Acrylamide in food products and the role of electrochemical biosensors in its detection: a comprehensive review.

In this review, the mechanisms of acrylamide formation in food, along with aspects related to its toxicity and associated consumption risks, are investigated, highlighting the potential impact on human health. European regulations regarding acrylamide content in food products are also addressed, emphasizing the importance of monitoring and detecting this substance in nutrition, by public health protection measures. The primary objective of the research is to explore and analyze innovative methods for detecting acrylamide in food, with a particular focus on electrochemical biosensors. This research direction is motivated by the need to develop rapid, sensitive, and efficient monitoring techniques for this toxic compound in food products, considering the associated consumption risks. The research has revealed several significant results. Studies have shown that electrochemical biosensors based on hemoglobin exhibited increased sensitivity and low detection limits, capable of detecting very low concentrations of acrylamide in processed foods. Additionally, it has been found that the use of functionalized nanomaterials, such as carbon nanotubes and gold nanoparticles, has led to the improvement of electrochemical biosensor performance in acrylamide detection. The integration of these technological innovations and functionalization strategies has enhanced the sensitivity, specificity, and stability of biosensors in measuring acrylamides. Thus, the results of this research offer promising perspectives for the development of precise and efficient methods for monitoring acrylamides in food, contributing to the improvement of food quality control and the protection of consumer health.

Electrochemical Detection of Cd2+, Pb2+, Cu2+ and Hg2+ with Sensors Based on Carbonaceous Nanomaterials and Fe3O4 Nanoparticles.

Two electrochemical sensors were developed in this study, with their preparations using two nanomaterials with remarkable properties, namely, carbon nanofibers (CNF) modified with Fe3O4 nanoparticles and multilayer carbon nanotubes (MWCNT) modified with Fe3O4 nanoparticles. The modified screen-printed electrodes (SPE) were thus named SPE/Fe3O4-CNF and SPE/Fe3O4-MWCNT and were used for the simultaneous detection of heavy metals (Cd2+, Pb2+, Cu2+ and Hg2+). The sensors have been spectrometrically and electrochemically characterized. The limits of detection of the SPE/Fe3O4-CNF sensor were 0.0615 µM, 0.0154 µM, 0.0320 µM and 0.0148 µM for Cd2+, Pb2+, Cu2+ and Hg2+, respectively, and 0.2719 µM, 0.3187 µM, 1.0436 µM and 0.9076 µM in the case of the SPE/ Fe3O4-MWCNT sensor (following optimization of the working parameters). Due to the modifying material, the results showed superior performance for the SPE/Fe3O4-CNF sensor, with extended linearity ranges and detection limits in the nanomolar range, compared to those of the SPE/Fe3O4-MWCNT sensor. For the quantification of heavy metal ions Cd2+, Pb2+, Cu2+ and Hg2+ with the SPE/Fe3O4-CNF sensor from real samples, the standard addition method was used because the values obtained for the recovery tests were good. The analysis of surface water samples from the Danube River has shown that the obtained values are significantly lower than the maximum limits allowed according to the quality standards specified by the United States Environmental Protection Agency (USEPA) and those of the World Health Organization (WHO). This research provides a complementary method based on electrochemical sensors for in situ monitoring of surface water quality, representing a useful tool in environmental studies.

Development of an Innovative Biosensor Based on Graphene/PEDOT/Tyrosinase for the Detection of Phenolic Compounds in River Waters.

Phenolic compounds, originating from industrial, agricultural, and urban sources, can leach into flowing waters, adversely affecting aquatic life, biodiversity, and compromising the quality of drinking water, posing potential health hazards to humans. Thus, monitoring and mitigating the presence of phenolic compounds in flowing waters are essential for preserving ecosystem integrity and safeguarding public health. This study explores the development and performance of an innovative sensor based on screen-printed electrode (SPE) modified with graphene (GPH), poly(3,4-ethylenedioxythiophene) (PEDOT), and tyrosinase (Ty), designed for water analysis, focusing on the manufacturing process and the obtained electroanalytical results. The proposed biosensor (SPE/GPH/PEDOT/Ty) was designed to achieve a high level of precision and sensitivity, as well as to allow efficient analytical recoveries. Special attention was given to the manufacturing process and optimization of the modifying elements' composition. This study highlights the potential of the biosensor as an efficient and reliable solution for water analysis. Modification with graphene, the synthesis and electropolymerization deposition of the PEDOT polymer, and tyrosinase immobilization contributed to obtaining a high-performance and robust biosensor, presenting promising perspectives in monitoring the quality of the aquatic environment. Regarding the electroanalytical experimental results, the detection limits (LODs) obtained with this biosensor are extremely low for all phenolic compounds (8.63 × 10-10 M for catechol, 7.72 × 10-10 M for 3-methoxycatechol, and 9.56 × 10-10 M for 4-methylcatechol), emphasizing its ability to accurately measure even subtle variations in the trace compound parameters. The enhanced sensitivity of the biosensor facilitates detection and quantification in river water samples. Analytical recovery is also an essential aspect, and the biosensor presents consistent and reproducible results. This feature significantly improves the reliability and usefulness of the biosensor in practical applications, making it suitable for monitoring industrial or river water.

