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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., BLURB FOR ETOC:Analysys and characterization of a Georgian red wine from Saperavi grape, obtained in Qvevri (Georgian traditional winemaking method), by using innovative techniques for the determination of the polyphenolic and key volatile compounds to produce traced quality wines and the valorisation of the Georgian wine sector.

BACKGROUND AND AIMS Saffron is the dried stigmas of Crocus sativus L. Crocus sativus L belongs to the family of Iridaceae and is mainly cultivated in several countries with mild and dry climates. It is one of the most expensive spices in the world and the main reason for its great cost is that saffron is still cultivated and harvested as it has been for millennia by hand. Saffron's name is derived from the Arab word for yellow, a name reflecting the high concentration of carotenoid pigments present in the saffron flowers’ stigmas which contribute most to the colour profile of this spice. Saffron contains numerous bioactive molecules known for their antioxidant activities, but also some such as picrocrocin, crocetin and Safranal (Sfr), Crocin (Crc) which are known to possess the ability to inhibit tumoral growth by inducing apoptosis. The aim of this study is to understand whether these four saffron-derived bioactive molecules display tumoral growth inhibition activity on kidney and bladder cancer cell lines. METHOD Saffron samples used for this purpose possess both different origins (Tuscany and Lombardy) and harvesting years (T_2017, T_2018, L_2017, L_2018) and, in addition, Sfr and Crc standard samples were also tested. Identification of Crocins, Safranal, Picrocrocin and Flavonoids was carried out by HPLC/DAD and HPLC/MS analysis. The identity of polyphenols was ascertained using data from HPLC/DAD analyses by comparison and combination of their retention times and UV/Vis spectra with those of authentic standards and previously reported data (Vignolini et al., 2008). The extract characterization is summarized in the table below. The extracts’ effects on cancer cell viability were evaluated through MTT assays and the tumoral cell lines chosen were Caki-1 and 786-O for the kidney and RT112 and RT4 for the bladder. MDCK cells were used as a healthy kidney model. RESULTS In the presence of both pure Sfr and Crc, post-hoc tests have shown a time and concentration-dependent decrease in viability for all the cancerous models in analysis, with more visible results observable for Crc, while having no effect on MDCK viability. Being Crc is the most represented molecule in all the extracts, before performing MTT assays, a normalization of this compound's concentration was carried out. L_2017, which contained the highest Sfr concentration among all the extracts tested, showed the highest inhibiting activity. Hypothesizing that Sfr was the molecule responsible for the decrease in cancer cell viability, all extracts and Sfr and Crc standards were tested on RT112 cells after normalizing the contents of these two molecules for each condition. We observed that Crc and Sfr alone and the Sfr-Crc mix had a small impact on cancer cell viability when compared with the effects of whole extracts. CONCLUSION Whole extracts demonstrated a higher impact on cancer cell viability compared with the standards tested both singularly and combined. This study, therefore, recognizes that numerous molecules in combination with Sfr and Crc have a role in reducing tumoral cell viability, demonstrating that the phyto-complex may possess higher therapeutic effects.

Hydroalcoholic extracts obtained from buds of

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-1β 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.

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.

Five quinoa (Chenopodium quinoa Willd.) genotypes; namely, KVL-SRA2, KVL-SRA3, Regalona, Q-37 and Q-52 were evaluated, under rainfed and irrigated conditions, for their allelopathic potential. Four concentrations of 1, 2, 4 and 8 g/100 ml of the aqueous extract of all plant parts were assessed for their effect on the germination and initial development of some weeds and crops. HPLC/DAD/MS analyses were conducted for genotypes with highly inhibitory effects to determine their contents of phenolic compounds and saponins. The estimated EC50 revealed that the aqueous extracts from genotypes grown under rainfed conditions had higher allelopathic activity than those genotypes grown under irrigated conditions. The suppressive effect of the extracts increased when the concentration of quinoa extracts increased. The highest reduction effects were achieved from Q-52; followed by Regalona and KVL-SRA2. Nevertheless, Q-37 and KVL-SRA3 caused minor amounts of inhibition in the tested plant traits. The negative influence of quinoa extracts was higher on monocotyledon plant species than on dicotyledon plant species. The highly susceptible plants were Hordeum vulgare, Allium cepa and Phalaris minor as compared to other tested plants. However, Vicia faba and Chenopodium album were slightly more susceptible plants. Qualitative-quantitative analysis showed sixteen flavonoids and three hydroxicinnamic acids (p-coumaroil derivatives); in particular, kaempferol dirhamnosyl-pentoside is the principal compound in Q-52 (11.25 mg/kg), while quercetin rhamnosyl-glucoside is the principal compound in Regalona (9.67 mg/kg). In quinoa, nine different aglycones, monodesmosidic, and bidesmosidic triterpene saponins were reported; in particular, in this work, hederagenin (hed), phytolaccagenic acid (PA), AG487, AG533 and AG515 derivatives were identified. Quinoa also contains biologically active phytoecdysteroids. Based on the bioassay results and its suppresive effect against weeds and crops, quinoa is considered as a crop with allelopathic potential that may have a negative impact on the growth of neighboring plants in the field and can be utilized for non chemical weed management.

