A novel,simple and robust method for the analysis of sulfonamides in fish farming water samples using column switching liquid chromatography

A novel method for the online extraction and preconcentration of four sulfonamides was developed using column switching liquid chromatography. Sulfadiazine, sulfathiazole, sulfamethoxypyridazine and sulfamethoxazole were analysed in water samples and preconcentrated in a C18 guard column. Suitable validation parameters were obtained, such as precision, accuracy and relative recovery, in accordance with the validation guidelines of the Food and Drug Administration. Low limits of detection (0.05–0.09 µg L^?1) and quantification (0.30 µg L^?1, for all of them) were obtained. The quadratic polynomial model was used to adjust the calibration data, and the coefficients of determination were higher than 0.999 for all the analytes. The method was shown to be robust to the assayed parameters according to Youden’s test. The proposed method was successfully used to determine sulfonamides in 11 different fish farming water samples, in which sulfadiazine (0.732 µg L^?1), sulfamethoxazole (0.531 µg L^?1), sulfathiazole (0.546–1.856 µg L^?1) and sulfamethoxypyridazine (0.369–1.509 µg L^?1) were found.     

Single-drop microextraction for the determination of manganese in seafood and water samples

We describe a method for single drop microextraction of manganese from fish, mollusk, and from natural waters using the reagent 1-(2-pyridylazo)-2-naphthol as the complexing agent and chloroform as the fluid extractor. After extraction, the analyte was directly submitted to graphite furnace electrothermal atomic absorption spectrometry. Once optimized, the method has a detection limit of 30 ng L^?1, a limit of quantification of 100 ng L^?1, and an enrichment factor of 16. Its accuracy was verified by applying the procedure to the following certified reference materials: apple leaves, spinach leaves, bovine liver, and mussel tissue. The procedure was also successfully applied to the determination of manganese in seafood and natural waters.

Production of recombinant bleaching enzymes from thermophilic microorganisms in fungal hosts

Cost-effective production of enzymes for industrial processes makes the appropriate selection of the host-vector expression system critical. We have developed two systems for the bulk production of bleaching enzymes from thermophiles. Kluyveromyces lactis has been developed as a secretion host employing expression vectors based on the 2μ-like plasmid pKD1 of Kluyveromyces drosophilarium. Our second system involves the filamentous fungus Trichoderma reesei. Fusion and nonfusion vectors have been constructed using the strong cellobiohydrolase 1 (cbh1) promoter. The KEX2 protease cleavage site and a 6 × HIS-tag have been incorporated to facilitate both cleavage and purification of the mature foreign proteins.     

Electrochemical Determination of Sunscreens Agents in Cosmetic Using Square Wave Voltammetry

Sunscreen agents and in particular UV filters are compounds added in different cosmetic formulations, that has the function of preventing damage caused by sun exposition. Therefore, this paper proposes the development of a simple, fast and reliable electroanalytical method utilizing square wave voltammetry (SWV) to the determination of Benzophenone‐3 (BZ‐3), camphor 4‐methylbenzylidene (MBC) and 2‐ethylhexyl‐4‐methoxycinnamate (OMC) in cosmetic samples. The electrochemical system consisted of a cell with three electrodes: work – gold electrode modified, reference – Ag/AgCl_(sat) and auxiliary – platinum, using as supporting electrolyte 4.0 mL of Britton Robinson Buffer 0.04 mol L^?1 (pH=4.0), 1.0 mL of methanol and 5.50×10^?4 mol L^?1 of cetyltrimethylammonium bromide (CTAB). The method was validated using three commercial sunscreen samples and the results showed recovery values between 83 and 98 %. The average values found for the analysed samples were 3.49 % m/m (728 mg L^?1) to BZ‐3, 0.56 % m/m (113 mg L^?1) to MBC and 0.99 % m/m (208 mg L^?1) to OMC. The detection (DL) and quantification (QL) limits were 0.47 mg L^?1 and 1.56 mg L^?1 to BZ‐3, 0.77 mg L^?1 and 2.58 mg L^?1 to MBC and 0.78 mg L^?1 and 2.59 mg L^?1 to OMC, respectively. The sunscreen protector samples were also evaluated by high‐performance liquid chromatography (HPLC) demonstrating a good correlation between the results and compared the results with allowed concentration.     

