Search Results (29)

There is a growing interest in using essential oils with phytoprotectant properties instead of synthetic pesticides to mitigate the risks of insect pesticide resistance, environmental harm, and adverse effects on non-target organisms and human health. This study focused on the effects of Coridothymus capitatus essential oil on host selection, settling behaviour, and survival of Macrosiphoniella sanborni in dual-choice and no-choice tests. The essential oil and methanol extract of C. capitatus were analyzed using Gas Chromatography–Mass Spectrometry (GC-MS) and Liquid Chromatography–Mass Spectrometry (LTQ-LC-MS Orbitrap), respectively. The antioxidant activity was also tested through the radical scavenging assay. The settling inhibitory activity in the dual-choice test increased dose-dependently from 60% to 72% for essential oil concentrations of 0.1 to 0.3% (v/v) for up to 120 min exposure, but decreased thereafter. However, under no-choice conditions, the inhibitory effect after 60 min of exposure was inversely proportional to the concentration but became proportional by the end of the experiment (72 h). After 72 h, both assays produced a mortality rate of 15% to 17%. C. capitatus was classified as a Carvacrol chemotype. Fifteen phenolic compounds were identified in the MeOH extract, and both the extract and essential oil exhibited substantial antioxidant activity. In conclusion, our findings indicate that C. capitatus essential oil affects the behaviour and survival of M. sanborni. Full article

Ginsenoside Re was the major bioactive component found rich in Panax ginseng C. A. Meyer, which exerted excellent cardiovascular protection, anti-inflammatory, and anti-oxidation effects. The generation of unexpected degradation products (DPs) may influence the therapeutic effect of Re, or even bring toxic effects to patients. However, to date, only a few reports were available about the stability of Re. The present study aims to systematically investigate the degradation behaviors of Re under different stress conditions, including hydrolysis (acidic, basic, and neutral), oxidation, humidity, thermal, and photolytic (ultraviolet and visible light) conditions. A total of thirteen DPs were putatively identified, and among them, nine were discovered for the first time in our study. The results showed that Re was sensitive to exposure to acidic, basic, and oxidation conditions. It underwent a series of chemical degradation reactions, including deglycosylation, dehydration, addition, oxidation at the double bond, and isomerization under various stress conditions. Structural characterization of these DPs was carried out by UHPLC-DAD-CAD and LC-LTQ/Orbitrap. A plausible mechanism of their formation was proposed to support the structures of all DPs of Re. In silico toxicity prediction and metabolism behavior assessment were done by Derek Nexus and Meteor Nexus software. Re and DP-1 to DP-6 were predicted to possess potential skin irritation/corrosion toxicity. DP-11 and DP-12 bear the potential for carcinogenicity, mutagenicity, irritation, hepatotoxicity, and skin sensitization. The observation of these DPs updates our knowledge regarding the stability of Re, which provides valuable information for quality control and to choose suitable storage conditions. Full article

Background and Objectives: In the undertaken study, proteomics alterations of blood-borne XDR S. Typhi isolated from Pakistan were investigated using mass spectrometry. Materials and Methods: MDR and XDR S. Typhi total protein lysates were fractionated, digested, and processed for nanoflow LC-LTQ-Orbitrap MS analysis. Results: Among the 1267 identified proteins, 37 were differentially regulated, of which 28 were up-regulated, and 9 were down-regulated in XDR S. Typhi as compared to MDR S. Typhi. Based on the functional annotation, proteins found up-regulated are involved mainly in metabolic pathways (ManA, FadB, DacC, GpmA, AphA, PfkB, TalA, FbaB, OtsA, 16504242), the biosynthesis of secondary metabolites (ManA, FadB, GlpB, GpmA, PfkB, TalA, FbaB, OtsA), microbial metabolism in diverse environments (FadB, GpmA, PfkB, NfnB, TalA, FbaB), and ABC transporters (PstS, YbeJ, MglB, RbsB, ArtJ). Proteins found down-regulated are involved mainly in carbon metabolism (FadB, GpmA, PfkB, FalA, FbaB) and the biosynthesis of amino acids (GpmA, PfkB, TalA, FbaB). Most of the identified differential proteins were predicted to be antigenic, and matched with resistome data. Conclusions: A total of 28 proteins were up-regulated, and 9 were down-regulated in XDR S. Typhi. Further characterization of the identified proteins will help in understanding the molecular signaling involved in the emergence of XDR S. Typhi. Full article

