Fulvio Mattivi

Flavonols are products of the flavonoid biosynthetic pathway, which also give rise to anthocyanins and condensed tannins in grapes. We investigated their presence in the berry skins of 91 grape varieties (Vitis vinifera L.), in order to produce a classification based on the flavonol profile. The presence of laricitrin 3-O-galactoside and syringetin 3-O-galactoside in red grapes is reported here for the first time. In red grapes, the main flavonol was quercetin (mean = 43.99%), followed by myricetin (36.81%), kaempferol (6.43%), laricitrin (5.65%), isorhamnetin (3.89%), and syringetin (3.22%). In white grapes, the main flavonol was quercetin (mean = 81.35%), followed by kaempferol (16.91%) and isorhamnetin (1.74%). The delphinidin-like flavonols myricetin, laricitrin, and syringetin were missing in all white varieties, indicating that the enzyme flavonoid 3‘,5‘-hydroxylase is not expressed in white grape varieties. The pattern of expression of flavonols and anthocyanins in red grapes was compared, in order to gain information on the substrate specificity of enzymes involved in flavonoid biosynthesis. Keywords: Vitis vinifera; grape; flavonols; anthocyanins; flavonoid 3‘-hydroxylase; flavonoid 3‘,5‘-hydroxylase; O-methyltransferase

Abstract Anthocyanins belong to the flavonoid group of polyphenolic compounds, which are responsible for the red and blue colors of plant organs such as fruits, flowers, and leaves. Due to their frequent presence in plants, particularly berry fruits, vegetables, and grapes, they are key components of the human diet. Interest in anthocyanins has increased widely during the past decade. Numerous studies have suggested that anthocyanins have a wide range of health‐promoting properties. These compounds are therefore considered to be a functional food factor, which may have important implications in the prevention of chronic diseases. The aim of this body of work is to investigate and review the current literature on anthocyanins, and particularly their pharmacokinetics and any health‐promoting properties, in order to summarize existing knowledge and highlight any aspects that require further study and analysis.

The gut is the most extensively studied niche of the human microbiome. The aim of this study was to characterise the initial gut microbiota development of a cohort of breastfed infants (n = 192) from 1 to 24 weeks of age.V4-V5 region 16S rRNA amplicon Illumina sequencing and, in parallel, bacteriological culture. The metabolomic profile of infant urine at 4 weeks of age was also examined by LC-MS.Full-term (FT), spontaneous vaginally delivered (SVD) infants' microbiota remained stable at both phylum and genus levels during the 24-week period examined. FT Caesarean section (CS) infants displayed an increased faecal abundance of Firmicutes (p < 0.01) and lower abundance of Actinobacteria (p < 0.001) after the first week of life compared to FT-SVD infants. FT-CS infants gradually progressed to harbouring a microbiota closely resembling FT-SVD (which remained stable) by week 8 of life, which was maintained at week 24. The gut microbiota of preterm (PT) infants displayed a significantly greater abundance of Proteobacteria compared to FT infants (p < 0.001) at week 1. Metabolomic analysis of urine at week 4 indicated PT-CS infants have a functionally different metabolite profile than FT (both CS and SVD) infants. Co-inertia analysis showed co-variation between the urine metabolome and the faecal microbiota of the infants. Tryptophan and tyrosine metabolic pathways, as well as fatty acid and bile acid metabolism, were found to be affected by delivery mode and gestational age.These findings confirm that mode of delivery and gestational age both have significant effects on early neonatal microbiota composition. There is also a significant difference between the metabolite profile of FT and PT infants. Prolonged breastfeeding was shown to have a significant effect on the microbiota composition of FT-CS infants at 24 weeks of age, but interestingly not on that of FT-SVD infants. Twins had more similar microbiota to one another than between two random infants, reflecting the influence of similarities in both host genetics and the environment on the microbiota..

Forty-one apple samples, representing eight of the most widely cultivated varieties in western Europe, were collected in Trentino, Italy. Samples were extracted from fresh fruit with a mixture of acetone/water to achieve a good extraction of polyphenols, including proanthocyanidin oligomers which were analyzed by normal-phase HPLC. Up to 20 compounds including catechin, epicatechin, B2 procyanidin, hydroxycinnamates, flavonols, anthocyanins, and dihydrochalcones were analyzed by reversed-phase HPLC and LC-MS. Total polyphenol content was independently measured with an optimized Folin−Ciocalteu assay. The mean content of total polyphenols lay between 66.2 and 211.9 mg/100 g of FW depending on the variety. With chromatographic analysis, it was possible to explain the whole amount of total polyphenols measured by the FC assay. Flavanols (catechin and proanthocyanidins) are the major class of apple polyphenols (71−90%), followed by hydroxycinnamates (4−18%), flavonols (1−11%), dihydrochalcones (2−6%), and in red apples anthocyanins (1−3%). Keywords: Apple; polyphenols; hydroxycinnamates; flavonols; dihydrochalcones; anthocyanidins

Compelling evidence of the health benefits of phenolic compounds and their impact on food quality have stimulated the development of analytical methods for the identification and quantification of these compounds in different matrices in recent years. A targeted metabolomics method has been developed for the quantification of 135 phenolics, such as benzoates, phenylpropanoids, coumarins, stilbenes, dihydrochalcones, and flavonoids, in fruit and tea extracts and wine using UPLC/QqQ-MS/MS. Chromatography was optimized to achieve separation of the compounds over a period of 15 min, and MRM transitions were selected for accurate quantification. The method was validated by studying the detection and quantification limits, the linearity ranges, and the intraday and interday repeatability of the analysis. The validated method was applied to the analysis of apples, berries, green tea, and red wine, providing a valuable tool for food quality evaluation and breeding studies.

Grape is qualitatively and quantitatively very rich in polyphenols. In particular, anthocyanins, flavonols and stilbene derivatives play very important roles in plant metabolism, thanks to their peculiar characteristics. Anthocyanins are responsible for the color of red grapes and wines and confer organoleptic characteristics on the wine. They are used for chemotaxonomic studies and to evaluate the polyphenolic ripening stage of grape. They are natural colorants, have antioxidant, antimicrobial and anticarcinogenic activity, exert protective effects on the human cardiovascular system, and are used in the food and pharmaceutical industries. Stilbenes are vine phytoalexins present in grape berries and associated with the beneficial effects of drinking wine. The principal stilbene, resveratrol, is characterized by anticancer, antioxidant, anti-inflammatory and cardioprotective activity. Resveratrol dimers and oligomers also occur in grape, and are synthetized by the vine as active defenses against exogenous attack, or produced by extracellular enzymes released from pathogens in an attempt to eliminate undesirable toxic compounds. Flavonols are a ubiquitous class of flavonoids with photo-protection and copigmentation (together with anthocyanins) functions. The lack of expression of the enzyme flavonoid 3',5'-hydroxylase in white grapes restricts the presence of these compounds to quercetin, kaempferol and isorhamnetin derivatives, whereas red grapes usually also contain myricetin, laricitrin and syringetin derivatives. In the last ten years, the technological development of analytical instrumentation, particularly mass spectrometry, has led to great improvements and further knowledge of the chemistry of these compounds. In this review, the biosynthesis and biological role of these grape polyphenols are briefly introduced, together with the latest knowledge of their chemistry.

Secondary metabolism contributes to the adaptation of a plant to its environment. In wine grapes, fruit secondary metabolism largely determines wine quality. Climate change is predicted to exacerbate drought events in several viticultural areas, potentially affecting the wine quality. In red grapes, water deficit modulates flavonoid accumulation, leading to major quantitative and compositional changes in the profile of the anthocyanin pigments; in white grapes, the effect of water deficit on secondary metabolism is still largely unknown. In this study we investigated the impact of water deficit on the secondary metabolism of white grapes using a large scale metabolite and transcript profiling approach in a season characterized by prolonged drought. Irrigated grapevines were compared to non-irrigated grapevines that suffered from water deficit from early stages of berry development to harvest. A large effect of water deficit on fruit secondary metabolism was observed. Increased concentrations of phenylpropanoids, monoterpenes, and tocopherols were detected, while carotenoid and flavonoid accumulations were differentially modulated by water deficit according to the berry developmental stage. The RNA-sequencing analysis carried out on berries collected at three developmental stages—before, at the onset, and at late ripening—indicated that water deficit affected the expression of 4,889 genes. The Gene Ontology category secondary metabolic process was overrepresented within up-regulated genes at all the stages of fruit development considered, and within down-regulated genes before ripening. Eighteen phenylpropanoid, 16 flavonoid, 9 carotenoid, and 16 terpenoid structural genes were modulated by water deficit, indicating the transcriptional regulation of these metabolic pathways in fruit exposed to water deficit. An integrated network and promoter analyses identified a transcriptional regulatory module that encompasses terpenoid genes, transcription factors, and enriched drought-responsive elements in the promoter regions of those genes as part of the grapes response to drought. Our study reveals that grapevine berries respond to drought by modulating several secondary metabolic pathways, and particularly, by stimulating the production of phenylpropanoids, the carotenoid zeaxanthin, and of volatile organic compounds such as monoterpenes, with potential effects on grape and wine antioxidant potential, composition, and sensory features.

The life sciences are currently being transformed by an unprecedented wave of developments in molecular analysis, which include important advances in instrumental analysis as well as biocomputing. In light of the central role played by metabolism in nutrition, metabolomics is rapidly being established as a key analytical tool in human nutritional studies. Consequently, an increasing number of nutritionists integrate metabolomics into their study designs. Within this dynamic landscape, the potential of nutritional metabolomics (nutrimetabolomics) to be translated into a science, which can impact on health policies, still needs to be realized. A key element to reach this goal is the ability of the research community to join, to collectively make the best use of the potential offered by nutritional metabolomics. This article, therefore, provides a methodological description of nutritional metabolomics that reflects on the state-of-the-art techniques used in the laboratories of the Food Biomarker Alliance (funded by the European Joint Programming Initiative "A Healthy Diet for a Healthy Life" (JPI HDHL)) as well as points of reflections to harmonize this field. It is not intended to be exhaustive but rather to present a pragmatic guidance on metabolomic methodologies, providing readers with useful "tips and tricks" along the analytical workflow.

Background: Obesity is a condition with a complex pathophysiology characterized by both chronic low-grade inflammation and changes in the gut microbial ecosystem. These alterations can affect the metabolism of tryptophan (TRP), an essential amino acid and precursor of serotonin (5-HT), kynurenine (KYN) and indoles. This study aimed to investigate alterations in KYN and microbiota-mediated indole routes of TRP metabolism in obese subjects relatively to non-obese controls and to determine their relationship with systemic inflammation. Methods: Eighty-five obese adults (avg. BMI=40.48) and forty-two non-obese control individuals (avg. BMI=24.03) were recruited. Plasma levels of TRP catabolites were assessed using Ultra High Performance Liquid Chromatography-ElectroSpray-Ionization-Tandem Mass Spectrometry. High-sensitive C-reactive protein (hsCRP) and high-sensitive interleukin 6 (hsIL-6) were measured in the serum as markers of systemic inflammation using enzyme-linked immunosorbent assay. Results: Both KYN and microbiota-mediated indole routes of TRP metabolism were altered in obese subjects, as reflected in higher KYN/TRP ratio and lower 5-HT and indoles levels, relatively to non-obese controls. HsIL-6 and hsCRP were increased in obesity and were overall associated with TRP metabolic pathways alterations. Conclusion: These results indicate for the first time that KYN and indole TRP metabolic pathways are concomitantly altered in obese subjects and highlight their respective associations with obesity-related systemic inflammation.

The ability of anthocyanins to permeate the gastric mucosa can be suggested as a possible explanation of the fast kinetics of plasma appearance of anthocyanins in rats and humans. This paper presents an in vivo experiment aimed to prove the involvement of the stomach in the absorption of grape anthocyanins in rats. The required analytical selectivity and sensitivity was achieved by high‐performance liquid chromatography, diode array detection and mass spectrometry. Malvidin 3‐glucoside appeared in both portal and systemic plasma after only 6 min. The average concentrations measured in portal and systemic plasma were 0.650±0.162 μM and 0.234±0.083 μM (mean±S.E.M.), respectively.

