Bromazepam determination in human plasma by high-performance liquid chromatography coupled to tandem mass spectrometry: a highly sensitive and specific tool for bioequivalence studies

A rapid, sensitive and specific method to quantify bromazepam in human plasma using diazepam as the internal standard (IS) is described. The analyte and the IS were extracted from plasma by liquid-liquid extraction using diethyl ether-hexane (80 : 20, v/v). The extracts were analyzed by high-performance liquid chromatography (HPLC) coupled to electrospray tandem mass spectrometry (MS/MS). Chromatography was performed isocratically on a Genesis C(18) analytical column (100 x 2.1 mm i.d., film thickness 4 microm). The method had a chromatographic run time of 5.0 min and a linear calibration curve over the range 5.0-150 ng ml(-1) (r(2) > 0.9952). The limit of quantification was 5 ng ml(-1). This HPLC/MS/MS procedure was used to assess the bioequivalence of two bromazepam 6 mg tablet formulations (bromazepam from Medley SA Indústria Farmacêutica as the test formulation and Lexotan from Produtos Roche Químico e Farmacêutico SA as the reference formulation). A single 6 mg dose of each formulation was administered to 24 healthy volunteers (12 males and 12 females). The study was conducted using an open, randomized, two-period crossover design with a 3 week washout interval. Since the 90% CI for C(max), AUC(last), AUC(0-240 h) (linear) and AUC((0- infinity )) ratios were all inside the 80-125% interval proposed by the US Food and Drug Administration, it was concluded that the bromazepam formulation from Medley is bioequivalent to the Lexotan formulation for both the rate and the extent of absorption.     

Instrumental neutron activation analysis for assessing homogeneity of a whole rice candidate reference material

A whole rice reference material was prepared for use in quality control of trace elements in rice and in similar matrices. This study aimed at evaluating the homogeneity of the material. As, Br, Co, Cs, Fe, K, Na, Rb and Zn were determined in the samples by instrumental neutron activation analysis. The material is homogeneous for the elements determined (p < 0.05), except for Co and Fe, which presented dispersion of results much higher than the predicted degree of repeatability.     

Analysis of low molecular weight aldehydes in air samples by capillary electrophoresis after derivatization with 4-hydrazinobenzoic acid

This work reports the analysis of selected aldehydes in air samples using capillary electrophoresis (CE). The method is based on the reaction of aldehydes with 4-hydrazinobenzoic acid (HBA) to give the corresponding hydrazones with maximum absorbance at 290 nm. Under optimized CE conditions, the HBA derivatives of four carbonyls (formaldehyde, acetaldehyde, propionaldehyde, and acrolein) were completely separated from one another, in less than 6 min, using a pH 9.3 tetraborate buffer at 0.040 mol L(-1) concentration as background electrolyte. A few method validation parameters were determined revealing good migration time repeatability (< 1.5% CV) and area repeatability (< 2% CV), excellent linearity (50-300 microg/L, r > 0.996) and adequate sensitivity for environmental applications. The limits of detection with respect to each single aldehyde were in the range of 2.7-8.8 ng L(-1). The methodology was applied to the determination of aldehydes indoors. Samples were collected in HBA impregnated octadecylsilica cartridges, at different times during the day. The most abundant carbonyls in the samples were acetaldehyde followed by formaldehyde, with estimated peak concentrations of 4.3 and 2.9 ppbv, respectively.     

Adding value to the Brazilian sisal: acid hydrolysis of its pulp seeking production of sugars and materials

The present work is inserted into the broad context of the upgrading of lignocellulosic fibers. Sisal was chosen in the present study because more than 50% of the world’s sisal is cultivated in Brazil, it has a short life cycle and its fiber has a high cellulose content. Specifically, in the present study, the subject addressed was the hydrolysis of the sisal pulp, using sulfuric acid as the catalyst. To assess the influence of parameters such as the concentration of the sulfuric acid and the temperature during this process, the pulp was hydrolyzed with various concentrations of sulfuric acid (30–50%) at 70 °C and with 30% acid (v/v) at various temperatures (60–100 °C). During hydrolysis, aliquots were withdrawn from the reaction media, and the solid (non-hydrolyzed pulp) was separated from the liquid (liquor) by filtering each aliquot. The sugar composition of the liquor was analyzed by HPLC, and the non-hydrolyzed pulps were characterized by viscometry (average molar mass), and X-ray diffraction (crystallinity). The results support the following conclusions: acid hydrolysis using 30% H₂SO₄ at 100 °C can produce sisal microcrystalline cellulose and the conditions that led to the largest glucose yield and lowest decomposition rate were 50% H₂SO₄ at 70 °C. In summary, the study of sisal pulp hydrolysis using concentrated acid showed that certain conditions are suitable for high recovery of xylose and good yield of glucose. Moreover, the unreacted cellulose can be targeted for different applications in bio-based materials. A kinetic study based on the glucose yield was performed for all reaction conditions using the kinetic model proposed by Saeman. The results showed that the model adjusted to all 30–35% H₂SO₄ reactions but not to greater concentrations of sulfuric acid. The present study is part of an ongoing research program, and the results reported here will be used as a comparison against the results obtained when using treated sisal pulp as the starting material.     