Chitosan-Based Electrochemical Sensors for Pharmaceuticals and Clinical Applications.

Chitosan (CTS), a biocompatible and multifunctional material derived from chitin, has caught researchers' attention in electrochemical detection due to its unique properties. This review paper provides a comprehensive overview of the recent progress and applications of CTS-based electrochemical sensors in the analysis of pharmaceutical products and other types of samples, with a particular focus on the detection of medicinal substances. The review covers studies and developments from 2003 to 2023, highlighting the remarkable properties of CTS, such as biocompatibility, chemical versatility, and large surface area, that make it an excellent candidate for sensor modification. Combining CTS with various nanomaterials significantly enhances the detection capabilities of electrochemical sensors. Various types of CTS-based sensors are analyzed, including those utilizing carbon nanomaterials, metallic nanoparticles, conducting polymers, and molecularly imprinted CTS. These sensors exhibit excellent sensitivity, selectivity, and stability, enabling the precise and reliable detection of medications. The manufacturing strategies used for the preparation of CTS-based sensors are described, the underlying detection mechanisms are elucidated, and the integration of CTS sensors with transducer systems is highlighted. The prospects of CTS-based electrochemical sensors are promising, with opportunities for miniaturization, simultaneous detection, and real-time monitoring applications.

Tyrosinase Immobilization Strategies for the Development of Electrochemical Biosensors-A Review.

The development of enzyme biosensors has successfully overcome various challenges such as enzyme instability, loss of enzyme activity or long response time. In the electroanalytical field, tyrosinase is used to develop biosensors that exploit its ability to catalyze the oxidation of numerous types of phenolic compounds with antioxidant and neurotransmitter roles. This review critically examines the main tyrosinase immobilization techniques for the development of sensitive electrochemical biosensors. Immobilization strategies are mainly classified according to the degree of reversibility/irreversibility of enzyme binding to the support material. Each tyrosinase immobilization method has advantages and limitations, and its selection depends mainly on the type of support electrode, electrode-modifying nanomaterials, cross-linking agent or surfactants used. Tyrosinase immobilization by cross-linking is characterized by very frequent use with outstanding performance of the developed biosensors. Additionally, research in recent years has focused on new immobilization strategies involving cross-linking, such as cross-linked enzyme aggregates (CLEAs) and magnetic cross-linked enzyme aggregates (mCLEAs). Therefore, it can be considered that cross-linking immobilization is the most feasible and economical approach, also providing the possibility of selecting the reagents used and the order of the immobilization steps, which favor the enhancement of biosensor performance characteristics.

Studies on the Detection of Oleuropein from Extra Virgin Olive Oils Using Enzymatic Biosensors.

Oleuropein (OLEU) is an important indicator of the quality and authenticity of extra virgin olive oils (EVOO). Electrochemical sensors and biosensors for the detection of oleuropein can be used to test the adulteration of extra virgin olive oils. The present study aimed at the qualitative and quantitative determination of oleuropein in commercial EVOO samples by applying electrochemical techniques, cyclic voltammetry (CV) and square wave voltammetry (SWV). The sensing devices used were two newly constructed enzyme biosensors, supported on single-layer carbon-nanotube-modified carbon screen-printed electrode (SPE/SWCNT) on whose surface tyrosinase (SPE/SWCNT/Tyr) and laccase (SPE/SWCNT/Lac) were immobilized, respectively. The active surfaces of the two biosensors were analyzed and characterized by different methods, cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS) and Fourier transform infrared spectroscopy (FTIR) and the results confirmed the efficient immobilization of the enzymes. SPE/SWCNT/Tyr was characterized by a low detection limit (LOD = 9.53 × 10-8 M) and a very good sensitivity (0.0718 µA·µM-1·cm-2) over a wide linearity range from 0.49 to 11.22 µM. The process occurring at the biosensor surface corresponds to kinetics (h = 0.90), and tyrosinase showed a high affinity towards OLEU. The tyrosinase-based biosensor was shown to have superior sensitive properties to the laccase-based one. Quantitative determination of OLEU in EVOOs was performed using SPE/SWCNT/Tyr and the results confirmed the presence of the compound in close amounts in the EVOOs analysed, proving that they have very good sensory properties.