ADVERTISEMENT RETURN TO ISSUEPREVArticleNEXTIsolation, Characterization, and Evolution in Red Wine Vinification of Resveratrol MonomersFulvio Mattivi, Fabiano Reniero, and Siegfried KorhammerCite this: J. Agric. Food Chem. 1995, 43, 7, 1820–1823Publication Date (Print):July 1, 1995Publication History Published online1 May 2002Published inissue 1 July 1995https://doi.org/10.1021/jf00055a013Request reuse permissionsArticle Views1379Altmetric-Citations201LEARN ABOUT THESE METRICSArticle Views are the COUNTER-compliant sum of full text article downloads since November 2008 (both PDF and HTML) across all institutions and individuals. These metrics are regularly updated to reflect usage leading up to the last few days.Citations are the number of other articles citing this article, calculated by Crossref and updated daily. Find more information about Crossref citation counts.The Altmetric Attention Score is a quantitative measure of the attention that a research article has received online. Clicking on the donut icon will load a page at altmetric.com with additional details about the score and the social media presence for the given article. Find more information on the Altmetric Attention Score and how the score is calculated. Share Add toView InAdd Full Text with ReferenceAdd Description ExportRISCitationCitation and abstractCitation and referencesMore Options Share onFacebookTwitterWechatLinked InRedditEmail Other access optionsGet e-Alertsclose Get e-Alerts

Bilitranslocase (TC 2.A.65.1.1) is an organic anion membrane carrier expressed at the sinusoidal domain of the liver plasma membrane and in epithelial cells of the gastric mucosa. Its substrates are sulfobromophthalein, bilirubin, and nicotinic acid. This work reports on the identification of a new class of bilitranslocase substrates, i.e., anthocyanins. Seventeen out thes 20 compounds tested behaved as competitive inhibitors of bilitranslocase transport activity (K(I)=1.4-22 microM). Their structure-activity relationship reveals that mono- and di-glucosyl anthocyanins, the anthocyanin species occurring in food, are better ligands than the corresponding aglycones. Moreover, the first interaction of anthocyanins with the carrier occurs through hydrophilic moieties, such as the 3-glucosyl moiety and the B ring for the monoglucosides, through the 5-glucosyl moiety and the A ring for the diglucosides, and through either the B or the A ring for the aglycones. These findings suggest that bilitranslocase could play a role in the bioavailability of anthocyanins.

Anthocyanins represent the main flavonoid pigments in red grape and wine, in red berries, and in many other fruits and vegetables and are widespread in the human diet. After ingestion, these complex, hydrophilic compounds quickly appear as intact molecules in the plasma. This study investigated their presence in the brain of anesthetized rats that received 8 mg/kg of body weight of a pure anthocyanin mixture extracted from Vitis vinifera grapes. The mixture was maintained in the stomach for 10 min. After this time, intact anthocyanins were detected by HPLC-DAD-MS not only in the plasma (176.4 +/- 50.5 ng/mL, mean +/- SEM) but also in the brain (192.2 +/- 57.5 ng/g). These results demonstrate for the first time that grape pigments can reach the mammalian brain within minutes from their introduction into the stomach.

In grapes, stilbene synthesis occurs in the skin, and it is induced by biotic and abiotic stresses. To date, experimental evidence of a constitutive production of resveratrols in healthy grape is scarce and not conclusive. The aim of the present work was to investigate stilbene biosynthesis in healthy grapes both at biochemical and molecular levels. By measuring the concentration of resveratrols in ripe berries of 78 Vitis vinifera varieties for 3 years, we could identify significant differences among genotypes, providing the first tentative varietal classification based on resveratrol content. Furthermore, an increasing stilbene accumulation from veraison to ripening phase was also observed. Using real-time reverse transcription polymerase chain reaction and a berry-specific cDNA array, gene expression analysis was carried out on two distinct pools of berries belonging to the high and low resveratrol producers and on three berry developmental stages. The stilbene synthase, phenylalanine ammonia-lyase, and 4-coumarate-CoA ligase expression profiles showed an increasing concentration of these transcripts from véraison to maturity and a higher accumulation in the grape of high resveratrol producers. Macroarray data analysis revealed that high resveratrol levels are also accompanied by the up-regulation of genes involved in plant defense and the concomitant underexpression of genes related to the ripening process and to indole alkaloid synthesis.

Background and Aims: Red wine is a very rich source of flavanols, consisting of catechins and oligomeric and polymeric proanthocyanidins (PAs). The aim of this study was to investigate the composition of the fraction of flavanols, which is extractable from the grape into the wine. Methods and Results: Using HPLC-DAD-MS analysis carried out before and after thioacidolysis of wine-like extracts obtained from skins and seeds, the flavanol composition of 20 samples derived from Cabernet Sauvignon, Carmenere, Marzemino, Merlot, Pinot Noir, Syrah and Teroldego was determined. Conclusions: It was confirmed that grape extracts are mainly rich in monomers and small oligomers (mean degree of polymerization <8). Grape seed extracts contained three monomers (catechin, epicatechin and epicatechin gallate) and procyanidin oligomers. Grape skin extracts contained four monomers (catechin, epicatechin, gallocatechin and epigallocatechin), procyanidins and prodelphinidins oligomers. Upper and extension units of PAs were constituted mainly of epicatechin units, with the co-presence of catechin and epicatechin gallate in the seed PAs, and of gallocatechin in the case of skin PAs. The terminal units of the grape PAs were a mixture of five compounds, the same found also as monomers. Catechin gallate, gallocatechin gallate and epigallocatechin gallate, which are found in other flavanol-rich, plant-derived foods and beverages, were not present in grape extracts. Significance of the Study: The grape variety determines both the amount and the structure of wine flavanols, leading to differences in the composition, which are likely to play an important role in the nutritional and sensorial properties of the wines.

The main metabolites of caffeic and ferulic acids (ferulic acid-4'-O-sulfate, caffeic acid-4'-O-sulfate, and caffeic acid-3'-O-sulfate), the most representative phenolic acids in fruits and vegetables, and the acyl glucuronide of ferulic acid were synthesized, purified, and tested for their antioxidant activity in comparison with those of their parent compounds and other related phenolics. Both the ferric reducing antioxidant power (FRAP) assay and the 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging method were used. Ferulic acid-4'-O-sulfate and ferulic acid-4'-O-glucuronide exhibited very low antioxidant activity, while the monosulfate derivatives of caffeic acid were 4-fold less efficient as the antioxidant than caffeic acid. The acyl glucuronide of ferulic acid showed strong antioxidant action. The antioxidant activity of caffeic acid-3'-O-glucuronide and caffeic acid-4'-O-glucuronide was also studied. Our results demonstrate that some of the products of phenolic acid metabolism still retain strong antioxidant properties. Moreover, we first demonstrate the ex vivo synthesis of the acyl glucuronide of ferulic acid by mouse liver microsomes, in addition to the phenyl glucuronide.

Accurate quantification and structural characterization of ellagitannins and ellagic acid conjugates in food, beverages, and food supplements are essential starting points for studying their effect on human health. However, accuracy is hindered both by the lack of pure standard compounds and by methods that maintain the compounds in their native form, avoiding any chemical modification of the structure. The objective of this work was to develop a new method for the purification, chromatographic separation, and accurate quantification of ellagitannins and ellagic acid conjugates to provide thorough characterization of the diversity in composition of 11 Rubus cultivars grown in Trentino, Italy. As such, two major steps were required: (i) the isolation and purification (with associated detailed structural characterization and determination of their molar extinction coefficients) of sanguiin H-6 and lambertianin C, providing essential data for their use, together with ellagic acid, as external standards, and (ii) the determination of the chemical structure of 20 novel minor ellagitannins and 4 ellagic acid conjugates on the basis of their Q-TOF-HDMS and DAD spectra. This survey of ellagitannins and ellagic acid conjugates provides evidence for the existence of significant differences in the pattern between and within blackberry and raspberry cultivars. To our knowledge, this is the first paper that has combined detailed metabolite profiling with accurate quantification of the main ellagitannins in Rubus using their respective standards.

Different polyphenol compounds are ingested when consuming a serving of fruits rich in polyphenols, spanning from one-phenol hydroxybenzoic acid to more complex polymeric compounds.Only a minor quantity of the polyphenols (5-10%) is absorbed.The remainder reaches the colon and is extensively metabolized by gut microbiota to low-molecular weight metabolites.Their subsequent tissue distribution is still undefined, though these microbial metabolites are currently believed to play a role in human health and disease states.To fill this knowledge gap, we performed a pharmacokinetics experiment in which a single bolus of 23 polyphenol microbial metabolites (total 2.7 μmol) was administered intravenously to rats to reliably reproduce a physiological post-absorption situation.Tissues and urine were collected shortly thereafter (15 s to 15 min) and were analyzed by UHPLC-MS/MS to quantitatively track these compounds.Remarkably, the brain was found to be a specific target organ for ten of the 23 polyphenol metabolites injected, which significantly increased in the treated animals.In most cases, their appearance in the brain was biphasic, with an early wave at 2 min (4 compounds) and a second wave starting at 5 min; at 15 min, 9 compounds were still detectable.Most compounds were excreted into the urine.The concentrations in the brain of the treated animals were compared against those of the control group by Student's t-test, with Pvalues < 0.1 considered statistically significant.These findings provide new perspectives for understanding the role of diet on brain chemistry.Our experimental approach has enabled us to obtain rich metabolomics information from a single experiment involving a limited number of animals.

Grapevine berries undergo complex biochemical changes during fruit maturation, many of which are dependent upon the variety and its environment. In order to elucidate the varietal dependent developmental regulation of primary and specialized metabolism, berry skins of Cabernet Sauvignon and Shiraz were subjected to gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-mass spectrometry (LC-MS) based metabolite profiling from pre-veraison to harvest. The generated dataset was augmented with transcript profiling using RNAseq.The analysis of the metabolite data revealed similar developmental patterns of change in primary metabolites between the two cultivars. Nevertheless, towards maturity the extent of change in the major organic acid and sugars (i.e. sucrose, trehalose, malate) and precursors of aromatic and phenolic compounds such as quinate and shikimate was greater in Shiraz compared to Cabernet Sauvignon. In contrast, distinct directional projections on the PCA plot of the two cultivars samples towards maturation when using the specialized metabolite profiles were apparent, suggesting a cultivar-dependent regulation of the specialized metabolism. Generally, Shiraz displayed greater upregulation of the entire polyphenol pathway and specifically higher accumulation of piceid and coumaroyl anthocyanin forms than Cabernet Sauvignon from veraison onwards. Transcript profiling revealed coordinated increased transcript abundance for genes encoding enzymes of committing steps in the phenylpropanoid pathway. The anthocyanin metabolite profile showed F3'5'H-mediated delphinidin-type anthocyanin enrichment in both varieties towards maturation, consistent with the transcript data, indicating that the F3'5'H-governed branching step dominates the anthocyanin profile at late berry development. Correlation analysis confirmed the tightly coordinated metabolic changes during development, and suggested a source-sink relation between the central and specialized metabolism, stronger in Shiraz than Cabernet Sauvignon. RNAseq analysis also revealed that the two cultivars exhibited distinct pattern of changes in genes related to abscisic acid (ABA) biosynthesis enzymes.Compared with CS, Shiraz showed higher number of significant correlations between metabolites, which together with the relatively higher expression of flavonoid genes supports the evidence of increased accumulation of coumaroyl anthocyanins in that cultivar. Enhanced stress related metabolism, e.g. trehalose, stilbene and ABA in Shiraz berry-skin are consistent with its relatively higher susceptibility to environmental cues.

Ellagitannins have shown anti-inflammatory and anti-Helicobacter pylori properties; however, their anti-inflammatory activity at gastric level was not previously investigated. The aim of this research was to evaluate the effects of ellagitannins from Rubus berries on gastric inflammation. Ellagitannin enriched extracts (ETs) were prepared from Rubus fruticosus L. (blackberry) and Rubus idaeus L. (raspberry). The anti-inflammatory activity was tested on gastric cell line AGS stimulated by TNF-α and IL-1β for evaluating the effect on NF-kB driven transcription, nuclear translocation and IL-8 secretion. In vivo the protective effect of ellagitannins was evaluated in a rat model of ethanol-induced gastric lesions. Rats were treated orally for ten days with 20 mg/kg/day of ETs, and ethanol was given one hour before the sacrifice. Gastric mucosa was isolated and used for the determination of IL-8 release, NF-kB nuclear translocation, Trolox equivalents, superoxide dismutase and catalase activities. In vitro, ETs inhibited TNF-α induced NF-kB driven transcription (IC₅₀: 0.67-1.73 µg/mL) and reduced TNF-α-induced NF-kB nuclear translocation (57%-67% at 2 µg/mL). ETs inhibited IL-8 secretion induced by TNF-α and IL-1β at low concentrations (IC₅₀ range of 0.7-4 µg/mL). Sanguiin H-6 and lambertianin C, the major ETs present in the extracts, were found to be responsible, at least in part, for the effect of the mixtures. ETs of blackberry and raspberry decreased Ulcer Index by 88% and 75% respectively and protected from the ethanol induced oxidative stress in rats. CINC-1 (the rat homologue of IL-8) secretion in the gastric mucosa was reduced in the animals receiving blackberry and raspberry ETs. The effect of ETs on CINC-1 was associated to a decrease of NF-κB nuclear translocation in ETs treated animals. The results of the present study report for the first time the preventing effect of ETs in gastric inflammation and support for their use in dietary regimens against peptic ulcer.

A hydrophilic interaction liquid chromatography (HILIC)-MS/MS profiling method was developed for the efficient separation and quantification of small polar molecules, mostly primary metabolites. The method was based on an ultrahigh performance liquid chromatography (UHPLC) separation system coupled with ESI mass spectrometry on a triple quadrupole mass spectrometer, operating in both positive and negative ionisation mode using rapid polarity switching. With the developed method quantitation of 135 compounds belonging in four major classes of polar compounds (sugars, aminoacids, organic acids and amines) was achieved in a single run of 30 min. The method was applied to grape extracts from different varieties and provided information on primary metabolite content. Multivariate statistical analysis was applied using the concentrations found, with the aim of investigating the differences in metabolite profiles. Classification of grapes according to their skin colour was carried out using principle component analysis based on the concentration variation of a number of the metabolites studied.

