Ultra-Fast Gradient LC Method for Omeprazole Analysis Using a Monolithic Column: Assay Development, Validation, and Application to the Quality Control of Omeprazole Enteric-Coated Pellets

A method was optimized for the analysis of omeprazole (OMZ) by ultra-high speed LC with diode array detection using a monolithic Chromolith Fast Gradient RP 18 endcapped column (50 x 2.0 mm id). The analyses were performed at 30 degrees C using a mobile phase consisting of 0.15% (v/v) trifluoroacetic acid (TFA) in water (solvent A) and 0.15% (v/v) TFA in acetonitrile (solvent B) under a linear gradient of 5 to 90% B in 1 min at a flow rate of 1.0 mL/min and detection at 220 nm. Under these conditions, OMZ retention time was approximately 0.74 min. Validation parameters, such as selectivity, linearity, precision, accuracy, and robustness, showed results within the acceptable criteria. The method developed was successfully applied to OMZ enteric-coated pellets, showing that this assay can be used in the pharmaceutical industry for routine QC analysis. Moreover, the analytical conditions established allow for the simultaneous analysis of OMZ metabolites, 5-hydroxyomeprazole and omeprazole sulfone, in the same run, showing that this method can be extended to other matrixes with adequate procedures for sample preparation.

Spectroscopic identification of a dinuclear metal centre in manganese(II)-activated aminopeptidase P from Escherichia coli: implications for human prolidase

Electron paramagnetic resonance (EPR) spectra and X-ray absorption (EXAFS and XANES) data have been recorded for the manganese enzyme aminopeptidase P (AMPP, PepP protein) from Escherichia coli. The biological function of the protein, a tetramer of 50-kDa subunits, is the hydrolysis of N-terminal Xaa-Pro peptide bonds. Activity assays confirm that the enzyme is activated by treatment with Mn2+. The EPR spectrum of Mn2+-activated AMPP at liquid-He temperature is characteristic of an exchange-coupled dinuclear Mn(II) site, the Mn-Mn separation calculated from the zero-field splitting D of the quintet state being 3.5 (+/- 0.1) Angstrom. In the X-ray absorption spectrum of Mn2+-activated AMPP at the Mn K edge, the near-edge features are consistent with octahedrally coordinated Mn atoms in oxidation state +2. EXAFS data, limited to k less than or equal to 12 Angstrom(-1) by traces of Fe in the protein, are consistent with a single coordination shell occupied predominantly by O donor atoms at an average Mn-ligand distance of 2.15 Angstrom, but the possibility of a mixture of O and N donor atoms is not excluded. The Mn-Mn interaction at 3.5 Angstrom, is not detected in the EXAFS, probably due to destructive interference from light outer-shell atoms. The biological function, amino acid sequence and metal-ion dependence of E. coli AMPP are closely related to those of human prolidase, an enzyme that specifically cleaves Xaa-Pro dipeptides. Mutations that lead to human prolidase deficiency and clinical symptoms have been identified. Several known inhibitors of prolidase also inhibit AMPP. When these inhibitors are added to Mn2+-activated AMPP, the EPR spectrum and EXAFS remain unchanged. It can be inferred that the inhibitors either do not bind directly to the Mn centres, or substitute for existing Mn ligands without a significant change in donor atoms or coordination geometry. The conclusions from the spectroscopic measurements on AMPP have been verified by, and complement, a recent crystal structure analysis.

Purification and Partial Characterization of an Exo-polygalacturonase from Paecilomyces variotii Liquid Cultures

An extracellular polygalacturonase (PG) produced from Paecilomyces variotii was purified to homogeneity through two chromatography steps using DEAE-Fractogel and Sephadex G-100. The molecular weight of P. variotii PG was 77,300 Da by gel filtration and SDS-PAGE. PG had isoelectric point of 4.37 and optimum pH 4.0. PG was very stable from pH 3.0 to 6.0. The extent of hydrolysis of different pectins by the purified enzyme was decreased with an increase in the degree of esterification. PG had no activity toward non-pectic polysaccharides. The apparent K (m) and V (max) values for hydrolyzing sodium polypectate were 1.84 mg/mL and 432 A mu mol/min/mg, respectively. PG was found to have temperature optimum at 65 A degrees C and was totally stable at 45 A degrees C for 90 min. Half-life at 55 A degrees C was 50.6 min. Almost all the examined metal cations showed partial inhibitory effects under enzymatic activity, except for Na(+1), K(+1), and Co(+2) (1 mM) and Cu(+2) (1 and 10 mM).

