Current Developments of Analytical Methodologies for Aflatoxins’ Determination in Food during the Last Decade (2013–2022), with a Particular Focus on Nuts and Nut Products

This review aims to provide a clear overview of the most important analytical development in aflatoxins analysis during the last decade (2013-2022) with a particular focus on nuts and nuts-related products. Aflatoxins (AFs), a group of mycotoxins produced mainly by certain strains of the genus Aspergillus fungi, are known to impose a serious threat to human health. Indeed, AFs are considered carcinogenic to humans, group 1, by the International Agency for Research on Cancer (IARC). Since these toxins can be found in different food commodities, food control organizations worldwide impose maximum levels of AFs for commodities affected by this threat. Thus, they represent a cumbersome issue in terms of quality control, analytical result reliability, and economical losses. It is, therefore, mandatory for food industries to perform analysis on potentially contaminated commodities before the trade. A full perspective of the whole analytical workflow, considering each crucial step during AFs investigation, namely sampling, sample preparation, separation, and detection, will be presented to the reader, focusing on the main challenges related to the topic. A discussion will be primarily held regarding sample preparation methodologies such as partitioning, solid phase extraction (SPE), and immunoaffinity (IA) related methods. This will be followed by an overview of the leading analytical techniques for the detection of aflatoxins, in particular liquid chromatography (LC) coupled to a fluorescence detector (FLD) and/or mass spectrometry (MS). Moreover, the focus on the analytical procedure will not be specific only to traditional methodologies, such as LC, but also to new direct approaches based on imaging and the ability to detect AFs, reducing the need for sample preparation and separative techniques

Sample preparation procedure for the determination of polycyclic aromatic hydrocarbons in petroleum vacuum residue and bitumen

This paper describes a novel method of sample preparation for the determination of trace concentrations of polycyclic aromatic hydrocarbons (PAHs) in high-boiling petroleum products. Limits of quantitation of the investigated PAHs in materials of this type range from tens of nanograms per kilogram to <20 μg/kg. The studies revealed that in order to separate most of interferences from the analytes without a significant loss of PAHs, it is necessary to use size exclusion chromatography as the first step of sample preparation, followed by adsorption using normal-phase liquid chromatography. The use of orthogonal separation procedure described in the paper allows the isolation of only a group of unsubstituted and substituted aromatic hydrocarbons with a specific range of molar mass. The lower the required limit of quantitation of PAHs, the larger is the scale of preparative liquid chromatography in both steps of sample preparation needed. The use of internal standard allows quantitative results to be corrected for the degree of recovery of PAHs during the sample preparation step. Final determination can be carried out using HPLC-FLD, GC-MS, or HPLC-UV–VIS/DAD. The last technique provides a degree of identification through the acquired UV–VIS spectra

Impact of the malaxation temperature on the phenolic profile of cv. Cobrançosa olive oils and assessment of the related health claim

Phenolic compounds contribute to the bioactive properties of olive oil. However, olive oils can only support a health claim concerning the protection against oxidative stress depending on the polyphenolic concentration, requiring effective measures during extraction to preserve/enhance their concentrations. The effect of the malaxation temperature (22, 28 and 34 °C) on the phenolic profile was studied for industrially extracted cv. Cobrançosa oils. Higher malaxation temperatures decreased the contents of the majority of the chromatographically detected compounds (P<0.05, one-way ANOVA), enabling oils differentiation. This decreasing trend was observed for hydroxytyrosol and tyrosol bound forms, determinant for the health claim, which were also negatively affected by temperature, despite revealing that all the industrially extracted oils tested supported the health claim. The observed constant free to bound forms ratio showed that the temperature range tested had a minor effect on bound-forms hydrolysis, being both free and bound forms equally affected by temperature.The authors are grateful to the Foundation for Science andTechnology (FCT, Portugal) forfinancial support by national fundsFCT/MCTES to CIMO (UIDB/00690/2020), to CEB (UIDB/04469/2020), to REQUIMTE-LAQV (UIDB/50006/2020) and to BioTecNorteoperation (NORTE‐01‐0145‐FEDER‐000004) funded by the EuropeanRegional Development Fund under the scope of Norte2020‐Programa Operacional Regional do Norte. Ítala Marx acknowledges the Ph.D.research grant (SFRH/BD/137283/2018) provided by FCT. NunoRodrigues thanks the National funding by FCT- Foundation for Scienceand Technology, P.I., through the institutional scientific employment program-contract.info:eu-repo/semantics/publishedVersio

Donepezil, Anti-Alzheimer's Disease Drug, Prevents Cardiac Rupture during Acute Phase of Myocardial Infarction in Mice

Background: We have previously demonstrated that the chronic intervention in the cholinergic system by donepezil, an acetylcholinesterase inhibitor, plays a beneficial role in suppressing long-term cardiac remodeling after myocardial infarction (MI). In comparison with such a chronic effect, however, the acute effect of donepezil during an acute phase of MI remains unclear. Noticing recent findings of a cholinergic mechanism for anti-inflammatory actions, we tested the hypothesis that donepezil attenuates an acute inflammatory tissue injury following MI. Methods and Results: In isolated and activated macrophages, donepezil significantly reduced intra- and extracellular matrix metalloproteinase-9 (MMP-9). In mice with MI, despite the comparable values of heart rate and blood pressure, the donepezil-treated group showed a significantly lower incidence of cardiac rupture than the untreated group during the acute phase of MI. Immunohistochemistry revealed that MMP-9 was localized at the infarct area where a large number of inflammatory cells including macrophages infiltrated, and the expression and the enzymatic activity of MMP-9 at the left ventricular infarct area was significantly reduced in the donepezil-treated group. Conclusion: The present study suggests that donepezil inhibits the MMP-9-related acute inflammatory tissue injury in the infarcted myocardium, thereby reduces the risk of left ventricular free wall rupture during the acute phase of MI