In the Vitaceae, viniferins represent a relatively restricted group of trans-resveratrol oligomers with antifungal properties, thus enabling plants to cope with pathogen attack. The aim of this study was to perform isolation and structural characterization of the whole class of viniferins accumulating in the leaves of hybrid Vitis vinifera (Merzling × Teroldego) genotypes infected with Plasmopara viticola . Infected leaves of resistant plants were collected 6 days after infection, extracted with methanol, and prepurified by flash chromatography using ENV+ and Toyopearl HW 40S resins. Further fractionation using normal-phase preparative chromatography and then reversed-phase preparative chromatography allowed isolation of 14 peaks. The isolated compounds were identified using advanced mass spectrometry techniques and extensive one- and two-dimensional nuclear magnetic resonance measurements, UV, CD, optical properties, and molecular mechanic calculations. The results demonstrated the presence in infected leaves of seven dimers (six stilbenes and one stilbenoid), of which four were new in grapevine (ampelopsin D, quadrangularin A, E-ω-viniferin, and Z-ω-viniferin), four trimers (three stilbenes and one stilbenoid), of which two (Z-miyabenol C and E-cis-miyabenol C) were new in grapevine, three tetramer stilbenoids, all new in grapevine, isohopeaphenol, ampelopsin H, and a vaticanol C-like isomer. The isolation of a dimer deriving from the condensation of (+)-catechin with trans-caffeic acid also indicated that other preformed phenolics are structurally modified in tissues infected with P. viticola.

Grapes are one of the major fruit crops and they are cultivated in many dry environments. This study comprehensively characterizes the metabolic response of grape berries exposed to water deficit at different developmental stages. Increases of proline, branched-chain amino acids, phenylpropanoids, anthocyanins, and free volatile organic compounds have been previously observed in grape berries exposed to water deficit. Integrating RNA-sequencing analysis of the transcriptome with large-scale analysis of central and specialized metabolites, we reveal that these increases occur via a coordinated regulation of key structural pathway genes. Water deficit-induced up-regulation of flavonoid genes is also coordinated with the down-regulation of many stilbene synthases and a consistent decrease in stilbenoid concentration. Water deficit activated both ABA-dependent and ABA-independent signal transduction pathways by modulating the expression of several transcription factors. Gene-gene and gene-metabolite network analyses showed that water deficit-responsive transcription factors such as bZIPs, AP2/ERFs, MYBs, and NACs are implicated in the regulation of stress-responsive metabolites. Enrichment of known and novel cis-regulatory elements in the promoters of several ripening-specific/water deficit-induced modules further affirms the involvement of a transcription factor cross-talk in the berry response to water deficit. Together, our integrated approaches show that water deficit-regulated gene modules are strongly linked to key fruit-quality metabolites and multiple signal transduction pathways may be critical to achieve a balance between the regulation of the stress-response and the berry ripening program. This study constitutes an invaluable resource for future discoveries and comparative studies, in grapes and other fruits, centered on reproductive tissue metabolism under abiotic stress.

FFQ, food diaries and 24 h recall methods represent the most commonly used dietary assessment tools in human studies on nutrition and health, but food intake biomarkers are assumed to provide a more objective reflection of intake. Unfortunately, very few of these biomarkers are sufficiently validated. This review provides an overview of food intake biomarker research and highlights present research efforts of the Joint Programming Initiative 'A Healthy Diet for a Healthy Life' (JPI-HDHL) Food Biomarkers Alliance (FoodBAll). In order to identify novel food intake biomarkers, the focus is on new food metabolomics techniques that allow the quantification of up to thousands of metabolites simultaneously, which may be applied in intervention and observational studies. As biomarkers are often influenced by various other factors than the food under investigation, FoodBAll developed a food intake biomarker quality and validity score aiming to assist the systematic evaluation of novel biomarkers. Moreover, to evaluate the applicability of nutritional biomarkers, studies are presently also focusing on associations between food intake biomarkers and diet-related disease risk. In order to be successful in these metabolomics studies, knowledge about available electronic metabolomics resources is necessary and further developments of these resources are essential. Ultimately, present efforts in this research area aim to advance quality control of traditional dietary assessment methods, advance compliance evaluation in nutritional intervention studies, and increase the significance of observational studies by investigating associations between nutrition and health.

Storage conditions and duration have a considerable influence on wine quality. Optimum temperature and humidity conditions may improve wine quality through ageing, while incorrect or excessively long storage leads to negative results. In order to evaluate the global effects of storage on red wine composition, 20 Sangiovese wines were stored in two different conditions (cellar or house) for a period of 2 years and analysed every 6 months. Untargeted LC–MS analysis showed various putative markers for the type and length of conservation (i.e. pigments, flavanols, pantothenic acid etc.), while targeted LC–MS confirmed and expanded these results within specific metabolic groups. The results of multivariate analysis showed that wines stored in the cellar changed little even after 2 years of storage, while wines stored in typical domestic conditions (house) developed approximately four times faster, reaching a composition similar to wines stored in the cellar for 2 years after just 6 months. The formation of several monosulfonated flavanols during domestic ageing provided the first evidence in wine of a reaction between wine tannins—both catechins and proanthocyanidins—and the exogenous antioxidant bisulfite. Moreover, ageing in domestic conditions appeared to induce an accelerated decrease in wine pigments, while specifically promoting the formation of pinotin A-like pigments and the hydrolysis of flavonol glycosides.

The abundance of lipids in plants is influenced by genotype and phenotype. Despite being a very important class of plant metabolites, knowledge of grape lipids is still very limited to date, with the exception of those located in seeds. Few investigations of grape lipids have shown that their profile depends on grape maturity, the variety and their location in the berry. Recent advances in liquid chromatography coupled to mass spectrometry have paved the way for faster analysis of lipids with minimal sample preparation. Here we describe a validation method for the extraction, identification and quantification of different classes of grape lipids: fatty acids, sterols, glycerolipids, glycerophospholipids and sphingolipids using liquid chromatographic electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS). The method was validated for 33 lipids, with linearity range (R(2)=0.95-1.00), LOQ (0.003-14.88 ng mL(-1)) and intraday and interday repeatability being evaluated for each lipid. The lipid profiling method developed was successfully applied to the analysis of 18 grape samples (10 red grape and 8 white grape varieties) from 4 different genetic groups: Vitis vinifera, Vitis non-vinifera, Muscat and hybrid; 33 lipids were identified and quantified. This method, which can be easily expanded to include further compounds and other plant tissues, is the starting point for analysis of the lipid profile in different grape tissues, an essential goal for better understanding the role of lipids in grape physiology.

Apples are one of the most commonly consumed fruits and their high polyphenol content is considered one of the most important determinants of their health-promoting activities. Here we studied the nutrikinetics of apple polyphenols by UHPLC-HRMS metabolite fingerprinting, comparing bioavailability when consumed in a natural or a polyphenol-enriched cloudy apple juice. Twelve men and women participated in an acute single blind controlled crossover study in which they consumed 250 mL of cloudy apple juice (CAJ), Crispy Pink apple variety, or 250 mL of the same juice enriched with 750 mg of an apple polyphenol extract (PAJ). Plasma and whole blood were collected at time 0, 1, 2, 3 and 5 h. Urine was collected at time 0 and 0-2, 2-5, 5-8, and 8-24 h after juice consumption. Faecal samples were collected from each individual during the study for 16S rRNA gene profiling. As many as 110 metabolites were significantly elevated following intake of polyphenol enriched cloudy apple juice, with large inter-individual variations. The comparison of the average area under the curve of circulating metabolites in plasma and in urine of volunteers consuming either the CAJ or the PAJ demonstrated a stable metabotype, suggesting that an increase in polyphenol concentration in fruit does not limit their bioavailability upon ingestion. Faecal bacteria were correlated with specific microbial catabolites derived from apple polyphenols. Human metabolism of apple polyphenols is a co-metabolic process between human encoded activities and those of our resident microbiota. Here we have identified specific blood and urine metabolic biomarkers of apple polyphenol intake and identified putative associations with specific genera of faecal bacteria, associations which now need confirmation in specifically designed mechanistic studies.

BACKGROUNDApples are rich in bioactive polyphenols and fiber. Evidence suggests that consumption of apples or their bioactive components is associated with beneficial effects on lipid metabolism and other markers of cardiovascular disease (CVD). However, adequately powered randomized controlled trials are necessary to confirm these data and explore the mechanisms.

The impact of diet on the microbiota composition and the role of diet in supporting optimal mental health have received much attention in the last decade. However, whether whole dietary approaches can exert psychobiotic effects is largely understudied. Thus, we investigated the influence of a psychobiotic diet (high in prebiotic and fermented foods) on the microbial profile and function as well as on mental health outcomes in a healthy human population. Forty-five adults were randomized into either a psychobiotic (n = 24) or control (n = 21) diet for 4 weeks. Fecal microbiota composition and function was characterized using shotgun sequencing. Stress, overall health and diet were assessed using validated questionnaires. Metabolic profiling of plasma, urine and fecal samples was performed. Intervention with a psychobiotic diet resulted in reductions of perceived stress (32% in diet vs. 17% in control group), but not between groups. Similarly, biological marker of stress were not affected. Additionally, higher adherence to the diet resulted in stronger decreases in perceived stress. While the dietary intervention elicited only subtle changes in microbial composition and function, significant changes in the level of 40 specific fecal lipids and urinary tryptophan metabolites were observed. Lastly, microbial volatility was linked to greater changes in perceived stress scores in those on the psychobiotic diet. These results highlight that dietary approaches can be used to reduce perceived stress in a human cohort. Using microbiota-targeted diets to positively modulate gut-brain communication holds possibilities for the reduction of stress and stress-associated disorders, but additional research is warranted to investigate underlying mechanisms, including the role of the microbiota.

This study aimed to establish relationships between wine composition and in-mouth sensory properties using a sensometabolomic approach. Forty-two red wines were sensorially assessed and chemically characterised using UPLC-QTOF-MS for targeted and untargeted analyses. Suitable partial least squares regression models were obtained for "dry", "sour", "oily", "prickly", and "unctuous". "Dry" was positively contributed by flavan-3-ols, anthocyanin derivatives (AntD), valine, gallic acid and its ethyl ester, and peptides, and negatively by sulfonated flavan-3-ols, anthocyanin-ethyl-flavan-3-ols, tartaric acid, flavonols (FOL), hydroxycinnamic acids (HA), protocatechuic ethyl ester, and proline. The "sour" model included molecules involved in "dry" and "bitter", ostensibly as a result of cognitive interactions. Derivatives of FOLs, epicatechin gallate, and N-acetyl-glucosamine phosphate contributed positively to "oily", as did vanillic acid, HAs, pyranoanthocyanins, and malvidin-flavan-3-ol derivatives for "prickly", and sugars, glutathione disulfide, AntD, FOL, and one HA for "unctuous". The presented approach offers an interesting tool for deciphering the sensory-active compounds involved in mouthfeel perception.

Highly reactive radicals, ROO•, were generated from 2,2‘-azobis[2-(2-imidazolin-2-yl)propane] and linoleic acid. The ROO• scavenging capacity of some Italian red wines was evaluated following the changes in oxygen consumption. Under the experimental conditions the time course of oxygen consumption shows two typical behaviors: trolox-like (class I) and gallic acid-like (class II). Usually the time course of wine was similar to that of gallic acid. The rate of oxygen consumption was found to decrease exponentially with the amount of wine or gallic acid added to the test solution. On this basis the capacity of red wines to scavenge peroxy radicals was expressed as content of gallic acid (SGA). The SGA values were found to be correlated to the amount of total proanthocyanidins and total polyphenols of some Italian red wines (p < 0.01). The proanthocyanidins extracted from seeds were shown to make a major contribution to the peroxy radical scavenging capacity of red wines, whereas, interestingly, the chemical class of the low molecular weight tannins reactive to vanillin did not correlate with the SGA values. Keywords: Wine; antioxidant capacity; lipid peroxidation; radical scavenging; proanthocyanidins; phenolics; anthocyanins

A headspace SPME GC–TOF-MS method was developed for the acquisition of metabolite profiles of apple volatiles. As a first step, an experimental design was applied to find out the most appropriate conditions for the extraction of apple volatile compounds by SPME. The selected SPME method was applied in profiling of four different apple varieties by GC–EI-TOF-MS. Full scan GC–MS data were processed by MarkerLynx software for peak picking, normalisation, alignment and feature extraction. Advanced chemometric/statistical techniques (PCA and PLS-DA) were used to explore data and extract useful information. Characteristic markers of each variety were successively identified using the NIST library thus providing useful information for variety classification. The developed HS-SPME sampling method is fully automated and proved useful in obtaining the fingerprint of the volatile content of the fruit. The described analytical protocol can aid in further studies of the apple metabolome.

