Effects of Growing Areas, Pruning Wound Protection Products, and Phenological Stage on the Stilbene Composition of Grapevine (Vitis vinifera L.) Canes.

Applying plant protection products (PPP) on grapevine pruning wounds is a viticultural practice used to mitigate the spread of grapevine tuck disease, which is posing serious economic losses in the vine-wine industry. However, the impact of PPP on woody tissues remains unclear. Our study, conducted in two European vineyards, investigated the effects of Cuprocol, Tessior, Esquive, and Bentogran on stilbenes, in canes of Cabernet sauvignon and Syrah, at three phenological stages. Main stilbenes, quantified by HPLC-UV-DAD (1260 Agilent Infinity System) and identified by HPLC-ESI/MS (Thermo Scientific LCQ FLEET system), included E-resveratrol, E-ε-viniferin, E-piceatannol, and E-polydatin. Canes exhibited varying proportions of individual stilbenes, reflecting differences based on climatic conditions and phenological phases, rather than on the application of specific PPP. Vines grown in cool-climate conditions exhibited higher levels of E-resveratrol, whereas vines from the Mediterranean climate area exhibited higher levels of E-ε-viniferin. We also observed divergences in the accumulation trend of wood stilbenes throughout the season in canes collected in the two different growing areas.

Non-destructive SPE-UPLC-based Quantification of Aflatoxins and Stilbenoid Phytoalexins in Single Peanut (Arachis spp.) Seeds.

Aflatoxins are highly carcinogenic secondary metabolites of some fungal species, particularly Aspergillus flavus. Aflatoxins often contaminate economically important agricultural commodities, including peanuts, posing a high risk to human and animal health. Due to the narrow genetic base, peanut cultivars demonstrate limited resistance to fungal pathogens. Therefore, numerous wild peanut species with tolerance to Aspergillus have received substantial consideration by scientists as sources of disease resistance. Exploring plant germplasm for resistance to aflatoxins is difficult since aflatoxin accumulation does not follow a normal distribution, which dictates the need for the analyses of thousands of single peanut seeds. Sufficiently hydrated peanut (Arachis spp.) seeds, when infected by Aspergillus species, are capable of producing biologically active stilbenes (stilbenoids) that are considered defensive phytoalexins. Peanut stilbenes inhibit fungal development and aflatoxin production. Therefore, it is crucial to analyze the same seeds for peanut stilbenoids to explain the nature of seed resistance/susceptibility to the Aspergillus invasion. None of the published methods offer single-seed analyses for aflatoxins and/or stilbene phytoalexins. We attempted to fulfill the demand for such a method that is environment-friendly, uses inexpensive consumables, and is sensitive and selective. In addition, the method is non-destructive since it uses only half of the seed and leaves the other half containing the embryonic axis intact. Such a technique allows germination and growth of the peanut plant to full maturity from the same seed used for the aflatoxin and stilbenoid analysis. The integrated part of this method, the manual challenging of the seeds with Aspergillus, is a limiting step that requires more time and labor compared to other steps in the method. The method has been used for the exploration of wild Arachis germplasm to identify species resistant to Aspergillus and to determine and characterize novel sources of genetic resistance to this fungal pathogen.

Polyphenolic Compounds in Extracts from Roasted Grapevine Canes: An Investigation for a Circular Approach to Increase Sustainability in the Viticulture Sectors.

