Biosensors for monitoring the isothermal breakdown kinetics of peanut oil heated at 180°C. Comparison with results obtained for extra virgin olive oil.

The present research was devoted to studying the kinetics of the artificial rancidification of peanut oil (PO) when a sample of this oil was isothermally heated at 180°C in an air stream. The formation of radical species due to heating was evaluated using a radical index whose value was determined using a biosensor method based on a superoxide dismutase (SOD), while the increasing toxicity was monitored using a suitable toxicity measuring probe based on the Clark electrode and immobilized yeast cells. An extra virgin olive oil was isothermally rancidified under the same experimental conditions and the corresponding data were used for the purpose of comparison. Both the so-called "model-fitting" and the classical kinetic methods were applied to the isothermal process biosensor data in order to obtain the kinetic constant rate value at 180°C.

Organophosphorus and carbamate pesticide analysis using an inhibition tyrosinase organic phase enzyme sensor; comparison by butyrylcholinesterase+choline oxidase opee and application to natural waters.

Recent research performed in our laboratory (using a butyrylcholinesterase+choline oxidase enzyme electrode) suggested the validity of the biosensor approach using enzyme inhibition OPEEs (i.e. enzyme electrodes working in organic phase) in the case of organophosphorus and carbamate pesticides, which are poorly soluble in aqueous solutions. Since these pesticides are generally much more soluble in chloroform than in water, the present research aimed at analysing this class of pesticides using a tyrosinase inhibition OPEE operating in water-saturated chloroform medium. The tyrosinase biosensor was assembled using an oxygen amperometric transducer coupled to the tyrosinase enzyme, immobilized in kappa-carrageenan gel. Lastly a detailed comparison between the inhibition monoenzymatic tyrosinase and inhibition bienzymatic (butyrylcholinesterase+choline oxidase) OPEEs was performed and discussed in this work.

Salicylic acid determination in cow urine and drugs using a bienzymatic sensor.

An enzymatic biosensor was developed for salicylic acid (salicylate ion) determined using a Clark type gas diffusion electrode and two enzymes (tyrosinase and salicylate hydroxylase) entrapped in a cellulose triacetate membrane. After optimization, the method was applied to the determination of salicylic acid in cow urine. Relatively good recoveries were achieved, between about 83% and 109%, using the calibration curve, and acceptable precision (R.S.D. about 8%). The method is now being tested for the determination of salicylic acid contained in commercially available drug specialities or galenic products. So far agreement with nominal values has been found to be between 75% and 110% with a R.S.D. of less than 8%.

Tyrosinase inhibition organic phase biosensor for triazinic and benzotriazinic pesticide analysis (part two).

Several triazine pesticides, such as atrazine, are much more soluble in several organic solvents, such as chloroform, than in water. Our recent research was aimed at analyzing this class of pesticides using tyrosinase OPEE (organic phase enzyme electrodes), exploiting their inhibiting action on the tyrosinase enzyme when operating in water-saturated chloroform medium. In this work we studied the response of a tyrosinase inhibition enzyme sensor to several triazinic (simazine, propazine, terbuthylazine) and benzotriazinic (azinphos-ethyl and azinphos-methyl) pesticides (LOD=0.5x10(-9) mol l(-1)). Recovery trials were also performed in vegetal matrixes (corn, barley, lentils). Lastly, the effect of the solvent (chloroform or water) on the inhibition process was investigated via Hill's equation and the diffusion of analyte from the solvent to the enzyme membrane.

Thermal analysis study of the interactions between acetaminophen and excipients in solid dosage forms and in some binary mixtures.

