Multivariate Optimization of Electrochemical Biosensors for the Determination of Compounds Related to Food Safety-A Review.

We summarize the application of multivariate optimization for the construction of electrochemical biosensors. The introduction provides an overview of electrochemical biosensing, which is classified into catalytic-based and affinity-based biosensors, and discusses the most recent published works in each category. We then explore the relevance of electrochemical biosensors for food safety analysis, taking into account analytes of different natures. Then, we describe the chemometrics tools used in the construction of electrochemical sensors/biosensors and provide examples from the literature. Finally, we carefully discuss the construction of electrochemical biosensors based on design of experiments, including the advantages, disadvantages, and future perspectives of using multivariate optimization in this field. The discussion section offers a comprehensive analysis of these topics.

Development of a voltammetric electronic tongue for the simultaneous determination of synthetic antioxidants in edible olive oils.

A voltammetric electronic tongue (E-tongue) is "a multisensor system, which consists of a number of low-selective sensors and uses advanced mathematical procedures for signal processing based on pattern recognition and/or data multivariate analysis such as artificial neural networks (ANNs), principal component analysis (PCA), among others". Thus, E-tongues in combination with chemometrics tools result in more accurate and selective analytical methods. In this work, we report results of a simple and reliable electroanalytical method to determine butyl hydroxyanisole (BHA), butyl hydroxytoluene (BHT) and propyl gallate (PG) in edible olive oils (EOO). Therefore, the square wave voltammetry (SWV) was used on platinum and carbon fiber disk ultramicroelectrodes (E-tongue configuration) combined with chemometrics tools to perform these studies. On the other hand, two data fusion strategies were used in order to combine electrochemical data obtained for each working electrode in the E-tongue: low-level data fusion (LLDF) and mid-level data fusion (MLDF). In addition, to reduce the dimensionality of the dataset in MLDF, the discrete wavelet transform (DWT) was used. Finally, to assert the predictive capability of the method for BHA, BHT, and PG determination in real samples, a recovery study for the antioxidants in EOO samples was performed, demonstrating the analytical accuracy of the proposed method. Moreover, from the comparison between the proposed electrochemical method with the AOAC reference method and others found in the literature in terms of the quality of the model (REP %) and the percent recovery assays (%) in different samples, our results were better than other reported previously for the simultaneous determination of BHA, BHT, and PG in real samples. Moreover, the percent recovery assays obtained with the proposed electrochemical method were in good agreement with those obtained by the chromatographic method.

Development of an enzymatic biosensor to determine eugenol in dental samples.

A GCE/CRGO-ßCD's/ADA-SPE/AuNPs biosensor was successfully developed to determine eugenol in dental samples. The optimal conditions to construct the biosensor were obtained from an experimental design based on the response surfaces methodology. The GCE/CRGO-ßCD/ADA-SPE/AuNPs biosensor exhibited a very good analytical performance for the quantification of eugenol. Thus, it shows a linear range between 1.3 × 10-8 and 1 × 10-5 mol L-1, with a sensitivity of (5.3 ± 0.3) x 10-3 A mol-1 L. The limits of detection and quantification were 4 × 10-9 mol L-1 and 1.3 × 10-8 mol L-1, respectively. Biosensors had an intraday and inter day reproducibility of 5% and 8%, respectively. The repeatability was of 3%, and the stability was 21 days (a decrease of 30% in current responses was observed after the fourth week). Recovery studies were performed in order to validate the proposed method. Recovery percentages were between 94 and 108%. A value of the apparent Michaellis-Menten constant, KMapp, of 3.1 × 10-6 mol L-1 was obtained using both Lineweaver-Burk and Eadi-Hofstee methods.

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 absorbance's 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 pH's from uv-vis spectrophotometric measurements. Values of k1 were (0.6 ±â€¯0.1) × 105 M-1 s-1, (2.0 ±â€¯0.2) × 105 M-1 s-1 and (7.0 ±â€¯0.5) × 106 M-1 s-1 and k3 (4.0 ±â€¯0.2) × 105 M-1 s-1, (6.0 ±â€¯0.6) × 105 M-1 s-1 and (6.0 ±â€¯0.9) × 106 M-1 s-1 for eugenol, isoeugenol and vanillin, respectively.