In this study, we compared the polyphenolic composition of the roasted grapevine wood chips of four Vitis vinifera cultivars-namely, Sorbara, Grasparossa, Malbo Gentile, and Spergola. These waste byproducts have the potential as infusion chips for the aging of alcoholic beverages and vinegars, contributing to an enriched sensory profile. Roasting amplifies aromatic nuances and triggers the depletion of crucial bioactive compounds, including polyphenols. We investigated the extent of polyphenolic loss in the ethanolic extract of roasted grapevine chips to repurpose this waste byproduct and assess its potential. We assessed the levels of trans-resveratrol, trans-ε-viniferin, trans-piceatannol, and the main resveratrol trimer. Our findings indicated a significant decrease in polyphenol content as the roasting temperature increased, from 16.85-21.12 mg GAE/g for grapevine chips roasted at 120 °C to 3.10-7.77 mg GAE/g for those roasted at 240 °C. This study also highlights notable genotypic differences in polyphenolic content. Among the red grape cultivars analyzed, Sorbara exhibited the highest levels (7.77-21.12 mg/GAEg), whereas the white grape cultivar Spergola showed the lowest polyphenolic content (3.10-16.85 mg/GAEg). These findings not only contribute to the scientific understanding of polyphenol stability but also hold practical implications for the enhancement of aged beverages, as well as advancing sustainable practices in the viticulture industries.

Profiling and Localization of Stilbene Phytoalexins Revealed by MALDI-MSI during the Grapevine-Botrytis cinerea Interaction.

Stilbene phytoalexins are among the most accumulated compounds during grapevine-pathogen interactions. However, their steady-state accumulation level and spatial distribution within the tissues to counteract Botrytis cinerea infection remain to be explored. In this work, matrix-assisted laser desorption/ionization-mass spectrometry imaging (MALDI-MSI) was used to determine the spatial distribution of different phytoalexins in grapevine leaves upon infection with B. cinerea. Ultraperformance liquid chromatography-fluorescence (UPLC-FL) was also employed to monitor the accumulation pattern of these phytoalexins. This study showed that stilbene compounds accumulate in areas close to the pathogen infection sites. It was also revealed that the accumulation patterns of the stilbene phytoalexins can vary from one time point postinfection to another with specific accumulation patterns within each time point. To the best of our knowledge, this is the first time that the separate localization of grapevine stilbene phytoalexins has been revealed following B. cinerea infection.

A highly efficient strategy for simultaneous analysis of eleven monomeric hydroxylated-stilbenes in grape wines and juices by a salting-out assisted liquid-liquid (SALLE) extraction combined with a high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS).

Wine and grape juice are known to have hydroxylated stilbenes, a group of phytoalexins, health-promoting compounds. The determination of stilbene species in wine and grape juices remains a challenging task. Here, we propose an efficient strategy for the simultaneous determination of eleven hydroxylated stilbenes in grape wines and grape juice by a SALLE in conjunction with an HPLC-ESI-MS/MS. SALLE and HPLC-MS/MS conditions were optimized. The proposed method was validated and found to be a simple, sensitive, and reliable measure for the determination of the stilbene species in wine and grape juice. The SALLE was fast and efficient, taking only 10 min. The HPLC-MS/MS was able to quickly separate and quantitate the eleven stilbenes (9 min running). This method was successfully applied to determine the contents of stilbene species in commercial wine and grape juice in Korea. This method is advantageous in reduced sample preparation time, low labor, and cost efficiency.

Chiral analysis of E-ε-viniferin enantiomers, towards a new chemotaxonomic marker of the vine.

BACKGROUND: The accurate characterization of grapevine cultivars (Vitis vinifera) is crucial for grape growers, winemakers, wine sellers, consumers and authorities, considering that mistakes could involve significant damage to the wine economic system. To avoid any misunderstanding, morphological, molecular and chemical tools are developed to positively identify grape varieties. RESULTS: E-ε-viniferin is a stilbene dimer mainly present in the woody part of grapevine and present as a mixture of two enantiomers: (7aR, 8aR)-(-)-E-ε-viniferin (1) and (7aS, 8aS)-(+)-E-ε-viniferin (2). In addition to phenotypic and genotypic approaches, a chemotaxonomic method using E-ε-viniferin enantiomers as chemical markers of grapevine cultivars was investigated. The isolation and purification of E-ε-viniferin enantiomers by preparative high-performance liquid chromatography (HPLC) and chiral HPLC from 14 red and eight white grapevine cane cultivars enabled us to determine the proportion of each enantiomer and therefore to calculate the enantiomeric excess for each variety. The relative abundance of each E-ε-viniferin enantiomer permitted us to distinguish grape varieties, as well as to establish cultivar relationships and patterns through statistical analysis. CONCLUSION: This pioneering work highlighting the enantiomeric excess of E-ε-viniferin as a chemical marker of grapevine paves the way for further studies to understand what mechanisms are involved in the production of these enantiomers in grapevine. © 2023 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Trans-gnetin H isolated from the seeds of Paeonia species induces autophagy via inhibiting mTORC1 signalling through AMPK activation.