Downy mildew, caused by the oomycete Plasmopara viticola, is a serious disease in Vitis vinifera, the most commonly cultivated grapevine species. Several wild Vitis species have instead been found to be resistant to this pathogen and have been used as a source to introgress resistance into a V. vinifera background. Stilbenoids represent the major phytoalexins in grapevine, and their toxicity is closely related to the specific compound. The aim of this study was to assess the resistance response to P. viticola of the Merzling × Teroldego cross by profiling the stilbenoid content of the leaves of an entire population and the transcriptome of resistant and susceptible individuals following infection.A three-year analysis of the population's response to artificial inoculation showed that individuals were distributed in nine classes ranging from total resistance to total susceptibility. In addition, quantitative metabolite profiling of stilbenoids in the population, carried out using HPLC-DAD-MS, identified three distinct groups differing according to the concentrations present and the complexity of their profiles. The high producers were characterized by the presence of trans-resveratrol, trans-piceid, trans-pterostilbene and up to thirteen different viniferins, nine of them new in grapevine.Accumulation of these compounds is consistent with a resistant phenotype and suggests that they may contribute to the resistance response.A preliminary transcriptional study using cDNA-AFLP selected a set of genes modulated by the oomycete in a resistant genotype. The expression of this set of genes in resistant and susceptible genotypes of the progeny population was then assessed by comparative microarray analysis.A group of 57 genes was found to be exclusively modulated in the resistant genotype suggesting that they are involved in the grapevine-P. viticola incompatible interaction. Functional annotation of these transcripts revealed that they belong to the categories defense response, photosynthesis, primary and secondary metabolism, signal transduction and transport.This study reports the results of a combined metabolic and transcriptional profiling of a grapevine population segregating for resistance to P. viticola. Some resistant individuals were identified and further characterized at the molecular level. These results will be valuable to future grapevine breeding programs.

The metabolic pigment composition of Sangiovese wines produced from grapes harvested at 20 different vineyards in Montalcino over three consecutive years (2008-2010) on a semi-industrial scale and of 55 commercial Brunello di Montalcino wines (2004-2007) was studied, using a targeted method capable of analyzing 90 pigments in an 11 min UHPLC-MS/MS chromatographic run. Interesting correlations were shown between various pigments formed during wine aging and those present in Sangiovese grapes. Vitisin B-like pigment and vitisin A-like pigment concentrations would seem to have a good correlation with ethyl-linked and direct-linked flavanol-anthocyanin concentrations, respectively. Moreover, the anthocyanic pattern recognition, genetically controlled by the plant variety, was shown to be inherited by the pigments formed during wine aging.

Wine micro-oxygenation is a globally used treatment and its effects were studied here by analysing by untargeted LC-MS the wine metabolomic fingerprint. Eight different procedural variations, marked by the addition of oxygen (four levels) and iron (two levels) were applied to Sangiovese wine, before and after malolactic fermentation. Data analysis using supervised and unsupervised multivariate methods highlighted some known candidate biomarkers, together with a number of metabolites which had never previously been considered as possible biomarkers for wine micro-oxygenation. Various pigments and tannins were identified among the known candidate biomarkers. Additional new information was obtained suggesting a correlation between oxygen doses and metal contents and changes in the concentration of primary metabolites such as arginine, proline, tryptophan and raffinose, and secondary metabolites such as succinic acid and xanthine. Based on these findings, new hypotheses regarding the formation and reactivity of wine pigment during micro-oxygenation have been proposed. This experiment highlights the feasibility of using unbiased, untargeted metabolomic fingerprinting to improve our understanding of wine chemistry.

Biosynthesis of the sesquiterpene rotundone in Vespolina grapes during berry ripening was investigated over two consecutive seasons, revealing that the compound accumulates from veraison to harvest and reaches relatively high concentrations (up to 5.44 μg/kg). Rotundone levels up to 1.91 μg/kg were also found in clones of Gruener Veltliner, a white grape variety known to give ‘peppery’ wines. These concentrations are higher than those reported for Syrah grapes and are similar to the levels found in some plants. Rotundone was shown to accumulate almost exclusively in berry exocarp, suggesting that skin contact during winemaking could be used to modulate the peppery character of red wine. However, rotundone yield after the winemaking process was relatively low. Indeed, only 10% of the rotundone present in grapes was extracted during fermentation, and only 6% was recovered in bottled wine. The results presented in this work provide key knowledge for manipulation of the peppery character of wine in order to optimize the intensity of this characteristic wine aroma.

Untargeted metabolomics are rapidly becoming an important tool for studying complex biological samples. Gas chromatography–mass spectrometry (GC–MS) is the most widely used analytical technology for metabolomic analysis of compounds that are volatile or can be chemically derivatised into volatile compounds. Unfortunately, data processing and analysis are not straightforward and the field is dominated by vendor-supplied software that does not always allow easy integration for large laboratories with different instruments. This paper presents an open-source pipeline for high-throughput GC–MS data processing, written in the R language and available as package metaMS. It features rapid annotation using in-house databases, and also provides support for building and validating such databases. The results are presented in simple-to-use tables, summarising the relative concentrations of identified compounds and unknowns in all samples. The use of the pipeline is illustrated using three experimental data sets.

pancreatic adenocarcinoma is the fourth leading cause of cancer related deaths due to its aggressive behavior and poor clinical outcome. There is a considerable variability in the frequency of serum tumor markers in cancer' patients. We performed a metabolomics screening in patients diagnosed with pancreatic cancer.Two targeted metabolomic assays were conducted on 40 serum samples of patients diagnosed with pancreatic cancer and 40 healthy controls. Multivariate methods and classification trees were performed.Sparse partial least squares discriminant analysis (SPLS-DA) was used to reduce the high dimensionality of a pancreatic cancer metabolomic dataset, differentiating between pancreatic cancer (PC) patients and healthy subjects. Using Random Forest analysis palmitic acid, 1,2-dioleoyl-sn-glycero-3-phospho-rac-glycerol, lanosterol, lignoceric acid, 1-monooleoyl-rac-glycerol, cholesterol 5α,6α epoxide, erucic acid and taurolithocholic acid (T-LCA), oleoyl-L-carnitine, oleanolic acid were identified among 206 metabolites as highly discriminating between disease states. Comparison between Receiver Operating Characteristic (ROC) curves for palmitic acid and CA 19-9 showed that the area under the ROC curve (AUC) of palmitic acid (AUC=1.000; 95% confidence interval) is significantly higher than CA 19-9 (AUC=0.963; 95% confidence interval: 0.896-1.000).Mass spectrometry-based metabolomic profiling of sera from pancreatic cancer patients and normal subjects showed significant alterations in the profiles of the metabolome of PC patients as compared to controls. These findings offer an information-rich matrix for discovering novel candidate biomarkers with diagnostic or prognostic potentials.

The impact of minute amounts of oxygen in the headspace on the post-bottling development of wine is generally considered to be very important, since oxygen can either damage or improve the quality of wine. This project aimed to gain new experimental evidence about the chemistry of the interaction between wine and oxygen. The experimental design included 216 bottles of 12 different white wines produced from 6 different cultivars (Inzolia, Muller Thurgau, Chardonnay, Grillo, Traminer and Pinot gris). Half of them were bottled using the standard industrial process with inert headspace and the other half without inert gas and with extra headspace. After 60 days of storage at room temperature, the wines were analysed using an untargeted LC-MS method. The use of a detailed holistic analysis workflow, with several levels of quality control and marker selection, gave 35 metabolites putatively induced by the different amounts of oxygen. These metabolite markers included ascorbic acid, tartaric acid and various sulfonated compounds observed in wine for the first time (e.g. S-sulfonated cysteine, glutathione and pantetheine; and sulfonated indole-3-lactic acid hexoside and tryptophol). The consumption of SO2 mediated by these sulfonation reactions was promoted by the presence of higher levels of oxygen on bottling.

Abstract Anthocyanins exert neuroprotection in various in vitro and in vivo experimental models. However, no details regarding their brain-related pharmacokinetics are so far available to support claims about their direct neuronal bioactivity as well as to design proper formulations of anthocyanin-based products. To gather this missing piece of knowledge, we intravenously administered a bolus of 668 nmol cyanidin 3-glucoside (C3G) in anaesthetized Wistar rats and shortly after (15 s to 20 min) we collected blood, brain, liver, kidneys and urine samples. Extracts thereof were analysed for C3G and its expected metabolites using UPLC/MS-MS. The data enabled to calculate a set of pharmacokinetics parameters. The main finding was the distinctive, rapid distribution of C3G in the brain, with an apparently constant plasma/brain ratio in the physiologically relevant plasma concentration range (19–355 nM). This is the first report that accurately determines the distribution pattern of C3G in the brain, paving the way to the rational design of future tests of neuroprotection by C3G in animal models and humans.

Every grape cultivar has its own unique genetic characteristics, leading to the production of a different secondary metabolite profile. Aroma is one of the most important aspects in terms of the quality of grapes and previous studies have assigned specific aromas to particular grape cultivars. In this study we present the molecular profiling of volatile aroma metabolites and their precursors in ten selected genotypes, including six Vitis vinifera cultivars, two American species (Arizonica Texas, Vitis cinerea) and two interspecific crosses. Chemical profiling was achieved through combined use of two orthogonal techniques, GC-MS and LC-HRMS, before and after enzymatic hydrolysis. The results show that both free and glycosidically bound aroma precursors behave differently in each different grape cultivar and species. As many as 66 free aroma volatile molecules (originally existing and released after hydrolysis) were profiled through GC-MS analysis, while 15 glycosylated precursors of volatiles were identified through LC-HRMS and correlation with GC-MS data.

In this study, we assessed the main bioactive compounds of a broad apple germplasm collection, composed by 247 accessions of wild (97) and domesticated (150) species. Among the stilbenes, trans- and cis-piceid were found to be ubiquitary components of both wild and cultivated apples. Apple was suggested to be the second dietary source of resveratrols. Results confirmed that the selection pressure of breeding and domestication did not uniformly affect all the phytochemicals contained in apples. For instance, organic acids (malic and ascorbic acid) and some phenolics (stilbenes, hydroxycinnamic acids, and dihydrochalcones) were significantly influenced by selection, while some relevant flavonoids (flavonols and flavan-3-ols) and triterpenoids (ursolic, oleanolic, and betulinic acids) were not. This comprehensive screening will assist in the selection of Malus accessions with specific nutraceutical traits suitable to establish innovative breeding strategies or to patent new functional foods and beverages.

We analyzed via untargeted UHPLC-ESI-Q-TOF-MS the metabolome of the berry tissues (skin, pulp, seeds) of some American Vitis species (Vitis cinerea, Vitis californica, Vitis arizonica), together with four interspecific hybrids, and seven Vitis vinifera cultivars, aiming to find differences in the metabolomes of the American Vitis sp. versus Vitis vinifera. Apart from the known differences, that is, more complex content of anthocyanins and stilbenoids in the American grapes, we observed higher procyanidin accumulation (tens to hundreds of times) in the vinifera skin and seeds in comparison to American berries, and we confirmed this result via phloroglucinolysis. In the American grapes considered, we did not detect the accumulation of pleasing aroma precursors (terpenoids, glycosides), whereas they are common in vinifera grapes. We also found accumulation of hydrolyzable tannins and their precursors in the skin of the wild American grapes, which has never been reported earlier in any of the species under investigation. Such information is needed to improve the design of new breeding programs, lowering the risk of retaining undesirable characteristics in the chemical phenotype of the offspring.

In this study, Vitis vinifera L wines cv. Cabernet Franc, Merlot, Sangiovese and Syrah, 2006 and 2007 vintages, produced in São Joaquim, a new wine-producing region in southern Brazil, were evaluated. As phenolic compound content is one of the most important parameters in assessing wine quality and is possibly partially responsible for the beneficial health properties of wines, in this paper the levels of the main anthocyanins, flavonols, hydroxycinnamic acid and hydroxybenzoic acid (HPLC-DAD and HPLC-DAD–MS analysis) and the in vivo antioxidant activity in mice are reported. The antioxidant capacity of plasma was assessed through the reduction of ferric iron (FRAP). Lipid peroxidation (TBARS), carbonyl protein (CP), reduced glutathione (GSH) levels and the catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GPx) activity were determined in livers of the test animals. The results for the phenolic compounds content of the wine samples were considered appropriate for quality red wines, and the wine consumption promoted a significant increase in FRAP and decreases in the TBARS and CP levels and in the CAT, SOD and GPx activity. Moreover, the phenolic content of the wines was positively correlated with the in vivo antioxidant capacity promoted by regular wine consumption.

The phenolic composition of grape berries grown in Italy (Cabernet Cortis, Johanniter, Solaris, Phoenix, and Regent) and in Germany (Regent and Phoenix) was analyzed using two complementary LC-MS/MS (Liquid Chromatography - Tandem Quadrupole Mass Spectrometry) methods. These five cultivars belong to fungus-resistant grape varieties (PIWI) and little is known about their phenolic profile, although phenolics such as phytoalexins have been recognized, playing a key role in the resistance mechanism. A triple quadruple mass spectrometer detector was used for both identification and quantification, and analytical tools from untargeted metabolomics were applied to check and control the quality of the results. Specifically, biological, technical, and instrumental replications were included in order to study the variability at different levels. The results allowed tuning of the sampling protocol and provided a rich phenolic metabolite profile for the aforementioned PIWI varieties, as compared with Vitis vinifera cultivars, especially in the class of stilbenes.

