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Lupin seeds are recognized for their nutritional value and potential health benefits, but they contain also a significant amount of alkaloids, an anti-nutritive class of compounds, which vary significantly in composition within and between species due to environmental factors. In this study, we developed a predictive multi-experiment approach using ultra-high performance liquid chromatography coupled with triple quadrupole with linear ionic trap tandem mass spectrometry (UHPLC-QqQ-LIT-MS/MS) for comprehensive alkaloid profiling and geographical classification of Lupinus albus L. samples originating from four different Italian regions. Six targeted quinolizidine alkaloids were detected and 21 other alkaloids were putatively identified. Hierarchical Cluster Analysis (HCA) and partial least squares discriminant analysis (PLS-DA) were applied to explore the data structure and successfully classify samples according to their geographical origin. The data demonstrate the efficacy of the developed approach in providing valuable insights in alkaloid profiles of lupin seeds and their potential as markers for geographical traceability.

Abstract Obsessive Compulsive Disorder (OCD) is listed as one of the top 10 most disabling neuropsychiatric conditions in the world. The neurobiology of OCD has not been completely understood and efforts are needed in order to develop new treatments. Beside the classical neurotransmitter systems and signalling pathways implicated in OCD, the possible involvement of the endocannabinoid system (ECS) has emerged in pathophysiology of OCD. We report here selective downregulation of the genes coding for enzymes allowing the synthesis of the endocannabinoids. We found reduced DAGLα and NAPE-PLD in blood samples of individuals with OCD (when compared to healthy controls) as well as in the amygdala complex and prefrontal cortex of dopamine transporter (DAT) heterozygous rats, manifesting compulsive behaviours. Also mRNA levels of the genes coding for cannabinoid receptors type 1 and type 2 resulted downregulated, respectively in the rat amygdala and in human blood. Moreover, NAPE-PLD changes in gene expression resulted to be associated with an increase in DNA methylation at gene promoter, and the modulation of this gene in OCD appears to be correlated to the progression of the disease. Finally, the alterations observed in ECS genes expression appears to be correlated with the modulation in oxytocin receptor gene expression, consistently with what recently reported. Overall, we confirm here a role for ECS in OCD at both preclinical and clinical level. Many potential biomarkers are suggested among its components, in particular NAPE-PLD, that might be of help for a prompt and clear diagnosis.

Phytosterols (PSs) are bioactive compounds in the sterol family, present in numerous complex food and plant matrices in free and conjugated forms. The interest in these compounds arises for phytotherapeutic purposes, particularly for their action on cholesterol metabolism and impact on cardiovascular diseases. There is a need to develop approaches that can selectively extract target analytes and accurately identify and quantify them with high precision. This work proposed the synthesis of molecularly imprinted polymers (MIPs) for PSs with a sonochemical approach, enabling a rapid polymerization step (5 min). This proposed MIP was able to extract 8 PSs (brassicasterol, stigmastanol, campesterol, campestanol, stigmasterol, β-sitosterol, Δ5-avenasterol, α-spinasterol) from a wide range of plant and food matrices belonging to different classes (Brassicaceae, dried fruits and Leguminosae) and was coupled to ultra-high liquid chromatography with tandem mass spectrometry (UHPLC-MS/MS). MIP based on dispersed solid phase extraction (dSPE-MIP) and targeted analysis has proven to be particularly effective in addressing the challenges associated with the complexity of plant-derived matrices, minimising interferences. This was demonstrated by the excellent control of the matrix effect, which was within ±15 %, ensuring the robustness and reliability of the method. The identification and quantification of 8 different PSs was successfully achieved with satisfactory recovery values ranging from 65 % to 100 %. The proposed strategy offers an affordable alternative to classical methods, providing enhanced sensitivity, selectivity and overall performance.

Phomopsin A (PHO-A), a potent mycotoxin produced by the pathogenic fungus Diaporthe toxica, is a growing international concern due to the high toxicity and prevalence in lupin plants, a vital source of food and feed. This hexapeptide mycotoxin has raised health concerns for both animals and humans, prompting regulatory bodies to set maximum allowable levels in lupin products.Different analytical methods have been developed for PHO-A detection, including Enzyme-Linked Immuno-Sorbent Assay (ELISA) and high-performance liquid chromatography (HPLC) coupled with different detectors; they often lack sensitivity and selectivity required to meet maximum regulatory limits (MRL). In this work, we present a robust and sensitive method for PHO-A quantification and determination, developed using ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS).This method incorporates a selective clean-up step using Micro Solid Phase Extraction (µ-SPE) to efficiently purify samples, ensuring a rapid, efficient, and selective recovery of PHO-A from the complex lupin matrix. The clean-up step not only helps in achieving lower detection limits but also minimizes matrix effects and contamination. The proposed method was validated following international guidelines, demonstrating reliable recovery, matrix effects, linearity, accuracy, and precision.Additionally, this method was successfully applied to artificially contaminated Lupinus albus L. samples, confirming the suitability for assessing PHO-A contamination. This analytical approach fills a critical gap in the literature by providing a specific, sensitive, and selective method for PHO-A analysis, contributing to the safety of lupin products and field monitoring.