Paeonia is a well-known species of ornamental plants, traditional Chinese medicines, and emerging oilseed crops. Apart from nutritional unsaturated fatty acids, the seeds of peonies are rich in stilbenes characterized by their wide-ranging health-promoting properties. Although the typical stilbene resveratrol has been widely reported for its multiple bioactivities, it remains uncertain whether the trimer of resveratrol trans-gnetin H has properties that regulate cancer cell viability, let alone the underlying mechanism. Autophagy regulated by trans-gnetin H was detected by western blotting, immunofluorescence, and quantitative real-time PCR. The effects of trans-gnetin H on apoptosis and proliferation were examined by flow cytometry, colony formation and Cell Counting Kit-8 assays. Trans-gnetin H significantly inhibits cancer cell viability through autophagy by suppressing the phosphorylation of TFEB and promoting its nuclear transport. Mechanistically, trans-gnetin H inhibits the activation and lysosome translocation of mTORC1 by inhibiting the activation of AMPK, indicating that AMPK is a checkpoint for mTORC1 inactivation induced by trans-gnetin H. Moreover, the binding of TSC2 to Rheb was markedly increased in response to trans-gnetin H stimulation. Similarly, trans-gnetin H inhibited the interaction between Raptor and RagC in an AMPK-dependent manner. More importantly, trans-gnetin H-mediated autophagy highly depends on the AMPK-mTORC1 axis. We propose a regulatory mechanism by which trans-gnetin H inhibits the activation of the mTORC1 pathway to control cell autophagy.

Sulfur dioxide-free Verdejo wines through the use of a pure stilbene extract: exploring possible synergistic effect with glutathione.

BACKGROUND: The search for alternatives to sulfur dioxide (SO2 ) in white wine production is a tough challenge for the oenological industry. The current article investigates the effectiveness of several preservatives in protecting Verdejo white wines. The following wines were elaborated: without preservative (CT wines), with SO2 (SO2 wines), with stilbene-enriched extract (ST99 wines), with glutathione (GSH wines) and finally with the combination stilbene extract plus glutathione (ST99-GSH wines). RESULTS: ST99 extract preserved the wines from malolactic fermentation (MLF), maintained the quality parameters of the wines, but affected their color from the beginning of the process. Synergistic effects between ST99 and GSH were found for oxygen consumption rates, related to oxidation processes. The content of amino acids and biogenic amines was slightly affected by the treatments, but the quality or safety of the wines was not compromised. However, after 12 months of ageing in bottle, the SO2 wines showed the highest score in the sensory analysis. The reasons for the evolution of treated wines and the implications of this study for maintaining the quality of free SO2 white wines are discussed. CONCLUSION: ST99 may be proposed as an alternative to SO2 in wines to be consumed in the short term. Its combination with GSH does not prolong its shelf-life. © 2022 Society of Chemical Industry.

Foliar application of methyl jasmonate and methyl jasmonate supported on nanoparticles: Incidence on grape phenolic composition over two seasons.

Tools to address the mismatch between technological and phenolic maturity of grapes are needed. Application of elicitors could be an effective alternative. This work compares the effect of the application of methyl jasmonate (MeJ) in conventional form and, as a novelty, in the form of MeJ-doped nanoparticles (ACP-MeJ) on the phenolic composition of Tempranillo grapes. Results showed that, regardless of season, both treatments increased the grape total phenols content. In 2019, most of the anthocyanins, and to a lesser extent the flavanols, increased with the application of MeJ, and several hydroxycinnamic acids increased in the grapes treated with ACP-MeJ, with dose 10 times lower than those of the MeJ conventional. In 2020, anthocyanins were not affected by the treatments, but total flavanols, flavonols, hydroxybenzoic acid, and stilbenes increased after ACP-MeJ application. Thus, foliar application of ACP-MeJ could serve to increase grape phenolic composition, reducing maturity decoupling and the environmental impact.

