Bio-phenols determination in olive oils: Recent mass spectrometry approaches.

Extra virgin olive oil (EVOO) is largely used in Mediterranean diet, and it is also worldwide apprised not only for its organoleptic properties but also for its healthy effects mainly attributed to the presence of several naturally occurring phenolic and polyphenolic compounds (bio-phenols). These compounds are characterized by the presence of multiple phenolic groups in more or less complex structures. Their content is fundamental in defining the healthy qualities of EVOO and consequently the analytical methods for their characterization and quantification are of current interest. Traditionally their determination has been conducted using a colorimetric assay based on the reaction of Folin-Ciocalteu (FC) reagent with the functional hydroxy groups of phenolic compounds. Identification and quantification of the bio-phenols in olive oils requires certainly more performing analytical methods. Chromatographic separation is now commonly achieved by HPLC, coupled with spectrometric devices as UV, FID, and MS. This last approach constitutes an actual cutting-edge application for bio-phenol determination in complex matrices as olive oils, mostly on the light of the development of mass analyzers and the achievement of high resolution and accurate mass measurement in more affordable instrument configurations. After a short survey of some rugged techniques used for bio-phenols determination, in this review have been described the most recent mass spectrometry-based methods, adopted for the analysis of the bio-phenols in EVOOs. In particular, the sample handling and the results of HPLC coupled with low- and high-resolution MS and MS/MS analyzers, of ion mobility mass spectrometry and ambient mass spectrometry have been reported and discussed.

Red Orange and Bitter Orange IntegroPectin: Structure and Main Functional Compounds.

DRIFT, HPLC-MS, and SPME-GC/MS analyses were used to unveil the structure and the main functional compounds of red (blood) orange (Citrus sinensis) and bitter orange (Citrus aurantium). The IntegroPectin samples show evidence that these new citrus pectins are comprised of pectin rich in RG-I hairy regions functionalized with citrus biophenols, chiefly flavonoids and volatile molecules, mostly terpenes. Remarkably, IntegroPectin from the peel of fresh bitter oranges is the first high methoxyl citrus pectin extracted via hydrodynamic cavitation, whereas the red orange IntegroPectin is a low methoxyl pectin. C. aurantium IntegroPectin has a uniquely high concentration of adsorbed flavonoids, especially the flavanone glycosides hesperidin, naringin, and eriocitrin.

BDE-47 exposure modulates cellular responses, oxidative stress and biotransformation related-genes in Mytilus galloprovincialis.

Polybrominated diphenyl ethers (PBDEs) are flame retardants, characterized by elevated stability in the marine environment, where are accumulated by organisms, inducing a wide panel of negative effects. In this study, some biochemical patterns related to toxicity, biotransformation and oxidative stress, were studied in the marine model system, Mytilus galloprovincialis, exposed to BDE-47. Mussels were fed with microalgae, previously treated with increasing concentrations of PBDEs (maximum dose 100 ng L-1 of BDE-47 per day). After 15 days of treatment, mussels were fed with the same diet without BDE-47, for additional 15 days. Gills and digestive glands were analyzed at T 0, at 15 and 30 days. Histopathological lesions were assessed in digestive glands of contaminated mussels, while expression of genes, related to cell cycle, multidrug resistance, oxidative stress and detoxification was evaluated on both gills and digestive glands. After 15 days, BDE-47 exposure significantly affected the cell activity in digestive gland and, at 30 days, only mussels exposed to the lower doses showed a certain recovery. Regarding the gene expression, both gills and digestive glands showed a significant down-regulation of the target genes at 15 days, although most of them were up-regulated at 30 days in digestive gland. The results on BDE-47 accumulation in mussels revealed a dose-dependent concentration in tissues, which remained elevated after further 15 days of depuration. This trend supports the responses of the biomarkers, indicating that exposure, at environmentally realistic concentrations of BDE-47, strongly modulates oxidative stress and related patterns of gene expression, suggesting concerns for long-term effect in the biota.

Volatile Compounds of Lemon and Grapefruit IntegroPectin.