Cocoa (Theobroma cacao, L.) represents an important market that gained relevance and became an esteemed commodity thanks to cocoa powder, chocolate, and other related products. This work analyzed 59 cocoa powder samples from the European market. Three distinct subgroups were identified: organic or conventional, alkalized or not alkalized, and raw or roasted processing. The impact of the technological process on their pH, color, and compositional traits, as well as their content of biogenic amines and salsolinol, was evaluated. The phenolic fraction was also investigated through both common and emerging methods. The results depict that the influence of the agronomical practices (organic/conventional) did not significantly (p < 0.05) affect the composition of the cocoa powders; similarly, the roasting process was not a determinant of the compounds traced. On the other hand, the alkalinization process greatly impacted color and pH, no matter the cocoa’s provenience or obtention or other processes, also resulting in reducing the phenolic fraction of the treated samples. Principal component analysis confirmed that the alkali process acts on pH, color, and phenolic composition but not on the content of other bioactive molecules (biogenic amines and salsolinol). All the samples were safe, while the alkalized powders saw a great reduction in beneficial biocompounds. A novel strategy could be to emphasize on the label whether cocoa powder is non-alkalized to meet the demand for more beneficial products.

Murraya koenigii (M. koenigii) is recognized as one of the most significant Indian medicinal plants, due to its therapeutic properties. In fact, various biological activities, including anti-inflammatory and antifungal, are attributed to carbazole alkaloids (CAs) and their metabolites, but it is still difficult to achieve a full classification of these compounds because of their heterogeneous structures. For this reason, the development of new strategies for their identification is necessary. In this work, a reliable method, including the simultaneous quantification of three main CAs of Murraya (Mahanimbine, Koenimbine and Koenigicine) and a putative identification for other compounds belonging to the huge family of CAs, was developed by means of HPLC-MS/MS. Also, an efficient extraction procedure followed by a suitable clean-up step was presented, in order to obtain reliable recoveries (resulted from 60 to 85% for all the analytes). The analyses were performed by using predictive multi experiment approach based on information-dependent acquisition (IDA), coupling multiple reaction monitoring (MRM) and precursor ion (PI) as survey scans, and enhanced product ion scan (EPI) as dependent scan. Competitive Fragmentation Modeling-ID (CFM-ID) was used to predict MS/MS spectra from the chemical structures of the compounds in order to create a suitable MRM inclusion list. The obtained results showed that this method can simultaneously provide quantitative information for the target analytes (Mahanimbine (7.22–5.62 mg/kg), Koenimbine (1.26–1.62 mg/kg) and Koenigicine (0.44–1.77 mg/kg)) and a putative identification for several compounds belonging to different classes of CAs which are not included in the target list, thanks to PI survey scan.

Nowadays, the adoption of In Vitro Fertilization (IVF) techniques is undergoing an impressive increase. In light of this, one of the most promising strategies is the novel use of non-physiological materials and naturally derived compounds for advanced sperm preparation methods. Here, sperm cells were exposed during capacitation to MoS2/Catechin nanoflakes and catechin (CT), a flavonoid with antioxidant properties, at concentrations of 10, 1, 0.1 ppm. The results showed no significant differences in terms of sperm membrane modifications or biochemical pathways among the groups, allowing the hypothesis that MoS2/CT nanoflakes do not induce any negative effect on the parameters evaluated related to sperm capacitation. Moreover, the addition of CT alone at a specific concentration (0.1 ppm) increased the spermatozoa fertilizing ability in an IVF assay by increasing the number of fertilized oocytes with respect to the control group. Our findings open interesting new perspectives regarding the use of catechins and new materials obtained using natural or bio compounds, which could be used to implement the current strategies for sperm capacitation.