Is It Possible to Improve the Bioavailability of Resveratrol and Polydatin Derived from Polygoni cuspidati Radix as a Result of Preparing Electrospun Nanofibers Based on Polyvinylpyrrolidone/Cyclodextrin?

The low bioavailability of resveratrol and polydatin obtained from Polygoni cuspidati extract limits the application of their pro-health properties. While nanofibers have attracted increasing attention in nutrition delivery due to their special properties, including an increase in the dissolution and permeability, which affects the bioavailability. Therefore, it is justified to obtain nanofibers from Polygoni cuspidati extract, which showed antioxidant and anti-inflammatory properties as a result of a presence of stilbene analogs in the Polygoni cuspidati extract (especially resveratrol and polydatin). In the first stage of the work, using the Design of Experiment (DoE) approach, the Polygoni cuspidati extract (70% of methanol, temperature 70 °C and 4 cycles) was obtained, which showed the best antioxidant and anti-inflammatory properties. Using the Polygoni cuspidati extract as a substrate, nanofibers were obtained by electrospinning. The identification of nanofibers was confirmed on the basis of the analysis of changes in XRPD diffractograms, SEM picture and FTIR-ATR spectra. Obtaining nanofibers from the Polygoni cuspidati extract significantly improved the solubility of resveratrol and polydatin (approx. 6-fold comparing to pure substance). As a consequence, the penetration coefficients of both tested resveratrol and polydatin also increased. The proposed strategy for the preparation of nanofibers from the Polygoni cuspidati extract is an innovative approach to better use the synergy of biological action of active compounds present in extracts. It is especially during the development of nutraceuticals based on the use of selected stilbenes.

Structures and Anti-Inflammatory Evaluation of Phenylpropanoid Derivatives from the Aerial Parts of Dioscorea polystachya.

Seven undescribed phenylpropanoid constituents, including three new bibenzyl derivatives (1-3) along with four new benzofuran stilbene derivatives (4-7), were isolated from the aerial parts of Dioscorea polystachya. The structures of these compounds were elucidated using a combination of spectroscopic analyses, including UV, IR, HRESIMS, 1D, and 2D NMR. Further, all the compounds were evaluated on the anti-inflammatory activity for their inhibition of nitric oxide (NO) production by RAW 264.7 macrophages cells, and some of them (1-3 and 6) displayed inhibitory activity with IC50 values in the range of 9.3-32.3 µM. Moreover, compound 3 decreased the expression of iNOS in Western blot analysis, suggesting compound 3 is mediated via the suppression of an LPS-induced NF-κB inflammasome pathway.

Induction of the Prenylated Stilbenoids Arachidin-1 and Arachidin-3 and Their Semi-Preparative Separation and Purification from Hairy Root Cultures of Peanut (Arachis hypogaea L.).

Prenylated stilbenoids such as arachidin-1 and arachidin-3 are stilbene derivatives that exhibit multiple pharmacological activities. We report an elicitation strategy using different combinations of cyclodextrin, hydrogen peroxide, methyl jasmonate and magnesium chloride to increase arachidin-1 and arachidin-3 production in peanut hairy root cultures. The treatment of hairy root cultures with cyclodextrin with hydrogen peroxide selectively enhanced arachidin-1 yield (132.6 ± 20.4 mg/L), which was 1.8-fold higher than arachidin-3. Similarly, cyclodextrin combined with methyl jasmonate selectively enhanced arachidin-3 yield (178.2 ± 6.8 mg/L), which was 5.5-fold higher than arachidin-1. Re-elicitation of the hairy root cultures further increased the levels of arachidin-1 and arachidin-3 by 24% and 42%, respectively. The ethyl acetate extract of the culture medium was consecutively fractionated by normal- and reversed-phase column chromatography, followed by semi-preparative HPLC purification on a C18 column to yield arachidin-1 with a recovery rate of 32% and arachidin-3 with a recovery rate of 39%, both at higher than 95% purity. This study provided a sustainable strategy to produce high-purity arachidin-1 and arachidin-3 using hairy root cultures of peanuts combined with column chromatography and semi-preparative HPLC.