An HS-SPME GC-MS analysis of the volatile compounds adsorbed at the outer surface of lemon and grapefruit pectins obtained via the hydrodynamic cavitation of industrial waste streams of lemon and grapefruit peels in water suggests important new findings en route to understanding the powerful and broad biological activity of these new pectic materials. In agreement with the ultralow degree of esterification of these pectins, the high amount of highly bioactive α-terpineol and terpinen-4-ol points to limonene (and linalool) decomposition catalyzed by residual citric acid in the citrus waste peel residue of the juice industrial production.

Antioxidant activity and phenolic composition in pomegranate (Punica granatum L.) genotypes from south Italy by UHPLC-Orbitrap-MS approach.

BACKGROUND: Pomegranate fruits are a rich source of polyphenols with numerous health-promoting effects. Pomegranate juices of five genotypes ('Mollar', 'Kingdom', 'Dente di Cavallo', and two old populations 'Francofonte' and 'Santa Tecla') were evaluated regarding anthocyanin and non-anthocyanin phenolic contents using ultrahigh performance liquid chromatography (UHPLC)-Orbitrap-mass spectrometry (MS). Moreover, total antioxidant activity (TAA) was evaluated using a 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) assay. RESULTS: Twenty-three phenolic compounds were identified. Cyanidin-3,5-O-diglucoside and pelargonidin-3,5-O-diglucoside were the most representative anthocyanins in all genotypes; the Santa Tecla population had the highest content of these anthocyanins, 97.64 mg L-1 and 40.29 mg L-1 respectively. In the Francofonte population, ferulic acid hexoside was the most abundant compound (391.18 mg L-1 ). TAA values ranged between 221.5 and 36.73 µmol Trolox equivalents/100 mL of juice. A high TAA value was recorded for the Santa Tecla pomegranate population. CONCLUSION: The UHPLC-Orbitrap-MS approach was employed for the first time to identify the phenolic compound profiling in five pomegranate genotypes. TAA was analysed using an ABTS assay, and the results showed a significant variability in nutraceutical potential of the pomegranate genotypes studied. The inclusion of phenolic information in the linear discriminant analysis allowed very good discriminations among genotypes to be obtained. © 2018 Society of Chemical Industry.

Electrospray ion mobility mass spectrometry of positively and negatively charged (1R,2S)-dodecyl(2-hydroxy-1-methyl-2-phenylethyl)dimethylammonium bromide aggregates.

RATIONALE: Self-assembling processes of surfactants in the gas phase constitute a developing research field of interest since they allow information to be gained on the peculiar structural organization of these aggregates, on their ability to incorporate from small molecules up to proteins and on their possible use as carriers of drugs in the gas phase or as cleaning agents and exotic reaction media. METHODS: The mass spectra of charged aggregates of the chiral surfactant (1R,2S)-dodecyl(2-hydroxy-1-methyl-2-phenylethyl)dimethylammonium bromide (DMEB) in the gas phase have been recorded using a Synapt G2-Si mass spectrometer in the positive and negative ion mode. For comparison purposes, the mass spectra of sodium bis(2-ethylhexyl)sulfosuccinate and sodium octane sulfonate aggregates have also been recorded under the same experimental conditions. The collisional cross sections of positively and negatively charged DMEB aggregates were obtained through an appropriate calibration of the measured drift times. RESULTS: For all the surfactants investigated, it has been found that there is a lowest and a highest limit of the aggregation number at each charge state: no aggregates are found outside this range. Moreover, the occurrence at each aggregation number and extra charge of a unique value of drift time points toward aggregates whose conformations do not show discernible shape change in the experiment time scale. The analysis of the collisional cross sections emphasizes that the DMEB aggregates are nearly spherical clusters somewhat affected by the charge state and constituted by interlaced polar and apolar domains. CONCLUSIONS: The analysis of all the experimental findings indicates that in the gas phase DMEB forms supramolecular aggregates characterized by an internal organization whose stability is triggered by the charge state. The comparison of the behavior of DMEB aggregates with that of sodium bis(2-ethylhexyl)sulfosuccinate and sodium octane sulfonate aggregates allows us to highlight the effects on the aggregate organization in gas phase due to nature of the head group and alkyl chain steric hindrance.