Ferromagnetism, paramagnetism, and a Curie-Weiss metal in an electron-doped Hubbard model on a triangular lattice

Motivated by the unconventional properties and rich phase diagram of NaxCoO2 we consider the electronic and magnetic properties of a two-dimensional Hubbard model on an isotropic triangular lattice doped with electrons away from half-filling. Dynamical mean-field theory (DMFT) calculations predict that for negative intersite hopping amplitudes (t < 0) and an on-site Coulomb repulsion, U, comparable to the bandwidth, the system displays properties typical of a weakly correlated metal. In contrast, for t > 0 a large enhancement of the effective mass, itinerant ferromagnetism, and a metallic phase with a Curie-Weiss magnetic susceptibility are found in a broad electron doping range. The different behavior encountered is a consequence of the larger noninteracting density of states (DOS) at the Fermi level for t > 0 than for t < 0, which effectively enhances the mass and the scattering amplitude of the quasiparticles. The shape of the DOS is crucial for the occurrence of ferromagnetism as for t > 0 the energy cost of polarizing the system is much smaller than for t < 0. Our observation of Nagaoka ferromagnetism is consistent with the A-type antiferromagnetism (i.e., ferromagnetic layers stacked antiferromagnetically) observed in neutron scattering experiments on NaxCoO2. The transport and magnetic properties measured in NaxCoO2 are consistent with DMFT predictions of a metal close to the Mott insulator and we discuss the role of Na ordering in driving the system towards the Mott transition. We propose that the Curie-Weiss metal phase observed in NaxCoO2 is a consequence of the crossover from a bad metal with incoherent quasiparticles at temperatures T > T-* and Fermi liquid behavior with enhanced parameters below T-*, where T-* is a low energy coherence scale induced by strong local Coulomb electron correlations. Our analysis also shows that the one band Hubbard model on a triangular lattice is not enough to describe the unusual properties of NaxCoO2 and is used to identify the simplest relevant model that captures the essential physics in NaxCoO2. We propose a model which allows for the Na ordering phenomena observed in the system which, we propose, drives the system close to the Mott insulating phase even at large dopings.

Improved VDC casting of aluminium alloys through an understanding of the surface properties of the molten metal

Vertical direct chill (VDC) casting of aluminium alloys is a mature process that has evolved over many decades through gradual change to both equipment design and casting practice. Today, air-pressurised, continuous lubrication, hot top mould systems with advanced station automation are selected as the process of choice for producing extrusion billet. Specific sets of operating parameters are employed on these stations for each alloy and size combination to produce optimal billet quality. The designs and parameters are largely derived from past experience and accumulated know-how. Recent experimental work at the University of Queensland has concentrated on understanding the way in which the surface properties of liquid aluminium alloys, e.g., surface tension, wetting angle and oxide skin strength, influence the size and shape of the naturally-stab le meniscus for a given alloy, temperature and atmosphere. The wide range of alloy-and condition-dependent values measured has led to the consideration of how these properties impact the stability of the enforced molten metal meniscus within the hot top mould cavity. The actual shape and position of the enforced meniscus is controlled by parameters such as the upstream conduction distance (UCD) from sub-mould cooling and the molten metal head. The degree of deviation of this actual meniscus from the predicted stable meniscus is considered to be a key driver in surface defect formation. This paper reports on liquid alloy property results and proposes how this knowledge might be used to better design VDC mould systems and casting practices.