In the last decade, great progress has been made in clarifying the main determinants of anthocyanin accumulation in grape berry skin. However, the molecular details of the fine variation among cultivars, which ultimately contributes to wine typicity, are still not completely understood. To shed light on this issue, the grapes of 170 F1 progeny from the cross 'Syrah'×'Pinot Noir' were characterized at the mature stage for the content of 15 anthocyanins during four growing seasons. This huge data set was used in combination with a dense genetic map to detect genomic regions controlling the anthocyanin pathway both at key enzymatic points and at particular branches. Genes putatively involved in fine tuning the global regulation of anthocyanin biosynthesis were identified by exploring the gene predictions in the QTL (quantitative trait locus) confidence intervals and their expression profile during berry development in offspring with contrasting anthocyanin accumulation. New information on some aspects which had scarcely been investigated so far, such as anthocyanin transport into the vacuole, or completely neglected, such as acylation, is provided. These genes represent a valuable resource in grapevine molecular-based breeding programmes to improve both fruit and wine quality and to tailor wine sensory properties according to consumer demand.

Downy mildew (Plasmopara viticola) is one of the most destructive diseases of the cultivated species Vitis vinifera. The use of resistant varieties, originally derived from backcrosses of North American Vitis spp., is a promising solution to reduce disease damage in the vineyards. To shed light on the type and the timing of pathogen-triggered resistance, this work aimed at discovering biomarkers for the defense response in the resistant variety Bianca, using leaf discs after infection with a suspension of P. viticola. We investigated primary and secondary metabolism at 12, 24, 48 and 96 hours post inoculation (hpi). We used methods of identification and quantification for lipids (LC-MS/MS), phenols (LC-MS/MS), primary compounds (GC-MS), and semi-quantification for volatile compounds (GC-MS). We were able to identify and quantify or semi-quantify 176 metabolites among which 53 were modulated in response to pathogen infection. The earliest changes occurred in primary metabolism at 24-48 hpi and involved lipid compounds, specifically unsaturated fatty acid and ceramide; amino acids, in particular proline; and some acids and sugars. At 48 hpi, we also found changes in volatile compounds and accumulation of benzaldehyde, a promoter of salicylic acid-mediated defense. Secondary metabolism was strongly induced only at later stages. The classes of compounds that increased at 96 hpi included phenylpropanoids, flavonols, stilbenes and stilbenoids. Among stilbenoids we found an accumulation of ampelopsin H+vaticanol C, pallidol, ampelopsin D +quadrangularin A, Z-miyabenol C and α-viniferin in inoculated samples. Some of these compounds are known as phytoalexins, while others are novel biomarkers for the defense response in Bianca. This work highlighted some important aspects of the host response to P. viticola in a commercial variety under controlled conditions, providing biomarkers for a better understanding of the mechanism of plant defense and a potential application in field studies of resistant varieties.

We carried out comprehensive mapping of volatile compounds in 70 wines, from 48 wineries and 6 vintages, representative of the two main production areas for Italian sparkling wines, by HS-SPME-GCxGC-TOF-MS and multivariate analysis. The final scope was to describe the metabolomics space of these wines, and to verify whether the grape cultivar signature, the pedoclimatic influence of the production area, and the complex technology were measurable in the final product. The wine chromatograms provided a wealth of information, with 1695 compounds being found. A large number of putative markers influenced by the cultivation area was observed. A subset of 196 biomarkers fully discriminated between the two types of sparkling wines investigated. Among the new compounds, safranal and α-isophorone were observed. We showed how correlation-based network analysis could be used as a tool to detect the differences in compound behaviour based on external/environmental influences.

Abstract Wine has one of the broadest chemical profiles, and the common oenological practice of adding the antioxidant and antimicrobial sulfur dioxide has a major impact on its metabolomic fingerprint. In this study, we investigated novel discovered oenological reactions primarily occurring between wine metabolites and sulfur dioxide. The sulfonated derivatives of epicatechin, procyanidin B2, indole acetic acid, indole lactic acid and tryptophol were synthesized and for the first time quantified in wine. Analysis of 32 metabolites in 195 commercial wines (1986–2016 vintages) suggested that sulfonation of tryptophan metabolites characterised white wines, in contrast to red wines, where sulfonation of flavanols was preferred. The chemical profile of the oldest wines was strongly characterised by sulfonated flavanols and indoles, indicating that could be fundamental metabolites in explaining quality in both red and white aged wines. These findings offer new prospects for more precise use of sulfur dioxide in winemaking.

The main aim of this work was to develop an untargeted normal phase LC-MS method, starting from a targeted method already validated for the analysis of 135 polar metabolites. Since the LC instrument and column were the same, most of the chromatographic conditions remained identical, while the adaptations focused on maintaining the ionic strength of the eluents constant. The sample preparation was simplified and the effectiveness of LC-MS for long batches was evaluated, in order to record the maximum number of metabolites with good chromatographic resolution and the best MS stability and accuracy. The method was applied to study the influence of storage conditions on wine composition. Slightly sub-optimum storage conditions had a major impact on the polar metabolite fingerprint of the red wines analysed and the markers revealed included phenolics, vitamins and metabolites indentified in wine for the first time (4-amino-heptanedioic acid and its ethyl ester).

Tryptophan and tyrosine metabolism has a major effect on human health, and disorders have been associated with the development of several pathologies. Recently, gut microbial metabolism was found to be important for maintaining correct physiology. Here, we describe the development and validation of a UHPLC-ESI-MS/MS method for targeted quantification of 39 metabolites related to tryptophan and tyrosine metabolism, branched chain amino acids and gut-derived metabolites in human plasma and urine. Extraction from plasma was optimised using 96-well plates, shown to be effective in removing phospholipids. Urine was filtered and diluted ten-fold. Metabolites were separated with reverse phase chromatography and detected using triple quadrupole MS. Linear ranges (from ppb to ppm) and correlation coefficients (r2 &gt; 0.990) were established for both matrices independently and the method was shown to be linear for all tested metabolites. At medium spiked concentration, recovery was over 80% in both matrices, while analytical precision was excellent (CV &lt; 15%). Matrix effects were minimal and retention time stability was excellent. The applicability of the methods was tested on biological samples, and metabolite concentrations were found to be in agreement with available data. The method allows the analysis of up to 96 samples per day and was demonstrated to be stable for up to three weeks from acquisition.

Nuts and vegetable oils are important sources of fat and of a wide variety of micronutrients and phytochemicals. Following their intake, several of their constituents, as well as their derived metabolites, are found in blood circulation and in urine. As a consequence, these could be used to assess the compliance to a dietary intervention or to determine habitual intake of nuts and vegetable oils. However, before these metabolites can be widely used as biomarkers of food intake (BFIs), several characteristics have to be considered, including specificity, dose response, time response, stability, and analytical performance. We have, therefore, conducted an extensive literature search to evaluate current knowledge about potential BFIs of nuts and vegetable oils. Once identified, the strengths and weaknesses of the most promising candidate BFIs have been summarized. Results from selected studies have provided a variety of compounds mainly derived from the fatty fraction of these foods, but also other components and derived metabolites related to their nutritional composition. In particular, α-linolenic acid, urolithins, and 5-hydroxyindole-3-acetic acid seem to be the most plausible candidate BFIs for walnuts, whereas for almonds they could be α-tocopherol and some catechin-derived metabolites. Similarly, several studies have reported a strong association between selenium levels and consumption of Brazil nuts. Intake of vegetable oils has been mainly assessed through the measurement of specific fatty acids in different blood fractions, such as oleic acid for olive oil, α-linolenic acid for flaxseed (linseed) and rapeseed (canola) oils, and linoleic acid for sunflower oil. Additionally, hydroxytyrosol and its metabolites were the most promising distinctive BFIs for (extra) virgin olive oil. However, most of these components lack sufficient specificity to serve as BFIs. Therefore, additional studies are necessary to discover new candidate BFIs, as well as to further evaluate the specificity, sensitivity, dose-response relationships, and reproducibility of these candidate biomarkers and to eventually validate them in other populations. For the discovery of new candidate BFIs, an untargeted metabolomics approach may be the most effective strategy, whereas for increasing the specificity of the evaluation of food consumption, this could be a combination of different metabolites.

Lipids are important constituents of plant and animal cells, being essential due to their biological functions. Despite the significant role of these compounds in grape berries, knowledge of grape lipid composition is still limited. This study addresses the free fatty acid composition and other lipids in skins and seeds of several Vitis vinifera grape cultivars using LC-MS/MS. A different profile of free fatty acids was determined in the grape tissues, showing a higher proportion of unsaturated fatty acids in seeds (ca. 60%). Lignoceric acid was one of the main free saturated fatty acids found in grape skins, together with palmitic and stearic acids. Berry skins showed similar ranges of C18-unsaturated fatty acids, whereas linoleic acid was predominant in the seed fatty acid composition. Higher content of glycerophospholipids was determined in grape seeds. Uvaol and oleanolic acid were only quantified in skins (1.5–3.9 and 38.6–57.6 mg/kg fresh weight, respectively). These preliminary results suggest a certain diversity in grape lipids according to their location in the berry tissues and cultivar, providing useful information for the fermentation process due to their role in wine sensory profile and yeast growth.

The aim of this project was to register, in a liquid chromatography–mass spectrometry-based untargeted single-batch analysis, the metabolome of 11 single-cultivar, single-vintage Italian red wines (Aglianico, Cannonau, Corvina, Montepulciano, Nebbiolo, Nerello, Primitivo, Raboso, Sagrantino, Sangiovese, and Teroldego) from 12 regions across Italy, each one produced in their terroirs under ad hoc legal frameworks to guarantee their quality and origin. The data provided indications regarding the similarity between the cultivars and highlighted a rich list of putative biomarkers of origin wines (pBOWs) characterizing each individual cultivar–terroir combination, where Primitivo, Teroldego, and Nebbiolo had the maximum number of unique pBOWs. The pBOWs included anthocyanins (Teroldego), flavanols (Aglianico, Sangiovese, Nerello, and Nebbiolo), amino acids and N-containing metabolites (Primitivo), hydroxycinnamates (Cannonau), and flavonols (Sangiovese). The raw data generated in this study are publicly available and, therefore, accessible and reusable as a baseline data set for future investigations.

Abstract Grapes and berries are two types of widely consumed fruits characterized by a high content in different phytochemicals. However, their accurate dietary assessment is particularly arduous, because of the already wide recognized bias associated with self-reporting methods, combined with the large range of species and cultivars and the fact that these fruits are popularly consumed not only in fresh and frozen forms but also as processed and derived products, including dried and canned fruits, beverages, jams, and jellies. Reporting precise type and/or quantity of grape and berries in FFQ or diaries can obviously be affected by errors. Recently, biomarkers of food intake (BFIs) rose as a promising tool to provide accurate information indicating consumption of certain food items. Protocols for performing systematic reviews in this field, as well as for assessing the validity of candidate BFIs have been developed within the Food Biomarker Alliance (FoodBAll) Project. This paper aims to evaluate the putative BIFs for blueberries, strawberries, raspberries, blackberries, cranberries, blackcurrant, and grapes. Candidate BFIs for grapes were resveratrol metabolites and tartaric acid. The metabolites considered as putative BFI for berries consumption were mostly anthocyanins derivatives together with several metabolites of ellagitannins and some aroma compounds. However, identification of BFIs for single berry types encountered more difficulties. In the absence of highly specific metabolites reported to date, we suggested some multi-metabolite panels that may be further investigated as putative biomarkers for some berry fruits.

Browning and over-oxidation are the most common alterations affecting white wines. The reduction of phenolic compounds by means of adsorbents is the method most frequently employed in oenology for countering these phenomena. The purpose of this study was to investigate the use of two types of adsorbents not as yet employed in oenology, chitin and chitosan, on a variety of white wines (Sauvignon, Pinot, Trebbiano, Albana, Verdicchio, Bianco di Custoza) and to compare their behaviour with that of two conventional adsorbents already used for these applications, potassium caseinate and polyvinyl pyrrolidone. The chitosans exhibited a high affinity to phenolic compounds (polyphenols, cinnamic acids, flavans, proanthocyanidins), in particular to cinnamic acids, and good stabilization to browning over time as determined by an accelerated test. Two of the chitosans tested were found to be comparable to the conventional adsorbents.

The inhibitory effect of anthocyanins has been investigated in the peroxidation of linoleic acid in micelles in the presence and in the absence of (+)-catechin. The peroxidation was initiated by thermal decomposition of 2,2′-azobis[2-(2-imidazolin-2-yl)propane], and the kinetics of peroxidation were followed by measuring the rate of oxygen consumption and the rate of disappearance of the antioxidant. The analysis of the antioxidant effect of various anthocyanins, alone or in the presence of catechin, demonstrates that catechin, which is relatively inefficient at inhibiting linoleic acid oxidation, regenerates the highly efficient antioxidant malvidin 3-glucoside and, at a lower extent, peonidin 3-glucoside. The malvidin 3-glucoside recycling by catechin strongly increases the antioxidant efficiency of these two antioxidants. This protective mechanism appears specific for malvidin and peonidin 3-glucosides. The high unpaired spin density of the phenolic O atoms in the radicals generated by these anthocyanins, calculated by the semiempirical quantum chemical AM1 method, may explain the observed behavior.

