The study of antioxidants and their implications in various fields, from food engineering to medicine and pharmacy, is of major interest to the scientific community. The present paper is a critical presentation of the most important tests used to determine the antioxidant activity, detection mechanism, applicability, advantages and disadvantages of these methods. Out of the tests based on the transfer of a hydrogen atom, the following were presented: the Oxygen Radical Absorption Capacity (ORAC) test, the Hydroxyl Radical Antioxidant Capacity (HORAC) test, the Total Peroxyl Radical Trapping Antioxidant Parameter (TRAP) test, and the Total Oxyradical Scavenging Capacity (TOSC) test. The tests based on the transfer of one electron include the Cupric Reducing Antioxidant Power (CUPRAC) test, the Ferric Reducing Antioxidant Power (FRAP) test, the Folin–Ciocalteu test. Mixed tests, including the transfer of both a hydrogen atom and an electron, include the 2,2′-Azinobis-(3-ethylbenzothiazoline-6-sulfonic acid (ABTS) test, and the [2,2-di(4-tert-octylphenyl)-1 -picrylhydrazyl] (DPPH) test. All these assays are based on chemical reactions and assessing the kinetics or reaching the equilibrium state relies on spectrophotometry, presupposing the occurrence of characteristic colours or the discolouration of the solutions to be analysed, which are processes monitored by specific wavelength adsorption. These assays were successfully applied in antioxidant analysis or the determination of the antioxidant capacity of complex samples. As a complementary method in such studies, one may use methods based on electrochemical (bio)sensors, requiring stages of calibration and validation. The use of chemical methods together with electrochemical methods may result in clarification of the operating mechanisms and kinetics of the processes involving several antioxidants.

Currently, there is growing interest in screening and quantifying antioxidants from biological samples in the quest for natural and effective antioxidants to combat free radical-related pathological complications. Antioxidants play an important role in human health and provide a defense against many diseases. Due to the valuable dietary role of these compounds, the analysis and determination of their amount in food is of particular importance. In recent years, many attempts have been made to provide simple, fast, and economical analytical approaches for the on-site detection and determination of antioxidant activity in food antioxidants. In this regard, electrochemical sensors and biosensors are considered promising tools for antioxidant research due to their high sensitivity, fast response time, and ease of miniaturization; thus, they are used in a variety of fields, including food analysis, drug screening, and toxicity research. Herein, we review the recent advances in sensors and biosensors for the detection of antioxidants, underlying principles, and emphasizing advantages, along with limitations regarding the ability to discriminate between the specific antioxidant or quantifying total antioxidant content. In this work, both direct and indirect methods for antioxidants detecting with electrochemical sensors and biosensors are analyzed in detail. This review aims to prove how electrochemical sensors and biosensors represent reliable alternatives to conventional methods for antioxidant analysis.

Chlorogenic acid (5-O-caffeoylquinic acid) is a phenolic compound from the hydroxycinnamic acid family. Epidemiological, biological, and biochemical studies concur to support the beneficial role of chlorogenic acid in human health, along with other dietary phenolic compounds. Thus, chlorogenic acid has been reported to exert inhibitory effects on carcinogenesis in the large intestine, liver, and tongue, and a protective action on oxidative stress in vivo, together with anti-inflammatory, antidiabetic and antihypertensive activities. It is also claimed to have antifungal, antibacterial and antiviral effects with relatively low toxicity and side effects, alongside properties that do not lead to antimicrobial resistance. Due to its importance, numerous methods for determining chlorogenic acid (CGA), as well as for its derivatives from coffee beans and other plants, were elaborated. The most frequently used methods are infrared spectroscopy, high performance liquid chromatography (HPLC), capillary electrophoresis, liquid chromatography-mass spectrometry and chemiluminescence. Although these methods proved to be efficient for quantifying CGA and its derived products, a number of deficiencies were identified: they are time consuming, laborious, and require expensive instruments. Therefore, electrochemical methods have been developed and used in the determination of CGA in different nutraceuticals or food products. The present review aims to present the main progresses and performance characteristics of electrochemical sensors and biosensors used to detect CGA, as it is reported in a high number of relevant scientific papers published mainly in the last decade.