Crystallization and characterization of an inflammatory lectin purified from the seeds of Dioclea wilsonii

DwL, a lectin extracted from the seeds of Dioclea wilsonii, is a metalloprotein with strong agglutinating activity against rabbit and ABO erythrocytes, inhibited by glucose and mannose. DwL was purified by affinity chromatography on a Sephadex G-50 column and ion exchange chromatography on a HiTrap SP XL column. SDS-PAGE revealed three electrophoretic bands corresponding to the α (25,634 ± 2 Da), β (12,873 ± 2 Da) and γ (12,779 ± 2 Da) chains. Protein sequencing was done by Tandem Mass Spectrometry. The primary sequence featured 237 amino acids and was highly homologous to other reported Diocleinae lectins. A complete X-ray dataset was collected at 2.0 ? for X-Man-complexed DWL crystals produced by the vapor diffusion method. The crystals were orthorhombic and belonged to the space group I222, with the unit-cell parameters a = 59.6, b = 67.9 and c = 109.0 ?. DWL differed in potency from other ConA-like lectins and was found to induce neutrophil migration in rats, making it particularly useful in structural/functional studies of this class of proteins.     

Transport properties of alkyltrimethylammonium bromide surfactants in aqueous solutions

Differential mutual diffusion coefficients of n-alkyltrimethylammonium bromides [CH₃(CH₂)_n–1N(CH₃)₃Br, C_nTAB] (n=10, 12, 14, 16) have been measured in aqueous solutions at 298.15 K using a conductimetric cell and an automatic apparatus to follow diffusion. The cell is based on an open-ended capillary, and the technique follows the diffusion process by measuring the resistance of a solution inside the capillaries at various times. The electrical conductances of those solutions have also been measured to calculate the critical micellar concentration (cmc). Thermodynamic analysis of the data suggests that the free ion concentration decreases at concentrations above the cmc, in agreement with theoretical predictions. The obtained values of the micellization parameters were used to model the mutual diffusion coefficients of C_nTAB aqueous solutions.     

Development of a rapid and specific assay for detection of busulfan in human plasma by high-performance liquid chromatography/electrospray ionization tandem mass spectrometry

A sensitive and specific assay for detection of busulfan in human plasma was developed. The assay is based on rapid isolation of busulfan by liquid-liquid extraction with ethyl acetate, and detection by high-performance liquid chromatography with electrospray ionization and tandem mass spectrometry. 1,6-Bis(methanesulfonyloxy)hexane, a synthesized analogue of busulfan, was used as the internal standard (IS). The acquisition was performed in the multiple reaction monitoring mode; busulfan and the IS were detected with no interferences from plasma matrix. The method was linear over the range 5-2500 ng mL(-1), with r2 > 0.99 and a run time of only 3.5 min. The intra- and inter-assay precisions were in the ranges 2.1-11.9% and 3.2-10.1%, respectively, and the intra- and inter-assay accuracies were 92.2-107.6% and 94.7-104.1%, respectively. The absolute recoveries were 82.0% (20 ng mL(-1)), 90.6% (1000 ng mL(-1)) and 80.0% (2000 ng mL(-1)) for busulfan, and 89.1% for the IS (1000 ng mL(-1)). The limits of detection and quantification were 2 and 5 ng mL(-1), respectively. The validated method was successfully applied to analyze plasma samples obtained from six adults receiving doses of 1 mg kg(-1) in a conditioning regimen prior to bone marrow transplantation. A marked intra-patient variation in busulfan concentrations during the steady state was observed, which limits the application of pharmacokinetic modeling and suggests that continuous therapeutic monitoring is necessary for adequate individualized dosing. In this regard, the present assay brings important advantages relative to other methods described in the literature, i.e., it is highly specific and simple to perform, with a rapid chromatographic run time (3.5 min), and the whole procedure can be completed in 4-5 h, which would permit dose corrections after the third dose allowing earlier and better dosing adjustments towards the target level of busulfan.     

Effect of the pore geometry in the characterization of the pore size distribution of activated carbons

In this work, the characterization of Activated Carbons (AC) by using the independent pore models is discussed, with special emphasis on the issue of how the assumed pore geometry can affect the resulting Pore Size Distribution (rPSD) and on the problem of the unicity of the PSD when different probe molecules are used in adsorption experiments. A theoretical test was performed using virtual solids based in the so-called Mixed Geometry Model (MGM) (Azevedo et al. 2010). The MGM uses a kernel of adsorption isotherms generated by GCMC for different pore sizes and two pore geometries: slit and triangular. The adsorption isotherms of a virtual MGM solid were fitted with both the traditional Slit Geometry Model (SGM) and the Mixed Geometry Model (MGM). It is demonstrated that, by assuming a different pore geometry model from that of the real sample, different PSDs may be obtained by fitting adsorption isotherms of different probe gases. Finally, experimental results are shown which both point toward the MGM as an acceptable extension of the SGM and confirm that the MGM is a closer representation of the actual porous structure of most activated carbons.