Two chemical classes of flavonoids, the flavan-3-ols (catechins and proanthocyanidins) and the anthocyanins, are the natural antioxidants present at the highest concentration in red grape and wine. In the berry, the anthocyanins are localized in the skins, similarly to other highly bioactive phenolics of grape such as the resveratrols and the flavonols, while the flavan-3-ols are contained both in the skins and seeds. During winemaking, only a fraction of the grape flavonoids are selectively extracted into the wine, with a time course and a final yield strongly depending on the grape variety. The knowledge of the diverse and cultivar-specific characteristics of the grape is therefore critical to the appropriate design of the winemaking process. By means of a selective extraction method specifically designed to mimic the winemaking process, it was possible to analyze the "phenolic potential of red grape," thus obtaining quantitative information about amount and localization of the extractable flavonoids in the grape. Twenty-five high-quality red grape cultivars (V. vinifera) were studied, including 4 of the worldwide leading cultivars and 21 Italian cultivars with the highest reputation for the production of both young and aged premium red wines. The results clearly indicate that the grape variety plays a central role in determining both the absolute amount of the flavonoids, and the distribution between the berry skin and seeds of the flavan-3-ols. The very high biodiversity of the red grape cultivars in terms of flavonoids indicate a largely under-exploited opportunity to produce a range of diverse premium wines with optimized levels of natural antioxidants.

Some recent studies have put forward the hypothesis that the presence of trans-resveratrol (trans-3,5,4'-trihydroxystilbene) in red wine may be related to some of its therapeutic properties. A fundamental step in view of this evaluation is the development of a method for the quick, accurate and precise analysis of this compound. Sample enrichment and purification can be obtained by solid-phase extraction using reverse-phase C18 cartridges. HPLC analysis carried out by means of a photodiode-array detector, with an internal standard method, allows the detection of up to 10 micrograms/L in wine, with a linear range between 0.6 and 300 ng injected and a precision of 3.3%. The results of the first analyses show that the concentrations of trans-resveratrol in wines might be much higher than so far reported in the literature.

A new direct HPLC method with fluorescence detection has been developed for the routine analysis of riboflavin, flavin mononucleotide and flavin–adenine dinucleotide, in wines and other beverages. These compounds are the main agents responsible for the “taste of light” that some white wines and other beverages develop when they are exposed to the light, due to the formation of sulfur compounds that produce an onion/garlic odor. A Hewlett-Packard 1100 gradient liquid chromatograph with 1046A fluorescence detector was used. To improve the selectivity, each compound was monitored to fit the best λexcitation/λemission (265/525 nm). A 500 nm cut-off filter was used. The column was a Hypersil C18 ODS, 200×2.1 mm, 5 μm particle size. The volume injected was 20 μl. A constant flow-rate of 0.6 ml/min was used with two solvents: solvent A, 0.05 M buffer NaH2PO4 at pH=3.0 with H3PO4 and solvent B, acetonitrile. The precision, linearity and sensitivity of this method have been established.

Although the composition of strawberry fruit has been extensively studied, especially for the most abundant phenolic compounds, agrimoniin has never been univocally identified as one of the most abundant phenolic compounds in the fruit. In this study agrimoniin was isolated in the fruit of Fragaria vesca and its structure characterized. Furthermore, its presence was definitively established to be the main ellagitannin in both F. vesca and Fragaria ananassa D. fruit. The presence of sanguiin H-6 and lambertianin C as minor compounds was confirmed in both F. vesca and F. ananassa D. samples. For the first time here is reported the full NMR assignments for agrimoniin. These data should represent a point of reference for NMR analysis of this and other structurally related ellagitannins. Finally, the establishment of an HPLC protocol for separation provided information making it possible to avoid confusion with sanguiin H-6, the main ellagitannin in Rubus species, which is also present in strawberries but at a much lower concentration.

Ellagitannins and ellagic acid conjugates are polyphenols present in the human diet, in particular strawberries (Fragaria spp.). The first aim of this study was isolation and structural characterization of casuarictin and 3-O-methyl ellagic acid 3′-O-α-rhamnopyranoside, which were found to be abundant in Fragaria spp., along with agrimoniin. The second aim was accurate profiling and quantification of 26 ellagitannins and ellagic acid conjugates in six Fragaria x ananassa cultivars and two Fragaria vesca species. The third aim was to describe the ellagitannins behavior during fruit ripening from the green stage to over-ripeness. It was shown that there are major qualitative and quantitative differences in the amount and profile of ellagitannins and ellagic acid conjugates between Fragaria spp. Genotype is a major factor in defining ellagitannin concentration and patterns between strawberries, and variable behavior of the genotypes was observed, in the context of a significant drop in ellagitannins during ripening.

Mass spectrometry-based imaging techniques applied to small molecules complement the growing research field of metabolomics and can be used to interpret many important biological processes occurring in plants. In untargeted imaging applications, chemical identification is a critical step since it cannot take advantage of separative techniques applied to neutral molecules (e.g. liquid chromatography). The use of high resolution spectrometers is of great help, but fragmentation experiments are often necessary. In many cases, the information on ion fragmentation is embedded in the data sets, because analytes break up during ionization, but the extraction of this information is not easy considering the complexity of the imaging data files. Here an approach is proposed for applying conventional untargeted MALDI (matrix-assisted laser desorption ionization) profiling and advanced data analysis to perform imaging of metabolites in apple tissues. The pipeline, based on intensity correlation analysis, is used to extract fragmentation information from untargeted, high resolution, wide range mass spectra and to reconstruct compound-specific images which can be used for interpretation purposes. The proposed approach was used to investigate the distribution of glycosylated flavonols and dihydrochalcones in Golden Delicious apples. The results indicate that the method is effective, showing a high potential for ascertaining detailed metabolite localization.

To asses the hypothesis that anthocyanins are rapidly taken up from the blood into tissues, where they accumulate up to their bioactivity threshold, an intravenous dose of cyanidin 3-glucoside (1) was administered to anaesthetized rats. Cyanidin 3-glucoside (1) and its metabolites were analyzed in the plasma, kidneys, liver, urine, and bile, using last-generation mass spectrometry. Compound 1 was found to rapidly disappear from plasma (t/2 = 0.36 min). As soon as 15 s after its administration, both 1 and its methylation product, peonidin 3-glucoside (2), were detected in the plasma, kidneys, and liver. At 1 min, both 1 and 2 had almost disappeared from the plasma, but attained their peak concentrations in the kidneys and in the liver. Compound 2 was rapidly excreted both in the bile and in the urine. Three additional methylated metabolites were detected in traces, namely, delphinidin 3-glucoside (3), petunidin 3-glucoside (4), and malvidin 3-glucoside (5). These data contribute to solving the paradox of the high bioactivity of anthocyanins in spite of their apparent low bioavailability.

Flavonoids are dietary components involved in decreasing oxidative stress in the vascular endothelium and thus the risk of endothelial dysfunction. However, their very low concentrations in plasma place this role in doubt. Thus, a relationship between the effective intracellular concentration of flavonoids and their bioactivity needs to be assessed. This study examined the uptake of physiological concentrations of cyanidin 3-glucoside, a widespread dietary flavonoid, into human vascular endothelial cells. Furthermore, the involvement of the membrane transporter bilitranslocase (TC No. 2.A.65.1.1) as the key underlying molecular mechanism for membrane transport was investigated by using purified anti-sequence antibodies binding at the extracellular domain of the protein. The experimental observations were carried out in isolated plasma membrane vesicles and intact endothelial cells from human endothelial cells (EA.hy926) and on an ischemia–reperfusion model in isolated rat hearts. Cyanidin 3-glucoside was transported via bilitranslocase into endothelial cells, where it acted as a powerful intracellular antioxidant and a cardioprotective agent in the reperfusion phase after ischemia. These findings suggest that dietary flavonoids, despite their limited oral bioavailability and very low postabsorption plasma concentrations, may provide protection against oxidative stress-based cardiovascular diseases. Bilitranslocase, by mediating the cellular uptake of some flavonoids, is thus a key factor in their protective activity on endothelial function.

The aim of this work was to characterise native flavonol profiles of different varieties of blueberries during different growing seasons. The analysis was performed using liquid chromatography–mass spectrometry (LC/MS) equipment, with a reversed phase column for flavonol separation. The identity of blueberry flavonols was also confirmed using high-resolution and high-accuracy mass spectrometry. Relatively large differences between different varieties of blueberries were found in terms of their flavonol content. The sum of flavonol glycosides varied between 137 mg/kg for the Duke variety to 272 mg/kg for the Simultan variety. Quercetin was the most abundant in all varieties (59.4% ± 8.7%), while the sequence in terms of the presence of other flavonols was variety-dependent (from 16.3% to 1.6%). The most significant conjugate sugars in blueberry extracts were galactoside (ranging from 35.8% to 72.1%) and glucoside (ranging from 12.1% to 27.1%). The study demonstrated that the percentage of different sugar glycosides depends on the variety. These data support the conclusion that cultivated blueberries are one of the most concentrated sources of dietary flavonol glycosides in the human diet. They can provide on average 196 mg/kg of a complex mixture of up to 23 different flavonol glycosides.

The contents of stilbene monomers, cis-resveratrol, trans-resveratrol, cis-piceid, trans-piceid, and tyrosol, were quantified in Vitis vinifera red wines, cvs. Cabernet Franc, Merlot, Sangiovese, and Syrah, 2006 and 2007 vintages, from the São Joaquim region, a new grape-growing region at southern Brazil. Moreover, the effect of chronic consumption of these wines on the antioxidant and hypolipidemic activities was monitored in C57BL6 LDL receptor knockout mice and treated with a hypercholesterolemic diet. Red wines from this region had substantial levels of resveratrols (the predominant forms were glycoside and trans) and tyrosol. Biomonitoring of antioxidant and hypolipidemic activities in vivo revealed that consumption of these wines increased the antioxidant capacity and reduced the hypercholesterolemia and hypertriglyceridemia promoted by the hypercholesterolemic diet. Significant correlations were found between the increase of antioxidant capacity markers, the decrease of lipid levels promoted by wine consumption, and the contents of stilbenes and tyrosol, supporting the important biological activity of these compounds.

Nowadays the trend in analytical chemistry is clearly towards the creation of multiple methods with extended coverage, enabling the determination of many different classes of compounds in a single analysis in which virtually all classes of different compounds are included in a single run. The aim of this study was to develop and validate a versatile and selective GC/MS/MS method for metabolite profiling of volatile compounds in apples, raspberries and grapes. Validation of the method was performed in terms of the limit of detection, limit of quantification, linearity range, and inter and intraday precision. Confirmation of the identity of the compounds in samples was carried out by checking compliance of the q/Q ratio of samples and reference standards. The multiple reaction monitoring with selection of two transition ions, one for quantification and one for confirmation, provided excellent selectivity and sensitivity, using the q/Q ratio as a confirmatory parameter. A multi target method was developed and validated for the simultaneous quantification and confirmation of 160 volatile compounds of raspberries, apples and grapes. The main classes were esters (42), alcohols (32), monoterpenes (31), aldehydes (17), ketones (12), norisoprenoids (8), acids (8), sesquiterpenes (7), pyrazines (3) and ethers (1) allowing the detection and quantification of 69 compounds in apples, 122 in grapes and 42 in raspberries. Moreover, the method developed can be easily extended to volatile compounds in other fruits and can therefore be widely used for quantification/profiling studies in the field of fruit aroma.

The free flavan-3-ol and proanthocyanidin (PA) profile and the antioxidant capacity of wines Vitis vinifera L., 2006 and 2007 vintages, from the São Joaquim region, at southern Brazil, are reported here for the first time. Catechin and epicatechin were the two main monomers in the wine samples, followed by gallocatechin and epigallocatechin; and the PA B1 was the main dimer. The terminal units of the PAs were constituted mainly by catechin units, with the co-presence of epicatechin, gallocatechin, epigallocatechin and traces of epicatechin gallate. The epicatechin and epigallocatechin units were the main constituents of the extension units of PAs with the co-presence of catechin and epicatechin gallate. The values for the mean degree of polymerisation ranged from 4.9 to 9.8. The wine samples demonstrated effective scavenging activity against DPPH and ABTS radicals and against lipid peroxidation in vitro. A positive correlation existed between flavan-3-ol content and antioxidant capacity in vitro.

An increasing number of studies have concerned the profiling of polyphenol microbial metabolites, especially in urine or plasma, but only a few have regarded their accurate quantification. This study reports on a new ultra-performance liquid chromatography tandem mass spectrometry method with electrospray ionisation (UHPLC–ESI-MS/MS) using a simple clean-up step with solid phase extraction (SPE) and validation on different biological matrices. The method was tested with spiked samples of liver, heart, kidneys, brain, blood and urine. The purification procedure, after the evaluation of three different cartridges, makes it possible to obtain cleaner samples and better quantification of putative trace metabolites, especially related to dietary studies, with concentrations below ng/g in tissue and for urine and blood, starting from ng/ml. Limits of detection and linear range were also assessed using mixed polyphenol metabolite standards. Short chromatographic separation was carried out for 23 target compounds related to the polyphenol microbial metabolism, coupled with a triple quadrupole mass spectrometer for their accurate quantification. By analysing different spiked biological samples we were able to test metabolite detection in the matrix and validate the overall recovery of the method, from purification to quantification. The method developed can be successfully applied and is suitable for high-throughput targeted metabolomics analysis related to nutritional intervention, or the study of the metabolic mechanism in response to a polyphenol-rich diet.