4‐Aminothiophenol Self‐Assembled Monolayer for the Development of a DNA Biosensor Aiming the Detection of Cylindrospermopsin Producing Cyanobacteria

The development of a DNA biosensor for the detection of cylindrospermopsin, based on self‐assembled monolayers (SAMs) of 4‐aminothiophenol, is investigated. SAMs were characterized by electrochemical reductive desorption. Detection of probe immobilization and hybridization has been achieved by cyclic and square‐wave voltammetry (SWV), using methylene blue (MB) as electroactive indicator. The SWV data obtained in phosphate buffer, with and without NaCl, after MB accumulation, revealed an increase of the redox indicator current peaks after the hybridization step. This behavior is consistent with MB intercalation into DNA, for high ionic strength media and attributed to electrostatic interactions in the absence of salt. Evidence for surface modification is also provided by atomic force microscopy and ellipsometry.     

Comparison of the endemic Azorean Hypericum foliosum with other Hypericum species: antioxidant activity and phenolic profile

Water extracts of Hypericum foliosum (HF) from five Azorean Islands were analysed for their antioxidant activity and total phenolic contents. The results were compared with those from medicinal Hypericum species (H. perforatum, H. androsaemum and H. undulatum) and also with Azorean green tea. HF exhibited strong scavenging activity (87-91%) and moderate inhibition of linoleic acid oxidation (56-72%), and presented no significant difference to the other studied plants; in contrast, the synthetic antioxidant butylated hydroxyltoluene showed antioxidant activity values of 22% and 88%, respectively. The average value of phenolics in HF was higher than that in the other Hypericum species and lower than that in tea. Additionally, the phenolic profiles of the Hypericum species were compared by reverse phase-high performance liquid chromatography/ultraviolet (RP-HPLC/UV). The method presented permits the simultaneous determination of phenolic acids, flavonoids, hypericin and hyperforin within 55 min. The most similar profile was observed between HF and H. androsaemum. This study indicates that HF is a source of bioactive compounds with potential health benefits.     

Influence of amino acid relative position on the oxidative modification of histidine and glycine peptides

The radical oxidation of isomeric peptides containing one reactive amino acid [histidine (H)] and another less reactive amino acid [glycine (G)] in the form of dipeptides (HG and GH) and tripeptides (HGG, GHG, and GGH) was studied by mass spectrometry coupled to liquid chromatography (LC-MS) for detection and LC-MSⁿ for structural characterization. The oxidation products identified were keto, hydroxy, keto-hydroxy, and hydroperoxide derivatives for both di- and tripeptides. Among these, it was found that insertion of oxygen atoms occurred at histidine for HG and HGG, and both histidine and glycine for GH, GHG, and GGH. In addition, oxidation products formed by alkoxyl rearrangement reactions with cleavage of the peptide chain were also identified for GH, GHG, and GGH, corroborating hydrogen abstraction step in G residues. These findings were supported through the identification of radical intermediate species formed and trapped with 5,5-dimethyl-1-pyrrolidine-N-oxide (DMPO) spin trap. The observation of DMPO adducts bearing two spin trap molecules reinforced the abstraction of two hydrogen atoms from the same molecule.     

Bioinspired systems for metal-ion sensing: new emissive peptide probes based on benzo[d]oxazole derivatives and their gold and silica nanoparticles

Seven new bioinspired chemosensors (2-4 and 7-10) based on fluorescent peptides were synthesized and characterized by elemental analysis, (1)H and (13)C NMR, melting point, matrix-assisted laser desorption-ionization time-of-flight mass spectrometry (MALDI-TOF-MS), and IR and UV-vis absorption and emission spectroscopy. The interaction with transition- and post-transition-metal ions (Cu(2+), Ni(2+), Ag(+), Zn(2+), Cd(2+), Hg(2+), Pb(2+), and Fe(3+)) has been explored by absorption and fluorescence emission spectroscopy and MALDI-TOF-MS. The reported fluorescent peptide systems, introducing biological molecules in the skeleton of the probes, enhance their sensitivity and confer them strong potential for applications in biological fields. Gold and silica nanoparticles functionalized with these peptides were also obtained. All nanoparticles were characterized by dynamic light scattering, transmission electron microscopy, and UV-vis absorption and fluorescence spectroscopy. Stable gold nanoparticles (diameter 2-10 nm) bearing ligands 1 and 4 were obtained by common reductive synthesis. Commercial silica nanoparticles were decorated at their surface using compounds 8-10, linked through a silane spacer. The same chemosensors were also taken into aqueous solutions through their dispersion in the outer layer of silica core/poly(ethylene glycol) shell nanoparticles. In both cases, these complex nanoarchitectures behaved as new sensitive materials for Ag(+) and Hg(2+) in water. The possibility of using these species in this solvent is particularly valuable because the impact on human health of heavy- and transition-metal-ion pollution is very severe, and all analytical and diagnostics investigations involve a water environment.