The analysis of antioxidants in different foodstuffs has become an active area of research, which has led to many recently developed antioxidant assays. Many antioxidants exhibit inherent electroactivity, and, therefore, the use of electrochemical methods could be a viable approach for evaluating the overall antioxidant activity of a matrix of nutraceuticals without the need for adding reactive species. Green tea is believed to be a healthy beverage due to a number of therapeutic benefits. Catechin, one of its constituents, is an important antioxidant and possesses free radical scavenging abilities. The present paper describes the electrochemical properties of three screen-printed electrodes (SPEs), the first one based on carbon nanotubes (CNTs), the second one based on gold nanoparticles (GNPs) and the third one based on carbon nanotubes and gold nanoparticles (CNTs-GNPs). All three electrodes were modified with the laccase (Lac) enzyme, using glutaraldehyde as a cross-linking agent between the amino groups on the laccase and aldehyde groups of the reticulation agent. As this enzyme is a thermostable catalyst, the performance of the biosensors has been greatly improved. Electro-oxidative properties of catechin were investigated using cyclic voltammetry (CV) and differential pulse voltammetry (DPV), and these demonstrated that the association of CNTs with GNPs significantly improved the sensitivity and selectivity of the biosensor. The corresponding limit of detection (LOD) was estimated to be 5.6 × 10−8 M catechin at the CNT-Lac/SPE, 1.3 × 10−7 M at the GNP-Lac/SPE and 4.9 × 10−8 M at the CNT-GNP-Lac/SPE. The biosensors were subjected to nutraceutical formulations containing green tea in order to study their catechin content, using CNT-GNP-Lac/SPE, through DPV. Using a paired t-test, the catechin content estimated was in agreement with the manufacturer’s specification. In addition, the relationship between the CNT-GNP-Lac/SPE response at a specific potential and the antioxidant activity of nutraceuticals, as determined by conventional spectrophotometric methods (DPPH, galvinoxyl and ABTS), is discussed in the context of developing a fast biosensor for the relative antioxidant activity quantification.

Peptides have been used as components in biological analysis and fabrication of novel sensors due to several reasons, including well-known synthesis protocols, diverse structures, and acting as highly selective substrates for enzymes. Bio-conjugation strategies can provide a simple and efficient way to convert peptide-analyte interaction information into a measurable signal, which can be further used for the manufacture of new peptide-based biosensors. This paper describes the sensitive properties of a peptide-modified graphene oxide screen-printed carbon electrode for accurate and sensitive detection of a natural polyphenol antioxidant compound, namely rosmarinic acid. Glutaraldehyde was chosen as the cross-linking agent because it is able to bind nonspecifically to the peptide. We demonstrated that the strong interaction between the immobilized peptide on the surface of the sensor and rosmarinic acid favors the addition of rosmarinic acid on the surface of the electrode, leading to an efficient preconcentration that determines a high sensitivity of the sensor for the detection of rosmarinic acid. The experimental conditions were optimized using different pH values and different amounts of peptide to modify the sensor surface, so that its analytical performances were optimal for rosmarinic acid detection. By using cyclic voltammetry (CV) as a detection method, a very low detection limit (0.0966 μM) and a vast linearity domain, ranging from 0.1 µM to 3.20 µM, were obtained. The novelty of this work is the development of a novel peptide-based sensor with improved performance characteristics for the quantification of rosmarinic acid in cosmetic products of complex composition. The FTIR method was used to validate the voltammetric method results.

