Comprehensive Two-Dimensional Gas Chromatography with a TOF MS Detector-An Effective Tool to Trace the Signature of Grape Varieties.

Varietal volatile compounds are characteristic of each variety of grapes and come from the skins of the grapes. This work focuses on the development of a methodology for the analysis of free compounds in grapes from Trincadeira, Cabernet Sauvignon, Syrah, Castelão and Tinta Barroca from the 2021 and 2022 harvests, using HS-SPME-GC × GC-TOFMS. To achieve this purpose, a previous optimization step of sample preparation was implemented, with the optimized conditions being 4 g of grapes, 2 g of NaCl, and 2 mL of H2O. The extraction conditions were also optimized, and it was observed that performing the extraction for 40 min at 60 °C was the best for identifying more varietal compounds. The fiber used was a triple fiber of carboxen/divinylbenzene/polydimethylsiloxane (CAR/DVB/PDMS). In addition to the sample preparation, the analytical conditions were also optimized, enabling the adequate separation of analytes. Using the optimized methodology, it was possible to identify fifty-two free volatile compounds, including seventeen monoterpenes, twenty-eight sesquiterpenes, and seven C13-norisoprenoids. It was observed that in 2021, more free varietal volatile compounds were identifiable compared to 2022. According to the results obtained through a linear discriminant analysis (LDA), the differences in volatile varietal signature are observed both among different grape varieties and across different years.

A Critical Review of Analytical Methods for the Quantification of Phthalates Esters in Two Important European Food Products: Olive Oil and Wine.

Phthalic acid esters (PAEs) are a class of chemicals widely used as plasticizers. These compounds, considered toxic, do not bond to the polymeric matrix of plastic and can, therefore, migrate into the surrounding environment, posing a risk to human health. The primary source of human exposure is food, which can become contaminated during cultivation, production, and packaging. Therefore, it is imperative to control and regulate this exposure. This review covers the analytical methods used for their determination in two economically significant products: olive oil and wine. Additionally, it provides a summary and analysis of information regarding the characteristics, toxicity, effects on human health, and current regulations pertaining to PAEs in food. Various approaches for the extraction, purification, and quantification of these analytes are highlighted. Solvent and sorbent-based extraction techniques are reviewed, as are the chromatographic separation and other methods currently applied in the analysis of PAEs in wines and olive oils. The analysis of these contaminants is challenging due to the complexities of the matrices and the widespread presence of PAEs in analytical laboratories, demanding the implementation of appropriate strategies.

A New Laboratory Scale Olive Oil Extraction Method with Comparative Characterization of Phenolic and Fatty Acid Composition.

The establishment of operation protocols for olive oil (OO) extraction at non-industrial scale is crucial for research purposes. Thus, the present study proposes a simple and cost-effective method for OO extraction at the laboratory scale (LS) level. To validate the proposed methodology, industrial OO extraction (IS) was performed in parallel, using the same cultivars 'Galega vulgar' (GV), 'Cobrançosa' (COB) and 'Arbequina' (ARB) collected from the same orchards, within the same period. Obtained results showed highest extractability for COB and ARB, of about 53%, while GAL showed 50%. All produced OO showed values lower than the regulated limits for the physicochemical parameters (acidity, K232, K268 and ΔK), classifying them as extra virgin OO (EVOO). Highest total phenolic content was observed for COB, with no significant differences (p-value > 0.05) between extraction methods. Regarding fatty acid composition, oleic acid (C18:1) showed the lowest percentage for ARB, with about 66% and 68%, for LS and IS, respectively, and the highest for GV with about 72% for both LS and IS. Furthermore, all samples from both extraction methods were compared to the European Community Regulation, with fatty acid composition within the regulated levels for EVOO. This work showed promising results regarding extraction yields and OO extractability, as well as its quality parameters.

New Insights into the Specificity, Authenticity, and Traceability Analysis of Olive Oils.

Olive oil is a traditional product of the Mediterranean diet [...].

Chemometric Discrimination of the Varietal Origin of Extra Virgin Olive Oils: Usefulness of 13C Distortionless Enhancement by Polarization Transfer Pulse Sequence and 1H Nuclear Magnetic Resonance Data and Effectiveness of Fusion with Mid-Infrared Spectroscopy Data.