Olea europaea L. fruit is a peculiar vegetal matrix containing high levels of fatty acids (98&ndash, 99% of the total weight of extra-virgin olive oil, EVOO) and low quantities (1&ndash, 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.

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 &amp, 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&times, 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, &alpha, pinene, &gamma, terpinene, p-cymene, terpinen-4-ol, &alpha, 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&ndash, 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&times, 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.

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&times, GC-TOF). Onion flakes showed a significantly higher flavonoid content (3.56 mg g&minus, 1) than onion rings (2.04 mg g&minus, 1). Onion flakes showed greater amount of volatile organic compounds (VOCs) (127.26 mg g&minus, 1) than onion rings (42.79 mg g&minus, 1), with different relative amounts of di- and trisulfides&mdash, 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&times, 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.

The review deals with polyphenol content of vegetables and fruits under different experimental conditions. The effect of fertilizers, mainly nitrogen containing fertilizers, on qualitative and especially quantitative content of the polyphenols mixture, was reviewed. Soil nitrogen affects both anthocyanins and flavonoids content, and generally, a higher polyphenolic content is observed when less nitrogen fertilizer is added to the soil. Also the effect of different agricultural management (conventional, organic, biodynamic, integrate) is reviewed with respect to polyphenols. In this case, a major effect has pointed out in the case of vegetables, while agricultural practice affects in a minimal way fruits polyphenols content. The effect of different management is, however, hardly pointed out, since many environmental factors are involved and affect polyphenols biosynthetic pathway.

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 β (ERβ) 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 ERβ (HCT8-β8). 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 17β-estradiol (17β-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 ERβ receptor, similar to what was observed after 17β-E2 challenge.

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. Copyright © 2013 John Wiley & Sons, Ltd.

This paper describes a circular economy process based on environmentally and economically sustainable procedures which was applied to the sector of olive oil processing on an industrial scale. Olea europaea L. tissues and by-products represent a renewable and low-cost source of polyphenols, in particular hydroxytyrosol (HTyr), a naturally occurring compound well known for its biological properties. Specifically, green leaves (GL), dried leaves (DL), and pitted olive pulp were treated with water in a pneumatic extractor to obtain the corresponding polyphenolic extracts. Three standardized fractions, named Soft Extract Olea GL, Soft Extract Olea DL, and Soft Extract Olea HTyr resulted after the following two steps: a separation process carried out by membrane technology, and a concentration step performed under reduced pressure and low temperature. The polyphenolic fractions showed antiradical activity and have potential industrial applications in the food, nutraceutical, pharmaceutical, feed, and agronomic fields. Novel functionalized extracts containing hydroxytyrosol methyl carbonate (HTyr-MC) were obtained from Soft Extract Olea HTyr through an innovative approach based on green chemistry procedures, which appear to be a promising tool to increase the applications of the polyphenolic extracts.

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%).

In the present study a single procedure to simultaneously determine phenolic acids, anthocyanins and glycoalkaloids was applied in 13 cultivars all with pigmented peel. A single exhaustive extraction for the three classes of compounds and a single analytical HPLC method were proposed. From a quantitative point of view, the RSD values from intra and inter-day assays were

HPLC/DAD/MS analysis of the phenolic acids and anthocyanin content of three cultivars of Solanum tuberosum L. (Vitelotte Noire, Highland Burgundy Red, with pigmented flesh, and Kennebec with white pulp) was performed. The analyses were carried out both on fresh tubers and after cooking treatments (boiling and microwaves). Starch digestibility and the % of resistant starch were also determined on cooked tubers by in vitro methods. For the pigmented potatoes, the heating treatment did not cause any changes in the phenolic acids content, while anthocyanins showed only a small decrement (16-29%). The cv. Highland Burgundy Red showed anthocyanins and phenolic acid concentrations close to 1 g/kg and more than 1.1 g/kg, respectively. Vitellotte Noire showed the highest amounts of resistant starch. Potato starch digestibility and % of resistant starch, considered as a component of dietary fiber, were affected both by cultivar and by heating/cooling treatments.