Aloin delivery on buccal mucosa: ex vivo studies and design of a new locoregional dosing system.

CONTEXT: Chemoprevention of potential malignant disorders or cancerous lesions that affect oral mucosae requires extended duration of treatment. Locoregional delivery of natural products could represent a promising strategy for this purpose. OBJECTIVE: To investigate the aptitude of aloin to permeate through, or accumulate in, the buccal mucosa and to develop a new prolonged oro-mucosal drug delivery system. MATERIALS AND METHODS: Permeation/accumulation of aloin from Curacao Aloe (containing 50% barbaloin) was evaluated ex vivo, using porcine buccal mucosa as the most useful model to simulate human epithelium. Oro-mucosal matrix tablets were prepared by dispersing aloin (10% w/w) in Eudragit® RS 100 as, biocompatible, low permeable, pH-independent, and non-swelling polymer. The prepared tablets were evaluated for drug-polymer compatibility, weight variation, drug uniformity content, diameter, thickness, hardness, friability, swelling, mucoadhesive strength, and drug release. RESULTS: Aloin has low tendency to cross buccal mucosa, permeation is marginal, and high drug amounts remain entrapped into the epithelium. Matrix tablets characteristics were in agreement with pharmacopoeial requirements. Drug release showed highly reproducible Higuchian profile. Delivery through matrix tablets promoted drug accumulation in the mucosal tissue. DISCUSSION AND CONCLUSION: Following application of matrix tablets on porcine buccal mucosa, the amount of discharged drug recovered in the tissue should be sufficient to produce the desired effects, providing therapeutic drug levels directly at the site of action. Aloin-loaded tablets are valid candidates for prevention/treatment of potentially malignant disorders and oral cancer and could potentially lead to clinically relevant drug delivery system as coadjuvant of conventional chemotherapy/radiation therapy.

HPLC/HRMS and GC/MS for Triacylglycerols Characterization of Tuna Fish Oils Obtained from Green Extraction.

BACKGROUND: Fish oil is one of the most common lipidic substances that is consumed as a dietary supplement. The high omega-3 fatty acid content in fish oil is responsible for its numerous health benefits. Fish species such as mackerel, herring, tuna, and salmon are particularly rich in these lipids, which contain two essential omega-3 fatty acids, known as eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). OBJECTIVES: Due to the scarcity of information in the literature, this study aimed to conduct a qualitative and quantitative characterization of triglycerides (TAGs) in crude tuna fish oil using HPLC/HRMS. Fatty acid (FA) determination was also performed using GC/MS. The tuna fish oils analyzed were produced using a green, low-temperature process from the remnants of fish production, avoiding the use of any extraction solvents. RESULTS: The analyses led to the tentative identification and semi-quantitation of 81 TAGs. In silico saponification and comparison with fatty acid methyl ester results helped to confirm the identified TAGs and their quantities. The study found that the produced oil is rich in EPA, DHA, and erucic acid, while the negligible isomerization of fatty acids to trans-derivatives was observed.

"Golden" Tomato Consumption Ameliorates Metabolic Syndrome: A Focus on the Redox Balance in the High-Fat-Diet-Fed Rat.

Tomato fruits defined as "golden" refer to a food product harvested at an incomplete ripening stage with respect to red tomatoes at full maturation. The aim of this study is to explore the putative influence of "golden tomato" (GT) on Metabolic Syndrome (MetS), especially focusing on the effects on redox homeostasis. Firstly, the differential chemical properties of the GT food matrix were characterized in terms of phytonutrient composition and antioxidant capacities with respect to red tomato (RT). Later, we assessed the biochemical, nutraceutical and eventually disease-modifying potential of GT in vivo in the high-fat-diet rat model of MetS. Our data revealed that GT oral supplementation is able to counterbalance MetS-induced biometric and metabolic modifications. Noteworthy is that this nutritional supplementation proved to reduce plasma oxidant status and improve the endogenous antioxidant barriers, assessed by strong systemic biomarkers. Furthermore, consistently with the reduction of hepatic reactive oxygen and nitrogen species (RONS) levels, treatment with GT markedly reduced the HFD-induced increase in hepatic lipid peroxidation and hepatic steatosis. This research elucidates the importance of food supplementation with GT in the prevention and management of MetS.