Fruit is a key component of a healthy diet. However, it is still not clear whether some classes of fruit may be more beneficial than others and whether all individuals whatever their age, gender, health status, genotype, or gut microbiota composition respond in the same way to fruit consumption. Such questions require further observational and intervention studies in which the intake of a specific fruit can be precisely assessed at the population and individual levels. Within the Food Biomarker Alliance Project (FoodBAll Project) under the Joint Programming Initiative "A Healthy Diet for a Healthy Life", an ambitious action was undertaken aiming at reviewing existent literature in a systematic way to identify validated and promising biomarkers of intake for all major food groups, including fruits. This paper belongs to a series of reviews following the same BFIRev protocol and is focusing on biomarkers of pome and stone fruit intake. Selected candidate biomarkers extracted from the literature search went through a validation process specifically developed for food intake biomarkers.

In order to differentiate the extra virgin olive oils (EVOO) of different origin of purchase, such as monovarietal Italian EVOO with protected denomination of origin (PDO) and commercial-blended EVOO purchased in supermarkets, a number of samples was subjected to the analysis of volatile aroma compounds by both targeted gas chromatography/mass spectrometry (GC-MS) and untargeted profiling by comprehensive two-dimensional gas chromatography/mass spectrometry (GC×GC-TOF-MS), analysis of phenols by targeted high-performance liquid chromatography/mass spectrometry (HPLC-DAD-ESI/MS), and quantitative descriptive sensory analysis. Monovarietal PDO EVOOs were characterized by notably higher amounts of positive LOX-derived C6 and C5 volatile compounds, which corresponded to the higher intensities of all the assessed positive fruity and green odor sensory attributes. Commercial-blended EVOOs had larger quantities of generally undesirable esters, alcohols, acids, and aldehydes, which coincided with the occurrence of sensory defects in many samples. Many minor volatile compounds that were identified by GC×GC-TOF-MS were found to differentiate each of the two investigated groups. The differences between the groups with respect to phenols and taste characteristics were evident, but less pronounced. The results that were obtained in this study have undoubtedly confirmed the existence of the large heterogeneity of oils that are sold declared as EVOO. It was shown that GC-MS, GC×GC-TOF-MS, and HPLC-DAD-ESI/MS analyses have complementary outputs, and that their use in combination has advantages in supporting the results of sensory analysis and objectively differentiating these groups of EVOO.

Dietary patterns, microbiome dysbiosis, and gut microbial metabolites (GMMs) have a pivotal role in the homeostasis of intestinal epithelial cells and in disease progression, such as that of colorectal cancer (CRC). Although GMMs and microorganisms have crucial roles in many biological activities, models for deciphering diet-microbiome-host relationships are largely limited to animal models. Thus, intestinal organoids (IOs) have provided unprecedented opportunities for the generation of in vitro platforms with the sufficient level of complexity to model physiological and pathological diet-microbiome-host conditions. Overall, IO responses to GMM metabolites and microorganisms can provide new insights into the mechanisms by which those agents may prevent or trigger diseases, significantly extending our knowledge of diet-microbiome-host interactions.

The development of a robust method to analyse the content of tryptophan and of indole-3-acetic acid at the microgram per litre level in must and wine is necessary in order to study the formation of 2-aminoacetophenone and of other indole compounds causing the 'untypical ageing off-flavour'. The present paper discusses the development and validation of a reversed-phase high-performance liquid chromatography method with fluorescence detection for the analysis of indole-3-acetic acid, tryptophan, tryptophol, indole and skatole in must and white wine. The required selectivity and sensitivity was gained through the solid-phase extraction on a polystyrene-based polymer column.

Abstract Background In response to pathogen attack, grapevine synthesizes phytoalexins belonging to the family of stilbenes. Grapevine cell cultures represent a good model system for studying the basic mechanisms of plant response to biotic and abiotic elicitors. Among these, modified β -cyclodextrins seem to act as true elicitors inducing strong production of the stilbene resveratrol. Results The transcriptome changes of Vitis riparia × Vitis berlandieri grapevine cells in response to the modified β -cyclodextrin, DIMEB, were analyzed 2 and 6 h after treatment using a suppression subtractive hybridization experiment and a microarray analysis respectively. At both time points, we identified a specific set of induced genes belonging to the general phenylpropanoid metabolism, including stilbenes and hydroxycinnamates, and to defence proteins such as PR proteins and chitinases. At 6 h we also observed a down-regulation of the genes involved in cell division and cell-wall loosening. Conclusions We report the first large-scale study of the molecular effects of DIMEB, a resveratrol inducer, on grapevine cell cultures. This molecule seems to mimic a defence elicitor which enhances the physical barriers of the cell, stops cell division and induces phytoalexin synthesis.

Despite the vast literature describing the biological effects of phenolic compounds, rather scarce data are available on their absorption from diet in humans. The present study focused on the absorption in humans of phenolic acids from white wine, particularly hydroxycinnamic acids and their esters with tartaric acid. The results obtained indicate that, following a single wine drink, hydroxycinnamic acids from white wine are absorbed from the gastrointestinal tract and circulate in the blood after being largely metabolized to the form of glucuronide and sulfate conjugates. Unmodified tartaric acid esters of hydroxycinnamic acids from wine are present in human plasma at low levels, if any. Wine hydroxycinnamic acids, although present in wine as conjugated forms, are still bioavailable to humans.

A four-year trial (1999–2002) was carried out in commercial vineyards in order to test the effect of manual leaf removal in the cluster zone at veraison on grape yield, berry composition, and stilbene concentration at harvest. Two <i>Vitis vinifera</i> L. red wine cultivars (Barbera and Croatina) and one white wine cultivar (Malvasia di Candia aromatica) were considered. The vines were Guyot trained, with 10 shoots per meter of row, and leaf removal accounted for approximately 22% of removed canopy surface. Meteorological data were recorded, as well as grape production, fruit composition, and stilbenes (<i>trans</i>-resveratrol, <i>trans</i>-piceid, <i>cis-</i>piceid) at harvest. Grape yield was not affected by leaf removal in any of the tested cultivars. Grape sugars and acidity were affected by leaf removal differently depending on meteorological conditions and cultivar. Leaf removal increased <i>trans</i>-piceid grape concentration in Barbera and decreased <i>trans-</i>resveratrol and <i>cis-</i>piceid in Croatina and Malvasia di Candia aromatica under cool meteorological conditions. Leaf removal had no effect on stilbene grape concentrations under warmer and drier climatic conditions.

Biomarker identification, i.e., finding those variables that indicate true differences between two or more populations, is an ever more important topic in the omics sciences. In most cases, the number of variables far exceeds the number of samples, making biomarker identification extremely difficult. We present a strategy based on the stability of putative biomarkers under perturbation of the data, and show that in several cases important gains can be achieved. The strategy is very general and can be applied with all common biomarker identification methods; it also has the advantage that it does not rely on error estimates from crossvalidation, that in this setting tend to be highly variable.

This paper presents methods for the definition of important analytical tools, such as the development of sensitive and rapid methods for analysing reduced and oxidised glutathione (GSH and GSSG), hydroxycinnamic acids (HCA), bound thiols (GSH-3MH and Cys-3MH) and free thiols (3MH and 3MHA), and their first application to evaluate the effect of reductive winemaking on the composition of Lugana juices and wines. Lugana is a traditional white wine from the Lake Garda region (Italy), produced using a local grape variety, Trebbiano di Lugana. An innovative winemaking procedure based on preliminary cooling of grape berries followed by crushing in an inert environment was implemented and explored on a winery scale. The effects of these procedures on hydroxycinnamic acids, GSH, GSSG, free and bound thiols and flavanols content were investigated. The juices and wines produced using different protocols were examined. Moreover, wines aged in tanks for 1, 2 and 3 months were analysed. The high level of GSH found in Lugana grapes, which can act as a natural antioxidant and be preserved in must and young wines, thus reducing the need of exogenous antioxidants, was particularly interesting. Moreover, it was clear that polyphenol concentrations (hydroxycinnamic acids and catechins) were strongly influenced by winemaking and pressing conditions, which required fine tuning of pressing. Above-threshold levels of 3-mercaptohexan-1-ol (3MH) and 3-mercaptohexyl acetate (3MHA) were found in the wines and changed according to the winemaking procedure applied. Interestingly, the evolution during the first three months also varied depending on the procedure adopted. Organic synthesis of cysteine and glutathione conjugates was carried out and juices and wines were subjected to LC-MS/MS analysis. These two molecules appeared to be strongly affected by the winemaking procedure, but did not show any significant change during the first 3 months of post-bottling ageing. This supports the theory, already proposed in the literature, that there are other synthetic pathways for free thiol formation.

Rotundone is an oxygenated sesquiterpene belonging to the family of guaianes, giving the 'peppery' aroma to white and black pepper and to red wines. Here we describe a novel, convenient protocol for the synthesis of rotundone, starting from a commercially available compound and requiring only two reaction steps, and an improved, faster method of GC separation (30 min) with selective quantisation of rotundone using tandem mass spectrometry in multiple reaction monitoring (MRM) mode with d(5)-rotundone as internal standard. With limits of detection (LODs) of 1.5 ng/L in white wine and 2.0 ng/L in red wine, intraday repeatability CV values of 6% and 5% at 50 ng/L and 500 ng/L and interday repeatability CV values of 13% and 6% at 50 ng/L and 500 ng/L, respectively, the improved protocol provides the desired sensitivity and selectivity for routine analysis of rotundone in both white and red wines. Initial application of this method highlighted the presence of unexpectedly high concentrations of rotundone, thus explaining the origin of the distinctive peppery aroma in Schioppettino and Vespolina red wines and in Gruener Veltliner white wines.

In the present study we chemically profiled tannin-enriched extracts from strawberries and tested their biological properties in a cell model of gastric inflammation. The chemical and biological features of strawberry tannins after in vitro simulated gastric digestion were investigated as well. The anti-inflammatory activities of pure strawberry tannins were assayed to get mechanistic insights. Tannin-enriched extracts from strawberries inhibit IL-8 secretion in TNFα-treated human gastric epithelial cells by dampening the NF-κB signaling. In vitro simulated gastric digestion slightly affected the chemical composition and the biological properties of strawberry tannins. By using pure compounds, we found that casuarictin may act as a pure NF-κB inhibitor while agrimoniin inhibits IL-8 secretion also acting on other biological targets; in our system procyanidin B1 prevents the TNFα-induced effects without interfering with the NF-κB pathway. We conclude that strawberry tannins, even after in vitro simulated gastric digestion, exert anti-inflammatory activities at nutritionally relevant concentrations.

Viticulture, like other fields of agriculture, is currently facing important challenges that will be addressed only through sustained, dedicated and coordinated research. Although the methods used in biology have evolved tremendously in recent years and now involve the routine production of large data sets of varied nature, in many domains of study, including grapevine research, there is a need to improve the findability, accessibility, interoperability and reusability (FAIR-ness) of these data. Considering the heterogeneous nature of the data produced, the transnational nature of the scientific community and the experience gained elsewhere, we have formed an open working group, in the framework of the International Grapevine Genome Program (www.vitaceae.org), to construct a coordinated federation of information systems holding grapevine data distributed around the world, providing an integrated set of interfaces supporting advanced data modeling, rich semantic integration and the next generation of data mining tools. To achieve this goal, it will be critical to develop, implement and adopt appropriate standards for data annotation and formatting. The development of this system, the GrapeIS, linking genotypes to phenotypes, and scientific research to agronomical and oeneological data, should provide new insights into grape biology, and allow the development of new varieties to meet the challenges of biotic and abiotic stress, environmental change, and consumer demand.

The multidisciplinary nature of nutrition research is one of its main strengths. At the same time, however, it presents a major obstacle to integrate data analysis, especially for the terminological and semantic interpretations that specific research fields or communities are used to. To date, a proper ontology to structure and formalize the concepts used for the description of nutritional studies is still lacking.We have developed the Ontology for Nutritional Studies (ONS) by harmonizing selected pre-existing de facto ontologies with novel health and nutritional terminology classifications. The ONS is the result of a scholarly consensus of 51 research centers in nine European countries. The ontology classes and relations are commonly encountered while conducting, storing, harmonizing, integrating, describing, and searching nutritional studies. The ONS facilitates the description and specification of complex nutritional studies as demonstrated with two application scenarios.The ONS is the first systematic effort to provide a solid and extensible formal ontology framework for nutritional studies. Integration of new information can be easily achieved by the addition of extra modules (i.e., nutrigenomics, metabolomics, nutrikinetics, and quality appraisal). The ONS provides a unified and standardized terminology for nutritional studies as a resource for nutrition researchers who might not necessarily be familiar with ontologies and standardization concepts.