Vicia faba L. is a leguminous plant with seeds rich in nutritional compounds, such as polyphenols and L-dopa, a dopamine precursor and first-line treatment for Parkinson’s symptoms. Recently, its by-products have been revalued as a sustainable source of bioactive compounds. In this study, aqueous extracts of Lucan broad bean pod valves (BPs) were characterized to evaluate their potential use as adjuvants in severe Parkinson’s disease. L-dopa content, quantified by LC-UV, was much higher in BPs than in seeds (28.65 mg/g dw compared to 0.76 mg/g dw). In addition, vicine and convicine, the metabolites responsible for favism, were not detected in pods. LC-ESI/LTQ-Orbitrap/MS² allowed the identification of the major polyphenolic compounds, including quercetin and catechin equivalents, that could ensure neuroprotection in Parkinson’s disease. ESI(±)-FT-ICR MS was used to build 2D van Krevelen diagrams; polyphenolic compounds and carbohydrates were the most representative classes. The neuroprotective activity of the extracts after MPP⁺-induced neurotoxicity in SH-SY5Y cells was also investigated. BP extracts were more effective than synthetic L-dopa, even at concentrations up to 100 µg/mL, due to the occurrence of antioxidants able to prevent oxidative stress. The stability and antioxidant component of the extracts were then emphasized by using naturally acidic solutions of Punica granatum L., Ribes rubrum L., and gooseberry (Phyllanthus emblica L.) as extraction solvents. Full article

The purple carrot cultivar ‘Purple Sun’ (Daucus carota L.) is characterized by a relevant content of phenolic compounds and anthocyanins, which may play an important role in reducing the risk of chronic diseases and in the treatment of metabolic syndrome. In the present study, the genetic diversity, phytochemical composition, and bioactivities of this outstanding variety were studied for the first time. Genetic analysis by molecular markers estimated the level of genetic purity of this carrot cultivar, whose purple-pigmented roots were used for obtaining the purple carrot ethanol extract (PCE). With the aim to identify specialized metabolites potentially responsible for the bioactivities, the analysis of the metabolite profile of PCE by LC-ESI/LTQ Orbitrap/MS/MS was carried out. LC-ESI/HRMS analysis allowed the assignment of twenty-eight compounds, putatively identified as isocitric acid (1), phenolic acid derivatives (2 and 6), hydroxycinnamic acid derivatives (9, 10, 12–14, 16, 17, 19, 22, and 23), anthocyanins (3–5, 7, 8, 11, and 18), flavanonols (15 and 21), flavonols (20 and 24), oxylipins (25, 26, and 28), and the sesquiterpene 11-acetyloxytorilolone (27); compound 26, corresponding to the primary metabolite trihydroxyoctanoic acid (TriHOME), was the most abundant compound in the LC-ESI/HRMS analysis of the PCE, and hydroxycinnamic acid derivatives followed by anthocyanins were the two most represented groups. The antioxidant activity of PCE, expressed in terms of reactive oxygen species (ROS) level and antioxidant enzymes activity, and its pro-metabolic effect were evaluated. Moreover, the antibacterial activity on Gram (−) and (+) bacterial strains was investigated. An increase in the activity of antioxidant enzymes (SOD, CAT, and GPx), reaching a maximum at 0.5 mg/mL of PCE with a plateau at higher PCE concentrations (1.25, 2.5, and 5.0 mg/mL), was observed. PCE induced an initial decrease in ROS levels at 0.1 and 0.25 mg/mL concentrations, reaching the ROS levels of control at 0.5 mg/mL of PCE with a plateau at higher PCE concentrations (1.25, 2.5, and 5.0 mg/mL). Moreover, significant antioxidant and pro-metabolic effects of PCE on myoblasts were shown by a reduction in ROS content and an increase in ATP production linked to the promotion of mitochondrial respiration. Finally, the bacteriostatic activity of PCE was shown on the different bacterial strains tested, while the bactericidal action of PCE was exclusively observed against the Gram (+) Staphylococcus aureus. The bioactivities of PCE were also investigated from cellular and molecular points of view in colon and hematological cancer cells. The results showed that PCE induces proliferative arrest and modulates the expression of important cell-cycle regulators. For all these health-promoting effects, also supported by initial computational predictions, ‘Purple Sun’ is a promising functional food and an optimal candidate for pharmaceutical and/or nutraceutical preparations. Full article