The purpose of our research was to develop a new enzymatic biosensor, GPH-MnPc-Tyr/SPE, using as a support screen-printed carbon electrode (SPE) modified with graphene, manganese phthalocyanine, and tyrosinase, with the aim of developing sensitive detection of chlorogenic acid (CGA). To immobilise tyrosinase on the sensor surface, crosslinking with the glutaraldehyde technique was used, thus increasing the enzyme bioactivity on this electrode. The modified electrode has a great catalytic effect on the electrochemical redox of chlorogenic acid, compared to the simple, unmodified SPE. The peak current response of the biosensor for CGA was linear in the range of 0.1–10.48 μM, obtaining a calibration curve using cyclic voltammetry (CV) and square-wave voltammetry (SWV). Subsequently, the detection limit (LOD) and the quantification limit (LOQ) were determined, obtaining low values, i.e., LOD = 1.40 × 10−6 M; LOQ = 4.69 × 10−6 M by cyclic voltammetry and LOD = 2.32 × 10−7 M; LOQ= 7.74 × 10−7 M, by square-wave voltammetry (SWV). These results demonstrate that the method is suitable for the detection of CGA in nutraceutical formulations. Therefore, GPH-MnPc-Tyr/SPE was used for the quantitative determination of CGA in three products, by means of cyclic voltammetry. The Folin–Ciocalteu spectrophotometric assay was used for the validation of the results, obtaining a good correlation between the voltammetric method and the spectrophotometric one, at a confidence level of 95%. Moreover, by means of the DPPH method, the antioxidant activity of the compound was determined, thus demonstrating the antioxidant effect of CGA in all nutraceuticals studied.

The classification of olive oils and the authentication of their biological or geographic origin are important issues for public health and for the olive oil market and related industries. The development of techniques for olive oil classification that are fast, easy to use, and suitable for online, in situ and remote operation is of high interest. In this study, the possibility of discriminating and classifying vegetable oils according to different criteria related to biological or geographical origin was assessed using cyclic voltammograms (CVs) as input data, obtained with electrochemical sensors based on carbonaceous nanomaterials and gold nanoparticles. In this context, 44 vegetable oil samples of different categories were analyzed and the capacity of the sensor array coupled with multivariate analysis was evaluated. The characteristics highlighted in voltammograms are related to the redox properties of the electroactive compounds, mainly phenolics, existing in the oils. Moreover, the antioxidant activity of the oils’ hydrophilic fraction was also estimated by conventional spectrophotometric methods (1,1-diphenyl-2-picrylhydrazyl (DPPH) and galvinoxyl) and correlated with the voltammetric responses of the sensors. The percentage of DPPH and galvinoxyl inhibition was accurately predicted from the voltammetric data, with a correlation coefficients greater than 0.97 both in calibration and in validation. The results indicate that this method allows for a clear discrimination of oils from different biological or geographic origins.

In addition to their antioxidant and antimicrobial action in functional foods, beverages, and in some dermato-cosmetic products, olive phenolic compounds are also recognized for their role in the prevention of diabetes and inflammation, treatment of heart disease and, consequently, of the numerous chronic diseases mediated by the free radicals. In recent years, attention has increased, in particular, regarding one of the most important compound in extra virgin olive oil (EVOO) having glycosidic structure, namely verbocoside, due to the existence in the literature of numerous studies demonstrating its remarkable contribution to the prophylaxis and treatment of various disorders of the human body. The purpose of this study was the qualitative and quantitative determination of verbascoside in commercial EVOOs from different regions by means of a newly developed sensor based on a screen-printed carbon electrode (SPCE) modified with graphene oxide (GPHOX), on the surface of which a pentapeptide was immobilized by means of glutaraldehyde as cross-linking agent. The modified electrode surface was investigated using both Fourier-transform infrared spectroscopy (FTIR) and scanning electron microscopy (SEM) methods. This newly developed sensor has shown a high sensibility compared to the unmodified electrode, a low detection limit (LOD) of up to 9.38 × 10−8 M, and a wide linearity range between 0.1 µM and 10.55 µM. The applicability of the modified sensor was confirmed by detecting verbascoside in ten different EVOOs samples using the cyclic voltammetry (CV) method, with very good results. The validation of the electroanalytical method was performed by using the standard addition method with very good recoveries in the range of 97.48–103.77%.