Sebastián Noel Robledo

Qualitative and quantitative electroanalysis of synthetic phenolic antioxidant mixtures in edible oils based on their acid-base properties

A simple electroanalytical method using square wave voltammetry at a Pt band ultramicroelectrode to perform a qualitative and quantitative analysis of different synthetic antioxidant mixtures permitted by official regulations in edible oils is proposed. The methodology was based on the comparison of voltammetric signals obtained in acetonitrile+0.1M (C4H9)4NF6P with those recorded in the same reaction medium when different aliquots of (C4H9)4NOH were added to allow a qualitative differentiation between antioxidants. Firstly, studies on solutions prepared from commercial reagents were carried out. Then, the results obtained were transferred to the analysis of a real matrix, i.e., an edible olive oil. From real samples spiked with a known amount of different synthetic antioxidant mixtures, we could deduce the presence of these antioxidants by comparing results obtained in the neutral medium with those obtained after the successive addition of base. The standard addition method was used to quantify the individually spiked synthetic antioxidants in the real sample. Recovery percentages were between 88% and 118%. The reproducibility was 1.5%, 3.1%, 4.1% and 4.1% in ACN+0.1M TBAHFP and 1.5%, 4.6%, 6.6% and 2.5% in Bz/EtOH (1:2)+0.1M H2SO4 for TBHQ, BHA, BHT and PG, respectively. The repeatability was 1% for PG in both media. These parameters show a good system performance.

Development of an amperometric biosensor based on peroxidases to quantify citrinin in rice samples.

An amperometric biosensor based on horseradish peroxidase (EC1.11.1.7,H2O2-oxide-reductases) to determine the content of citrinin mycotoxin in rice samples is proposed by the first time. The method uses carbon paste electrodes filled up with multi-walled carbon nanotubes embedded in a mineral oil, horseradish peroxidase, and ferrocene as a redox mediator. The biosensor is covered externally with a dialysis membrane, which is fixed to the body side of the electrode with a Teflon laboratory film, and an O-ring. The reproducibility and the repeatability were of 7.0% and 3.0%, respectively, showing a very good biosensor performance. The calibration curve was linear in a concentration range from 1 to 11.6nM. The limits of detection and quantification were 0.25nM and 0.75nM, respectively. For comparison, the citrinin content in rice samples was also determined by fluorimetric measurements. A very good correlation was obtained between the electrochemical and spectrophotometric methods.

Quantitative determination of tocopherols in edible vegetable oils using electrochemical ultra-microsensors combined with chemometric tools

We have developed an electroanalytical method to quantify different isomers of tocopherols in edible vegetable oils. The method uses the square wave voltammetry on a carbon fiber disk ultramicroelectrode in benzene/ethanol+0.1 mol L(-1)H2SO4. Because the oxidation peaks of these natural antioxidants show an important overlapping, we have used two chemometric tools to obtain the multivariate calibration model. One method was the multivariate curve resolution-alternating least square (MCR-ALS), which assumes a linear behavior, i.e., the total signal is the sum of individual signals of components, and another nonlinear method such as artificial neuronal networks (ANNs). From the accuracy and precision analysis between nominal and estimated concentrations by both methods, we could infer that the ANNs method was a good model to quantify tocopherols in edible oil samples. Recovery percentages were between 94% and 99%. In addition, we found a difference of 1.4-6.8% between the total content of tocopherols in edible oil samples and the vitamin E content declared by the manufacturers.

Square wave voltammetry with multivariate calibration tools for determination of eugenol, carvacrol and thymol in honey.

The simultaneous determination of eugenol (EU), thymol (Ty) and carvacrol (CA) in honey samples, employing square wave voltammetry (SWV) and chemometrics tools, is informed for the first time. For this purpose, a glassy carbon electrode (GCE) was used as working electrode. The operating conditions and influencing parameters (involving several chemical and instrumental parameters) were first optimized by cyclic voltammetry (CV). Thus, the effects of the scan rate, pH and analyte concentration on the electrochemical response of the above mentioned molecules were studied. The results show that the electrochemical responses of the three compounds are very similar and that the voltammetric traces present a high degree of overlap under all the experimental conditions used in this study. Therefore, two chemometric tools were tested to obtain the multivariate calibration model. One method was the partial least squares regression (PLS-1), which assumes a linear behaviour. The other nonlinear method was an artificial neural network (ANN). In this last case we used a supervised, feed-forward network with Levenberg-Marquardt back propagation training. From the accuracies and precisions analysis between nominal and estimated concentrations calculated by using both methods, it was inferred that the ANN method was a good model to quantify EU, Ty and CA in honey samples. Recovery percentages were between 87% and 104%, except for two samples whose values were 136% and 72%. The analytical methodology was simple, fast and accurate.