Olive oil and herbs, two key components of the Mediterranean diet, are known for their beneficial effects on humans. In our study, we incorporated aromatic and medicinal herbs into local monovarietal olive oils via maceration procedures for enrichment. We identified the herbal-derived ingredients that migrate to olive oils and contribute positively to their total phenolic content and functional properties, such as radical scavenging activity. Thus, we characterized the essential oil composition of the aromatic herbs (GC-MS), and we determined the phenolic content and antioxidant capacity of the additives and the virgin olive oils before and after enrichment. The herbal phenolic compounds were analyzed by LC-LTQ/Orbitrap HRMS. We found that olive oils infused with Origanum vulgare ssp. hirtum, Rosmarinus officinalis and Salvia triloba obtained an increased phenolic content, by approximately 1.3 to 3.4 times, in comparison with the untreated ones. Infusion with S. triloba led to a significantly higher antioxidant capacity. Rosmarinic acid, as well as phenolic glucosides, identified in the aromatic herbs, were not incorporated into olive oils due to their high polarity. In contrast, phenolic aglycones and diterpenes from R. officinalis and S. triloba migrated to the enriched olive oils, leading to a significant increase in their phenolic content and to an improvement in their free radical scavenging capacity. Full article

Chestnut shells (CS), the principal by-product of the chestnut processing industry, contain high concentrations of flavonoids and other polyphenols with huge interest for the nutraceuticals field. Nonetheless, the bioaccessibility and bioactivity of phytochemicals can be influenced by their digestibility, making it imperative to evaluate these activities prior to application of CS as a nutraceutical ingredient. This work aims to appraise the effects of in vitro simulated gastrointestinal digestion on the bioaccessibility, bioactivity, and metabolic profiling of CS. An increase in the total phenolic and flavonoid contents, antioxidant/antiradical properties, radical scavenging capacity, and inhibition on acetylcholinesterase activity was evidenced during in vitro simulated digestion. Metabolomic profiling by LC-ESI-LTQ-Orbitrap-MS revealed changes during the simulated digestion, particularly in phenolic compounds (46% of total compounds annotated), lipids (22%), phenylpropanoids (9%), organic acids (7%), carbohydrates (5%), nucleosides (5%), amino acids (4%), and alcohols (1%). Phenolic acids (gallic acid, syringic acid, and hydroxyphenylacetic acid) and flavonoids (epicatechin) were the major polyphenolic classes identified. The heatmap-positive correlations highlighted that the bioactivity of CS is closely related to the phenolic compounds and their bioaccessibility. These findings suggest the reuse of CS as a potential nutraceutical ingredient with antioxidant and neuroprotective effects, encouraging the use of appropriate extraction and/or encapsulation techniques to enhance the bioaccessibility of phenolic compounds. Full article

Traditionally, antibiotics have been used to treat human and animal diseases caused by pathogenic bacteria. The aquaculture industry, which is massively expanding currently, also makes use of several antibiotic classes, resulting in potential antibiotic residues in the surrounding aquatic environment, as well as the cultured products raising bacterial resistance. The aim of this study was the optimization, validation, and application of a solid-phase extraction (SPE) method in combination with liquid chromatography (LC)-LTQ/Orbitrap mass spectrometry in order to determine the most commonly used antibiotics in waters sampled from fish farms, both saltwater and freshwater, located in Greece. Under optimum conditions, the method was validated, achieving recoveries in the range of 57.7% (for sulfamethoxazole in river water) to 95.8% (for florfenicol in river water). The method quantification limits were within the range of 0.25 and 10 ng·L⁻¹ in all cases, with relative standard deviations (RSDs) < 15.9%. The application of the proposed methodology revealed the presence of oxytetracycline and trimethoprim traces. Finally, an assessment of the environmental risk posed by the detected antibiotics was performed, calculating either the risk quotient (RQ) for three trophic levels (8.013 × 10⁻⁶ < RQ < 0.496) or the mixture RQ (0.005 < RQ < 0.682), proving that in all cases, the risk was medium to low. Full article