Determination of trace levels of organic fining agents in wines: Latest and relevant findings.

The production of red wine plays a key role in the local and international economies of several nations. During the winemaking process, to clarify the final product, before bottling, and to remove undesired substances (proteins, phenols, and tannins), fining agents are commonly added to wines. These substances have different origins (animal and vegetable proteins or mineral compounds), and they show a potential risk for the health of allergic subjects. For these reasons, the residues of fining agents, constituted by exogenous proteins based on gluten, egg, and milk proteins, should not be present in the final product and their trace residues should be quantified with accuracy. In the last decade, several analytical approaches have been developed for their quantitative determination using different sample treatment protocols and analytical techniques. These methods are based on liquid chromatography coupled with mass spectrometry or enzyme-linked immunosorbent assays (ELISAs). Recently, biosensors have been proposed as a potential alternative to immunoassay approaches, allowing rapid, cheap, and simple multi-residue detection. This short review aimed to report the most recent and relevant findings in the field.

Chemical Characterization, Antibacterial Activity, and Embryo Acute Toxicity of Rhus coriaria L. Genotype from Sicily (Italy).

This study reports a full characterization of the Sicilian sumac, Rhus coriaria L. This fruit represents a potential source of fiber (33.21 ± 1.02%) and unsaturated fatty acids, being the contents of linoleic and α-linolenic acids, 30.82 ± 1.21% and 1.85 ± 0.07%, respectively. In addition, the content of phenolic and total anthocyanin was 71.69 ± 1.23 mg/g as gallic acid equivalents, and 6.71 ± 0.12 mg/g as cyanidin-3-O-glucoside equivalents, respectively. The high content in mineral elements, consisting mainly of potassium, calcium, magnesium, and phosphorus, followed by aluminum, iron, sodium, boron, and zinc, was detected by inductively coupled plasma mass spectrometry (ICP-MS). Moreover, its antimicrobial activity was evaluated against multidrug resistant (MDR) microorganisms, represented by Escherichia coli and Klebsiella pneumoniae strains isolated from poultry. The activity of seven different sumac fruit extracts obtained using the following solvents-ethanol (SE), methanol (SM), acetone (SA), ethanol and water (SEW), methanol and water (SMW), acetone and water (SAW), water (SW)-was evaluated. The polyphenol profile of SM extract, which showed better activity, was analyzed by ultra-high performance liquid chromatography coupled with mass spectrometry (UHPLC-MS). The major component identified was gallic acid, followed by quercetin, methyl digallate, pentagalloyl-hexoside, and kaempferol 3-O-glucoside. The non-toxicity of Sicilian R. coriaria was confirmed by testing the effect of the same extract on zebrafish embryos.

Diuron environmental levels effects on marine nematodes: Assessment of ecological indices, taxonomic diversity, and functional traits.

Coastal marine systems are the most sensitive zones to emerging pollutants. The present study aims to investigate the effect of Diuron on the meiofaunal assemblages, collected from the Bizerte channel (Tunisia). Microcosm experiments were set up using four increasing Diuron concentrations [D1 (10 ng g-1 dry weight (DW)), D2 (50 ng g-1 DW), D3 (250 ng g-1 DW) and (1250 ng g-1 DW)] compared to non-contaminated sediments (controls) and all plots were incubated for 30 days. Our results show that Diuron-supplemented sediments provoked the significant decrease of meiofaunal abundance as well as a change in nematodes' diversity and structure composition. All univariate indices, as well as the cumulative k-dominance, were lower in the Diuron than the control plot. Additionally, the ordination of treatments according to the two-dimensional nMDS plots analysis showed a clear structural separation of the Diuron treated replicates from the controls based on the functional groups lists. These current data emphasize the utility of the use of biological traits in the detection of disturbances in the aquatic biotope.