The label authentication of monovarietal extra virgin olives is of great relevance from a socio-economical point of view. This work aims to gain insights into the prediction of the varietal origin of extra virgin olive oil (EVOO) samples obtained from single olive cultivars, French cultivars Olivière, Salonenque, and Tanche and Portuguese cultivars Blanqueta, Carrasquenha, and Galega Vulgar, collected in 2016-2017 and 2017-2018 harvest seasons. To pursue this study, spectroscopic approaches based on one-dimensional nuclear magnetic resonance (1D NMR) spectroscopy, namely, 1H and 13C NMR distortionless enhancement by polarization transfer (DEPT) 45 pulse sequence, and Fourier transform mid-infrared spectroscopy (FT-MIR) are used in combination with partial least square discriminant analysis (PLS1-DA). The results obtained by PLS1-DA models using 1H and 13C NMR DEPT 45 data are compared to those of PLS1-DA models using MIR data. The application of a control chart method allows for the optimization of the interpretation of the PLS1-DA results, and an efficient two-step strategy is proposed to improve the discrimination of the six studied cultivars. Then, NMR and MIR data are combined by either a mid- or high-level data fusion approach to further improve the discrimination. The models are also tested on samples from other cultivars to check their ability to reject varieties that were not considered in the calibration process.

Seeking a Fast Screening Method of the Varietal Origin of Olive Oil: The Usefulness of an NMR-Based Approach.

This work encompasses the use of 1D multinuclear NMR spectroscopy, namely, 1H NMR and 13C NMR DEPT 45, combined with a multivariate statistical analysis to characterize olive oils produced from nine different varieties: Galega Vulgar, Cobrançosa, Cordovil de Serpa, Blanqueta, Madural, Verdeal Alentejana, Arbequina, Picual and Carrasquenha. Thus, the suitability of an NMR-based spectroscopic tool to discriminate olive oils according to their varietal origin is addressed. The results obtained show that the model based on 13C NMR DEPT 45 data has a stronger performance than the model based on 1H NMR data, proving to be promising in the discrimination of the olive oils under study based on their varietal origin, being particularly relevant for olive oils of the Galega Vulgar variety.

Revealing the yeast modulation potential on amino acid composition and volatile profile of Arinto white wines by a combined chromatographic-based approach.

The importance of yeasts in aroma production during wine fermentation is a significant concern for obtaining a wine that appraises a broad number of consumers. For wine producers, wine aroma modulation is an essential issue where the yeasts used during the winemaking process represents a feasible way to improve the complexity and enhance wines specific characteristics. During the fermentation process of wines, yeasts convert grapes sugars into alcohol, carbon dioxide and a large number of secondary metabolites, depending on yeast metabolism, affecting the wine composition, namely its aroma and amino acids (AAs) composition. So, the present work aims to study the effect of different Saccharomyces-type yeasts on the AAs composition and volatile profile of Arinto white wines. To pursue this goal, four white wines from Arinto grapes were fermented with three different commercial yeasts (Saccharomyces bayanus EC1118, Saccharomyces cerevisiae CY3079, Saccharomyces bayanus QA23) and one Native yeast. Arinto wines AAs composition was quantified by HPLC-DAD, after a derivatization step to obtain the aminoenone derivatives. The volatile content of Arinto wines was determined by GC/MS, after an HS-SPME extraction. Results showed significant differences among the AAs content and volatile profile in the Arinto wines. The higher AAs content was found in the Arinto wines fermented with the CY3079 yeast (470.74 mg•L-1), and the lowest content of AAs in the Arinto wines fermented with EC1118 yeast (343.06 mg•L-1). Native yeast results in wines with a volatile profile richer in esters compared to the other sample wines. Principal component analysis (PCA) obtained with combined data of AAs and volatile compounds, after normalization, for each Arinto wine samples, shows a clear separation of wines fermented with Native and CY3079 yeasts in relation to QA23 and EC1118 fermented wines . The first and second principal components are responsible for 44.40% and 32.20%, respectively, of the system's variance, which clearly showed a differentiation among wines.

Combination of Stable Isotope Analysis and Chemometrics to Discriminate Geoclimatically and Temporally the Virgin Olive Oils from Three Mediterranean Countries.

The knowledge of the isotopic composition of virgin olive oil (VOO) allows the evaluation of authenticity and geographical origin, being an important tool against fraud. This study aimed to assess if VOOs produced in three Mediterranean regions could be discriminated on the basis of multivariate statistical analysis of geoclimatic and isotopic data. A total of 138 geo-referenced VOO samples from Portugal, France and Turkey from two different cultivation years were collected. The isotopic composition (δ13C, δ2H and δ18O) of VOOs was obtained using an elemental analyzer coupled to an isotope ratio mass spectrometer (EA-IRMS). One-way analysis of variance for δ13C, δ2H and δ18O showed some significant differences either between crop years or geoclimatic conditions. Based on multiple regression analyses using meteorological and geographical parameters, a meteoric water line for olive oil from Portugal, France and Turkey, in two harvest years, were created to assess the impact of climate change on their δ2H and δ18O values. Principal component analysis and Linear Discriminant Analysis, used to sort samples according to geoclimatic origin, performed best for French and Portuguese olive oils. In light of the results, multivariate isotopic analysis of VOO samples may discriminate not only between geoclimatic regions but also among cultivation years.