Liquid phase sintering of aluminium alloys

The principle that alloys are designed to accommodate the manufacture of goods made from them as much as the properties required of them in service has not been widely applied to pressed and sintered P/M aluminium alloys. Most commercial alloys made from mixed elemental blends are identical to standard wrought alloys. Alternatively, alloys can be designed systematically using the phase diagram characteristics of ideal liquid phase sintering systems. This requires consideration of the solubilities of the alloying elements in aluminium, the melting points of the elements, the eutectics they form with aluminium and the nature of the liquid phase. The relative diffusivities are also important. Here we show that Al-Sn, which closely follows these ideal characteristics, has a much stronger sintering response than either Al-Cu or Al-Zn, both of which have at least one non-ideal characteristic. (C) 2001 Elsevier Science B.V. All rights reserved.

Identification of slow and fast-acting toxins in a highly ciguatoxic barracuda (Sphyraena barracuda) by HPLC/MS and radiolabelled ligand binding

A barracuda implicated in ciguatera fish poisoning in Guadeloupe was estimated to have an overall flesh toxicity of 15 MUg/g using mouse bioassay. A lipid soluble extract was separated into two toxic fractions, FrA and FrB, on a LH20 Sephadex column eluted with dichloromethane/methanol (1:1). When intraperitoneal injected into mice, FrA provoked symptoms characteristic of slow-acting ciguatoxins, whereas FrB produced symptoms indicative of fast-acting toxins (FAT). High performance liquid chromatography/mass spectrometry/radio-ligand binding (HPLC/MS/RLB) analysis confirmed the two fractions were distinct, because only a weak overlap of some compounds was observed. HPLC/MS/RLB analysis revealed C-CTX-1 as the potent toxin present in FrA, and two coeluting active compounds at m/z 809.43 and 857.42 in FrB, all displaying the characteristic pattern of ion formation for hydroxy-polyethers. Other C-CTX congeners and putative hydroxy-polyether-like compounds were detected in both fractions, however, the RLB found them inactive. C-CTX-1 accounted for >90% of total toxicity in this barracuda and was confirmed to be a competitive inhibitor of brevetoxin binding to voltage-sensitive sodium channels (VSSCs) with a potency two-times lower than P-CTX-1. However, FAT active on VSSCs and

Stability of extemporaneously prepared captopril oral liquid formulation for paediatric patients

Captopril, an inhibitor of angiotensin converting enzyme (ACE), is used to treat medical conditions like hypertension and heart failure, and it is usually administered in tablet form for adults. Since this dosage form is not recommended for infants and children up to 6 years, hospital pharmacies have to prepare liquid formulations for oral administration of captopril. Traditionally, concentration of captopril used in the formulations is 1mg/ml. The problem is that captopril is prone to oxidation, and its stability in solution is affected by pH, concentration of captopril, the presence of oxygen or metal ions. The influence of different formulation ingredients on the properties of physical and chemical stability of captopril in liquid preparations has been evaluated. Main of the study: to evaluate the stability of captopril for 30 days when formulated in a 1 mg/ml suspension adjuvanted with citric acid.