Persistent Organic Pollutants and Fatty Acid Profile in a Typical Cheese from Extensive Farms: First Assessment of Human Exposure by Dietary Intake.

Dairy products represent an important source of beneficial substances for humans. At the same time, they can expose the consumers to environmental contaminants ingested by animals through their diet, influencing their health negatively. This experiment aims to evaluate the risk and benefits related to the consumption of typical stretched cheeses, considering their fatty acid (FA) profile and persistent organic pollutants (POPs) content. Six representative farms, two of them organic, raising Cinisara cattle were selected, considering the typical extensive management systems, based on feeding of natural pasture integrated with concentrate and hay depending on the availability of forage on pastures. A total of 18 cheeses produced in winter, spring and summer with bulk milk of each farm were sampled and analyzed. The chemical composition of cheeses was influenced by farm management, and the FA profile mainly by the season. In particular, cheeses made in spring showed a healthier FA profile with the content of polyunsaturated fatty acids (PUFA), of omega3-PUFA and omega6/omega3 ratio pair to 7.29%, 1.44% and 1.32, respectively, while in winter 5.44%, 0.98% and 2.55, respectively, and in summer 4.77% 0.49% and 3.04, respectively. Due to high levels of feeding integration, cheese made in winter presented unhealthier characteristics compared to the cheeses made in spring and summer, showing high levels of saturated FA (66.2%, 64.2% and 65.5%, respectively), and large contents of polycyclic aromatic hydrocarbons (PAH) (57.07 ng/g fat, 36.25 ng/g fat and 10.22 ng/g fat, respectively) and polychlorinated biphenyls (PCBs) (36.19 ng/g fat, 4.68 ng/g fat and 3.73 ng/g fat, respectively), mainly in those from non-organic farms. Levels of PCBs considered to be hazardous to human health were found in nine samples.

Flavonoids in Lemon and Grapefruit IntegroPectin*.

Following the analysis of terpenes present in new lemon and grapefruit "IntegroPectin" pectins obtained via the hydrodynamic cavitation of industrial lemon and grapefruit processing waste, the HPLC-MS analysis of flavonoid and other phenolic compounds reveals the presence of eriocitrin, naringin, hesperidin and kaempferol typical of the respective citrus fruits. The pectic fibers rich in rhamnogalacturonan-I regions act as chemical sponges adsorbing and concentrating at their outer surface highly bioactive citrus flavonoids and terpenes. These findings, together with the unique molecular structure of these new whole citrus pectins, provide preliminary insight into the broad-scope biological activity of these new biomaterials. Numerous new biomedical applications are anticipated, including likely use in the prevention and treatment of microbial infections and neurodegenerative disease.

Study of green Sicilian table olive fermentations through microbiological, chemical and sensory analyses.

The production of five different green table olive cultivars was studied by a combined strategy consisting of chemical, microbiological and sensory analyses. Cultivable microflora of samples collected during processing was monitored by plate counts on seven synthetic culture media. In all samples Enterobacteriaceae, Pseudomonaceae, staphylococci, lactic acid bacteria and spore-forming bacteria were undetectable. Yeasts and moulds were countable from the day 42 (2 log CFU/ml) till the end of fermentation (6 log CFU/ml). The use of three different approaches for microorganism detection, including a culture-independent methodology, revealed the presence of barely three yeast species during the entire fermentation period: Candida parapsilosis, Pichia guilliermondii and Pichia kluyveri. Biochemical features of technological interest were evaluated for 94 strains in order to investigate their potential role in fermentation of green Sicilian table olives. Olive drupes sampled at picking and periodically during fermentation were also carpologically analyzed, revealing that all the cultivars were suitable for table olive fermentation process. After 120 days of fermentation all products met acceptable commercial standards, although GC-MS analysis evidenced several differences among varieties in terms of aroma components. Results from sensory evaluation led to the conclusion that a revision of technological procedures may improve the final quality of product.

Nile Red lifetime reveals microplastic identity.

