Quantitative Determination of Amlodipine Besylate in Tablets by High Performance Liquid Chromatography and UV Spectrophotometry

The purpose of this study was to develop and validate analytical methods for determination of amlodipine besylate in tablets. Simple, accurate and precise liquid chromatographic and spectrophotometric methods are proposed. For the chromatographic method, the conditions were: a LiChrospher (R) 100 RP-18 Merck (R) (125 mm x 4.6 mm, 5 mu m) column; methanol/water containing 1 % of trietylamine adjusted to pH 5.0 with phosphoric acid (35:65) as mobile phase; a flow rate of 1.0 mL/min and UV detector at 238 nm. Linearity was in the range of 50.0 - 350.0 mu g/mL with a correlation coefficient (r) = 0.9999. For the spectrophotometric method, the first dilutions of samples were performed in methanol and the consecutives in ultrapure water. The quantitation was made at 364.4 nm. Linearity was determined within the range of 41.0 - 61.0 mu g/mL with a correlation coefficient (r) = 0.9996. Our results demonstrate that both methods can be used in routine analysis for quality control of tablets containing amlodipine besylate.

LC-MS-MS determination of ibuprofen, 2-hydroxyibuprofen enantiomers, and carboxyibuprofen stereoisomers for application in biotransformation studies employing endophytic fungi

The purpose of this study was the development and validation of an LC-MS-MS method for simultaneous analysis of ibuprofen (IBP), 2-hydroxyibuprofen (2-OH-IBP) enantiomers, and carboxyibuprofen (COOH-IBP) stereoisomers in fungi culture medium, to investigate the ability of some endophytic fungi to biotransform the chiral drug IBP into its metabolites. Resolution of IBP and the stereoisomers of its main metabolites was achieved by use of a Chiralpak AS-H column (150 x 4.6 mm, 5 mu m particle size), column temperature 8 degrees C, and the mobile phase hexane-isopropanol-trifluoroacetic acid (95: 5: 0.1, v/v) at a flow rate of 1.2 mL min(-1). Post-column infusion with 10 mmol L(-1) ammonium acetate in methanol at a flow rate of 0.3 mL min(-1) was performed to enhance MS detection (positive electrospray ionization). Liquid-liquid extraction was used for sample preparation with hexane-ethyl acetate (1:1, v/v) as extraction solvent. Linearity was obtained in the range 0.1-20 mu g mL(-1) for IBP, 0.05-7.5 mu g mL(-1) for each 2-OH-IBP enantiomer, and 0.025-5.0 mu g mL(-1) for each COOH-IBP stereoisomer (r >= 0.99). The coefficients of variation and relative errors obtained in precision and accuracy studies (within-day and between-day) were below 15%. The stability studies showed that the samples were stable (p > 0.05) during freeze and thaw cycles, short-term exposure to room temperature, storage at -20 degrees C, and biotransformation conditions. Among the six fungi studied, only the strains Nigrospora sphaerica (SS67) and Chaetomium globosum (VR10) biotransformed IBP enantioselectively, with greater formation of the metabolite (+)-(S)-2-OH-IBP. Formation of the COOH-IBP stereoisomers, which involves hydroxylation at C3 and further oxidation to form the carboxyl group, was not observed.

LC-MS-MS Identification and Determination of the Flavone-C-Glucoside Vicenin-2 in Rat Plasma Samples Following Intraperitoneal Administration of Lychnophora Extract

The flavone C-glucoside, vicenin-2, in semi-purified extracts of the leaves of Lychnophora ericoides was quantified in rat plasma samples using a method based on reversed-phase high performance liquid chromatography coupled to tandem mass spectrometry. Vicenin-2 was analyzed on a LiChrospher (R) RP18 column using an isocratic mobile phase consisting of a mixture of methanol: water (30:70, v/v) plus 2.0% glacial acetic acid at a flow rate of 0.8 mL min(-1). Genistein was used as internal standard. The mass spectrometer was operated in positive ionization mode and analytes were quantified by multiple reaction monitoring at m/z 595 > 457 for vicenin-2 and m/z 271 > 153 for internal standard. Prior to the analysis, each rat plasma sample was acidified with 200 mu L of 50 mmol L(-1) acetic acid solution and extracted by solid-phase extraction using a C18 cartridge. The absolute recoveries were reproducible and the coefficients of variation values were lower than 5.2%. The method was linear over the 12.5 - 1500 ng mL(-1) concentration range and the quantification limit was 12.5 ng mL(-1). Within-day and between-day assay precision and accuracy were studied at three concentration levels (40, 400 and 800 ng mL(-1)) and were lower than 15%. The developed and validated method seems to be suitable for analysis of vicenin-2 in plasma samples obtained from rats that receive a single i.p. dose of 200 mg kg(-1) vicenin-2 extract.