Computational Analysis and Biological Activities of Oxyresveratrol Analogues, the Putative Cyclooxygenase-2 Inhibitors.

Polyphenols are a large family of naturally occurring phytochemicals. Herein, oxyresveratrol was isolated from ethanolic crude extracts of Artocarpus lacucha Buch.-Ham., and chemically modified to derive its lipophilic analogues. Biological screening assays showed their inhibitory potency against cyclooxygenase-2 (COX-2) with very low cytotoxicity to the MRC-5 normal cell lines. At the catalytic site of COX-2, docking protocols with ChemPLP, GoldScore and AutoDock scoring functions were carried out to reveal hydrogen bonding interactions with key polar contacts and hydrophobic pi-interactions. For more accurate binding energetics, COX-2/ligand complexes at the binding region were computed in vacuo and implicit aqueous solvation using M06-2X density functional with 6-31G+(d,p) basis set. Our computational results confirmed that dihydrooxyresveratrol (4) is the putative inhibitor of human COX-2 with the highest inhibitory activity (IC50 of 11.50 ± 1.54 µM) among studied non-fluorinated analogues for further lead optimization. Selective substitution of fluorine provides a stronger binding affinity; however, lowering the cytotoxicity of a fluorinated analogue to a normal cell is challenging. The consensus among biological activities, ChemPLP docking score and the binding energies computed at the quantum mechanical level is obviously helpful for identification of oxyresveratrol analogues as a putative anti-inflammatory agent.

Variability of Constitutive Stilbenoid Levels and Profiles in Grape Cane (Vitis vinifera L.) Depending upon Variety and Clone, Location in the Vineyard, Pruning Time, and Vintage.

Stilbenoids in grape cane (Vitis vinifera L.) are bioactive compounds relevant for plant defense and the potential valorization of this byproduct. Our screening of grape cane from 102 varieties showed constitutive stilbenoid levels in a wide range (557-7748 mg/kg of dry weight). Analyses of genetically distinct clones of selected cultivars unraveled that intravarietal variability (e.g., cv. Riesling, 3236-6541 mg/kg) was higher than that across samples from a single clone but different vineyard positions (3017-3710 mg/kg). Furthermore, stilbenoid levels in samples obtained in October, December, and February (3 years, 2017-2019) showed pronounced quantitative and qualitative variability and the highest yields upon December pruning. For instance, vitisin B and ε-viniferin in cv. Pinot Noir and Accent were predominant in 2017 and 2019 (both >90% of total stilbenoids) but not in 2018 (both <55%) when temperatures were high and precipitation low. In brief, we report the variability of stilbenoid levels in grape cane depending upon genetic and environmental factors.

Conductive polymer sorbent for extraction and determination of resveratrol and polydatin in Polygonum cuspidatum root samples.

INTRODUCTION: The quantitative analysis of trace resveratrol and polydatin in plant tissues is suitable for elucidation of the compounds' mechanisms of action. OBJECTIVES: The main objective of this work was to develop a feasible and effective sample pretreatment method to measure the concentrations of resveratrol and polydatin in complex samples. METHODOLOGY: A polymer sorbent, poly(2-mercaptobenzimidazole), was electrochemically prepared and utilized for selective extraction, while resveratrol and polydatin were used as target analytes. The sorbent was characterized by cyclic voltammetry, scanning electron microscopy and Fourier transform infrared spectroscopy. After extraction and elution, the analytes were analyzed by a Thermo U3000 HPLC system. Several affecting parameters, including the volume of elution solution, sample pH value, sample flow rate and sample volume, were evaluated and optimized. RESULTS: The proposed method showed good linearity with low limits of detection (from 0.5 to 0.8 ng·mL-1 ) and ideal accuracy with spiked recoveries from 81.30% to 99.16%. A good enrichment factor (more than 200-fold) together with good sensitivity was obtained with this method. Analysis of resveratrol and polydatin in Polygonum cuspidatum samples by this method is efficient. CONCLUSION: The method developed in this work exhibits several significant merits, including easy operation and high extraction efficiency, indicating that electrochemically prepared polymer sorbent is useful for sample pretreatment and analysis of traditional Chinese medicine samples.