Electrochemical Reduction of the Mycotoxin Citrinin at Bare and Modified with Multi-Walled Carbon Nanotubes Glassy Carbon Electrodes in a Non-Aqueous Reaction Medium

-1 tanto em eletrodo de carbono vitreo (GC) nao modificado quanto em eletrodo GC modificado com nanotubos de carbono de paredes multiplas foi investigado por voltametria ciclica e eletrolise a potencial controlado. Os resultados obtidos permitiram inferir um mecanismo de eletroreducao complexo com etapas quimicas e eletroquimicas, acopladas a reacao inicial de transferencia de eletron. Citrinina apresentou pico catodico unico que corresponde no minimo a duas etapas de reducao envolvendo um eletron e duas reacoes quimicas homogeneas, segundo mecanismo de autoprotonacao do tipo ECEC (transferencia de eletron acoplada com aceitacao de proton). As reacoes quimicas se originam nas transferencias de proton intermolecular do substrato para os seus produtos intermedios basicos de reducao, caracterizando um mecanismo de autoprotonacao. Analise cinetica por procedimentos de simulacao de resultados voltametricos permitiu uma caracterizacao completa do mecanismo de reducao heterogeneo de citrinina. Electrochemical reduction mechanism of citrinin in acetonitrile plus 0.1 mol L -1 (C4H9)4NClO4 at both bare and modified with multi-walled carbon nanotube glassy carbon (GC) electrodes was investigated by cyclic voltammetry and controlled potential electrolysis. Results allowed to infer a complex electroreduction mechanism with chemical and electrochemical steps coupled to the initial electron transfer reaction. Citrinin shows a single cathodic peak that corresponds at least to two electron reduction steps and two homogeneous chemical reactions, conforming to an ECEC (electron-transfer coupled with proton acceptance) self-protonation mechanism. The chemical reactions are originated in the intermolecular proton transfers from the substrate to its basic reduction intermediates, featuring a self-protonation mechanism. Kinetics analysis by simulation procedures of voltammetric results permitted a fully characterization of the mechanism of citrinin heterogeneous reduction.

Determination of kinetic parameters of the enzymatic reaction between soybean peroxidase and natural antioxidants using chemometric tools

The oxidation of eugenol, isoeugenol and vanillin natural antioxidants catalyzed by the soybean peroxidase enzyme was studied using uv-vis spectroscopy. An experimental design was used to optimize the different variables. The multivariate curve resolution method was used to obtain the profiles of antioxidant absorbances as a function of time due to uv-vis absorption bands of both antioxidants and the enzymatic reaction product/s show a strong overlap. From these results, apparent Michaelis-Menten constants as well as the kinetic parameters k1 and k3 involved in the catalytic cycle of peroxidases were calculated. The antioxidant apparent acidity constants were also determined at different pHs from uv-vis spectrophotometric measurements. Values of k1 were (0.6 ± 0.1) × 105 M-1 s-1, (2.0 ± 0.2) × 105 M-1 s-1 and (7.0 ± 0.5) × 106 M-1 s-1 and k3 (4.0 ± 0.2) × 105 M-1 s-1, (6.0 ± 0.6) × 105 M-1 s-1 and (6.0 ± 0.9) × 106 M-1 s-1 for eugenol, isoeugenol and vanillin, respectively.

Development of an electrochemical biosensor for the determination of triglycerides in serum samples based on a lipase/magnetite-chitosan/copper oxide nanoparticles/multiwalled carbon nanotubes/pectin composite

A very sensitive electrochemical biosensor to determine totals triglycerides (TGs) in serum samples has been developed. It is based on the electrochemical oxidation of glycerol at glassy carbon electrodes modified with magnetic nanoparticles bonded to lipase enzyme and copper oxide nanoparticles, both supported on a multiwalled carbon nanotubes/pectin dispersion. Glycerol is produced by enzymatic reaction between the TGs present in samples and the lipase immobilized. The quantification of triglycerides was performed by amperometric measurements. The proposed electrochemical biosensor improves the performance of others methods developed for the TGs quantification. The determination of TGs does not need a pretreatment of serum samples. The PLS-1 algorithm was used for the quantification of TGs. According to this algorithm, the of detection and quantification limits were from 3.2 × 10-3 g L-1 to 3.6 × 10-3 g L-1, and from 9.6 × 10-3 to 1.1 × 10-2 g L-1, respectively. The sensitivity was 1.64 × 10-6 A L g-1. The proposed electrochemical biosensor exhibited a very good performance, a stability of 20 days, very good reproducibility and repeatability, and it is presented as a very good alternative for the determination of TGs in human serum clinical samples.