A modified, quick, easy, cheap, effective, rugged, and safe (QuEChERS) extraction procedure combined with sonication and Ultra-High Performance Liquid Chromatography–Orbitrap-Mass Spectrometry (UHPLC–Orbitrap-MS) was developed as a sensitive and reliable methodology for the determination of multiclass pesticides in full-fat milk. Different amounts of EMR-lipid sorbent were assayed for the cleanup step in order to achieve both acceptably high recoveries and low co-extractives in the final extracts. Accurate mass measurements of the analyte’s pseudo-molecular ions and tandem MS fragmentation were used to quantify and identify the target pesticides. Analytical performance characteristics of the method, such as linearity, recovery, precision, the limit of detection (LOD) and quantification (LOQ), matrix effects (ME), and expanded uncertainty, have been determined for method validation fulfilling all criteria for its use as a validated routine method. The method was successfully applied to real samples (by local farms and commercial), revealing the presence of carbendazim in one milk sample at a concentration level below the maximum residue limits. Full article

The objectives of the present work were to evaluate the semen of dogs by means of proteomics methods and to compare with proteomics results of the blood of the animals, in order to increase available knowledge on the topic and present relevant reference values for semen samples. Semen samples were collected from five Beagle-breed dogs. Reproductive assessment of the animals by means of clinical, ultrasonographic and seminological examinations confirmed their reproductive health. The sperm-rich fraction and the prostatic fraction of semen were processed for proteomics evaluation. LC-MS/MS analysis was performed by means of a LTQ Orbitrap Elite system. The technology combines high separation capacity and strong qualitative ability of proteins in biological samples that require deep proteome coverage. Protein classification was performed based on their functional annotations using Gene Ontology (GO). In blood plasma, semen sperm-rich fraction, and semen prostatic fraction, 59, 42 and 43 proteins, respectively, were detected. Two proteins were identified simultaneously in plasma and the semen sperm-rich fraction, 11 proteins in plasma and the semen prostatic fraction, and three proteins in the semen sperm-rich and prostatic fractions. In semen samples, most proteins were related to cell organization and biogenesis, metabolic processes or transport of ions and molecules. Most proteins were located in the cell membrane, the cytosol or the nucleus. Finally, most proteins performed functions related to binding or enzyme regulation. There were no differences between the semen sperm-rich fraction and prostatic fractions in terms of the clustering of proteins. In conclusion, a baseline reference for proteins in the semen of Beagle-breed dogs is provided. These proteins are involved mostly in supporting spermatozoan maturation, survival and motility, enhancing the reproductive performance of male animals. There appears potential for the proteomics examination of semen to become a tool in semen evaluation. This analysis may potentially identify biomarkers for reproductive disorders. This can be particularly useful in stud animals, also given its advantage as a non-invasive method. Full article

The presence of organic pollutants such as pesticides and pharmaceuticals in the aquatic environment, and especially in regions where fish farms are installed, is a matter of major importance due to their possible risks to ecosystems and public health. The necessity of their detection leads to the development of sensitive, reliable, economical and environmentally friendly analytical methods for controlling their residue in various environmental substrates. In the present work, a solid-phase extraction method was developed, optimized and validated for the analysis of 7 pesticides and 25 pharmaceuticals in seawater using LC-HR-LTQ/Orbitrap-MS. The method was then applied in seawater samples collected from an aquaculture farm located in the Ionian Sea, Greece, in order to evaluate environmental pollution levels. None of the pesticides were detected, while paracetamol was the only pharmaceutical compound that was found (at trace levels). At the same time, passive sampling was conducted as an alternative screening technique, showing the presence of contaminants that were not detected with spot sampling. Among them, irgarol was detected and as far as pharmaceuticals is concerned, trimethoprim and sulfadiazine were found; however, all positive findings were at the very low ppt levels posing no threat to the aquatic environment. Full article

A simple methodology was developed for the determination of four Endocrine Disrupting Chemicals (EDCs) in dairy products. The EDCs included alkylphenols (4-tert-octylphenol, technical nonylphenol isomers, 4-nonylphenol) and bisphenol-A. The methodology consisted of a quick, easy, cheap, effective, rugged and safe (QuEChERS) extraction followed by liquid chromatography (LC) coupled to the hybrid LTQ/Orbitrap mass spectrometer (MS). The high resolution (HR) analysis provided the required selectivity demonstrating excellent sensitivity and enabled the high-mass accuracy of the analytes within short time of analysis, after a chemometric optimization of the instrument parameters. An experimental design was employed for the estimation of the effect of different parameters on the QuEChERS extraction efficiency to obtain the optimum conditions. Method validation proved that analysis exhibited excellent linearity (R² > 0.9966), low enough precision (0.6 to 13.3%) and recoveries in the range of 91 to 108%. Limits of detection (LOD < 6.5 ng g⁻¹) and quantification (LOQ < 20 ng g⁻¹) as well as matrix effects (ME) were also evaluated. The optimized method was successfully applied to analyze dairy commodities varying in fat content and packaging material including milk, yogurts and infant formulae. Detected concentration levels (MDL-10.4 ng g⁻¹) for bisphenol-A BPA in milk samples resulted in 0.36% of TDI for the medium case (average BPA concentrations) and 1.15% of TDI for the worst case (maximum BPA concentration). Full article