Microplastic pollution is recognized as a worldwide environmental problem. The increasing daily use and release of plastics into the environment have led to the accumulation of fragmented microplastics, with potentially awful consequences for the environment, and animal and human health. The detection and identification of microplastics are of utmost importance, but available methods are still limited. In this work, a new approach is presented for the analysis of microplastics based on hydrophobic fluorescence staining with Nile Red, using spectrally resolved confocal fluorescence microscopy and fluorescence lifetime imaging microscopy (FLIM). Significant differences were observed in the emission spectra and fluorescence lifetimes of the analyzed microplastics. Nile Red fluorescence shows determinable behavior based on the polymer matrix and provides a fingerprint for the identification of fragments from different types of plastics. Lifetime imaging coupled with phasor analysis constitutes a fast, robust, and straightforward method for mapping and identifying different microplastics within the same sample in an aquatic environment.

High Yields of Shrimp Oil Rich in Omega-3 and Natural Astaxanthin from Shrimp Waste.

A valued marine oil rich in omega-3 lipids and natural astaxanthin is obtained with remarkably high yield (up to 5 wt %) extending to pink shrimp waste (head and carapace) using the approach to extract fish oil from fish processing byproducts using d-limonene. Biobased limonene is an excellent solvent for both unsaturated lipids and astaxanthin-based carotenoids preventing oxidative degradation during the extraction cycle including solvent separation at 85 °C. Explaining the deep red color of the shrimp oil obtained, computational simulation suggests that d-limonene is also a good solvent for natural astaxanthin abundant in shrimp.

Changes in the proteome of sea urchin Paracentrotus lividus coelomocytes in response to LPS injection into the body cavity.

BACKGROUND: The immune system of echinoderm sea urchins is characterised by a high degree of complexity that is not completely understood. The Mediterranean sea urchin Paracentrotus lividus coelomocytes mediate immune responses through phagocytosis, encapsulation of non-self particles, and production of diffusible factors including antimicrobial molecules. Details of these processes, and molecular pathways driving these mechanisms, are still to be fully elucidated. PRINCIPAL FINDINGS: In the present study we treated the sea urchin P. lividus with the bacterial lipopolysaccharide (LPS) and collected coelomocytes at different time-points (1, 3, 6 and 24 hours). We have shown, using label-free quantitative mass spectrometry, how LPS is able to modulate the coelomocyte proteome and to effect cellular pathways, such as endocytosis and phagocytosis, as soon as the immunomodulating agent is injected. The present study has also shown that treatment can modulate various cellular processes such as cytoskeleton reorganisation, and stress and energetic homeostasis. CONCLUSIONS: Our data demonstrates, through mass spectrometry and the following functional annotation bioinformatics analysis, how the bacterial wall constituent is sufficient to set off an immune response inducing cytoskeleton reorganisation, the appearance of clusters of heat shock proteins (Hsp) and histone proteins and the activation of the endocytic and phagocytic pathways. Data are available via ProteomeXchange with identifier PXD008439.

Economic and Technical Feasibility of Betanin and Pectin Extraction from Opuntia ficus-indica Peel via Microwave-Assisted Hydrodiffusion.

Investigating the feasibility of betanin and pectin extraction from Opuntia ficus-indica peel via microwave-assisted hydrodiffusion and gravity, this study identifies selected important economic and technical aspects associated with this innovative production route starting from prickly pear fruit discards. Which benefits would be derived from this process? Would production be limited to Opuntia-growing countries or, likewise to what happens with dried lemon peel chiefly imported from Argentina, would production take place abroad also? Can distributed manufacturing based on clean extraction technology compete with centralized production using conventional chemical processes?

Omega-3 Extraction from Anchovy Fillet Leftovers with Limonene: Chemical, Economic, and Technical Aspects.

We investigate selected chemical, technical, and economic aspects of the production of fish oil rich in polyunsaturated omega-3 fatty acids from anchovy filleting leftovers using d-limonene as the extraction solvent at ambient temperature and pressure. Entirely derived from the orange peel prior to orange squeezing for juice production, the bio-based solvent is easily recovered, affording a circular economy process with significant potential for practical applications.