissociation (CTD) mass spectrometry of peptide cations using kiloelectronvolt helium cations.

A kiloelectronvolt beam of helium ions is used to ionize and fragment precursor peptide ions starting in the 1+ charge state. The electron affinity of helium cations (24.6 eV) exceeds the ionization potential of protonated peptides and can therefore be used to abstract an electron from--or charge exchange with--the isolated precursor ions. Kiloelectronvolt energies are used, (1) to overcome the Coulombic repulsion barrier between the cationic reactants, (2) to overcome ion-defocussing effects in the ion trap, and (3) to provide additional activation energy. Charge transfer dissociation (CTD) of the [M+H](+) precursor of Substance P gives product ions such as [M+H](2+•) and a dominant series of a ions in both the 1+ and 2+ charge states. These observations, along with the less-abundant a + 1 ions, are consistent with ultraviolet photodissociation (UVPD) results of others and indicate that C-C(α) cleavages are possible through charge exchange with helium ions. Although the efficiencies and timescale of CTD are not yet suitable for on-line chromatography, this new approach to ion activation provides an additional potential tool for the interrogation of gas phase ions.

Rapid and simultaneous determination of sulfonate ester genotoxic impurities in drug substance by liquid chromatography coupled to tandem mass spectrometry: comparison of different ionization modes.

Two ionization techniques for liquid chromatography-mass spectrometry (LC-MS) determination of sulfonate ester potentially genotoxic impurities (PGIs) were evaluated. Twelve PGIs including methyl, ethyl, propyl and isopropyl esters of methanesulfonate, benzenesulfonate and p-toluenesulfonate were studied in this research. Electrospray ionization (ESI) and atmospheric pressure chemical ionization (APCI) sources were compared in terms of performance and quality parameters for detection of the twelve PGIs. Their mass spectra obtained by APCI and ESI were very different in both fragment ions and relative abundances. In APCI negative ion mode the twelve sulfonate esters showed their stable precursor ions of [M-alkyl](-), which readily yielded product ions of [M-alkyl-CH3](-) (for aliphatic sulfonate esters) or [M-alkyl-SO2](-) (for aromatic sulfonate esters) with collision-induced dissociation (CID) applied; and working in selected reaction monitoring (SRM) mode has allowed limits of detection to be decreased. In the case of ESI ionization, these compounds showed their precursor ions [M+H](+), but their abundance was easily competed by formation of ions [M+NH4](+) and/or [M+Na](+), which led to poor analytical sensitivity and reproducibility. Although mobile phase additives could enhance the responses of adduct ions like [M+NH4](+) and [M+Na](+), no improvement was obtained when using SRM mode. Twelve sulfonate esters were systematically compared and APCI was shown to be a better ionization technique for rapid and sensitive determination of these PGIs. Performance of the developed approach for rapid determination of 12 PGIs was also evaluated. Quality parameters were established and good precision (relative standard deviations <8%) and very low limits (2-4ng/mL) of detection were obtained, mainly when using APCI in SRM mode.

Hemocompatibility assessment of two siRNA nanocarrier formulations.

PURPOSE: Since the discovery of RNAi and its therapeutic potential, carrier systems have been developed to deliver small RNAs (particularly siRNA) for modulation of gene expression at the post-transcriptional level. An important factor determining the fate and usability of these systems in vivo is interaction with blood components, blood cells, and the immune system. In this study, a lipid-based and a polymer-based carrier system containing siRNA have been investigated in vitro in terms of their hemocompatibility. METHODS: The nanocomplexes studied were Angiplex, a targeted lipid-based system, and pHPMA-MPPM polyplex, a formulation based on a cationic polymer. siVEGFR-2 was encapsulated in both carriers and activation of platelets, coagulation, and complement cascade as well as induction of platelet aggregation were evaluated in vitro. RESULTS: Both systems had been shown before to cause significant silencing in vitro. Our findings indicated that pHPMA-MPPM polyplex triggered high platelet activation and aggregation although it did not stimulate coagulation substantially. Angiplex, on the other hand, provoked insignificant activation and aggregation of platelets and activated coagulation minimally. Complement system activation by Angiplex was in general low but stronger than pHPMA-MPPM polyplex. CONCLUSIONS: Taken together, these in vitro assays may help the selection of suitable carriers for systemic delivery of siRNA in early preclinical investigations and reduce the use of laboratory animals significantly.

Novel cation selective exhaustive injection-sweeping procedure for 5-nitroimidazole determination in waters by micellar electrokinetic chromatography using dispersive liquid-liquid microextraction.

A novel method consisting of cation-selective exhaustive injection and sweeping (CSEI-sweeping) as on-line preconcentration followed by a micellar electrokinetic chromatography (MEKC) separation has been developed for the determination of 5-nitroimidazoles (5-NDZ) in environmental waters. Moreover, dispersive liquid-liquid microextraction (DLLME) has been proposed for first time as sample treatment technique prior to CSEI-sweeping-MEKC. DLLME was applied to 5mL of sample. Dibromomethane (1156µL) and 2-butanol (1363µL) were employed as extractant and dispersive solvents, respectively. Salting-out effect was achieved by the addition of 16% (w/v) NaCl to the samples. After DLLME and organic solvent evaporation, the residue was redissolved in a low conductivity solvent (5mM phosphoric acid with 5% of methanol) and electrokinetically injected at 9.8kV for 632s in a bare fused-silica capillary (57.2cm, 50µm I.D.). Prior to the injection, the capillary was rinsed with 50mM phosphate buffer pH 2.5, followed by a plug of a higher conductivity buffer (100mM phosphate pH 2.5, 50mbar, 264s) and a plug of water (50mbar, 2s). Separation was carried out applying -30kV at 20°C in 44mM phosphate buffer pH 2.5, containing 8% tetrahydrofuran and 123mM sodium dodecyl sulfate. Analytical signals were monitored at 276nm. Validation was performed in river and well waters, obtaining satisfactory results in terms of linearity, precision (% RSD generally lower than 10%) and trueness (recoveries higher than 70% in almost all cases). LODs ranged from 0.61 to 2.44ng/mL. The combination of this microextraction technique with the proposed capillary electrophoresis methodology supposes a simple, sensitive and cheap alternative for 5-NDZ analyses, in accordance with the aims of green chemistry.