The transition to a sustainable economic and environmental management of olive oil sector needs to be implemented in both national and regional territories through the introduction and development of innovative growing systems and variety. In this study, the olive oil quality parameters of local and allochthonous varieties cultivated in different orchards located in the Abruzzo region (Italy), using traditional and super high-density systems, were analyzed. Frantene, Lecciana, Koroneiki, and a mix of Arbequina and Lecciana provided olive oils rich in flavonoids and secoiridoids compounds with respect to the local varieties Frantoio, Leccino, and a mix of Dritta, Leccino, and Pendolino. Oleic/linoleic ratio was influenced by cultivar and training systems with super high-density olive oils rich in oleic acid. Frantene showed a peculiar fatty acid profile different from cultivars grown in the same location; moreover, interesting similarities were found between Frantene and the mix of Dritta, Leccino, and Pendolino in terms of health-related compounds. The potential development of innovative sustainable training system to improve olive oil quality was highlighted. The study’s results identify olive varieties suitable for super high-density systems spread in the Abruzzo region, representing a valid alternative for the olive growers to improve both the quality of the olive oil, as well as the company’s income.

Lupin alkaloids (LAs) represent a class of toxic secondary metabolites in plants, in particular in Lupinus spp.; they are produced as a defense mechanism due to their strong bitter taste and are very dangerous for human and animals. In this work, a sensitive and reliable high performance liquid chromatography—tandem mass spectrometry (HPLC-MS/MS) analytical method for the identification and quantification of thirteen lupin alkaloids was developed and validated according to FDA guidelines. Efficient extraction and clean-up steps, carried out by solid-phase extraction, were finely tuned on the basis of the characteristics of the analytes and lupin samples, providing good selectivity with minimized matrix interference. The effectiveness of the method was proven by the satisfactory recovery values obtained for most of the analytes and a matrix effect ≤23% for all tested levels. In addition, a sensitive and reliable determination of the target compounds was obtained; LOQs were between 1 and 25 µg Kg−1, i.e., below the requested maximum levels (−1). The method was applied to evaluate the LAs profile in different batches of raw L. albus L. samples, varying in size and across farming treatments.

The demand for next-generation multifunctional nanovectors, combining therapeutic effects with specific cellular targeting, has significantly grown during the last few years, pursuing less invasive therapy strategies. Polyphenol-conjugated silver nanoparticles (AgNPs) appear as potential multifunctional nanovectors, integrating the biorecognition capability and the antioxidant power of polyphenols, the antimicrobial activity of silver, and the drug delivery capability of NPs. We present a spectroscopic and microscopic investigation on polyphenol-synthesized AgNPs, selecting caffeic acid (CA) and catechol (CT) as model polyphenols and using them as reducing agents for the AgNP green synthesis, both in the presence and in the absence of a capping agent. We exploit the plasmonic properties of AgNPs to collect Surface-Enhanced Raman Scattering (SERS) spectra from the nanosized region next to the Ag surface and to characterize the molecular environment in the proximity of the NP, assessing the orientation and tunable deprotonation level of CA, depending on the synthesis conditions. Our results suggest that the SERS investigation of such nanovectors can provide crucial information for their perspective biomedical application.

Aflatoxins (AFs) are fungi secondary metabolites produced by the Aspergillus family. These compounds can enter the food chain through food contamination, representing a risk to human health. Commercial immunoaffinity columns are widely used for the extraction and cleanup of AFs from food samples; however, their high cost and large solvent consumption create a need for alternative strategies. In this work, an alternative strategy for producing molecularly imprinted polymers (MIPs) was proposed to extract aflatoxins AFB1, AFB2, AFG1, and AFG2 from complex food samples, using liquid chromatography coupled with tandem mass spectrometry (LC–MS/MS). The MIPs were synthesized via a low-cost and rapid (5 min) sonochemical free-radical polymerization, using 1-hydroxy-2-naphthoic acid as a dummy template. MIPs-based solid phase extraction performance was tested on 17 dietary supplements (vegetables, fruits, and cereals), obtaining appreciable recovery rates (65–90%) and good reproducibility (RSD ≤ 6%, n = 3); the selectivity towards other mycotoxins was proved and the data obtained compared with commercial immunoaffinity columns. The proposed strategy can be considered an alternative affordable approach to the classical immunoaffinity columns, since it is more selective and better performing.

The rapid introduction of new psychoactive substances (NPS) has definitively changed the drug market. Among the several NPS that were identified in the last decades, fentanyl and its analogues deserve special attention. These are synthetic opioids with high potency and are associated with increasing number of deaths; for this reason, forensic toxicologists are paying close attention to these analytes and sensitive analytical methods for their detection in biological samples of drug users are needed.The aim of this study was the development of a LC–MS/MS method for the determination of fentanyl, 23 analogues and metabolites in urine by exploiting parallel artificial liquid membrane extraction (PALME). This technique was shown to be particularly suitable for fentanyl extraction and allowed to obtain a high enrichment factor by using a few microliters of organic solvent (1-octanol) immobilized into a polyvinylidene fluoride (PVDF) membrane. The extraction was carried out on a 96 well plate providing high laboratory throughput.The applied strategy allowed to measure concentrations ranging from 0.1 ng mL−1 for fentanyl and most analogues to 5 ng mL−1 for metabolites, by using an entry level mass spectrometer. Because of the different concentration levels generally found in real samples, linearity was studied in different ranges i.e. LOQ to 50 ng mL−1 for parent drugs and LOQ to 200 ng mL−1 for metabolites. All the validation parameters were found within the imposed limits, and notably matrix effect was not significant for all the analytes, showing the selectivity achieved by PALME extraction.