Development and Validation of a Simple Method for Routine Analysis of Ractopamine Hydrochloride in Raw Material and Feed Additives by HPLC

A simple method was optimized and validated for determination of ractopamine hydrochloride (RAC) in raw material and feed additives by HPLC for use in quality control in veterinary industries. The best-optimized conditions were a C8 column (250 x 4.6 mm id, 5.0 mu m particle size) at room temperature with acetonitrile-100 mM sodium acetate buffer (pH 5.0; 75 + 25, v/v) mobile phase at a flow rate of 1.0 mL/min and UV detection at 275 nm. With these conditions, the retention time of RAC was around 5.2 min, and standard curves were linear in the concentration range of 160-240 mu g/mL (correlation coefficient >= 0.999). Validation parameters, such as selectivity, linearity, limit of detection (ranged from 1.60 to 2.05 mu g/mL), limit of quantification (ranged from 4.26 to 6.84 mu g/mL), precision (relative standard deviation <= 1.87%), accuracy (ranged from 96.97 to 100.54%), and robustness, gave results within acceptable ranges. Therefore, the developed method can be successfully applied for the routine quality control analysis of raw material and feed additives.

Comparison of the effects of pilocarpine and cevimeline on salivary flow

Objective: The aim of the present study was to compare the effect of low-dose pilocarpine and cevimeline as stimulants for salivary flow in healthy subjects. Methods: In this cross-over clinical trial with a 1-week washout period, 40 male volunteers were submitted to an oral dose of pilocarpine 1% (Salagen (TM)) -60 mu g kg(-1) body-weight (Group 1) or Cevimeline (Evoxac (TM)) -30 mg (Group 2). Saliva samples were collected and the salivary flow rate was measured (ml min(-1)) at baseline and 20, 40, 60, 80, 140 and 200 min after administration of drugs. In addition, salivary secretion was also measured under mechanical stimulation to observe salivary gland function. Results: The data were analyzed by Friedman and Wilcoxon signed-rank tests (significance level = 5%). Pilocarpine and cevimeline significantly increased salivary flow 140 min after intake. There was a significant higher secretion with cevimeline 140 and 200 min after administration. There were no differences seen among subjects in the salivary glands function by mechanical stimulation. Conclusion: Both drugs showed efficacy in increasing the salivary flow in healthy volunteers, but cevimeline was more effective than pilocarpine.

Diffusion modeling of percutaneous absorption kinetics: 2. Finite vehicle volume and solvent deposited solids

The diffusion model for percutaneous absorption is developed for the specific case of delivery to the skin being limited by the application of a finite amount of solute. Two cases are considered; in the first, there is an application of a finite donor (vehicle) volume, and in the second, there are solvent-deposited solids and a thin vehicle with a high partition coefficient. In both cases, the potential effect of an interfacial resistance at the stratum corneum surface is also considered. As in the previous paper, which was concerned with the application of a constant donor concentration, clearance limitations due to the viable eqidermis, the in vitro sampling rate, or perfusion rate in vivo are included. Numerical inversion of the Laplace domain solutions was used for simulations of solute flux and cumulative amount absorbed and to model specific examples of percutaneous absorption of solvent-deposited solids. It was concluded that numerical inversions of the Laplace domain solutions for a diffusion model of the percutaneous absorption, using standard scientific software (such as SCIENTIST, MicroMath Scientific software) on modern personal computers, is a practical alternative to computation of infinite series solutions. Limits of the Laplace domain solutions were used to define the moments of the flux-time profiles for finite donor volumes and the slope of the terminal log flux-time profile. The mean transit time could be related to the diffusion time through stratum corneum, viable epidermal, and donor diffusion layer resistances and clearance from the receptor phase. Approximate expressions for the time to reach maximum flux (peak time) and maximum flux were also derived. The model was then validated using reported amount-time and flux-time profiles for finite doses applied to the skin. It was concluded that for very small donor phase volume or for very large stratum corneum-vehicle partitioning coefficients (e.g., for solvent deposited solids), the flux and amount of solute absorbed are affected by receptor conditions to a lesser extent than is obvious for a constant donor constant donor concentrations. (C) 2001 Wiley-Liss, Inc. and the American Pharmaceutical Association J Pharm Sci 90:504-520, 2001.

