Characterization of Volatile and Flavonoid Composition of Different Cuts of Dried Onion (Allium cepa L.) by HS-SPME-GC-MS, HS-SPME-GC×GC-TOF and HPLC-DAD.

Onion is widely used worldwide in various forms for both food and medicinal applications, thanks to its high content of phytonutrients, such as flavonoids and volatile sulfur compounds. Fresh onion is very perishable and drying is widely applied for extending shelf-life, thus obtaining a very easy-to-use functional food ingredient. The flavonoid and volatile fractions of different onion cuts (flakes, rings) prepared through different drying cycles in a static oven, were characterized by high-performance liquid chromatography with a diode-array detector HPLC-DAD, Head Space-Solid Phase Micro Extraction followed by Gas Chromatography coupled with Mass Spectrometry (HS-SPME-GC-MS) and Head-Space Solid Phase Micro Extraction followed by comprehensive two-dimensional Gas-Chromatography (HS-SPME-GC×GC-TOF). Onion flakes showed a significantly higher flavonoid content (3.56 mg g-1) than onion rings (2.04 mg g-1). Onion flakes showed greater amount of volatile organic compounds (VOCs) (127.26 mg g-1) than onion rings (42.79 mg g-1), with different relative amounts of di- and trisulfides-disulfides largely predominate the volatile fraction (amounts over 60% on the total volatile content), followed by trisulfides and dipropyl disulfide and dipropyl trisulfide were the most abundant VOCs. HS-SPME-GC×GC-TOF allowed for the detection of the presence of allylthiol, diethanol sulfide, 4,6-diethyl1,2,3,5-tetrathiolane, not detected by HS-SPME-GC-MS, and provided a fast and direct visualization and comparison of different samples. These results highlight different nutraceutical properties of dried onion samples processed otherwise, only differing in shape and size, thus pointing out potentially different uses as functional ingredients.

GC-MS and HS-SPME-GC×GC-TOFMS Determination of the Volatile Composition of Essential Oils and Hydrosols (By-Products) from Four Eucalyptus Species Cultivated in Tuscany.

Essential oils are widely used as functional ingredients for potential multi-purpose functional uses. Hydrosols, co-products of the distillation of plant material, are used in food and cosmetic industries and in biological agriculture, but their volatile composition is poorly investigated. The volatile fractions of essential oils and hydrosols from four less-studied 1,8-cineol-rich Eucalyptus species (E. parvula L.A.S. Johnson & K.D. Hill, E. cinerea F. Muell, E. pulverulenta Sims and E. pulverulenta baby blue Sims), cultivated in Tuscany in a system of organic farming, were characterized by solvent dilution (essential oils) or extraction (hydrosols) followed by GC-MS and by HS-SPME-GC×GC-TOFMS analysis. GC-MS analysis showed that essential oils were mainly constituted by oxygenated monoterpenes, particularly 1,8-cineole, with monoterpenes hydrocarbons up to 10.8%. Relative differences in the abundance of minor terpenes as limonene, α-pinene, γ-terpinene, p-cymene, terpinen-4-ol, α-terpineol, and alloaromandrene were pointed out and seem to be suitable for differentiation among EOs of the four different Eucalyptus species. Hydrosols of these species were characterized for the first time: they were mainly constituted by oxygenated monoterpenes (97.6⁻98.9%), with 1,8-cineole up to 1.6 g/L, while monoterpene and sesquiterpene hydrocarbons were detected only in traces. HS-SPME-GC×GC-TOFMS analysis also allowed providing metabolic profiling of hydrosols for the direct comparison and visualization of volatile components, pointing out the potentially different uses of these products as functional ingredients in food, beverage, and cosmetic industries.

In vitro effects of extracts of extra virgin olive oil on human colon cancer cells.