Monitoring synthetic colorants in foods is important due to their potential toxicity and pathogenicity. We propose here a new and simple method for the extraction and determination of erythrosine B (ERT-B) in food samples. A composite of polydopamine-based molecularly imprinted polymers coating magnetic nanoparticles (Fe3O4@PDA@MIP) was synthesized using a green approach and exploited for the magnetic dispersive solid-phase extraction (MDSPE) of ERT-B. Fe3O4@PDA@MIP provides a rapid extraction of ERT-B, exhibiting good reusability and preconcentration ability. Moreover, the MIP showed a relatively good imprinting factor (3.0 ± 0.05), demonstrating excellent selectivity against patent blue (an interfering dye) and other food matrix components. The proposed MDSPE was coupled to colorimetric smartphone-based detection that allowed us to obtain similar performances of UV–Vis spectroscopy detection. The smartphone-based optical detection facilitated the determination of ERT-B in the 0.5–10 mg/L range, with a limit of detection of 0.04 mg/L. The developed method was successfully employed to determine ERT-B in food samples (juice, candy, and candied cherries) with good recovery values (82–97%).

A new sustainable route to nanodispersed and functionalized carbon black in water phase (W-CB) is proposed. The sonochemical strategy exploits ultrasounds to disaggregate the CB, while two selected functional naturally derived compounds, sodium cholate (SC) and rosmarinic acid (RA), act as stabilizing agents ensuring dispersibility in water adhering onto the CB nanoparticles’ surface. Strategically, the CB-RA compound is used to drive the AuNPs self-assembling at room temperature, resulting in a CB surface that is nanodecorated; further, this is achieved without the need for additional reagents. Electrochemical sensors based on the proposed nanomaterials are realized and characterized both morphologically and electrochemically. The W-CBs’ electroanalytical potential is proved in the anodic and cathodic window using caffeic acid (CF) and hydroquinone (HQ), two antioxidant compounds that are significant for food and the environment. For both antioxidants, repeatable (RSD ≤ 3.3%; n = 10) and reproducible (RSD ≤ 3.8%; n = 3) electroanalysis results were obtained, achieving nanomolar detection limits (CF: 29 nM; HQ: 44 nM). CF and HQ are successfully determined in food and environmental samples (recoveries 97–113%), and also in the presence of other phenolic classes and HQ structural isomers. The water dispersibility of the proposed materials can be an opportunity for (bio) sensor fabrication and sustainable device realization.

Polyphenols (PCs) are a numerous class of bioactive molecules and are known for their antioxidant activity. In this work, the potential of the quadrupole/linear ion trap hybrid mass spectrometer (LIT-QqQ) was exploited to develop a semi-untargeted method for the identification of polyphenols in different food matrices: green coffee, Crocus sativus L. (saffron) and Humulus lupulus L. (hop). Several conjugate forms of flavonoids and hydroxycinnamic acid were detected using neutral loss (NL) as a survey scan coupled with dependent scans with enhanced product ion (EPI) based on information-dependent acquisition (IDA) criteria. The presented approach is focused on a specific class of molecules and provides comprehensive information on the different conjugation models that are related to specific base molecules, thus allowing a quick and effective identification of all possible combinations, such as mono-, di-, or tri-glycosylation or another type of conjugation such as quinic acid esters.

New Psychoactive Substances (NPS) are a global concern since they are spreading at an unprecedented rate. Despite their commerce still being limited compared to traditional illicit drugs, the identification of NPS in seizures may represent a challenge because of the variety of possible structures. In this study we report the successful application of molecular networking (MN) to identify unexpected fentanyl analogs in two seizures. The samples were extracted with 1 mL of methanol and analyzed with an untargeted data-dependent acquisition approach by LC–HRMS. The obtained data were examined using the MN workflow within the Global Natural Product Search (GNPS). A job was submitted to GNPS by including both seizures and standard mixtures containing synthetic cannabinoids and fentanyls raw files; spectra obtained from standards were used to establish representative networks for both molecular classes. All synthetic cannabinoids in the mixture were linked together resulting in a molecular network despite their different fragmentation spectra. Looking at fentanyls, all the molecules with the typical 188.143 and 105.070 fragments were combined in a representative network. By exploiting the standard networks two unexpected fentanyls were found in the analyzed seizures and were putatively annotated as para-fluorofuranylfentanyl and (iso)butyrylfentanyl. The identity of these two fentanyl analogs was confirmed by NMR analysis. Other m/z ratios in the seizures were compatible with fentanyl derivatives; however, they appeared to be minor constituents, probably impurities or synthetic byproducts. The latter might be of interest for investigations of common fingerprints among different seizures.