Flavan-3-ols are dietary bioactive molecules that have beneficial effects on human health and reduce the risk of various diseases. Monomeric flavan-3-ols are rapidly absorbed in the small intestine and released in the blood stream as phase II conjugates. Polymeric flavan-3-ols are extensively metabolized by colonic gut microbiota into phenyl-γ-valerolactones and their related phenylvaleric acids. These molecules are the main circulating metabolites in humans after the ingestion of flavan-3-ol rich-products; nevertheless, they have received less attention and their role is not understood yet. Here, we describe the quantification of 8 phenyl-γ-valerolactones and 3 phenylvaleric acids in the urine of 11 subjects on consumption of apples by using UHPLC-ESI-Triple Quad-MS with pure reference compounds. Phenyl-γ-valerolactones, mainly as sulfate and glucuronic acid conjugates, reached maximum excretion between 6 and 12 after apple consumption, with a decline thereafter. Significant differences were detected in the cumulative excretion rates within subjects and in the ratio of dihydroxyphenyl-γ-valerolactone sulfate to glucuronide conjugates. This work observed for the first time the presence of two distinct metabotypes with regards to the excretion of phenyl-γ-valerolactone phase II conjugates.

Scope To identify reliable biomarkers of food intake (BFIs) of pulses. Methods and results A randomized crossover postprandial intervention study is conducted on 11 volunteers who consumed lentils, chickpeas, and white beans. Urine and serum samples are collected at distinct postprandial time points up to 48 h, and analyzed by LC‐HR‐MS untargeted metabolomics. Hypaphorine, trigonelline, several small peptides, and polyphenol‐derived metabolites prove to be the most discriminating urinary metabolites. Two arginine‐related compounds, dopamine sulfate and epicatechin metabolites, with their microbial derivatives, are identified only after intake of lentils, whereas protocatechuic acid is identified only after consumption of chickpeas. Urinary hydroxyjasmonic and hydroxydihydrojasmonic acids, as well as serum pipecolic acid and methylcysteine, are found after white bean consumption. Most of the metabolites identified in the postprandial study are replicated as discriminants in 24 h urine samples, demonstrating that in this case the use of a single, noninvasive sample is suitable for revealing the consumption of pulses. Conclusions The results of the present untargeted metabolomics work reveals a broad list of metabolites that are candidates for use as biomarkers of pulse intake. Further studies are needed to validate these BFIs and to find the best combinations of them to boost their specificity.

An extensive survey was conducted on 110 Italian monovarietal red wines from a single vintage to determine their standard compositional, color, and phenolic characteristics, analysing more than 35 parameters evaluated through methods commonly used in the wine industry. ‘Primitivo’ achieved the highest average alcohol strength (15.4% v/v) and dry extract values, while ‘Cannonau’ showed the lowest total acidity. ‘Corvina’ had the lowest phenolic content (1065 mg/L by Folin-Ciocalteu assay), remarkably different from the highest found in ‘Sagrantino’ (3578 mg/L), the latter being also the richest variety in both proanthocyanidins and vanillin-reactive flavanols. ‘Teroldego’ wines were the richest in both total and monomeric anthocyanins (702 and 315 mg/L, respectively), followed by ‘Aglianico’ and ‘Raboso Piave’, while ‘Corvina’, ‘Nebbiolo’, and ‘Nerello Mascalese’ were the poorest. ‘Montepulciano’ and ‘Sangiovese’ showed intermediate values for the majority of the parameters analyzed. A multivariate PCA-DA approach allowed achieving both a classification of the different wines as well as the discrimination of ‘Sangiovese’ wines produced in two regions (Emilia Romagna and Toscana) that returned a 42–66% success rate depending on the zone considered. Taking into account the number and diversity of the wines analyzed, a correlation study helped in better understanding the underlying relations between the most common and widespread analytical techniques for phenolic and color determinations.

Converging data support the role of chronic low-grade inflammation in depressive symptomatology in obesity. One mechanism likely to be involved relies on the effects of inflammation on tryptophan (TRP) metabolism. While recent data document alterations in the indole pathway of TRP metabolism in obesity, the relevance of this mechanism to obesity-related depressive symptoms has not been investigated. The aim of this preliminary study was to assess the association between plasma levels of TRP and indole metabolites and depressive symptoms in 44 subjects with severe or morbid obesity, free of clinically relevant neuropsychiatric disorders. The interaction effect of inflammation, reflected in serum high-sensitive C-reactive protein (hsCRP) levels, and indoles on depressive symptoms was also determined. Higher serum levels of hsCRP and lower concentrations of TRP and indoles, particularly indole-3-carboxaldehyde (IAld), correlated with more severe depressive symptoms. Interestingly, the effect of high hsCRP levels in predicting greater depressive symptoms was potentiated by low IAld levels. These results comfort the link between inflammation, the indole pathway of TRP metabolism, and obesity-related depressive symptoms.

The aim of this study was the optimization and validation of a green, robust, and comprehensive method for the determination of volatile carbonyl compounds (VCCs) in wines that could be added as a new quality control tool for the evaluation of a complete fermentation, correct winemaking style, and proper bottling and storage. A HS-SPME-GC-MS/MS method was optimized and automated using the autosampler to improve overall performance. A solvent-less technique and a strong minimization of all volumes were implemented to comply with the green analytical chemistry principles. There were as many as 44 VCC (mainly linear aldehydes, Strecker aldehydes, unsaturated aldehydes, ketones, and many other) analytes under investigation. All compounds showed a good linearity, and the LOQs were abundantly under the relevant perception thresholds. Intraday, 5-day interday repeatability, and recovery performances in a spiked real sample were evaluated showing satisfactory results. The method was applied to determine the evolution of VCCs in white and red wines after accelerated aging for 5 weeks at 50 °C. Furans and linear and Strecker aldehydes were the compounds that showed the most important variation; many VCCs increased in both classes of samples, whereas some showed different behaviors between white and red cultivars. The obtained results are in strong accordance with the latest models on carbonyl evolution related to wine aging.

This research paper describes a new application in the field of quality control of white wine. A high-performance liquid chromatography analysis of riboflavin was used together with a simple sensorial test to produce a classification of wines, rating their susceptibility to exposure to light. Based on a wide survey covering 85 commercial white wines of different varieties in three countries (Italy, Spain and Slovenia), an average value of riboflavin of 98.63 microg/l with a rather high standard deviation of 41.91 microg/l, and a normal distribution was obtained. Our statistical study of the frequency distribution of the sensorial scores of light-exposed wines by means of the Expectation-Maximization algorithm demonstrated that a large majority (71%) of these products were susceptible to the light, 31% of them belonging to the most severely affected group. Content of riboflavin was correlated with severity of appearance of the off-flavor in light-exposed white wines. These methods are directly applicable in the quality control of wine, being a valuable aid for oenologists in choosing appropriate fining protocols to reduce the appearance of the undesired "sunlight flavor" in bottled products.

A moderate consumption of red wine may reduce the risk of cardiovascular diseases via wine-derived phenolic compounds. A variety of biological mechanisms have been proposed for wine-derived phenolic compounds including nitric oxide-mediated vasorelaxation. This study examined whether the vasodilating effect of wine-derived phenolic compounds was associated with the inhibition of phosphodiesterases (PDEs) and, in particular, PDE5. For this purpose, human recombinant PDE5A1 isoform was prepared by expression of the full-length cDNA of PDE5A1 into COS-7 cells. Red wine and the extracts from grape skin inhibited PDE5A1 activity, whereas the seed extracts had a negligible effect. The mixture of anthocyanins inhibited the enzyme activity (IC50 = 11.6 microM), with malvidin-3-O-beta-glucoside (IC50 = 35.4 microM) and malvidin (IC50 = 24.9 microM) the most potent among the monoglucosides and aglycons, respectively. trans-Resveratrol and trans-piceid exhibited negligible effects, whereas hydroxycinnamates were completely inactive. These results indicate that polyphenols-induced vasorelaxation may also be sustained by smooth muscle PDE inhibition by anthocyanins present in red wines and grapes.

Forty-one samples of apples (peel plus pulp), obtained from eight cultivars, were examined for concentration of some important phytochemicals and for antioxidant activity expressed as peroxyl radical trapping efficiency. Five major polyphenolic groups plus ascorbate were identified and quantified by HPLC in the apple varieties. Oligomeric and polymeric proanthocyanidins were found to be about two-thirds of total polyphenols. The antioxidant efficiency of the apple extracts and of representative pure compounds for each group of phytochemicals was measured in a micellar system mimicking lipid peroxidation in human plasma. Although the amount of polyphenols measured by HPLC is similar to that measured by standard methods, the antioxidant efficiency calculated on the basis of the contribution of the pure compounds was lower than the antioxidant efficiency of the apple extracts. The higher efficiency of apples appears to be strictly related to the overwhelming presence of oligomeric proanthocyanidins.

Ellagitannins are a major class of phenolics largely responsible for the astringent and antioxidant properties of raspberries and blackberries. The Rubus ellagitannins constitute a complex mixture of monomeric and oligomeric tannins. Rubus oligomeric ellagitannins contain, beside the well-known ellagic acid and gallic acid moieties, the sanguisorboyl linking ester group. When exposed to acids or bases, ester bonds are hydrolyzed and the hexahydroxydiphenic acid spontaneously cyclizes into ellagic acid. This study describes a new, rapid procedure for the acid hydrolysis of Rubus ellagitannins in methanol, which results in maximal yield and enables the quantification of all the major reaction products. Additionally, the method provides the rationale for estimating the mean degree of polymerization of Rubus ellagitannins. Keywords: Rubus; ellagitannins; ellagic acid; sanguisorbic acid; gallic acid; hexahydroxydiphenic acid

Plant volatiles are signals used by herbivorous insects to locate host plants and select oviposition sites. Whether such volatiles are used as indicators of plant quality by adult insects in search of host plants has been rarely tested. We tested whether volatiles indicate plant quality by studying the oviposition of the grapevine moth Lobesia botrana on the grapevine plant Vitis vinifera. Host plants were infected with a variety of microorganisms, and larval fitness was correlated to the infected state of the substrate. Our results show an oviposition preference for volatiles that is significantly correlated with the fitness of the substrate. The chemical profiles of the bouquets from each V. vinifera-microorganism system are clearly differentiated in a PCA analysis. Both the volatile signal and the quality of the plant as larval food were affected by the introduction of microorganisms. Our study represents a broad approach to the study of plant-insect interactions by considering not only the direct effect of the plant but also the effect of plant-microorganism interactions on insect population dynamics.

Moderate wine consumption has been shown to lower cardiovascular risk. One of the mechanisms could involve the control of postprandial hyperlipaemia, a well-defined risk factor for atherosclerosis, reasonably by reducing the absorption of lipid oxidised species from the meal. The objective of the present study was to investigate whether wine consumption with the meal is able to reduce the postprandial increase in plasma lipid hydroperoxides and cholesterol oxidation products, in human subjects. In two different study sessions, twelve healthy volunteers consumed the same test meal rich in oxidised and oxidisable lipids (a double cheeseburger), with 300 ml of water (control) or with 300 ml of red wine (wine). The postprandial plasma concentration of cholesterol oxidation products was measured by GC-MS. The control meal induced a significant increase in the plasma concentration of lipid hydroperoxides and of two cholesterol oxidation products, 7-β-hydroxycholesterol and 7-ketocholesterol. The postprandial increase in lipid hydroperoxides and cholesterol oxidation products was fully prevented by wine when consumed with the meal. In conclusion, the present study provides evidence that consumption of wine with the meal could prevent the postprandial increase in plasma cholesterol oxidation products.

The development and the validation of innovative approaches for biomarker selection are of paramount importance in many ‐ omics technologies. Unfortunately, the actual testing of new methods on real data is difficult, because in real data sets, one can never be sure about the “true” biomarkers. In this paper, we present a publicly available metabolomic ultra performance liquid chromatography–mass spectrometry spike‐in data set for apples. The data set consists of 10 control samples and three spiked sets of the same size, where naturally occurring compounds are added in different concentrations. In this sense, the data set can serve as a test bed to assess the performance of new algorithms and compare them with previously published results. We illustrate some of the possibilities provided by this spike‐in data set by comparing the performance of two popular biomarker‐selection methods, the univariate t ‐test and the multivariate variable importance in projection. To promote a widespread use of the data, raw data files as well as preprocessed peak lists are made available. Copyright © 2012 John Wiley &amp; Sons, Ltd.

Anthocyanins are natural coloured pigments representing part of the protective mechanism of many plants, including Vitis vinifera L., and they have a considerable influence on wine quality. Moreover, the presence of anthocyanins in ripe berries is used as the accepted qualitative parameter for distinguishing red from white grapes, since these pigments are known to be present only in red berries. On the other hand, pyranoanthocyanins are important pigments for the colour stability of red wines and are known to be formed after berry crushing, during vinification and wine ageing. In this work, for the first time we provide clear evidence that the skin of international white grape cultivars (Chardonnay, Sauvignon Blanc and Riesling) contains measurable traces of anthocyanins. In addition, for the first time we report clear proofs about the presence of pyranomalvidin 3-O-glucoside (B-type vitisin) and carboxypyranomalvidin 3-O-glucoside (A-type vitisin) in fresh grapes and we quantify them.