The aim of the present study was to verify the distribution of lignans in pulp and stones of olive fruits. Analyses were carried out by HPLC/DAD/MS on four cultivars: Frantoio and Taggiasca from Italy, Arbequina and Picual from Spain. The main results can be summarized as follows: (a) lignans were demonstrated to be present in the stone; (b) acetoxypinoresinol, pinoresinol, previously detected in the respective extra virgin olive oils, and OH-pinoresinol were found in the samples; (c) the total lignan content ranged between 0.1 and 0.29 mg/g of dried stone weight. These values agree with the average lignan content reported in literature for the respective extra virgin olive oils.

The aim of the present study was to compare the phenolic content in selected monocultivar extra virgin olive oils. Analyses were carried out by HPLC/DAD/MS on Picual, Picuda, Arbequina and Hojiblanca oils from Spain and Seggianese and Taggiasca oils from Italy. Oils from cultivar Picual showed similar characteristics to those of Seggianese oils, with total amounts of secoiridoids of 498.7 and 619.2mg/L, respectively. The phenolic composition of Arbequina oils is close to that of the Taggiasca variety with lignans among the main compounds. The determination of free and linked OH-Tyr, by way of an acid hydrolysis, represents a rapid and suitable method, especially when standards are not available, to determine antioxidant potentialities in terms of MPC, particularly for fresh extra virgin olive oils rich in secoiridoidic derivatives.

Liquid preparations such as tinctures and “bud extracts” are typical products widely used in European countries but which have been poorly studied to date. Our research was focused on obtaining a phytochemical characterization of secondary metabolites of black currant, dog rose and silver linden bud extracts and evaluating the changes of selected chemical markers over time. 16 different preparations have been analyzed by the use of HPLC-DAD-MS. Several flavonols were identified in black currant samples including quercetin derivatives (6.7–30.4 mg/L). Also hydroxycinnamic acids, mainly p- coumaric acid derivatives (4.1–38.9 mg/L) were identified. Flavonols such as glycosides of quercetin and kaempferol, hydroxycinnamic acids, and ellagitannins were detected in dog rose samples, with gallotannins being the main components (up to 1.7 g/L). The Tilia tomentosa extracts contained flavonols (quercetin, kaempferol, apigenin derivatives) as principal components with the exception of a single commercial extract with hydroxycinnamic acids as the most abundant metabolites. In this study, without applying accelerated ageing protocols, the stability over time of these liquid preparations was evaluated for up to 10 months and demonstrated negligible variations.

We describe chemical hydrolytic procedures to evaluate the total amount of tyrosol and hydroxy-tyrosol free and/or linked to secoiridoidic molecules (acid hydrolysis). At the same time a rapid determination of the lignans in complex minor polar compound (MPC) extracts is proposed (alkaline hydrolysis). High-performance liquid chromatography/diode array detection (HPLC/DAD) and HPLC/MS were applied as reference methods to evaluate the quantitative results from the hydrolysis experiments. The optimized acid hydrolysis procedures were first applied to an oleuropein standard and then to MPC fractions extracted from several commercial extra virgin olive oils. The results confirm the applicability of the method, consisting in the acid hydrolysis of complex mixtures of secoiridoidic derivatives, to determine the antioxidant potential in terms of MPC. These data can contribute to forecasting the potential ageing resistance of an extra virgin olive oil in terms of antioxidant potency. Finally, alkaline hydrolysis allows confirmation and easy determination of the amount of lignans, especially in those MPC fractions which are particularly complex. Copyright © 2006 Society of Chemical Industry

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.

Minor polar compounds of 88 extra virgin olive oils were analyzed by HPLC-DAD-MS (high-performance liquid chromatography-diode array detector-mass spectrometry) and by the Folin-Ciocalteu (FC) spectrophotometric method, to validate and evaluate, for olive oils, the linear association between FC and HPLC data. The Pearson correlation coefficients were calculated between HPLC and FC results. The highest, positive R were related with deacetoxyoleuropein aglycone (R = 0.93) and oleuropein aglycone (R = 0.93) as single compounds and with the sum of orthodiphenols (R = 0.94) and the sum of all compounds (R = 0.95), showing that both estimations of total phenols content are reliably correlated, regardless for the absolute contents and are independent of the relative composition of the phenolic fraction. On the other hand the HPLC quantifications of apigenin and lignans showed no significant correlation with FC. These results, supported also by principal component analysis, may suggest caution about the interpretation of FC results to compare olive oils with very different phenolic profiles.