In this work, we present a comprehensive study comparing the electrochemical performance of MX2 nanosheet-based electrodes (where M: Mo or W; X: S or Se). The nanosheets, obtained via a sonochemical strategy, were characterized using SEM and Raman spectroscopy to confirm the success of the exfoliation process. The electrochemical properties have been studied using a redox probe and flavonoids with an o-diphenol target moiety since the latter is involved in redox processes of high significance. It was revealed that selenides exhibit a better performance employing redox probes (higher k0 and lower RCT) and higher oxidation currents for oxidation of catechol-containing flavonoids (CCF). It is worthy of note that MX2 materials also have enhanced antifouling properties toward flavonoids compared with carbon-based electrodes.

Abstract Background Nano-PCR is a recent tool that is used in viral diseases diagnosis. The technique depends on the fundamental effects of gold nanoparticles (AuNPs) and is considered a very effective and sensitive tool in the diagnosis of different diseases including viral diseases. Although several techniques are currently available to diagnose foot and mouth disease virus (FMDV), a highly sensitive, highly specific technique is needed for specific diagnosis of the disease. In the present work, a novel AuNPs biosensor has been designed using thiol-linked oligonucleotides that recognize the conserved 3D gene of FMDV. Results The AuNPs-FMDV biosensor specifically recognizes RNA standards of FMDV, but not that of swine vesicular disease virus (SVDV) isolates. The analytical sensitivity of the AuNPs-FMDV biosensor was 10 copy number RNA standards in RT-PCR and 1 copy number RNA standard in real-time rRT-PCR with a 94.5% efficiency, 0.989 R2, a − 3.544 slope and 100% specificity (no cross-reactivity with SVDV). These findings were confirmed by the specific and sensitive recognition of 31 Egyptian FMDV clinical isolates that represents the three FMDV serotypes (O, A, and SAT2). Conclusions The AuNPs-FMDV biosensor presents in this study demonstrates a superior analytical and clinical performance for FMDV diagnosis. In addition, this biosensor has a simple workflow and accelerates epidemiological surveillance, hence, it is qualified as an efficient FMDV diagnosis tool for quarantine stations and farms particularly in FMDV endemic areas.

Detection of relevant contaminants using screening approaches is a key issue to ensure food safety and respect for the regulatory limits established. Electrochemical sensors present several advantages such as rapidity; ease of use; possibility of on-site analysis and low cost. The lack of selectivity for electrochemical sensors working in complex samples as food may be overcome by coupling them with molecularly imprinted polymers (MIPs). MIPs are synthetic materials that mimic biological receptors and are produced by the polymerization of functional monomers in presence of a target analyte. This paper critically reviews and discusses the recent progress in MIP-based electrochemical sensors for food safety. A brief introduction on MIPs and electrochemical sensors is given; followed by a discussion of the recent achievements for various MIPs-based electrochemical sensors for food contaminants analysis. Both electropolymerization and chemical synthesis of MIP-based electrochemical sensing are discussed as well as the relevant applications of MIPs used in sample preparation and then coupled to electrochemical analysis. Future perspectives and challenges have been eventually given.

A hairpin DNA (hpDNA) piezoelectric gas sensors array with heptamer loops as sensing elements was designed, realized, and challenged with pure volatile organic compounds VOCs and real samples (beer). The virtual binding versus five chemical classes (alcohols, aldehydes, esters, hydrocarbons, and ketones) of the entire combinatorial library of heptamer loops (16,384 elements) was studied by molecular modelling. Six heptamer loops, having the largest variance in binding the chemical classes, were selected to build the array. The six gas sensors were realized by immobilizing onto gold nanoparticles (AuNPs) via a thiol spacer the hpDNA constituted by the heptamer loops and the same double helix stem of four base pairs (GAAG at 5′ and CTTC at 3′ end). The HpDNA-AuNP was used to modify the surface of 20 MHz quartz crystal microbalances (QCMs). The realized E-nose was able to clearly discriminate among 15 pure VOCs of different chemical classes, as demonstrated by hierarchical cluster analysis. The analysis of real beer samples during fermentation was also carried out. In such a challenging matrix consisting of 23 different VOCs, the hpDNA E-nose with heptamer loops was able to discriminate among different fermentation times with high success rate. Class assignment using the Bayes theorem gave an excellent 98% correct beer samples classification in cross-validation.

