Determination of salicylate in blood serum using an amperometric biosensor based on salicylate hydroxylase immobilized in a polypyrrole-glutaraldehyde matrix

The use of an amperometric biosensor for the salicylate determination in blood serum is described. The biosensor is based on salicylate hydroxylase (EC 1.14.13.1) electropolymerized onto a glassy carbon-working electrode with polypyrrole and glutaraldehyde, to improve the biosensor lifetime. The hexacyanoferrate (II) was also incorporated to work as a redox mediator to minimize possible interferences. The salicylate is enzymatically converted to catechol, which is monitored amperometrically by its electrooxidation at+0.170 V versus SCE (saturated calomel electrode). Salicylate determination was carried out maintaining the ratio between beta-NADH and salicylate at 4:1 (30 degrees C). The amperometric response of the biosensor was linearly proportional to the salicylate concentration between 2.3x10(-6) and 1.4x10(-5) mol l(-1), in 0.1 mol l(-1) phosphate buffer (pH 7.8), containing 0.1 mol l(-1) KCl and 5.0x10(-4) mol l(-1) Na(2)H(2)EDTA, as supporting electrolyte. The recovery studies, in the presence of several interfering compounds, showed recoveries between 96.4 and 104.8%. The useful lifetime of the biosensor in the concentration range evaluated was at least 40 days, in continuous use. Blood serum samples analyzed by this biosensor showed a good correlation compared to the spectrophotometric method (Trinder) used as reference, presenting relative deviations lower than 7.0%.     

Trace elements in a profile of the unsaturated zone of the São Paulo Basin

This study comprises Tertiary sediments (clays, silts, and fine clayey sands) from the unsaturated zone of the Experimental Station in the University of São Paulo, taken from a nine meters depth profile; ten samples were separated in bulk and <53 m fractions. Chemical composition was determined by INAA to ascertain the distribution of trace elements throughout the sediment deposition, and to establish the background parameters, useful for mobilization studies of such elements after industrial waste disposal.     

Spectrophotometric flow injection determination of total phosphorus in beer using on-line UV/thermal induced digestion

A flow injection system for the automatic determination of total phosphorus in beer is described. The developed manifold uses a two-stage photooxidation/thermal digestion procedure together with oxidizing and hydrolyzing reagents to convert all forms of phosphorus compounds to orthophosphate. Polyphosphates are hydrolyzed by acid and heat, and organo-phosphates are digested by UV-catalyzed peroxodisulfate oxidation. The orthophosphate formed is then spectrophotometrically determined by the phosphomolybdenum blue reaction, using stannous chloride as reducing agent. The results obtained for a set of 19 beer samples (with concentrations from 120 to 735 mg P/L) were in good agreement with the reference method, the maximum relative deviation found being 4.7%. Relative standard deviations for ten consecutive determinations were lower than 1.5%, and a detection limit of 1 mg P/L was achieved.     

Natural radionuclide-free phosphoric acid production

High-uranium phosphate rock from Itataia, Brazil, was milled for wet-process phosphoric acid production using the dihydrate method. Uranium contained in the phosphoric acid was recovered by solvent extraction. The distribution of long-lived natural radionuclides of the ²³⁸U and ²³²Th decay series involved in these operations was evaluated. ²²⁶Ra, ²²⁸Ra and ²¹⁰Pb were found to predominate in the phosphogypsum, while ²²⁸Th, ²³⁰Th and ²³²Th in the uranium-free phosphoric acid. Thorium is removed from the phosphoric acid by solvent extraction to produce a NORM-free phosphoric acid.     

Characterization of Lippia sidoides oil extract-b-cyclodextrin complexes using combined thermoanalytical techniques

Thermoanalytical techniques, being rapid and un-expensive have been used for the investigation of the cyclodextrin inclusion complexes for three decades. The conventional thermoanalytical techniques (TG and DTA/DSC) follow the thermal properties of the uncomplexed compounds. Consequently, the inclusion complex formation as well as the liberation of the entrapped guest cannot be followed. Monitoring the products of the thermal fragmentation of parent cyclodextrin and the included molecule(s), applying TG-MS combined technique provides evidence concerning the inclusion complex formation, and besides, gives selective signal to follow the decomposition of the cyclodextrin inclusion complexes. b-cyclodextrin inclusion complexes of Thymol and Lippia sidoides Cham essential oil extract have been prepared and investigated using conventional and combined (TG-MS) thermoanalytical techniques. The evolved gas analysis proved the inclusion complex formation between the host and guests. By the evaluation of the experimental results the elaboration of the entrapped guests from the cyclodextrin cavity could be followed. This revised version was published online in July 2006 with corrections to the Cover Date.     

WAXS study on orthorhombic crystalline phases in hot-drawn high density polyethylene: POLYMER CHARACTERISATION

The temperature dependence in the range 23–135°C of hot-drawn HDPE plates maintained free from contraction was investigated by WAXS and DSC. A specially designed cylindrical chamber was used for the WAXS patterns. The results confirm the coexistence of two populations of orthorhombic crystals which have different lattice and thermal behaviour. The crystal lattice parameter a shows expansion, while b exhibits expansion up to 90°C, followed by contraction as the temperature is raised to melting. On the other hand, the c parameter is practically constant.     