Thermogravimetry (TG) and differential scanning calorimetry (DSC) were used to assess the compatibility between acetaminophen (Ac) and some excipients (polyvinylpyrrolidone (P), magnesium stearate (M), citric acid (C), aspartame (As), mannitol (Mn), cellulose (Cll) and starch (S)) in several of the more commercially available pharmaceutical formulations and in solid binary mixtures. The present study compared thermodynamic data on acetaminophen melting and vaporization processes of pure acetaminophen with those found for several solid mixtures and in some commercially available acetaminophen-based dosage forms. Appreciable modifications occur only for solid mixtures with high content of excipient. Acetaminophen-based dosage forms and its solid binary mixtures usually show "additivity" of calorimetric peaks number of pure components in their calorimetric curve profiles, thus revealing a good thermoanalytical compatibility between acetaminophen and the excipients examined, except for samples containing appreciable content of mannitol.

Further applications of a new biosensor method for dating cellulosic finds.

Using a cellulosic material dating method recently proposed by three of the authors of the present article further applications to real samples are discussed. In the first instance, to wood samples, that is, to a type of sample for which the method was specifically developed but with the samples differing widely in age, and then to textile or paper samples. Of course in the latter two cases the results obtained are still quite preliminary, above all because of the difficulty of procuring certainly dated samples of this type.

Antioxidant capacity of the algae using a biosensor method.

Three different methods, i.e. a biosensor method, a voltammetric method and a spectrophotometric method, have been used to evaluate the total antioxidant capacity of certain types of algae. In the final evaluation of the data also the variation in time of the antioxidant capacity of cultivated algae was considered and some experimental factors, such as the use of different solvent mixtures to extract the antioxidant substances contained in the algae, were discussed.

Comparison of fluorimetric, voltammetric and biosensor methods for the determination of total antioxidant capacity of drug products containing acetylsalicylic acid.

Antioxidant capacity of several drug specialities containing as mean component acetylsalicylic acid were experimentally evaluated using an enzymatic electrode, recently developed by the present authors, based on superoxide dismutase (SOD) enzyme. The precision of this method of analysis was found to be good (for drug samples RSD < or = 5%). The results were also compared with those ones by a traditional spectrofluorimetric method and by two other methods, respectively, based on cyclic and pulsed voltammetry, recently trialled by the present authors.

Biosensors for determination of total antioxidant capacity of phytotherapeutic integrators: comparison with other spectrophotometric, fluorimetric and voltammetric methods.

Enzymatic electrodes based on superoxide dismutase (SOD) biosensors, working both in aqueous and non-aqueous solutions, recently developed by the present authors, were used to experimentally evaluate the antioxidant capacity of several phytotherapeutic diet integrators. The precision of this method of analysis was found to be reasonable (R. S. D. < or = 10%). The results were also compared with those obtained using a traditional spectrophotometric method as well as a spectrofluorimetric method described in literature. Lastly, the comparison was extended to another method based on cyclic voltammetry currently being trialled by the present authors.

Determination of choline containing phospholipids in serum, bile and amniotic fluids by the derivative enzymatic-spectrophotometric method.

A comparison was performed of the analytical results obtained by applying a well-known enzymatic-spectrophotometric method for the determination of choline containing phospholipids in biological human fluids (sera, bile and amniotic fluids) following the standard procedure method, or else using the first or second derivative methods. In terms of result reproducibility the comparison was extended to include also a biosensor-based method developed in recent years by the present authors. Some advantages are associated with using the first derivative method in the case of serum samples containing blood traces, especially via the 'Gran's plot' method.

Biosensors for determination of total and natural antioxidant capacity of red and white wines: comparison with other spectrophotometric and fluorimetric methods.

Research was carried out to experimentally evaluate the antioxidant capacity of several red and white wines using a superoxide dismutase (SOD) biosensor recently developed by the present authors. Measurements were performed by comparing the biosensor response to increasing concentration of the superoxide radical produced in solution by the xanthine/xanthine oxidase system, both in the presence and absence of the test sample.The results were compared with those of two traditional spectrophotometric methods and of a spectrofluorimetric method described in literature.Lastly, also the polyphenol, sulfite and ascorbic acid contents of the different wine samples examined were measured using a tyrosinase biosensor, a sulfite oxidase biosensor and an ascorbate oxidase biosensor, respectively.