The Mediterranean diet is associated with a lower incidence of atherosclerosis, cardiovascular diseases, and some types of cancer. Recent interest has been focused on the biological activity of phenolic compounds present in extra virgin olive oils (EVOOs). Both in vivo and in vitro studies have shown that EVOO components have positive effects on metabolic parameters, such as plasma lipoproteins, oxidative damage, inflammatory markers, platelet function, and antimicrobial activity. We have investigated the possible interactions between 2 extracts of extra virgin olive oil and estrogen receptor ß (ERß) in an in vitro model of colon cancer. The qualification and quantification of the components of the 2 samples tested showed that phenolic compounds-hydroxytyrosol, secoiridoids, and lignans-are the major represented compounds. EVOO extracts were tested on a colon cancer cell line engineered to overexpress ERß (HCT8-ß8). By using custom made Oligo microarray, gene expression profiles of colon cancer cells challenged with EVOO-T extracts when compared with those of cells exposed to 17ß-estradiol (17ß-E2). This study demonstrated that the EVOO extracts tested showed an antiproliferative effect on colon cancer cells through the interaction with estrogen-dependent signals involved in tumor cell growth. Specifically, the ability of EVOO extracts to inhibit cell proliferation was superimposable to the activation of the ERß receptor, similar to what was observed after 17ß-E2 challenge.

Polyphenol-Rich Extracts from Agroindustrial Waste and Byproducts: Results and Perspectives According to the Green Chemistry and Circular Economy.

Polyphenols are natural secondary metabolites found in plants endowed with multiple biological activities (antioxidant, anti-inflammatory, antimicrobial, cardioprotective, and anticancer). In view of these properties, they find many applications and are used as active ingredients in nutraceutical, food, pharmaceutical, and cosmetic formulations. In accordance with green chemistry and circular economy strategies, they can also be recovered from agroindustrial waste and reused in various sectors, promoting sustainable processes. This review described structural characteristics, methods for extraction, biological properties, and applications of polyphenolic extracts obtained from two selected plant materials of the Mediterranean area as olive (Olea europaea L.) and pomegranate (Punica granatum L.) based on recent literature, highlighting future research perspectives.

Phenolic distribution in liquid preparations of Vaccinium myrtillus L. and Vaccinium vitis idaea L.

INTRODUCTION: Liquid preparations such as tinctures and 'bud extracts' of bilberry and lingonberry are typical products used in some European countries but they have not been widely studied to date. OBJECTIVE: Our aim was to define a phytochemical profile in terms of phenolic compounds for liquid preparations of Vaccinium myrtillus L. (bilberry) and Vaccinium vitis idaea L. (lingonberry) foliar tissues, and evaluate their stability. METHODS: Up to 17 extracts of lingonberry, and bilberry have been analysed by HPLC equipped with a diode-array detector (DAD) in combination with MS. Some of these samples were freshly prepared (laboratory extracts). RESULTS: Phenolic profiles of the bud extracts and tinctures of bilberry and lingonberry were obtained at 330 nm, typical of the cinnamoyl compounds and suitable to distinguish these two species. Analogously, the profiles at 280 nm were used to detect the arbutin derivatives recognised as the typical markers of lingonberry leaves only. Among the cinnamoyl derivatives, chlorogenic acid was the main compound found in bilberry samples, while caffeoyl arbutin was the principal marker in lingonberry samples. In quantitative terms, the hydroxycinnamic acids were the main class in bilberry bud extracts ranging between 4.67 and 6.81 mg/L (from 52 to 84% of the total phenols). The young sprouts of this species, independently from the extraction mixture, show a higher phenolic content than adult leaves of the same plant. CONCLUSIONS: The analytical method allowed to define fingerprints of bilberry and lingonberry liquid preparations. Moreover, without applying accelerated ageing protocols, the stability over time of 11 samples was evaluated up to 6 months, demonstrating negligible variations.

Evaluation of the Fruit Quality and Phytochemical Compounds in Peach and Nectarine Cultivars.

Qualitative traits and chemical properties of 32 peach cultivars (yellow flesh and white flesh fruits) and 52 nectarine cultivars (yellow flesh and white flesh fruits) of different pomological characteristics is performed, and the correlation between cultivars and chemical characteristics is analyzed. Yellow nectarines have a higher variability in soluble solids concentration (SSC) and titratable acidity (TA) values. Evaluation of color parameters (a*, b*, L*) shows a significant interaction between pulp color (white vs. yellow) and types (peaches vs. nectarines) of fruit. The difference between yellow and white fruits is stronger in nectarines than in peaches. Sucrose is the main sugar detected in peach fruits, with a percentage content of 78.37% and 76.70% of the total sugar content in yellow and white peaches, respectively, and 78.29% and 78.12% in yellow and white nectarines, respectively. Variability is found among cultivars for the chemical compounds analyzed. The yellow flesh has higher amounts of total carotenoids and TPC, while white-flesh fruits present an average antioxidant value higher than yellow-flesh fruits. No significant correlation is found for polyphenol content and DPPH, while an interaction (p < 0.005) between neochlorogenic acid content and peaches and nectarines is evidenced, with a neochlorogenic acid content higher in nectarines than in peaches.