Mixed extractant systems for metal recovery

Hoje em dia, a prevenção dos resíduos de metais é uma questão muito importante para um grande número de empresas, pois necessitam optimizar o seu sistema de tratamento de águas residuais a fim de alcançarem os limites legais dos teores em iões metálicos e poderem efectuar a descarga das águas residuais no domínio hídrico público. Devido a esta problemática foram efectuados estudos inovadores relacionados com a remoção de iões metálicos de águas residuais, verificando-se que as tecnologias de membrana oferecem uma série de vantagens para o efeito. Uma dessas tecnologias, referida como Membrana Líquida de Suporte (SLM), é baseada num mecanismo de extracção. A membrana hidrofóbica, impregnada com uma solução extractora, funciona como barreira entre a água residual e uma solução, geralmente ácida. A diferença de pH entre a água residual e a solução actua como força motriz para o transporte de iões metálicos da água residual para a referida solução. Poderá ocorrer um problema de falta de estabilidade, resultante da possível fuga da solução extractora para fora dos poros das membranas. Estudos anteriores mostraram que os ácidos alquilfosfóricos ou ácidos fosfónicos, como os reagentes D2EHPA e CYANEX e hidroxioximas como o LIX 860-I podem ser muito úteis para a extração de iões metálicos como ferro, cobre, níquel, zinco e outros. A clássica extracção líquido-líquido também tem mostrado que a mistura de diferentes extractores pode ter um efeito sinergético. No entanto, não é claro que haja um efeito óptimo da razão de extractor ou que tipo de complexo é formado durante o processo de extracção. O objectivo deste projecto é investigar este comportamento sinergético e as complexas formações por meio de um método espectrofotométrico, o “Job’s method” e “Mole-ratio method”. Estes métodos são utilizados para estimar a estequiometria dos vários complexos entre dois solutos, a partir da variação de absorvância dos complexos quando comparado com a absorvância do soluto. Com este projecto, o Job’s method e mole-ratio method serão aplicados a um sistema de três componentes, para conseguir mais informações sobre a complexação de níquel (II) e a fim de determinar a razão extractor: metal dos complexos formados durante a aplicação de mistura de extractores D2EHPA e LIX 860-I. Segundo Job’s method a elavada absorvância situa-se na região de 0,015-0,040 M de LIX 860-I e uma baixa concentração de D2EHPA. Quando as diferentes experiências são encontradas num conjunto experimental foram avaliadas de acordo com o método de trabalho, o valor máximo do gráfico foi encontrado para uma baixa fração molar do ião metálico e uma maior concentração de D2EHPA. Esta mudança foi encontrado de 0,50 até 0,30, que poderia apontar para a direção da formação de diferentes complexos. Para o Mole-Ratio method, a estequiometria dos complexos metal pode ser determinada a partir do ponto de intersecção das linhas tangente do gráfico da absorbância versus a concentração do ligante. Em todos os casos, o máximo foi obtido em torno de uma concentração total de 0,010 M. Quando D2EHPA foi aplicado sozinho, absorvâncias muito baixos foram obtidas.

Neural-network approach to modeling liquid crystals in complex confinement

Finding the structure of a confined liquid crystal is a difficult task since both the density and order parameter profiles are nonuniform. Starting from a microscopic model and density-functional theory, one has to either (i) solve a nonlinear, integral Euler-Lagrange equation, or (ii) perform a direct multidimensional free energy minimization. The traditional implementations of both approaches are computationally expensive and plagued with convergence problems. Here, as an alternative, we introduce an unsupervised variant of the multilayer perceptron (MLP) artificial neural network for minimizing the free energy of a fluid of hard nonspherical particles confined between planar substrates of variable penetrability. We then test our algorithm by comparing its results for the structure (density-orientation profiles) and equilibrium free energy with those obtained by standard iterative solution of the Euler-Lagrange equations and with Monte Carlo simulation results. Very good agreement is found and the MLP method proves competitively fast, flexible, and refinable. Furthermore, it can be readily generalized to the richer experimental patterned-substrate geometries that are now experimentally realizable but very problematic to conventional theoretical treatments.

Monitorização do adoçante ciclamato de sódio através de sensor óptico

O presente trabalho pretendeu desenvolver e testar um sensor óptico para detectar ciclamato de sódio, um adoçante artificial utilizado nas bebidas em geral. A primeira abordagem neste sentido baseou-se na preparação de um sensor óptico através da formação de complexos corados entre o ciclamato e várias espécies metálicas, nomeadamente Hg(II), Ba(II), Fe(II), Ag(II), Pb(II), Cd(II), Mn (II), Ni(II), Cu(II), Co(II), Sn(II) e Mg(II). Perante a ausência de resultados satisfatórios optou-se por explorar a acção do ciclamato de sódio na transferência/partilha de um corante entre duas fases líquidas imiscíveis. As fases líquidas utilizadas foram a água e o clorofórmio. Testaram-se várias famílias de corantes mas só uma classe se mostrou com as características apropriadas para o objectivo pretendido. Dentro dessa família de corantes, seleccionou-se aquele que, à partida, garantiu o melhor desempenho. O sensor foi testado em diferentes condições de pH e também na presença de potenciais interferentes de forma a estabelecer as melhores condições de utilização. O método mostrou-se bastante simples de executar, rápido na obtenção de resultados e com boas características para ser avaliado visualmente, mas sempre de acordo com os critérios de objectividade que um trabalho deste tipo requer. Além o disso permitiu ser calibrado de uma forma rápida e simples, características essenciais para a aplicação deste método na despistagem de ciclamato em análises de rotina. O método desenvolvido foi ainda aplicado à análise de vinho dopado com diferentes concentrações de ciclamato de sódio. Destes testes verificou-se a necessidade de optimização do método através da introdução de outras substâncias na fase não aquosa diminuindo a vulnerabilidade do sensor a outros interferentes. Como conclusão, o método correspondeu às expectativas, mostrando-se viável para aplicação à análise de vinhos, ainda com uma margem significativa de desenvolvimento no sentido de o tornar mais fiável e preciso.