Food industries produce a high amount of waste every year. These wastes represent a source of bioactive compounds to be used to produce cosmetic and nutraceutical products. In this study, the possibility to retrain food waste as a potential source of bioactive metabolites is evaluated. In particular, metabolite profiles of different parts (bulb, leaves, stems and little stems) of fennel waste were investigated by liquid chromatography coupled with mass spectrometry (LC-ESI/LTQ Orbitrap MS). To discriminate the different plant parts, a Multivariate Data Analysis approach was developed. Metabolomic analysis allowed the identification of different metabolites mainly belonging to hydroxycinnamic acid derivatives, flavonoid glycosides, flavonoid aglycons, phenolic acids, iridoid derivatives and lignans. The identification of compounds was based on retention times, accurate mass measurements, MS/MS data, exploration on specific metabolites database and comparison with data reported in the literature for F. vulgare. Moreover, the presence of different oxylipins was relieved; these metabolites for the first time were identified in fennel. Most of the metabolites identified in F. vulgare possess anti-inflammatory, antioxidant and/or immunomodulatory properties. Considering that polyphenols are described to possess antioxidant activity, spectrophotometric tests were performed to evaluate the antioxidant activity of each part of the fennel. Full article

Grape canes, the main byproducts of the viticulture industry, contain high-value bioactive phenolic compounds, whose application is limited by their instability and poorly solubility in water. Encapsulation in cyclodextrins allows these drawbacks to be overcome. In this work, a grape cane pilot-plant extract (GC_PPE) was encapsulated in hydroxypropyl beta-cyclodextrin (HP-β-CD) by a spray-drying technique and the formation of an inclusion complex was confirmed by microscopy and infrared spectroscopy. The phenolic profile of the complex was analyzed by LC-ESI-LTQ-Orbitrap-MS and the encapsulation efficiency of the phenolic compounds was determined. A total of 42 compounds were identified, including stilbenes, flavonoids, and phenolic acids, and a complex of (epi)catechin with β-CD was detected, confirming the interaction between polyphenols and cyclodextrin. The encapsulation efficiency for the total extract was 80.5 ± 1.1%, with restrytisol showing the highest value (97.0 ± 0.6%) and (E)-resveratrol (32.7 ± 2.8%) the lowest value. The antioxidant capacity of the inclusion complex, determined by ORAC-FL, was 5300 ± 472 µmol TE/g DW, which was similar to the value obtained for the unencapsulated extract. This formulation might be used to improve the stability, solubility, and bioavailability of phenolic compounds of the GC_PPE for water-soluble food and pharmaceutical applications. Full article

The purpose of this work was to study the distribution of oleacein (OLEA) and its metabolites in rat plasma and different tissues, namely brain, heart, kidney, liver, lung, small intestine, spleen, stomach, skin, and thyroid, following the acute intake of a refined olive oil containing 0.3 mg/mL of OLEA. For this purpose, a distribution kinetics study was carried out. The plasma and tissues were collected at 1, 2, and 4.5 h after the intervention, and analyzed by LC-ESI-LTQ-Orbitrap-MS. Unmetabolized OLEA was detected in the stomach, small intestine, liver, plasma and, most notably, the heart. This finding may be useful for the development of new applications of OLEA for cardiovascular disease prevention. Noteworthy are also the high levels of hydroxytyrosol (OH-TY) and OLEA + CH₃ found in the small intestine, liver, and plasma, and the detection of nine OLEA metabolites, five of them arising from conjugation reactions. Liver, heart, spleen, and lungs were the target tissues where the metabolites were most distributed. However, it is important to note that OH-TY, in our experimental conditions, was not detected in any target tissue (heart, spleen, thyroids, lungs, brain, and skin). These results shed further light on the metabolism and tissue distribution of OLEA and contribute to understanding the mechanisms underlying its effect in human health. Full article