Unsaturated liquid percolation flow through nonwetted packed beds

The unsaturated flow of liquid through packed beds of large particles was studied using six different liquids, all with contact angles greater than 90degrees on the bed packing (wax spheres of 9, 15 and 19.4 mm diameter). The liquid flow was discrete in nature, as drops for low flow rates and rivulets for high flow rates. For unsaturated liquid flows, the actual percolation velocity, not superficial velocity, should be used to characterize the flow. The percolation velocity did not vary with packed-bed depth, but was a strong function of liquid flow rate, liquid and particle properties. Effects of liquid and particle properties (but not flow rate) are well captured by a simple correlation between the liquid-particle friction factor and Reynolds number based on actual percolation velocities. Liquid dispersion, characterized by the maximum dispersion angle, varies significantly with liquid and particle properties. The tentative correlation suggested here needs further validation for a wider range of conditions.

Metodologia para obtenção do hidrograma de escoamento superficial em encostas e canais. Parte II: modelo computacional e análise de sensibilidade

Estimativas de vazão máxima de escoamento superficial são necessárias para o projeto de obras hidráulicas em bacias urbanas e rurais. A dificuldade em aplicar os procedimentos disponíveis para calcular a variação do escoamento superficial com o tempo e de seu valor máximo deve-se à inexatidão dos métodos usados para esse objetivo e à variabilidade nos resultados que podem ser obtidos por profissionais que usem o mesmo procedimento. Dessa forma, a investigação de um método que produza estimativas confiáveis da vazão máxima e do hidrograma de escoamento superficial é de grande interesse. Neste trabalho, desenvolveu-se e avaliou-se a sensibilidade de um software (HIDROGRAMA 2.1) que permite a obtenção do hidrograma de escoamento superficial, da vazão máxima e seu tempo de ocorrência, da altura e da velocidade máximas do escoamento, do volume e da lâmina de escoamento superficial em encosta e em canais. O modelo apresentou grande sensibilidade ao período de retorno, à taxa de infiltração estável e ao comprimento da encosta e do canal.

Estudo da implementação de um sistema de vigilância e alarme num recurso hídrico

Neste trabalho estudou-se a implementação de um sistema de vigilância e alerta da qualidade da água de um recurso hídrico, para um possível caso de poluição. Em 25 de Agosto de 2008 foram derramadas 4 toneladas de ácido clorídrico acidentalmente para as águas do rio Febros. Este rio situa-se no concelho de Vila Nova de Gaia e é um pequeno afluente do rio Douro, tendo cerca de 14 km de extensão e tem a particularidade de atravessar o Parque Biológico de Gaia. A falta de uma rápida intervenção e da existência de um plano de ação levou a que parte da fauna e flora fosse destruída. Por este motivo realizou-se este estudo que se baseou na criação de um sistema de vigilância e alerta a ser implementado neste rio. A informação da hidrogeometria do rio e da capacidade de transporte e dispersão de poluentes é indispensável para o bom funcionamento deste sistema. O coeficiente de dispersão longitudinal é um parâmetro muito importante no estudo da qualidade da água. Recorreu-se à utilização da Rodamina WT como marcador, determinando assim a evolução da sua concentração ao longo do tempo e espaço. No cálculo do coeficiente de dispersão foi utilizado o modelo Transient Storage, que demonstrou ser um bom modelo de ajuste aproximando-se dos valores medidos em campo. Para três estações diferentes com distâncias de 290, 390 e 1100 metros do ponto de injeção, obtiveram-se valores de coeficiente de dispersão de 0,18, 0,15 e 0,39 m2/s respetivamente. Os valores do ajuste expressos sob a forma de coeficiente de correlação foram 0,988, 0,998 e 0,986, para a mesma ordem de estações. A constante de rearejamento do rio foi também determinada recorrendo ao método dos marcadores inertes, utilizando o propano como marcador gasoso. A constante determinada próximo de Casal Drijo, entre 2 estações de amostragem a 140 e 290 m do local de injeção, foi de 13,4 dia-1. Com os resultados do coeficiente de dispersão e da constante de rearejamento para além da velocidade e caudal da corrente do rio conseguir-se-á construir o modelo de simulação e previsão de um possível poluente. O sistema de vigilância a implementar sugere-se assim que seja construído por duas partes, uma de análise de evolução da nuvem de poluição e plano de ação outra de monitorização contínua e emissão de alerta. Após uma análise do investimento à implementação deste sistema chegou-se à conclusão que o valor de investimento é de 15.182,00 €.