Cooperative effects in the detection of a nitroaliphatic liquid explosive and na explosive taggant in the vapor phase by calix[4]arene-based carbazole-containing conjugated polymers

Two fluorescent molecular receptor based conjugated polymers were used in the detection of a nitroaliphatic liquid explosive (nitromethane) and an explosive taggant (2,3-dimethyl-2,3-dinitrobutane) in the vapor phase. Results have shown that thin films of both polymers display remarkably high sensitivity and selectivity toward these analytes. Very fast, reproducible, and reversible responses were found. The unique behavior of these supramolecular host systems is ascribed to cooperativity effects developed between the calix[4] arene hosts and the phenylene ethynylene-carbazolylene main chains. The calix[4]-arene hosts create a plethora of host-guest binding sites along the polymer backbone, either in their bowl-shaped cavities or between the outer walls of the cavity, to direct guests to the area of the transduction centers (main chain) at which favorable photoinduced electron transfer to the guest molecules occurs and leads to the observed fluorescence quenching. The high tridimensional porous nature of the polymers imparted by the bis-calixarene moieties concomitantly allows fast diffusion of guest molecules into the polymer thin films.

Neural-network approach to modeling liquid crystals in complex confinement

Finding the structure of a confined liquid crystal is a difficult task since both the density and order parameter profiles are nonuniform. Starting from a microscopic model and density-functional theory, one has to either (i) solve a nonlinear, integral Euler-Lagrange equation, or (ii) perform a direct multidimensional free energy minimization. The traditional implementations of both approaches are computationally expensive and plagued with convergence problems. Here, as an alternative, we introduce an unsupervised variant of the multilayer perceptron (MLP) artificial neural network for minimizing the free energy of a fluid of hard nonspherical particles confined between planar substrates of variable penetrability. We then test our algorithm by comparing its results for the structure (density-orientation profiles) and equilibrium free energy with those obtained by standard iterative solution of the Euler-Lagrange equations and with Monte Carlo simulation results. Very good agreement is found and the MLP method proves competitively fast, flexible, and refinable. Furthermore, it can be readily generalized to the richer experimental patterned-substrate geometries that are now experimentally realizable but very problematic to conventional theoretical treatments.

Rheo-optical characterization of liquid crystalline acetoxypropylcellulose melt undergoing large shear flow and relaxation after flow cessation

The rheological and structural characteristics of acetoxypropylcellulose (APC) nematic melt are studied at shear rates ranging from 10 s(-1) to 1000 s(-1) which are relevant to extrusion based processes. APC shows a monotonic shear thinning behavior over the range of shear rates tested. The negative extrudate-swell shows a minimum when a critical shear rate (gamma) over dot(c) is reached. For shear rates smaller than (gamma) over dot(c), the flow-induced texture consists of two set of bands aligned parallel and normal to the flow direction. At shear rates larger than (gamma) over dot(c), the flow induced texture is reminiscent of a 2 fluids structure. Close to the shearing walls, domains elongated along the flow direction and stacked along the vorticity are imaged with POM, whereas SALS patterns indicate that the bulk of the sheared APC is made of elliptical domains oriented along the vorticity. No full nematic alignment is achieved at the largest shear rate tested. Below (gamma) over dot(c), the stress relaxation is described by a stretched exponential. Above (gamma) over dot(c), the stress relaxation is described by a fast and a slow process. The latter coincides with the growth of normal bands thicknesses, as the APC texture after flow cessation consists of two types of bands with parallel and normal orientations relative to the flow direction. Both bands thicknesses do not depend on the applied shear rate, in contrast to their orientation. (C) 2015 Elsevier Ltd. All rights reserved.