Sensitive Detection of Hydroxytyrosol in Extra Virgin Olive Oils with a Novel Biosensor Based on Single-Walled Carbon Nanotubes and Tyrosinase.

Hydroxytyrosol (HT) is an important marker for the authenticity and quality assessment of extra virgin olive oils (EVOO). The aim of the study was the qualitative and quantitative determination of hydroxytyrosol in commercial extra virgin olive oils of different origins and varieties using a newly developed biosensor based on a screen-printed electrode modified with single-layer carbon nanotubes and tyrosinase (SPE-SWCNT-Ty). The enzyme was immobilized on a carbon-based screen-printed electrode previously modified with single-layer carbon nanotubes (SPE-SWCNT-Ty) by the drop-and-dry method, followed by cross-linking with glutaraldehyde. The modified electrode surface was characterized by different methods, including electrochemical (cyclic voltammetry (CV), differential pulse voltammetry (DPV), electrochemical impedance spectroscopy (EIS)) and spectrometric (Fourier transform infrared (FTIR) spectroscopy) methods. Cyclic voltammetry was used for the quantitative determination of HT, obtaining a detection limit of 3.49 × 10-8 M and a quantification limit of 1.0 × 10-7 M, with a wide linearity range (0.49-15.602 µM). The electrochemical performance of the SPE-SWCNT-Ty biosensor was compared with that of the modified SPE-SWCNT sensor, and the results showed increased selectivity and sensitivity of the biosensor due to the electrocatalytic activity of tyrosinase. The results obtained from the quantitative determination of HT showed that commercial EVOOs contain significant amounts of HT, proving the high quality of the finished products. The determination of the antiradical activity of HT was carried out spectrophotometrically using the free reagent galvinoxyl. The results showed that there is a very good correlation between the antiradical capacity of EVOOs, the voltammetric response and implicitly the increased concentration of HT. SPE-SWCNT-Ty has multiple advantages such as sensitivity, selectivity, feasibility and low cost and could be used in routine analysis for quality control of food products such as vegetable oils.

Simultaneous Determination of Caffeic Acid and Ferulic Acid Using a Carbon Nanofiber-Based Screen-Printed Sensor.

This work aims to achieve the simultaneous qualitative and quantitative determination of two hydroxycinnamic acids (ferulic acid and caffeic acid) from standard solutions and from a phyto-homeopathic product using a carbon nanofiber-based screen-printed sensor (CNF/SPE). The two compounds are mentioned in the manufacturer's specifications but without indicating their concentrations. The stability and reproducibility of the CNF/SPE were found to be effective and the sensitivity was high for both caffeic acid-CA (limit of detection 2.39 × 10-7 M) and ferrulic acid-FA (limit of detection 2.33 × 10-7 M). The antioxidant capacity of the compounds in the analyzed product was also determined by the DPPH (2,2-diphenyl-1-picrylhydrazyl) method. The electrochemical method was efficient and less expensive than other analytical methods; therefore, its use can be extended for the detection of these phenolic compounds in various dietary supplements or pharmaceutical products.

Evaluation of Olive Oil Quality with Electrochemical Sensors and Biosensors: A Review.

Electrochemical sensors, sensor arrays and biosensors, alongside chemometric instruments, have progressed remarkably of late, being used on a wide scale in the qualitative and quantitative evaluation of olive oil. Olive oil is a natural product of significant importance, since it is a rich source of bioactive compounds with nutritional and therapeutic properties, and its quality is important both for consumers and for distributors. This review aims at analysing the progress reported in the literature regarding the use of devices based on electrochemical (bio)sensors to evaluate the bioactive compounds in olive oil. The main advantages and limitations of these approaches on construction technique, analysed compounds, calculus models, as well as results obtained, are discussed in view of estimation of future progress related to achieving a portable, practical and rapid miniature device for analysing the quality of virgin olive oil (VOO) at different stages in the manufacturing process.

Development of a Novel Electrochemical Biosensor Based on Carbon Nanofibers-Cobalt Phthalocyanine-Laccase for the Detection of p-Coumaric Acid in Phytoproducts.