Sex Differences in X-ray-Induced Endothelial Damage: Effect of Taurine and N-Acetylcysteine.

Ionizing radiation (IR) can induce some associated pathological conditions due to numerous cell damages. The influence of sex is scarcely known, and even less known is whether the effect of antioxidants is sex-dependent. Given the increased use of IR, we investigated whether male human umbilical vein endothelial cells (MHUVECs) and female human umbilical vein endothelial cells (FHUVECs) respond differently to IR exposure and whether the antioxidants 10 mM taurine (TAU) and 5 mM N-acetylcysteine (NAC) can prevent IR-induced damage in a sex-dependent way. In untreated cells, sex differences were observed only during autophagy, which was higher in FHUVECs. In non-irradiated cells, preincubation with TAU and NAC did not modify viability, lactate dehydrogenase (LDH) release, migration, or autophagy, whereas only NAC increased malondialdehyde (MDA) levels in FHUVECs. X-ray irradiation increased LDH release and reduced viability and migration in a sex-independent manner. TAU and NAC did not affect viability while reduced LDH release in irradiated cells: they have the same protective effect in FHUVECs, while, TAU was more protective than NAC in male cells.. Moreover, TAU and NAC significantly promoted the closure of wounds in both sexes in irradiated cells, but NAC was more effective at doing this in FHUVECs. In irradiated cells, TAU did not change autophagy, while NAC attenuated the differences between the sexes. Finally, NAC significantly decreased MDA in MHUVECs and increased MDA in FHUVECs. In conclusion, FHUVECs appear to be more susceptible to IR damage, and the effects of the two antioxidants present some sex differences, suggesting the need to study the influence of sex in radiation mitigators.

Perspectives on Populus spp. (Salicaceae) bud extracts as antioxidant and anti-inflammatory agents.

Hydroalcoholic extracts obtained from buds of P. nigra, P. deltoides and P. trichocarpa were characterized by HPLC-DAD-MS analysis and subsequently evaluated in vitro for their antioxidant and anti-inflammatory activities. ABTS and DPPH assays evidenced that P. nigra showed the best antioxidant activity in line with its highest total phenolic content. The analysis of the anti-inflammatory activity clearly demonstrated that all extracts suppressed the production of key pro-inflammatory cytokines (IL-6, Il-1ß and TNF-α) and HMGB1 inflammatory danger signal. These results show antioxidant and critical anti-inflammatory activities mediated by the extracts, emphasising their potentiality as therapeutic agents.

Extra Virgin Olive Oil and Cardiovascular Protection in Chronic Kidney Disease.

The high mortality related to chronic kidney disease (CKD) is not only due to the disease itself; in fact, CKD also represents an important risk factor for cardiovascular (CV) morbidity and mortality. Among the functional foods that seems to have cardioprotective action, extra virgin olive oil (EVOO) plays a pivotal health-promoting role. The aim of this study was to evaluate the possible cardioprotective effects of an EVOO containing a very high content (>900 ppm) of minor phenolic compounds (MPCs). The selected EVOO was analyzed by HPLC-DAD-MS to establish the MPC content. The Olea extract obtained from the selected EVOO was tested against the RAW 264.7 cell line in order to investigate its anti-inflammatory activity. We enrolled 40 CKD patients under conservative therapy for in vivo clinical testing. All CKD patients consumed 40 mL/day of raw EVOO for 9 weeks (T1). At baseline (T0) and at T1, we monitored the patients' blood and urinary parameters. The patients' body composition was assessed using bioelectrical impedance analysis and the carotid intima-media thickness (CIMT) using ultrasound imaging. At T1, we observed a decrease in inflammatory parameters, CIMT, and oxidative stress biomarkers. We also noticed improvements in lipid and purine metabolism, atherogenic indices, and body composition. Thus, this study highlighted the cardioprotective action of EVOO in nephropathic patients.