Appropriate and standardized techniques for the extraction of secondary metabolites with interesting biological activity from plants are required. In this work, a comparison of different conventional and unconventional extraction techniques (maceration—M, Soxhlet—S, ultrasound assisted extraction—UAE, and rapid solid-liquid dynamic extraction—RSLDE) was investigated. Bioactive compounds were extracted from Thymus vulgaris L. (thyme), Cannabis sativa L. (industrial hemp) and Coriandrum sativum L. (coriander) and chemically characterized for their volatile fraction and polyphenolic content by means of gas chromatography-mass spectrometry (GC-MS) and high performance liquid chromatography-ultraviolet (HPLC-UV). Linalool (48.19%, RSLDE) and carvacrol (21.30%, M) for thyme, caryophyllene (54.78%, S) and humulene (14.13%, S) for hemp, and linalool (84.16%, RSLDE) for coriander seeds were the main compounds among terpenes, while thyme was the richest source of polyphenols with rosmarinic acid (51.7 mg/g dry extract-S), apigenin (7.6 mg/g dry extract-S), and luteolin (4.1 mg/g dry extract-UAE) being the most abundant. In order to shed light on their potential as natural food preservatives, the biological activity of the extracts was assessed in terms of antioxidant activity (2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid—ABTS˙+, ferric reducing antioxidant power—FRAP, 2,2-diphenyl-1-picrylhydrazyl—DPPH˙ assays) and phenolic content (Folin–Ciocâlteu method). For thyme, Soxhlet extracts showed best performances in FRAP and ABTS˙+ assays (74 mg TE/g dry extract and 134 mg TE/g dry extract, respectively), while Soxhlet and RSLDE extracts recorded similar activity in DPPH˙ (107–109 mg TE/g dry extract). For hemp and coriander, indeed, RSLDE extracts accounted for higher antioxidant activity as evidenced by FRAP (80 mg TE/g dry extract and 18 mg TE/g dry extract, respectively) and ABTS˙+ (557 mg TE/g dry extract and 48 mg TE/g dry extract, respectively) assays. With respect to DPPH˙, the best results were observed for UAE extracts (45 mg TE/g dry extract and 220 mg TE/g dry extract, respectively). Our findings suggest that all the investigated techniques are valid extraction methods to retain bioactive compounds and preserve their activity for application in food and pharmaceutical formulations. Among them, the innovative RSLDE stands out for the slightly higher antioxidant performances of the extracts, coupled with the facility of use and standardization of the extraction process.

Detection and monitoring of volatiles is a challenging and fascinating issue in environmental analysis, agriculture and food quality, process control in industry, as well as in ‘point of care’ diagnostics. Gas chromatographic approaches remain the reference method for the analysis of volatile organic compounds (VOCs); however, gas sensors (GSs), with their advantages of low cost and no or very little sample preparation, have become a reality. Gas sensors can be used singularly or in array format (e.g., e-noses); coupling data output with multivariate statical treatment allows un-target analysis of samples headspace. Within this frame, the use of new binding elements as recognition/interaction elements in gas sensing is a challenging hot-topic that allowed unexpected advancement. In this review, the latest development of gas sensors and gas sensor arrays, realized using peptides, molecularly imprinted polymers and DNA is reported. This work is focused on the description of the strategies used for the GSs development, the sensing elements function, the sensors array set-up, and the application in real cases.

In this work a peptide based gas sensor array based of ZnO nanoparticles (ZnONPs) has been realized. Four different pentapeptides molecularly modeled for alcohols and esters having cysteine as a common spacer have been immobilized onto ZnONPs. ZnONPs have been morphologically and spectroscopically characterized. Modified nanoparticles have been then deposited onto quartz crystal microbalances (QCMs) and used as gas sensors with nitrogen as carrier gas. Analysis of the pure compounds modeled demonstrated a nice fitting of modeling with real data. The peptide based ZnONPs had very low sensitivity to water, compared to previously studied AuNPs peptide based gas sensors allowing the use of the array on samples with high water content. Real samples of fruit juices have been assayed; stability of the signal, good repeatability, and discrimination ability of the array was achieved.