Maximização da produção de paraxileno

Pretendeu-se, com o presente trabalho, efectuar um estudo sobre a possibilidade de aumentar a produção de paraxileno na unidade processual existente na refinaria da Galp no Porto. Foram estudados o conjunto de colunas T-0306 e T-0307 e a coluna T-0303, recorrendo ao uso do AspenPlus, na realização da simulação do processo nas condições actuais e para a realização da análise de sensibilidade, que permitiu verificar se o equipamento existente possui capacidade para satisfazer as necessidades exigidas pelo processo quando se pretende aumentar a produção. Com a simulação do processo, foi verificado que este já se encontra a laborar em condições óptimas. Na realização da análise de sensibilidade foram variados para o conjunto T-0306 e T- 0307, o caudal de alimentação entre 62780 e 95000 kg/h, o caudal de destilado da T-0306 entre 15833 e 23911 kg/h e o caudal de destilado da T-0307 entre 139 e os 235 kg/h. No caso da T-0303, foi variado o caudal de refinado entre 201240 e 304600 kg/h, o caudal de destilado e o caudal da corrente lateral, variam entre 77290 a 116930 kg/h. Pretendeu-se obter soluções que conseguissem cumprir as especificações relativas aos produtos obtidos, nomeadamente paraxileno com uma pureza de 99,6% e uma recuperação de 99%, tolueno com uma pureza de 97% e uma recuperação de 90% (T-0306 e T-0307). No caso da T- 0303, obter paraxileno com uma pureza de 2,23% e paradietilbenzeno com uma pureza de 99,0% e ambos com uma recuperação de 99%. Para as simulações que cumpriram as especificações anteriores, verificou-se se a energia transferida em aeroarrefecedores, fornalhas e permutadores excedem em 25% o seu valor de projecto. Em caso afirmativo isso significou o acréscimo de uma peça de equipamento idêntica à existente. A análise de sensibilidade permitiu concluir a separação é possível para os caudais actuais de extracto e refinado e para um aumento destes de 32, 40 e 50%. Foi ainda possível concluir que para os caudais actuais é necessária a colocação de novos aeroarrefecedores para as colunas T-0306 e T-0303 e de uma nova fornalha para a coluna T-0303. Para um aumento dos caudais de 32% para além das alterações referidas anteriormente é necessária a colocação de um novo aeroarrefecedor na coluna T-0307 e de uma nova fornalha da coluna T-0306. Se aumentarmos os caudais em 40% será também necessário o acréscimo de permutadores para a coluna T-0307. Para o aumento de 50%, os calores de permuta são excedidos em bastante mais do que 25%, em todos os aeroarrefecedores, permutadores e fornalhas. Uma vez que todas as colunas se encontram a funcionar acima da sua capacidade, concluiu-se que em ambos os casos seria necessária a montagem de uma segunda linha em paralelo e igual à existente. Implicando com esta alteração um investimento total de 10.439.574,36€.

Rapid determination of tartaric acid in wines

A flow-spectrophotometric method is proposed for the routine determination of tartaric acid in wines. The reaction between tartaric acid and vanadate in acetic media is carried out in flowing conditions and the subsequent colored complex is monitored at 475 nm. The stability of the complex and the corresponding formation constant are presented. The effect of wavelength and pH was evaluated by batch experiments. The selected conditions were transposed to a flowinjection analytical system. Optimization of several flow parameters such as reactor lengths, flow-rate and injection volume was carried out. Using optimized conditions, a linear behavior was observed up to 1000 µg mL-1 tartaric acid, with a molar extinction coefficient of 450 L mg-1 cm-1 and ± 1 % repeatability. Sample throughput was 25 samples per hour. The flow-spectrophotometric method was satisfactorily applied to the quantification of tartaric acid (TA) in wines from different sources. Its accuracy was confirmed by statistical comparison to the conventional Rebelein procedure and to a certified analytical method carried out in a routine laboratory.