Characterization of Tempranillo negro (VN21), a high phenolic content grapevine Tempranillo clone, through UHPLC-QqQ-MS/MS polyphenol profiling.

Grapevine cultivar and clone genotype is an important factor in the phenolic composition of wine. In this study, a new intense dark black berry color variant of Tempranillo, known as Tempranillo negro or VN21, is described. A targeted chromatographic approach based on UHPLC-QqQ-MS/MS was used to study the anthocyanins and non-colored phenols of the grape berry (skin and seeds) and wine. RJ43, one of the most cultivated clones in D.O.Ca. Rioja (Spain), was analyzed for comparison. Results suggest that the unique color of the grape skin in Tempranillo negro could be explained by higher concentrations of peonidin and cyanidin derivatives. This genotype accumulated anthocyanins in the seeds. Those differences in the berry were enhanced in the VN21 wines, which displayed notably higher concentrations of anthocyanins, and significantly increased contents of proanthocyanidins and stilbenes. This study exemplifies the application of phenol chromatographic analyses of spontaneous somatic variants to grapevine clonal selection.

Phytochemistry and pharmacological activities of Arundina graminifolia (D.Don) Hochr. And other common Orchidaceae medicinal plants.

ETHNOPHARMACOLOGICAL RELEVANCE: Arundina graminifolia (Orchidaceae) has been widely used for heat clearance and detoxification, anti-inflammatory diuretic, and anti-microbes for two thousand years in national minorities, especially among the Dai people. It was known as "Zhuyelan" (Chinese: ), "Wenshanghai" (Chinese: ) and "Baiyangjie" (Chinese: ) in the Dai nationality, and mainly used as antidote, which is characterized by "relieving the poison before getting sick and treating illness". Therefore, it has been typically applied in the treatment of food poisoning, snake bites, rheumatism, stomachache and traumatic injuries. It is also used to treat bronchitis, tuberculosis and pneumonia in the Bulang and the Wa ethnic people. AIM OF THIS REVIEW: This review aims to provide up-to-date information about the botanical characterization, traditional uses, phytochemistry, and pharmacology of A. graminifolia, and the related importantly medicinal plants (e.g. Bletilla striata, Cremastra appendiculata, and Dendrobium officinale) of the same Orchidaceae family. Our work aims to further promote scientific cognition, basic research and in-depth discovery of potential drug candidates for Minority Medicine of our nation. MATERIALS AND METHODS: Relevant information was obtained via piles of resources including classic books about Chinese herbal medicine, and worldwide accepted scientific databases including Web of Science, Pubmed, Google Scholar, SciFinder, Elsevier, Springer, NCBI, ACS Publications, CNKI and Wanfang data. RESULTS: Phytochemical investigations have been intensively performed for these two decades, over 134 compounds, mainly include stilbenoids, phenanthrenes, quinones, ketones, phenolic acids, and glycosides, have been isolated and identified from A. graminifolia. The literature surveys exhibited that the ethnomedicinal uses of A. graminifolia, such as detoxification, anti-inflammation and the ability to cure trauma and pain associated with infections, are correlated with its modern pharmacological activities, including antibacterial, anti-oxidation, anti-lipid peroxidation. Stilbenoids and phenanthrenes have been regarded as the main active substances, among which stilbenoids with open ring style have superior antitumor activity. Furthermore, phytochemical investigations, biological activities, as well as the main molecular mechanism involved the coexisted stilbenoids and phenanthrenes from other most common used medicinal plants of the same Orchidaceae family were presented, compared and discussed together. CONCLUSIONS: This review presents the current research findings of A. graminifolia and three other related medicinal plants of the same family. Some of the traditional uses of A. graminifolia have been assessed by pharmacological studies. Despite A. graminifolia is used as an antidote and anti-aging dote, a few unsolved problems including the molecular mechanism underlying biological activities, pharmacokinetics, and in vivo detoxification tests still need to be settled extensively. Therefore, it is necessary to conduct a comprehensive survey and collect investigation information on A. graminifolia.