Determination of the antioxidant capacity of samples of different types of tea, or of beverages based on tea or other herbal products, using a superoxide dismutase biosensor.

Research was performed to experimentally evaluate the antioxidant capacity of different plant products sold by herbalists (ginger, dog rose, ginseng and camomile) and of several types of tea (ordinary tea, green tea, detheinated tea, lemon and peach flavoured tea) using a superoxide dismutase (SOD) biosensor recently developed by the present authors. Measurements were carried out by comparing biosensor response to the superoxide radical produced in solution using the xanthine-xanthine oxidase system, both in the presence and absence of the antioxidant sample considered. Precision of antioxidant capacity measures for herbal products and for non diluted samples was good, generally with a R.S.D.%< or =10% and a LOD value about 0.1 for relative antioxidant capacity. Also a "pool" of polyphenols from different tea samples was measured using a tyrosinase biosensor (LOD approximately 2 microM).

Determination of hydrogen peroxide in disinfectant solutions using a biosensor with two antagonist enzymes.

The development and characterisation of a new biosensor for hydroperoxides is described, which is obtained by combining an oxygen gas diffusion amperometric electrode and two immobilized enzymes (peroxidase and tyrosinase) working in parallel and competing for the same substrate (catechol). The response of the biosensor to several hydroperoxides was investigated (LOD=0.5.10(-4) M for hydrogen peroxide). It was experimentally found that the biosensor is able to respond also to aqueous solutions of ionic peroxides (LOD=0.2.10(-4) M for potassium peroxidisulphate). The biosensor was applied to the determination of the hydrogen peroxide content of pharmaceutical products, i.e. aqueous disinfectant solutions (RSD% < or =0.5; recoveries by standard addition method between 96.0 and 98.5%).

Determination of antioxidant properties of aromatic herbs, olives and fresh fruit using an enzymatic sensor.

The aim was to experimentally evaluate the antioxidant capacity of different fresh aromatic herbs (field balm, marjoram, parsley, rosemary, sage, sweet basil), several varieties of olives from Central Italy ('Carboncello', 'Rosciolo', 'Olivastro', 'Coratello', 'Leccino', 'Frantoio') and several types of fresh fruit (apple, apricot, banana, cherry, fig, grape, medlar, melon, peach, pear, pineapple, plum, water melon, yellow plum) using a superoxide dismutase (SOD) biosensor developed by the present authors. Measurements were carried out by comparing the biosensor response to the concentration of superoxide radical produced in solution using a xanthine/xanthine oxidase system in the presence and absence of the antioxidant sample considered. Tests carried out on different samples of fruit and aromatic herbs showed that the homogenised samples had better antioxidant properties than the centrifuged ones (obtained by centrifuging the homogenate), which sometimes gave extremely low antioxidant capacity values. The reliability of the proposed method was checked by comparing the trend of some experimental results found using the SOD biosensor with those reported in the literature obtained using the classic (ORAC) method. The precision of this method of analysis was found to be good for samples of aromatic herbs (RSD%

Kinetic analysis of thermal decomposition for penicillin sodium salts: model-fitting and model-free methods.

A kinetic study on decomposition processes of some penicillin salts was carried out. Both isothermal and dynamic thermogravimetric curves were used. As expected by their complex structures, several steps with different energies were involved in decomposition processes. Model-fitting and -free kinetic approaches were applied to nonisothermal and isothermal data. The kinetic triplet (f(alpha),A and E(a)) related to model-fitting method that defines a single step reaction resulted to be at variance with the multi-step nature of salts-decomposition. The model-free approach represented by the isothermal and nonisothermal isoconversional methods, gave different dependencies of the activation energy on the extent of conversion. The complex nature of the multi-step process of the studied compounds was more easily revealed using a broader temperature range in nonisothermal isoconversional method. The failure in the model-fitting method did not allow calculating shelf life and half-life times.