Atomistic Modeling of Silica Based Sol-Gel Processes

Density Functional Theory is used to study water, methanol, ethanol, TMOS, and TEOS molecules and the most important silica clusters participating in sol-gel processes. Calculated bond lengths, bond angles and electric dipole moments compare well with experimental data. The energy of these molecules is reported and used to discuss the energetics of the hydrolysis and condensation reactions. Molecular Dynamics is employed to simulate liquid water, methanol, ethanol, TMOS, TEOS and experimental sol-gel solutions. Calculated densities and enthalpies of vaporisation compare well with experimental data. Preliminary results are presented for MD simulations of sol-gel solutions.     

Thermogravimetric and differential scanning calorimetric investigations of manganese–glycine interactions

Thermogravimetric (t.g.) and differential scanning calorimetric (d.s.c.) data have been used to study metal–amino acid interactions in adducts of general formula MnCl₂ · ngly (gly = glycine, n = 0.7, 2.0, 4.0 and 5.0). All the prepared adducts exhibit only a one step mass loss associated with the release of glycine molecules, except for the 0.7gly adduct, which exhibits two glycine mass loss steps. From d.s.c. data, the enthalpy values associated with the glycine mass loss can be calculated: MnCl₂ · 0.7gly = 409 and 399 kJ mol^–1, MnCl₂ · 2.0gly = 216 kJ mol^–1, MnCl₂ · 4.0gly = 326 kJ mol^–1 and MnCl₂ · 5.0gly = 423 kJ mol^–1, respectively. The enthalpy associated with the ligand loss, plotted as function of the number of ligands for the n = 2.0, 4.0 and 5.0 adducts, gave a linear correlation, fitting the equation: H (ligand loss)/kJ mol^–1 = 67 × (number of ligands, n) + 76. A similar result was achieved when the enthalpy associated with the ligand loss was plotted as a function of the _a(COO^–) bands associated with the coordination through the carboxylate group, 1571, 1575 and 1577 cm^–1, respectively, for the n = 2.0, 4.0 and 5.0 adducts, giving the equation H (ligand loss) /kJ mol^–1 = 33.5 × _a(COO^–) /cm^–1 – 52418.5. This simple equation provides evidence for the enthalpy associated with the ligand loss being very closely related to the electronic density associated with the metal–amino acid bonds.     

Automatic flow procedure for the determination of ethanol in wine exploiting multicommutation and enzymatic reaction with detection by chemiluminescence

An automatic procedure for the determination of ethanol in wines using a flow system based on multicommutation and enzymatic reaction is described. Alcohol oxidase was immobilized on aminopropyl glass beads and packed in an acrylic column. The peroxide due to enzymatic reaction with ethanol reacted with luminol and generated the chemiluminescence radiation that was monitored by using a laboratory-made detector based on photodiodes. The system manifold comprised a set of 3-way solenoid valves controlled by a microcomputer furnished with electronic interfaces, which ran on software written in Quick BASIC 4.5 to provide facilities to perform on-line sample dilution, reagent addition, and data acquisition. After system parameters optimization, ethanol samples were processed without prior pretreatment. The following suitable features were achieved: linear response ranging from 2.5 to 25% (v/v) ethanol, relative standard deviation of 1.8% (n = 10), detection limit of 0.3% (v/v) ethanol, sampling rate of 23 determinations per hour, and low reagent consumption of 0.23 mg luminol and 7 mg hexacyanoferrate (III) per determination. When the results were compared with those obtained using the AOAC Official Method, no significant difference at the 90% confidence level was observed.     

Reagentless biosensor for isocitrate using one step modified Pt-Ir microelectrode

The Pt-Ir microelectrode modified through one step electropolymerization is proposed for the isocitrate amperometric biosensor construction. The enzyme (isocitrate dehydrogenase-ICDH), coenzyme (NADP(+)) and mediator (Meldola's Blue) were immobilized onto the microelectrode surface in one step from a PIPES buffer solution containing pyrrole. The optimized experimental conditions were 25 cycles of cyclic voltammetric in a solution containing 3.58 10(-5) mol l(-1) of mediator, 3.51 10(-4) mol l(-1) of coenzyme and 2.68 U ml(-1) of enzyme. In contrast to the biosensor for isocitrate reported in literature, just one enzyme was immobilized and no coenzyme addition in the solution of analysis was necessary. Catalytic currents were proportional to the isocitrate concentration between 7.7 10(-6) and 1.04 10(-4) mol l(-1), showing good repeatability. The detection limit of the proposed biosensor was 3.50 10(-6) mol l(-1), the response time was lower than 20 s, the lifetime was about 30 determinations and no significant interference of sugars and citric acid was verified. Orange juice samples were analysed by both methodology biosensor and spectrophotometric commercial kit, and the obtained results presented a good correlation. The data demonstrated that the developed biosensor is suitable for isocitrate determination in orange juice without matrix interferences.