Determination of polyphenols in wines by reaction with 4-aminoantipyrine and photometric flow-injection analysis

A new flow-injection analytical procedure is proposed for the determination of the total amount of polyphenols in wines; the method is based on the formation of a colored complex between 4-aminoantipyrine and phenols, in the presence of an oxidizing reagent. The oxidizing agents hexacyanoferrate(III), peroxodisulfate, and tetroxoiodate(VII) were tested. Batch trials were first performed to select appropriate oxidizing agents, pH, and concentration ratios of reagents, on the basis of their effect on the stability of the colored complex. Conditions selected as a result of these trials were implemented in a flow-injection analytical system in which the influence of injection volume, flow rate, and reaction- coil length, was evaluated. Under the optimum conditions the total amount of polyphenols, expressed as gallic acid, could be determined within a concentration range of 36 to 544 mg L–1, and with a sensitivity of 344 L mol–1 cm–1 and an RSD <1.1%. The reproducibility of analytical readings was indicative of standard deviations <2%. Interference from sugars, tartaric acid, ascorbic acid, methanol, ammonium sulfate, and potassium chloride was negligible. The proposed system was applied to the determination of total polyphenols in red wines, and enabled analysis of approximately 55 samples h–1. Results were usually precise and accurate; the RSD was <3.9% and relative errors, by the Folin–Ciocalteu method, <5.1%.

Study of the voltammetric behaviour of metam and its application to an amperometric flow system

The electrochemical behaviour of the pesticide metam (MT) at a glassy carbon working electrode (GCE) and at a hanging mercury drop electrode (HMDE) was investigated. Different voltammetric techniques, including cyclic voltammetry (CV) and square wave voltammetry (SWV), were used. An anodic peak (independent of pH) at +1.46 V vs AgCl/Ag was observed in MTaqueous solution using the GCE. SWV calibration curves were plotted under optimized conditions (pH 2.5 and frequency 50 Hz), which showed a linear response for 17–29 mg L−1. Electrochemical reduction was also explored, using the HMDE. A well defined cathodic peak was recorded at −0.72 V vs AgCl/ Ag, dependent on pH. After optimizing the operating conditions (pH 10.1, frequency 150 Hz, potential deposition −0.20 V for 10 s), calibration curves was measured in the concentration range 2.5×10−1 to 1.0 mg L−1 using SWV. The electrochemical behaviour of this compound facilitated the development of a flow injection analysis (FIA) system with amperometric detection for the quantification of MT in commercial formulations and spiked water samples. An assessment of the optimal FIA conditions indicated that the best analytical results were obtained at a potential of +1.30 V, an injection volume of 207 μL and an overall flow rate of 2.4 ml min−1. Real samples were analysed via calibration curves over the concentration range 1.3×10−2 to 1.3 mg L−1. Recoveries from the real samples (spiked waters and commercial formulations) were between 97.4 and 105.5%. The precision of the proposed method was evaluated by assessing the relative standard deviation (RSD %) of ten consecutive determinations of one sample (1.0 mg L−1), and the value obtained was 1.5%.

O alisamento dos resultados numa empresa: o caso do Grupo SONANGOL

Dissertação de Mestrado apresentada ao Instituto de Contabilidade e Administração do Porto para a obtenção do grau de Mestre em Contabilidade e Finanças sob orientação de Professor Doutor Adalmiro Alvaro Malheiro de Castro Andrade Pereira

Oxygen mass transfer impact on citric acid production by Yarrowia lipolytica from crude glycerol

Production of citric acid from crude glycerol from biodiesel industry, in batch cultures of Yarrowia lipolytica W29 was performed in a lab-scale stirred tank bioreactor in order to assess the effect of oxygen mass transfer rate in this bioprocess. An empirical correlation was proposed to describe oxygen volumetric mass transfer coefficient (kLa) as a function of operating conditions (stirring speed and specific air flow rate) and cellular density. kLa increased according with a power function with specific power input and superficial gas velocity, and slightly decreased with cellular density. The increase of initial kLa from 7 h-1 to 55 h-1 led to 7.8-fold increase of citric acid final concentration. Experiments were also performed at controlled dissolved oxygen (DO) and citric acid concentration increased with DO up to 60% of saturation. Thus, due to the simpler operation setting an optimal kLa than at controlled DO, it can be concluded that kLa is an adequate parameter for the optimization of citric acid production from crude glycerol by Y. lipolytica and to be considered in bioprocess scale-up. Our empirical correlation, considering the operating conditions and cellular density, will be a valid tool for this purpose.