Determination of lecithin drug specialties and diet integrators; by means of first or second derivative enzymatic-spectrophotometric analysis.

The possibility has been investigated of applying derivative analysis to a classical enzymatic-spectrophotometric method for lecithin determination for the purpose of developing an analytical direct method that does not require long pretreatment of the test sample even in the case of turbid samples. Several samples of drugs and food integrators containing lecithin were thus tested using both the standard and the derivative method and then comparing the results obtained. The RSD% values of measurements on real (food or drugs) samples were always <2.5. Using a first derivative spectrophotometric method, average recovery was always between 102 and 105%.

Superoxide dismutase biosensors working in non-aqueous solvent.

Enzymatic electrodes based on superoxide dismutase enzyme were developed. Using the superoxide dismutase enzyme sensor assembled according to the classical model, poor results were obtained. Results were improved by adopting a new way of assembling the biosensor using a cellulose triacetate layer in which the SOD enzyme is entrapped and sandwiched between two gas-permeable membranes, or using a kappa-carrageenan gel layer entrapping the enzyme, sandwiched between an external gas permeable membrane and an internal cellulose acetate membrane, coupled in each case to the oxygen amperometric transducer. Results obtained by applying the newly developed biosensor to assaying hydrophobic compounds showing radical scavenging properties, operating in dimethylsulfoxide, were also satisfactory.

Evaluation of radical scavenging properties of several plants, fresh or from a herbalist's, using a superoxide dismutase biosensor.

The experimental evaluation of properties of defence against free radicals represents an extremely interesting heuristic and applicational objective. Research was carried out to experimentally evaluate the scavenging properties of several fruits and plants, fresh or from a herbalist's using an amperometric superoxide dismutase (SOD) biosensor recently developed by the present authors. The superoxide radical was produced in solution using the xanthine/xanthine oxidase system; measurements were carried out by comparing biosensor response to superoxide radical both in the presence and absence of the sample considered.

Thermal stability of disodium and calcium phosphomycin and the effects of the excipients evaluated by thermal analysis.

In the present study, Thermogravimetry (TG) and Differential Scanning Calorimetry (DSC) are simultaneously applied to determine the thermal properties of two antibiotic salts, disodium and calcium phosphomycin, used either pure or in association with several excipients. This study was carried out kinetically as well by mathematical elaboration of the TG curves performed according to an isothermal procedure applied at different fixed temperatures. Kinetic parameters showing agreement with those produced by the isothermal method were also obtained by means of a non-isothermal method using a dynamic TG curve alone. The main aims of the work were to provide reliable kinetic parameters (kinetic constant k, activation energy E(a) and pre-exponential factor A) to evaluate the thermal stability of phosphomycin salts in the presence and absence of the excipients generally contained in the phosphomycin-based pharmaceutical forms available on the market, and to obtain information concerning compatibility towards the active components. These kinetic parameters were then used to extrapolate shelf-life and half-life values at room temperature for pure active components in the solid state and for their pharmaceutical derivatives.

Identification of different types of imperial age marble finds using instrumental chemical analysis and pattern recognition analysis.

A physical-chemical characterisation of several marbles frequently used in ancient times for artistic or decorative purposes was performed in support the work of historians and restorers. The data were obtained using several different types of instrumental chemical methods (Thermogravimetry, Differential Thermal Analysis, X-ray Diffractometry and ICP Plasma Emission Spectroscopy) and have been summarised in short tables. The data have already proved useful in the identification of a small number of finds (statues or architectonic elements) from Ancient Rome (Imperial Age, 2nd-3nd cent. A.D.) for the purpose of which also a well-known pattern recognition analysis software package was used for data processing. In practice, the research showed that an organised set of chemical data obtained using several modern instrumental methods can provide a valid basis for the reasonably rapid and reliable identification of the type of marble used to make artistic artifacts that have not yet been subjected to typological study.