In this study samples of feedstuffs were collected from different feed mills and animal farms located in central Italy and analyzed for ionophore coccidiostat residues at carry-over levels by liquid chromatography-mass spectrometry. Since unavoidable cross-contamination of feedstuffs may occur during their production as well as distribution and storage, the collection of samples covered all these different stages. Residues of lasalocid, monensin, salinomycin and maduramicin were detected in 32.4% of samples, both at production and storage level. The maximum content for unavoidable carry-over set by Regulation (EU) No 574/2011 was exceeded in 11.3% of samples. The variability of the results highlighted the different approach of each investigated feed business operator to avoid any cross-contamination in non-target feed. The method developed in this study can be able to detect ionophore coccidiostats at low concentrations consequent to carry-over.

The aim of this research was to determine the effect of rearing systems for pig production, as concerns performance, meat lipid content, the fatty acid profile, histidinic antioxidants, coenzyme Q10, and TBARs. One hundred pigs were assigned to one of three treatments: intensively reared commercial hybrid pig (I), free range commercial hybrid pig (FR) or organically reared crossbred pig (O), according to organic EU Regulations. I pigs showed the best productive performance, but FR and O increased: C20:1n9, Δ9-desaturase (C18) and thioesterase indices in meat. Lipid, dipeptides and CoQ10 appeared correlated to glycolytic and oxidative metabolic pathways. We can conclude that all studied parameters were influenced by the rearing system used, and that differences were particularly evident in the O system, which produced leaner meat with higher oxidative stability. In this respect, the organic pig rearing system promotes and enhances biodiversity, environmental sustainability and food quality.

Herein, and in contrast to current production of anti-Zika virus antibodies, we propose a semi-combinatorial virtual strategy to select short peptides as biomimetic antibodies/binding agents for the detection of intact Zika virus (ZIKV) particles. The virtual approach was based on generating different docking cycles of tetra, penta, hexa, and heptapeptide libraries by maximizing the discrimination between the amino acid motif in the ZIKV and dengue virus (DENV) envelope protein glycosylation site. Eight peptides, two for each length (tetra, penta, hexa, and heptapeptide) were then synthesized and tested vs. intact ZIKV particles by using a direct enzyme linked immunosorbent assay (ELISA). As a reference, we employed a well-established anti-ZIKV antibody, the antibody 4G2. Three peptide-based assays had good detection limits with dynamic range starting from 105 copies/mL of intact ZIKV particles; this was one order magnitude lower than the other peptides or antibodies. These three peptides showed slight cross-reactivity against the three serotypes of DENV (DENV-1, -2, and -3) at a concentration of 106 copies/mL of intact virus particles, but the discrimination between the DENV and ZIKV was lost when the coating concentration was increased to 107 copies/mL of the virus. The sensitivity of the peptides was tested in the presence of two biological matrices, serum and urine diluted 1:10 and 1:1, respectively. The detection limits decreased about one order of magnitude for ZIKV detection in serum or urine, albeit still having for two of the three peptides tested a distinct analytical signal starting from 106 copies/mL, the concentration of ZIKV in acute infection.

The performances of a quartz crystal microbalances (QCMs) based on an electronic nose (E-nose), modified with hairpin-DNA (hpDNA) for carrot aroma profiling has been evaluated. Solid phase micro-extraction (SPME) headspace sampling, combined with gas chromatography (GC), was used as a reference method. The changes in carrot aroma profiles stored at different temperatures (−18 °C, 4 °C, 25 °C, and 40 °C) were monitored during time up to 26 days. The principal component analysis of the data evidenced the different aroma patterns related to the presence of different key compounds. The output data achieved with the hpDNA-based E-nose were able to detect aroma patterns similar to gas chromatography with mass spectrometry (GC-MS). This work demonstrates that hpDNA has different sizes of loops that can be used for the development of sensor arrays able to detect aroma patterns in food and their changes with advantages in terms of easiness of usage, rapidity, and cost of analysis versus classical methods.

In recent years, agricultural and industrial residues have attracted a lot of interest in the recovery of phytochemicals used in the food, pharmaceutical, and cosmetic industries. In this paper, a study on the recovery of phenol compounds from Lycium spp. leaves is presented. Ultrasound-assisted extraction (UAE) and microwave-assisted extraction (MAE) have been used with alcoholic and hydroalcoholic solvents. Methanolic UAE was the most successful technique for extracting phenols from Lycium leaves, and we used on leaves from L. barbarum and L. chinense cultivated in Italy. The extracts were then characterized as regards to the antioxidant properties by in vitro assays and the phenol profiling by a high performance liquid chromatography-diode array detector (HPLC-DAD). Chlorogenic acid and rutin were the main phenol compounds, but considerable differences have been observed between the samples of the two Lycium species. For example, cryptochlorogenic acid was found only in L. barbarum samples, while quercetin-3-O-rutinoside-7-O-glucoside and quercetin-3-O-sophoroside-7-O-rhamnoside only in L. chinense leaves. Finally, multivariate statistical analysis techniques applied to the phenol content allowed us to differentiate samples from different Lycium spp. The results of this study confirm that the extraction is a crucial step in the analytical procedure and show that Lycium leaves represent an interesting source of antioxidant compounds, with potential use in the nutraceutical field.