Assessment of Dimethoate in Olive Oil Samples Using a Dual Responsive Molecularly Imprinting-Based Approach.

A new generation of advanced materials developed by molecular imprinting technology showing a stimuli-responsive functionality are emerging. The switchable ability to control the uptake/release of the target analyte by action of external stimulus combined with a remarkable selectivity and specificity, makes these functional materials very attractive for sample preparation purposes. In this work, the usefulness of a sample preparation tool for the selective enrichment/pre-concentration of dimethoate from olive oil spiked samples based on "tailor-made" dual responsive magnetic and photonic molecularly imprinted polymers as sorbents is explored. To achieve this goal, a smart molecularly imprinted polymer (MIP) possessing magnetic and photonic responsiveness was successfully synthesized, and its physico-chemical and morphological characterization was assessed. Further, the trace analysis of dimethoate in spiked olive oil samples was validated and successfully implemented using smart-MIPs as sorbents in the sample preparation step, with high recoveries (83.5 ± 0.3%) and low detection limit (0.03µg·mL-1).

Revisiting 3D van Krevelen diagrams as a tool for the visualization of volatile profile of varietal olive oils from Alentejo region, Portugal.

This paper aims to study the volatile composition of monovarietal extra-virgin olive oils from different varieties from Alentejo region (Portugal). Volatile profile of 82 olive oils was performed by head-space solid phase micro-extraction hyphenated with gas chromatography/mass spectrometry (HS-SPME-GC/MS). A total of 107 volatile compounds, belonging to several class of compounds, like aldehydes, hydrocarbons, alcohols, terpenoids, ketones, sulphurous compounds, acids and esters were identified, assigned to a specific biogenic origin and total abundances were plotted in modified 3D van Krevelen diagrams. The 3D plots showed that there are compositional differences among olive oil varieties. Discriminant analysis confirmed that the pattern of volatile compounds includes enough information to group sample variety amongst discrete monovarietal olive oil. Combining statistical analysis (t-Student) and graphical-tools (van Krevelen diagram) seems to be a powerful tool to determine what molecular families were characteristic of each olive oil variety, contributing to the valorization of monovarietal Portuguese olive oil.

Winemaking with vine-shoots. Modulating the composition of wines by using their own resources.

Vine-shoots from two important Vitis vinifera, Airén and Cencibel, have been prepared in two different formats (chip and granule) and added (12 g/L) in their own wines in different winemaking steps. Results have shown significant differences depending in all conditions tested, and wine chemical composition was modulated while in contact with vine-shoots. Compounds such as trans-resveratrol, increased its concentration up to 4 mg/L in Airén white wines. In Cencibel red wines, vanillin was found in a concentration four times above its odor threshold and independently of the vine-shoot variety, format and moment of addition, compounds such as (-)-epicatechin and (+)-catechin increased the concentration with respect to the control wine. When vine-shoots were added after fermentation, ß-ionol appeared for first time in all wines. In case of guaiacol, the higher increment was observed for Airén wines. Therefore, it is possible to elaborate distinctive wines using their own resources.

A photoswitchable "host-guest" approach for the selective enrichment of dimethoate from olive oil.

This work describes the development of a new selective photocontrollable molecularly imprinted-based sorbent for the selective enrichment/pre-concentration of dimethoate from spiked olive oil samples. To achieve this goal an improved molecularly imprinted strategy relying on the embedding of a functional monomer containing an azobenzene chromophore as light-responsive element, on the crosslinked tridimensional molecular imprinted network, has been assessed. To address the mechanisms underlying template recognition and uptake/release of the analyte from the functional imprinted material, computational studies using a quantum chemical approach, have been explored. This new functional sorbent provides a straightforward controllable uptake/release of the target template using light as the stimuli tool, which is highly advantageous due to light manipulation characteristics, such as superior clean, precision and remote controllable properties. In general, this work will contribute to the implementation of a photoswitchable analytical methodology that proves to be suitable for the selective isolation and further quantification of dimethoate from olive oil matrices at levels similar to the maximum residues limits imposed by the legislation. The limits of detection, calculated based on 3σ, was 1.6 mgL-1 and the limit of quantification, based on 10σ, was 5.2 mgL-1. The implemented sample preparation shows high reproducibility and recoveries (93.3 ±â€¯0.4%).