Expanded encyclopaedias of DNA elements in the human and mouse genomes

All data are available on the ENCODE data portal: www.encodeproject. org. All code is available on GitHub from the links provided in the methods section. Code related to the Registry of cCREs can be found at https:// github.com/weng-lab/ENCODE-cCREs. Code related to SCREEN can be found at https://github.com/weng-lab/SCREEN.© The Author(s) 2020. The human and mouse genomes contain instructions that specify RNAs and proteins and govern the timing, magnitude, and cellular context of their production. To better delineate these elements, phase III of the Encyclopedia of DNA Elements (ENCODE) Project has expanded analysis of the cell and tissue repertoires of RNA transcription, chromatin structure and modification, DNA methylation, chromatin looping, and occupancy by transcription factors and RNA-binding proteins. Here we summarize these efforts, which have produced 5,992 new experimental datasets, including systematic determinations across mouse fetal development. All data are available through the ENCODE data portal (https://www.encodeproject.org), including phase II ENCODE1 and Roadmap Epigenomics2 data. We have developed a registry of 926,535 human and 339,815 mouse candidate cis-regulatory elements, covering 7.9 and 3.4% of their respective genomes, by integrating selected datatypes associated with gene regulation, and constructed a web-based server (SCREEN; http://screen.encodeproject.org) to provide flexible, user-defined access to this resource. Collectively, the ENCODE data and registry provide an expansive resource for the scientific community to build a better understanding of the organization and function of the human and mouse genomes.This work was supported by grants from the NIH under U01HG007019, U01HG007033, U01HG007036, U01HG007037, U41HG006992, U41HG006993, U41HG006994, U41HG006995, U41HG006996, U41HG006997, U41HG006998, U41HG006999, U41HG007000, U41HG007001, U41HG007002, U41HG007003, U54HG006991, U54HG006997, U54HG006998, U54HG007004, U54HG007005, U54HG007010 and UM1HG009442

Advanced Analytical Techniques in Food Analysis

The necessity of more effective and powerful analytical tools and methods for the assessment of food quality and authenticity was highlighted in 2014 by the European Parliament resolution on the food crisis (1). This is not limited to the use of innovative analytical instrumentation but extends to the application of more comprehensive approaches (e.g., fingerprinting) and advanced data handling. This highlight has accelerated the paradigm change in food analysis. Indeed, food analytical scientists are now coupling the use of advanced analytical techniques with approaches borrowed from metabolomics, particularly in data handling. The application of modern, advanced, and innovative techniques are playing a fundamental role in this new scenario, increasing the overall knowledge in the food field and supporting the resolution of complex problems and fraud detection

Exploring 20 eV electron impact ionization in gas chromatography-tandem mass spectrometry for the determination of estrogenic compounds

In electron ionization mass spectrometry (MS), the generation of characteristic fragmentation patterns allows reliable and sensitive identification of compounds. However, loss or a less intense signal of the molecular ion (or more diagnostic ions) can often be observed, which can be detrimental for identification and/or sensitivity, even when MS/MS approaches are applied for quantification. The benefits of applying lower ionization energy (i.e., 20 eV compared to 70 eV) using a gas chromatography (GC) - tandem MS (MS/MS) instrument were investigated in the detection of three estrogenic compounds, namely estrone (E1), 17β-estradiol (E2), and 17α-ethynylestradiol (EE2), emerging aquatic pollutants included in the European Commission Watch List. As expected, the relative intensity of molecular ions (M+.) or high-mass fragments closely related (M+.−CH3) increased significantly at 20 eV compared to 70 eV (from 4.6 % to 35.0 % for EE2, from 22.5 % to 87.3 % for E2, and from 76 % to 100 % for E1). This change in the spectrum profile led to an overall increase in the sensitivity of the compounds when detected using the multiple reaction monitoring mode. These results were compared with the instrumental limit of quantification obtained in liquid chromatography – MS/MS showing a limit of quantification of about 100-folds lower for GC-MS/MS and a completely neglectable matrix effect, thus posing the base for the development of a miniaturized sample preparation method (with an overall lower concentration factor) to achieve the challenging low limits of detection required by the EU regulation for estrogenic compounds

Evolution of hyphenated techniques for mineral oil analysis in food

The occurrence of mineral oil in food is known since the early 1990s, and it was discovered by chance in one of the first applications using the hyphenated LC\u2013GC system. Since then, the relationship between hyphenated techniques and mineral oil analysis has been tightly interrelated and successful. This review aims to show and explain how this mutual interaction has driven the development of the hyphenated techniques on one side and has supported the increase of knowledge on the other, supporting the complex task of mineral oil determination in food. The paper presents the background of the mineral oil problem in food (a brief history of its finding, toxicology, and occurrence), moving then toward the analytical determination. The development of different hyphenated techniques in relation to mineral oil determination is discussed, focusing mainly on 2D techniques, such as LC\u2013GC. The necessity of additional dimensions, such as LC\u2013LC\u2013GC and comprehensive approaches, such as GC 7 GC and LC 7 GC, is also discussed. Finally, the role of the hyphenation with MS is presented