Analysis of aroma and polyphenolic compounds in Saperavi red wine vinified in Qvevri.

The purpose of this study is to analyze and characterize a Georgian red wine from Saperavi grape, obtained in Qvevri (Georgian traditional winemaking method), by using innovative techniques for the determination of the polyphenolic content, aroma, and its correlation to the sensory characteristics. This peculiar red wine, after high-performance liquid chromatography with diode-array detection and mass spectrometry (HPLC-DAD-MS), headspace solid-phase microextraction-gas chromatography-mass spectrometry (HS-SPME-GC-MS), and HS-SPME-GCxGC-MS/TOF (two-dimensional gas chromatography) chemical characterization showed a high polyphenol content (19.6 × 102 mg/L, 38.4% anthocyanins) and a wide range of volatile compounds, among which terpenes were associated with the aroma of flowers, lemongrass, and wood. Analyses were also conducted to determine the total polyphenol content correlated to antioxidant activity with the Folin-Ciocalteu spectrophotometric in vitro method (4.650 g GAE/L). In conclusion, for the first time on Saperavi wine, innovative techniques such as HPLC-DAD-MS, GC-MS, and GCxGC-MS/TOF were simultaneously applied in association with the traditional analytic techniques to perform a complete chemical characterization. These activities are part of a project about circular viticulture in the Georgian territory that will lead the production of traced quality wines and the valorization of the Georgian wine sector.

Health Effects of Phenolic Compounds Found in Extra-Virgin Olive Oil, By-Products, and Leaf of Olea europaea L.

Olea europaea L. fruit is a peculiar vegetal matrix containing high levels of fatty acids (98-99% of the total weight of extra-virgin olive oil, EVOO) and low quantities (1-2%) of phenolics, phytosterols, tocopherols, and squalene. Among these minor components, phenolics are relevant molecules for human health. This review is focused on their beneficial activity, in particular of hydroxytyrosol (HT), oleuropein (OLE), oleocanthal (OLC), and lignans found in EVOO, olive oil by-products and leaves. Specifically, the cardioprotective properties of the Mediterranean diet (MD) related to olive oil consumption, and the biological activities of polyphenols recovered from olive oil by-products and leaves were described. Recent European projects such as EPIC (European Prospective Investigation into Cancer and Nutrition) and EPICOR (long-term follow-up of antithrombotic management patterns in acute coronary syndrome patients) have demonstrated the functional and preventive activities of EVOO showing the relation both between cancer and nutrition and between consumption of EVOO, vegetables, and fruit and the incidence of coronary heart disease. The data reported in this review demonstrate that EVOO, one of the pillars of the MD, is the main product of Olea europaea L. fruits; leaves and by-products are secondary but precious products from which bioactive compounds can be recovered by green technologies and reused for food, agronomic, nutraceutical, and biomedical applications according to the circular economy strategy.

Biogenic Amines, Phenolic, and Aroma-Related Compounds of Unroasted and Roasted Cocoa Beans with Different Origin.

Biogenic amines (BAs), polyphenols, and aroma compounds were determined by chromatographic techniques in cocoa beans of different geographical origin, also considering the effect of roasting (95, 110, and 125 °C). In all samples, methylxantines (2.22-12.3 mg kg-1) were the most abundant followed by procyanidins (0.69-9.39 mg kg-1) and epicatechin (0.16-3.12 mg kg-1), all reduced by heat treatments. Volatile organic compounds and BAs showed variable levels and distributions. Although showing the highest BAs total content (28.8 mg kg-1), Criollo variety presented a good aroma profile, suggesting a possible processing without roasting. Heat treatments influenced the aroma compounds especially for Nicaragua sample, increasing more than two-fold desirable aldehydes and pyrazines formed during the Maillard cascade and the Strecker degradation. As the temperature increased, the concentration of BAs already present in raw samples increased as well, although never reaching hazardous levels.