A magnetic controllable tool for the selective enrichment of dimethoate from olive oil samples: A responsive molecular imprinting-based approach.

Aiming to develop a straightforward magnetic-based sample preparation methodology for the selective extraction of dimethoate from olive oil, the synthesis of dimethoate-imprinted polymer on the surface of modified magnetic nanoparticles has been attempted. Molecular recognition assays have proven their suitability for the selective pre-concentration of dimethoate. Mechanistic basis for template selective recognition has been explored using a quantum chemical approach, providing new insights about the mechanisms underlying template recognition. Thus, a magnetic molecularly imprinted solid-phase extraction method was developed allowing the extraction of dimethoate from spiked olive oil samples, at levels similar to the maximum residue limits imposed by legislation, followed by the quantification of their levels by high-performance liquid chromatography with diode-array detection. Recoveries of 94.55% were obtained, with relative standard deviations lower than 0.53% (n = 3). The developed sample preparation technique enables a selective pre-concentration/enrichment of dimethoate from olive oil matrix with minimum handling and less solvent consumption.

"On-off" switchable tool for food sample preparation: merging molecularly imprinting technology with stimuli-responsive blocks. Current status, challenges and highlighted applications.

Sample preparation still remains a great challenge in the analytical workflow representing the most time-consuming and laborious step in analytical procedures. Ideally, sample pre-treatment procedures must be more selective, cheap, quick and environmental friendly. Molecular imprinting technology is a powerful tool in the development of highly selective sample preparation methodologies enabling to preconcentrate the analytes from a complex food matrix. Actually, the design and development of molecularly imprinted polymers-based functional materials that merge an enhancement of selectivity with a controllable and switchable mode of action by means of specific stimulus constitutes a hot research topic in the field of food analysis. Thus, combining the stimuli responsive mechanism and imprinting technology a new generation of materials are emerging. The application of these smart materials in sample preparation is in early stage of development, nevertheless new improvements will promote a new driven in the demanding field of food sample preparation. The new trends in the advancement of food sample preparation using these smart materials will be presented in this review and highlighted the most relevant applications in this particular area of knowledge.

Influence on wine biogenic amine composition of modifications to soil N availability and grapevine N by cover crops.

BACKGROUND: Vineyard soil management can modify the nitrogen soil availability and, therefore, grape amino acid content. These compounds are precursors of biogenic amines, which have negative effects on wine quality and human health. The objective was to study whether the effect of conventional tillage and two cover crops (barley and clover) on grapevine nitrogen status could be related to wine biogenic amines. Over 4 years, soil NO3- -N, nitrogen content in leaf and wine biogenic amine concentration were determined. RESULTS: Barley reduced soil NO3- -N availability and clover increased it. In 2011, at bloom, nitrogen content decreased with barley treatment in both blade and petiole. In 2012, nitrogen content in both leaf tissues at bloom was greater with clover than with tillage and barley treatments. Also, total biogenic amines decreased in barley with respect to tillage and clover treatments. There were correlations between some individual and total biogenic amine concentrations with respect to nitrogen content in leaf tissues. CONCLUSION: Wine biogenic amine concentration can be affected by the grapevine nitrogen status, provoked by changes in the soil NO3- -N availability with both cover crop treatments. © 2017 Society of Chemical Industry.

Impact of salt reduction on biogenic amines, fatty acids, microbiota, texture and sensory profile in traditional blood dry-cured sausages.

Meat industry needs to reduce salt in their products due to health issues. The present study evaluated the effect of salt reduction from 6% to 3% in two Portuguese traditional blood dry-cured sausages. Physicochemical and microbiological parameters, biogenic amines, fatty acids and texture profiles and sensory panel evaluations were considered. Differences due to salt reduction were perceptible in a faint decline of water activity, which slightly favoured microbial growth. Total biogenic amines content ranged from 88.86 to 796.68mgkg-1 fresh matter, with higher amounts, particularly of cadaverine, histamine and tyramine, in low-salt products. Still, histamine and other vasoactive amines remained at low levels, thus not affecting consumers' health. Regarding fatty acids, no significant differences were observed due to salt. However, texture profile analysis revealed lower resilience and cohesiveness in low-salt products, although no textural changes were observed by the sensory panel. Nevertheless, low-salt sausages were clearly preferred by panellists.

Dual-layer solid-phase extraction based on molecular imprinting technology: Seeking a route to enhance selectivity for trace analysis of pesticide residues in olive oil.