The present paper developed a new enzymatic biosensor whose support is a screen-printed electrode based on carbon nanofibers modified with cobalt phthalocyanine and laccase (CNF-CoPc-Lac/SPE) to determine the p-coumaric acid (PCA) content by cyclic voltammetry and square wave voltammetry. Sensor modification was achieved by the casting and cross-linking technique, using glutaraldehyde as a reticulation agent. The biosensor's response showed the PCA redox processes in a very stable and sensitive manner. The calibration curve was developed for the concentration range of p-coumaric acid of 0.1-202.5 µM, using cyclic voltammetry and chronoamperometry. The biosensor yielded optimal results for the linearity range 0.4-6.4 µM and stood out by low LOD and LOQ values, i.e., 4.83 × 10-7 M and 1.61 × 10-6 M, respectively. PCA was successfully determined in three phytoproducts of complex composition. The results obtained by the voltammetric method were compared to the ones obtained by the FTIR method. The amount of p-coumaric acid determined by means of CNF-CoPc-Lac/SPE was close to the one obtained by the standard spectrometric method.

Laccase and Tyrosinase Biosensors Used in the Determination of Hydroxycinnamic Acids.

In recent years, researchers have focused on developing simple and efficient methods based on electrochemical biosensors to determine hydroxycinnamic acids from various real samples (wine, beer, propolis, tea, and coffee). Enzymatic biosensors represent a promising, low-cost technology for the direct monitoring of these biologically important compounds, which implies a fast response and simple sample processing procedures. The present review aims at highlighting the structural features of this class of compounds and the importance of hydroxycinnamic acids for the human body, as well as presenting a series of enzymatic biosensors commonly used to quantify these phenolic compounds. Enzyme immobilization techniques on support electrodes are very important for their stability and for obtaining adequate results. The following sections of this review will briefly describe some of the laccase (Lac) and tyrosinase (Tyr) biosensors used for determining the main hydroxycinnamic acids of interest in the food or cosmetics industry. Considering relevant studies in the field, the fact has been noticed that there is a greater number of studies on laccase-based biosensors as compared to those based on tyrosinase for the detection of hydroxycinnamic acids. Significant progress has been made in relation to using the synergy of nanomaterials and nanocomposites for more stable and efficient enzyme immobilization. These nanomaterials are mainly carbon- and/or polymer-based nanostructures and metallic nanoparticles which provide a suitable environment for maintaining the biocatalytic activity of the enzyme and for increasing the rate of electron transport.

Development of a Novel Electrochemical Biosensor Based on Carbon Nanofibers-Gold Nanoparticles-Tyrosinase for the Detection of Ferulic Acid in Cosmetics.

The present paper deals with the electrochemical behavior of three types of sensors based on modified screen-printed electrodes (SPEs): a sensor based on carbon nanofibers (CNF/SPE), a sensor based on nanofibers of carbon modified with gold nanoparticles (CNF-GNP/SPE) and a biosensor based on nanofibers of carbon modified with gold nanoparticles and tyrosinase (CNF-GNP-Ty/SPE). To prepare the biosensor, the tyrosinase (Ty) was immobilized on the surface of the electrode already modified with carbon nanofibers and gold nanoparticles, by the drop-and-dry technique. The electrochemical properties of the three electrodes were studied by cyclic voltammetry in electroactive solutions, and the position and shape of the active redox peaks are according to the nature of the materials modifying the electrodes. In the case of ferulic acid, a series of characteristic peaks were observed, the processes being more intense for the biosensor, with the higher sensitivity and selectivity being due to the immobilization of tyrosinase, a specific enzyme for phenolic compounds. The calibration curve was subsequently created using CNF-GNP-Ty/SPE in ferulic acid solutions of various concentrations in the range 0.1-129.6 µM. This new biosensor allowed low values of the detection threshold and quantification limit, 2.89 × 10-9 mol·L-1 and 9.64 × 10-9 mol·L-1, respectively, which shows that the electroanalytical method is feasible for quantifying ferulic acid in real samples. The ferulic acid was quantitatively determined in three cosmetic products by means of the CNF-GNP-Ty/SPE biosensor. The results obtained were validated by means of the spectrometric method in the infrared range, the differences between the values of the ferulic acid concentrations obtained by the two methods being under 5%.

Voltamperometric Sensors and Biosensors Based on Carbon Nanomaterials Used for Detecting Caffeic Acid-A Review.

Caffeic acid is one of the most important hydroxycinnamic acids found in various foods and plant products. It has multiple beneficial effects in the human body such as antioxidant, antibacterial, anti-inflammatory, and antineoplastic. Since overdoses of caffeic acid may have negative effects, the quality and quantity of this acid in foods, pharmaceuticals, food supplements, etc., needs to be accurately determined. The present paper analyzes the most representative scientific papers published mostly in the last 10 years which describe the development and characterization of voltamperometric sensors or biosensors based on carbon nanomaterials and/or enzyme commonly used for detecting caffeic acid and a series of methods which may improve the performance characteristics of such sensors.