Administration of minor polar compound-enriched extra virgin olive oil decreases platelet aggregation and the plasma concentration of reduced homocysteine in rats.

We investigated the effect of extra virgin olive oil (EVOO) on platelet aggregation and plasma concentrations of homocysteine (Hcy) redox forms in rats in relation to the minor polar compound (MPC) concentration of EVOO. We used 3 olive oil samples with similar fatty acid but different MPC concentrations: refined olive oil (RF) with traces of MPC (control oil), native EVOO with low MPC concentration (LC), and EVOO with high MPC concentration (HC) enriching LC with its own MPC. Oil samples were administered to rats by gavage (1.25 mL/kg body weight) using 2 experimental designs: acute (24-h food deprivation and killed 1 h after EVOO administration) and subacute (12-d treatment, a daily dose of oil for 12 d, and killed after 24 h of food deprivation). Platelet aggregation was induced by ADP (ex vivo tests) and a reduction in platelet reactivity occurred in cells from rats given LC in the subacute study and in cells from rats administered HC in both studies as indicated by an increase in the agonist half maximal effective concentration. HC inhibited platelet aggregation induced by low ADP doses (reversible aggregation) in cells of rats in both the acute and subacute studies, whereas LC had this effect only in the subacute experiment. Moreover, in rats administered HC in both experiments, the plasma concentration of free reduced Hcy (rHcy) was lower and Hcy bound to protein by disulfide bonds (bHcy) was greater than in RF-treated rats. bHcy was also greater in rats given LC than in RF-treated rats in the subacute experiment. Plasma free-oxidized Hcy was greater in rats given LC and HC than in those administered RF only in the subacute experiment. In conclusion, these results show that MPC in EVOO inhibit platelet aggregation and reduce the plasma rHcy concentration, effects that may be associated with cardiovascular protection.

Evolution of minor polar compounds and antioxidant capacity during storage of bottled extra virgin olive oil.

We characterized "Olivastra Seggianese" extra virgin olive oil (EVOO) and evaluated its chemical and sensory characteristics and antioxidant and antiradical activities during storage under novel conditions. Two oils (A and B) were analyzed for the commodity characteristics at blending (t0) and after 9, 12, and 18 months; panel tests were performed and minor polar compounds (MPC) content was assessed at blending (t0) and after 6, 9, 12, and 18 months. Antioxidant and antiradical activities in vitro were evaluated at t0 and after 12 months, by human low density lipoprotein (LDL) and 1,1-diphenyl-2-picrylhydrazil radical (DPPH*) tests. Oil A, which had an initially higher MPC content, possessed "harder" organoleptic characteristics than oil B, which had a lower MPC content and was endowed with a "smoother" taste profile. Statistical analyses showed that secoiridoids, particularly deacetoxy-oleuropein aglycone, should be quantified to evaluate EVOO stability during storage. The antioxidant activity toward human LDL was linked to MPC content and to storage time. The tests on the stable free radical DPPH* confirmed the results on human LDL. We propose this as an additional parameter to evaluate olive oil quality and stability over time.

Flavonoids and biflavonoids in Tuscan berries of Juniperus communis L.: detection and quantitation by HPLC/DAD/ESI/MS.

The aim of the present work was to develop a quali-quantitative investigation, using HPLC/DAD and HPLC/ESI/MS techniques, of the phenolic composition of berries collected from wild Tuscan plants of Juniperus communis L. and grown in three different geographical zones. The applied chromatographic elution method made it possible to well separate up to 16 different compounds belonging to flavonoids, such as isoscutellarein and 8-hydroxyluteolin or hypolaetin glycosides, and six biflavonoids, among them amentoflavone, hynokiflavone, cupressoflavone, and methyl-biflavones. To the best of the authors' knowledge this is the first report on the presence of these compounds in juniper berries. The flavonoidic content in the analyzed berries ranged between 1.46 and 3.79 mg/g of fresh pulp, whereas the amount of the biflavonoids was always lower, varying between 0.14 and 1.38 mg/g of fresh weight.

HPLC-DAD/MS characterization of flavonoids and hydroxycinnamic derivatives in turnip tops (Brassica rapa L. Subsp. sylvestris L.).