Aiming to introduce a multiresidue analysis for the trace detection of pesticide residues belonging to organophosphorus and triazine classes from olive oil samples, a new sample preparation methodology comprising the use of a dual layer of "tailor-made" molecularly imprinted polymers (MIPs) SPE for the simultaneous extraction of both pesticides in a single procedure has been attempted. This work has focused on the implementation of a dual MIP-layer SPE procedure (DL-MISPE) encompassing the use of two MIP layers as specific sorbents. In order to achieve higher recovery rates, the amount of MIP layers has been optimized as well as the influence of MIP packaging order. The optimized DL-MISPE approach has been used in the preconcentration of spiked organic olive oil samples with concentrations of dimethoate and terbuthylazine similar to the maximum residue limits and further quantification by HPLC. High recovery rates for dimethoate (95%) and terbuthylazine (94%) have been achieved with good accuracy and precision. Overall, this work constitutes the first attempt on the development of a dual pesticide residue methodology for the trace analysis of pesticide residues based on molecular imprinting technology. Thus, DL-MISPE constitutes a reliable, robust, and sensitive sample preparation methodology that enables preconcentration of the target pesticides in complex olive oil samples, even at levels similar to the maximum residue limits enforced by the legislation.

Characterisation of "Catalão" and "Salsichão" Portuguese traditional sausages with salt reduction.

The present study evaluated the effect of salt reduction on traditional dry-cured sausages' safety, quality and product acceptance, comprising physicochemical and microbiological parameters, biogenic amines, fatty acids, texture profile and sensory analysis. According to our results, salt content had a major effect on microbiological counts, although not compromising the products' safety. Marked differences were identified regarding biogenic amines, in particular for histamine, tyramine and cadaverine, which were detected in larger amounts in products with 3%. Moreover, significant differences in the fatty acids profile have also been found, but only in less abundant components such as linoleic, lauric and heneicosanoic acids. Texture profile analysis of low-salt products, revealed a decrease in hardness and chewiness, along with an increase in adhesiveness values. Sensory evaluations revealed that despite the less intense aroma, products with 3% salt, had a more balanced salt perception. Our results suggest that salt content may be reduced to 50% in dry-cured products, with the obvious health-related advantages.

Antagonistic activity of fungi of Olea europaea L. against Colletotrichum acutatum.

Fungi naturally present in olive trees were identified and tested for their antagonistic potential against Colletotrichum acutatum. A total of 14 isolates were identified, 12 belonged to genera Alternaria, Epicoccum, Fusarium, Aspergillus, Anthrinium, Chaetomium, Diaporthe, Nigrospora, one to family Xylariaceae and one was unclassified. All fungal isolates showed some inhibitory action over the growth of C. acutatum during dual culture growth, however, when agar-diffusible tests were performed only five fungal isolates caused C. acutatum growth inhibition: Alternaria sp. isolate 2 (26.8%), the fungus from Xylariaceae family (14.3%), Alternaria sp. isolate 1 (10.7%); Diaporthe sp. (10.7%), Nigrospora oryzae (3.5%). Volatile substances produced by these isolates were identified through gas-chromatography techniques, as phenylethyl alcohol, 4-methylquinazoline, benzothiazole, benzyl alcohol, lilial, galaxolide, among others. These inhibitory volatiles could play a significant role in reduction of C. acutatum expansion in olive and their study as potential biocontrol agents should be further explored.

Design and development of molecularly imprinted polymers for the selective extraction of deltamethrin in olive oil: An integrated computational-assisted approach.

This work firstly addresses the design and development of molecularly imprinted systems selective for deltamethrin aiming to provide a suitable sorbent for solid phase (SPE) extraction that will be further used for the implementation of an analytical methodology for the trace analysis of the target pesticide in spiked olive oil samples. To achieve this goal, a preliminary evaluation of the molecular recognition and selectivity of the molecularly imprinted polymers has been performed. In order to investigate the complexity of the mechanistic basis for template selective recognition in these polymeric matrices, the use of a quantum chemical approach has been attempted providing new insights about the mechanisms underlying template recognition, and in particular the crucial role of the crosslinker agent and the solvent used. Thus, DFT calculations corroborate the results obtained by experimental molecular recognition assays enabling one to select the most suitable imprinting system for MISPE extraction technique which encompasses acrylamide as functional monomer and ethylene glycol dimethacrylate as crosslinker. Furthermore, an analytical methodology comprising a sample preparation step based on solid phase extraction has been implemented using this "tailor made" imprinting system as sorbent, for the selective isolation/pre-concentration of deltamethrin from olive oil samples. Molecularly imprinted solid phase extraction (MISPE) methodology was successfully applied for the clean-up of spiked olive oil samples, with recovery rates up to 94%.