Sterigmatocystin: A mycotoxin to be seriously considered.

Sterigmatocystin is a carcinogenic compound that affects several species of crops and several species of experimental animals. The sterigmatocystin biosynthetic pathway is the best known and most studied. The International Agency for Research on Cancer classifies sterigmatocystin in the Group 2B. Three groups of analytical methods to determine sterigmatocystin in food can be found: chromatographic, ELISA immunoassays and chemical sensors. In addition, sterigmatocystin is a precursor of aflatoxin B1 in those cases where cereals and/or food are contaminated with fungi capable of producing aflatoxins. Chemical structures of sterigmatocystin and aflatoxin B1 are similar. These mycotoxins are pathogens of animals and cereals, producing a major economic impact on biotechnology and agricultural and food industries. This review summarizes different aspects related to sterigmatocystin such as its biosynthesis, toxicological studies and analytical methods for its determination.

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.

Citrinin (CIT) determination in rice samples using a micro fluidic electrochemical immunosensor.

The development of an electrochemical immunosensor incorporated in a micro fluidic cell for quantification of citrinin (CIT) mycotoxin in rice samples is described for the first time. Both CIT present in rice samples and immobilized on a gold surface electrodeposited on a glassy carbon (GC) electrode modified with a cysteamine self-assembled monolayer were allowed to compete for the monoclonal mouse anti-CIT IgG antibody (mAb-CIT) present in solution. Then, an excess of rabbit anti mouse IgG (H+L) labelled with the horseradish peroxidase (secAb-HRP) was added, which reacts with the mAb-CIT which is in the immuno-complex formed with the immobilized CIT on the electrode surface. The HPR, in the presence of hydrogen peroxide (H(2)O(2)) catalyzes the oxidation of catechol (H(2)Q) whose back electrochemical reduction was detected on a GC electrode at -0.15 V vs Ag/AgCl by amperometric measurements. The current measured is proportional to the enzymatic activity and inversely proportional to the amount of CIT present in the rice samples. This immunosensor for CIT showed a range of work between 0.5 and 50 ng mL(-1). The detection (LOD) and the quantification (LOQ) limits were 0.1 and 0.5 ng mL(-1), respectively. The coefficients of variation intra- and inter-assays were less than 6%. The electrochemical detection could be done within 2 min and the assay total time was 45 min. The immunosensor was provided to undertake at least 80 determinations for different samples with a minimum previous pre-treatment. Our electrochemical immunosensor showed a higher sensitivity and reduced analysis time compared to other analytical methods such as chromatographic methods. This methodology is fast, selective and very sensitive. Thus, the immunosensor showed to be a very useful tool to determine CIT in samples of cereals, mainly rice samples.

An amperometric biosensor based on peroxidases from Brassica napus for the determination of the total polyphenolic content in wine and tea samples.

An amperometric biosensor based on peroxidases from Brassica napus hairy roots (PBHR) used to determine the total polyphenolic content in wine and tea samples is proposed by the first time. The method employs carbon paste (CP) electrodes filled up with PBHR, ferrocene (Fc), and multi-walled carbon nanotubes embedded in a mineral oil (MWCNT+MO) at a given composition (PBHR-Fc-MWCNT+MO). The biosensor was covered externally with a dialysis membrane, which was fixed at the electrode body side part with a Teflon laboratory film and an O-ring. Calibration curves obtained from steady-state currents as a function of the concentration of a polyphenolic standard reference compound such as t-resveratrol (t-Res) or caffeic acid (CA) were then used to estimate the total polyphenolic content in real samples. The reproducibility and the repeatability were of 7.0% and 4.1% for t-Res (8.4% and 5.2% for CA), respectively, showing a good biosensor performance. The calibration curves were linear in a concentration range from 0.05 to 52 mg L(-1) and 0.06 to 69 mg L(-1) for t-Res and CA, respectively. The lowest polyphenolic compound concentration values measured experimentally for a signal to noise ratio of 3:1 were 0.023 mg L(-1) and 0.020 mg L(-1) for t-Res and CA, respectively.