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.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.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).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.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.

In the fresh tissues of Prunus mahaleb L., three classes of phenolics were characterised: phenolic acid derivatives (main compound being o-coumaric acid glucoside), quercetin glycosides, and anthocyanins (cyanidin 3,5-diglucoside, cyanidin 3-sambubioside, cyanidin 3-xylosyl-rutinoside and cyanidin 3-rutinoside). Coumarin was also identified. The kernels showed a high content of coumarin (0.87 mgg(-1)) which is the main class of metabolites in this sample, but present in pitted berries as well (0.63 mgg(-1)). Flavonoids are mainly concentrated in the skin and pulp (0.55 mgg(-1)). In 'Mirinello di Torremaggiore' liqueur, produced from P. mahaleb L. in accordance with traditional procedures, anthocyanins make up 16.5%, phenolic acids 43.3%, coumarin 36.2% and flavonoids 4% of total compounds. Anthocyanins are the main class in solid residues from liqueur production (70%). These findings point out that solid residues of P. mahaleb can be considered an interesting and innovative source of appreciable amounts of cyanidin glycosides (3.3 mgg(-1)).

Publisher Summary The cardio-protective role of olive oil has been highlighted in several studies and beneficial health effects have been attributed, at least in part, to the presence of non-nutrient minor polar compounds (MPC). As an example, phenolic compounds in virgin olive oil improve endothelial function in hypercholesterolemic patients and inhibit low-density lipoprotein (LDL) oxidation in humans and animals, in vivo and in vitro, hydroxytyrosol and oleuropein being also potent scavengers of several free radicals. After consumption of 25 mL extra virgin olive oil, plasma concentrations of hydroxytyrosol range from 50 to 160 nM. Hydroxytyrosol affects oxidative stress as well as arachidonic acid mobilization and metabolism by macrophage RAW 264.7 cells. In the same cell line, it also reduces iNOS (inducible nitric oxide synthase) and COX-2 (cyclo-oxygenase 2) gene expression, mainly by preventing the activation of NF-κB (nuclear factor-kappa B), STAT-1α (signal transducer and activator of transcription-1α), and IRF-1 (interferon regulatory factor-1). At nutritionally relevant concentrations, hydroxytyrosol reduces monocyte adhesion to human endothelial cells, as well as the expression of adhesion molecules such as VCAM-1 (vascular cell adhesion molecule-1), an effect that seems to be mediated by NF-κB and activator protein-1 (AP-1). The redox-sensitive transcription factor NF-κB can be activated by various stimuli, including reactive oxygen species, oxidized LDL, hypoxia/anoxia, cytokines, and bacterial and viral products, and is largely recognized as a therapeutic target in inflammation and atherosclerosis.

Hydrolysable tannins (HTs), secondary metabolites widely distributed in the plant kingdom, are generally multiple esters of gallic acid with glucose. HTs have been shown to be effective antagonists against viruses, bacteria and eukaryotic microorganisms. The present review examines the antimicrobial and antiviral activity of HTs, the mechanism(s) of action, and some structure-activity relationships.

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.

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.

Epidemiological studies demonstrate that the Mediterranean diet, in which olive oil is the major source of fat, reduces the risk of coronary heart disease and cancer. It has been proposed that the beneficial effects of olive oil not only depend on oleic acid, but are also associated with minor polar compounds (MPC). A positive correlation between inflammation and cardiovascular diseases has long been described, monocyte/macrophages and NF-kappaB playing a pivotal role. The aim of this work was to investigate the effects of an extra-virgin olive oil extract (MPC-OOE), particularly rich in MPC and prepared by some of us, on NF-kappaB translocation in monocytes and monocyte-derived macrophages (MDM) isolated from healthy volunteers. In a concentration-dependent manner, MPC-OOE inhibited p50 and p65 NF-kappaB translocation in both un-stimulated and phorbol-myristate acetate (PMA)-challenged cells, being particularly effective on the p50 subunit. Interestingly, this effect occurred at concentrations found in human plasma after nutritional ingestion of virgin olive oil and was quantitatively similar to the effect exerted by ciglitazone, a PPAR-gamma ligand. However, MPC-OOE did not affect PPAR-gamma expression in monocytes and MDM. These data provide further evidence of the beneficial effects of extra-virgin olive oil by indicating its ability to inhibit NF-kappaB activation in human monocyte/macrophages.