Flavonoids and hydroxycinnamic derivatives of turnip tops (Brassica rapa L. subsp. sylvestris L.) were characterized for the first time in four samples from different origins. Turnip tops exhibit a high polyphenols content (ranging from 107 to 191 mg/100 g, fresh weight) and a good antiradical activity, determined with the DPPH* test. After a liquid-liquid extraction and fractionation procedures, most flavonoids (isorhamnetin, kaempferol, and quercetin glycosides) and hydroxycinnamic derivatives were identified by means of HPLC-DAD/MS techniques. Isorhamnetin glycosides were the main flavonoid derivatives, differing from that found in the vegetables belonging to the Brassica oleracea group.

Antioxidant effect of two virgin olive oils depends on the concentration and composition of minor polar compounds.

In vitro studies show that some individual minor polar phenolic compounds (MPC) present in virgin olive oil prevent oxidation of human low-density lipoproteins (LDL), but few data are available on the antioxidant effect of whole oil extract. Thus, whole virgin olive extracts were studied to determine whether they maintain the antioxidant activity and whether this last is linked to MPC composition of a single virgin oil. Using HPLC-DAD the MPC content in Taggiasca and Seggianese virgin olive oils was measured. Taggiasca oil was less rich in total MPC (208.5 mg/L) than Seggianese oil (441.9 mg/L). In addition, the major compounds of Taggiasca oil were lignan derivatives, whereas the major compounds in Seggianese oils were secoiridoid derivatives. Moreover, Taggiasca oil was practically free of 5-hydroxytyrosol and 5-hydroxytyrosol derivatives, deacetoxy-oleuropein aglycone and oleuropein aglycone. The antioxidant activity of the oils on human LDL was evaluated by measuring malondialdehyde and conjugate diene generation induced by copper ions. In both tests, the oil extracts dose-dependently reduced malondialdehyde and conjugate diene generation. Moreover, antioxidant potency correlated with total MPC; thus, Seggianese extract was more active. The two oils differed quantitatively and qualitatively, and these differences influenced their biological activities; thus clinical trials focused on studying the effects of olive oils should specify the oils used.

Polyphenols and Volatile Compounds in Commercial Chokeberry (Aronia melanocarpa) Products.

Aronia melanocarpa (Michx.) Elliott commercial products (dried fruit, juice and compote) were analyzed for their polyphenol content by chromatographic and spectrophotometric analyses in order to ascertain the fate of this group of compounds when fresh fruit is processed and sold in different forms on the market. Different classes of polyphenols were investigated: hydroxycinnamic derivatives ranged from 0.65 mg/g to 4.30 mg/g, flavonoids from 0.36 mg/g to 1.12 mg/g, and anthocyanins from 0.65 to 7.08 mg/g sample. 4-O-Caffeoyl-quinic acid was tentatively identified for the first time in Aronia. In order to characterize better chokeberry juice, a GC profile of aroma compounds was obtained. The aroma juice compounds belong mainly to the chemical classes of alcohols (48.9%) and ketones (30.28%). The most abundant compound is 3-penthen-2-one (23.6%).

Evaluation of the phenolic content in the aerial parts of different varieties of Cichorium intybus L.

Fresh aerial parts of different chicory varieties: green chicory (c.v. "Catalogna"), two red chicory varieties ("radicchio rosso di Chioggia" and "radicchio rosso di Treviso"), and Witloof or Belgian endive were analyzed by HPLC/DAD/MS. The chromatographic fingerprint was diagnostic for each variety. A monocaffeoyl tartaric acid, chlorogenic acid, and chicoric acid were detected in all the varieties, while cyanidin 3-O-glucoside, delphinidin 3-O-(6'' malonyl) glucoside, and cyanidin 3-O-(6'' malonyl) glucoside were the main phenolic compounds in the red varieties. The flavonoidic compounds, quercetin 3-O-glucuronide and luteolin 7-O-glucuronide, were absent in the Witloof sample. The phenolic compounds from total leaves were the same as those obtained from only the colored parts; nevertheless, the total amount was remarkably lower with a decrease of up to 80% for Belgian endive. Chemical stability at high temperature was observed for the phenolic fraction from the green variety after decoction at 100 degrees C for 30 min.