This is a review of recent affinity-based approaches that detect pesticides in food. The importance of the quantification and monitoring of pesticides is firstly discussed, followed by a description of the different approaches reported in the literature. The different sensing approaches are reported according to the different recognition element used: antibodies, aptamers, or molecularly imprinted polymers. Schemes of detection and the main features of the assays are reported and commented upon. The large number of affinity sensors recently developed and tested on real samples demonstrate that this approach is ready to be validated to monitor the amount of pesticides used in food commodities.

Polyphenolic compounds (PCs) have received exceptional attention at the end of the past millennium and as much at the beginning of the new one. Undoubtedly, these compounds in foodstuffs provide added value for their well-known health benefits, for their technological role and also marketing. Many efforts have been made to provide simple, effective and user friendly analytical methods for the determination and antioxidant capacity (AOC) evaluation of food polyphenols. In a parallel track, over the last twenty years, nanomaterials (NMs) have made their entry in the analytical chemistry domain; NMs have, in fact, opened new paths for the development of analytical methods with the common aim to improve analytical performance and sustainability, becoming new tools in quality assurance of food and beverages. The aim of this review is to provide information on the most recent developments of new NMs-based tools and strategies for total polyphenols (TP) determination and AOC evaluation in food. In this review optical, electrochemical and bioelectrochemical approaches have been reviewed. The use of nanoparticles, quantum dots, carbon nanomaterials and hybrid materials for the detection of polyphenols is the main subject of the works reported. However, particular attention has been paid to the success of the application in real samples, in addition to the NMs. In particular, the discussion has been focused on methods/devices presenting, in the opinion of the authors, clear advancement in the fields, in terms of simplicity, rapidity and usability. This review aims to demonstrate how the NM-based approaches represent valid alternatives to classical methods for polyphenols analysis, and are mature to be integrated for the rapid quality assessment of food quality in lab or directly in the field.

A virtual development of hexapeptide receptors bioinspired by the acetylcholinesterase enzyme active site is proposed. A semicombinatorial approach was applied to generate a virtual hexapeptides library with different affinity properties towards organophosphate and carbamate pesticides. The virtual screening process was addressed to obtain peptides able to separate pesticide subclasses in the experimental work. Three hexapeptides, two generated by molecular modeling and one having a scrambled sequence, were used as selective sorbent materials for pesticides in preanalytical solid-phase extraction (SPE) method. Selective adsorption and cross-reactivity were tested directly on a mix of four pesticides (carbaryl, chlorpyrifos-ethyl, malathion, and thiabendazole) having different structures and physico-chemical properties, at a total concentration of 120 ppb (each pesticide at concentration of 30 ppb). The results were compared to traditional sorbent material such as C-18 and strata-X. Data showed that only one of the hexapeptides virtually designed had significant differences in competitive absorption between aliphatic pesticide malathion, fungicide thiabendazole chosen as negative control, and aromatic pesticides. These results partially supported the simulated strategy.

A rapid and cost-effective computational methodology for designing and rationalizing the selection of small peptides as receptors for dioxin-like compounds was proposed. The backbone of the dioxin Ah receptor binding site was used to design a series of penta- and hexapeptide libraries, with 1400 elements in total. Peptide flexibility was considered and 10 conformers were found to be a good option to represent peptide conformational space with fair speed-accuracy ratio. Each peptide conformer was treated as a possible receptor, generating a dedicated box and then running a docking process using as ligands a family of 76 dibenzo-p-dioxins and 113 dibenzofurans mono- and polychlorinated. Significant predictions were confirmed by comparing primary structure of top and bottom ranked peptides binding dioxins confirming that scrambled positions of the same amino acids gave completely different predicted binding. The hexapeptide EWFQPW, with the best binding score, was chosen as selective sorbent material in solid-phase extraction. The retention performances were tested using the 2,3,7,8-tetrachlorodibenzo-p-dioxin and two polychlorinated biphenyls in order to verify the hexapeptide specificity. The solid-phase extraction experimental procedure was optimized, and analytical parameters of hexapeptide sorbent material were compared with the resin without hexapeptide and a commercial reversed phase cartridge.