Pterostilbene modifies triglyceride metabolism in hepatic steatosis induced by high-fat high-fructose feeding: a comparison with its analog resveratrol.

The use of phenolic compounds as a new therapeutic approach against NAFLD has emerged recently. In the present study, we aim to study the effect of pterostilbene in the prevention of liver steatosis developed as a consequence of high-fat (saturated) high-fructose feeding, by analysing the changes induced in metabolic pathways involved in triglyceride accumulation. Interestingly, a comparison with the anti-steatotic effect of its parent compound resveratrol will be made for the first time. Rats were distributed into 5 experimental groups and fed either a standard laboratory diet or a high-fat high-fructose diet supplemented with or without pterostilbene (15 or 30 mg per kg per d) or resveratrol (30 mg per kg per d) for 8 weeks. Serum triglyceride, cholesterol, NEFA and transaminase levels were quantified. Liver histological analysis was carried out by haematoxylin-eosin staining. Different pathways involved in liver triglyceride metabolism, including fatty acid synthesis, uptake and oxidation, triglyceride assembly and triglyceride release, were studied. Pterostilbene was shown to partially prevent high-fat high-fructose feeding induced liver steatosis in rats, demonstrating a dose-response pattern. In this dietary model, it acts mainly by reducing de novo lipogenesis and increasing triglyceride assembly and release. Improvement in mitochondrial functionality was also appreciated. At the same dose, the magnitude of pterostilbene and resveratrol induced effects, as well as the involved mechanisms of action, were similar.

High-resolution mass spectrometry-based methodology for the identification of the metabolites of pterostilbene produced by rat, dog and human hepatocytes.

Pterostilbene, a natural bibenzjyl compound, has been demonstrated to have pleiotropic anticancer effects against a variety of cancer types. The aim of this study was carried out to disclose the metabolic profiles of pterostilbene using rat, dog and human hepatocytes. Metabolites were characterized by ultra-high-performance liquid chromatography/quadrupole Orbitrap mass spectrometry with electrospray ionization interface operating in positive ion mode. The structures of the metabolites were proposed by accurate MS, MS/MS spectra and based on their fragmentation patterns. A total of 12 metabolites, including six new ones, were detected and identified. M10 and M12 were unambiguously identified as pinostilbene and 3'-hydroxy-pterostilbene, respectively, by using reference standards. Our results revealed that pterostilbene was metabolized through the following pathways: (a) hydroxylation to form 3'-hydroxy-pterostilbene (M12), which further undergoes glucuronidation (M9), demethylation (M7) and GSH conjugation through the ortho-quinone intermediate; (b) demethylation to produce desmethyl-pterostilbene (M10), which is further subject to glucuronidation (M4); (c) direct conjugation with glucuronide (M11); and (d) direct sulfation (M8). Among the tested species, no significant species difference was observed. The current study provides valuable information on the metabolism of pterostilbene, which is helpful for us to understand the action of this compound.

Development of an UHPLC-diode arrays detector (DAD) method for the analysis of polydatin in human plasma.

A new chromatographic method by Ultra High Performance Liquid Chromatographic (UHPLC) technology, has been developed and validated for the determination of polydatin and resveratrol, as potential metabolite, in human plasma. After the optimization of the chromatographic conditions, the method has been validated on spiked human plasma samples. The optimized extraction allowed to obtain analytes recovery up to 98.48 ± 4.03 %. Then, the isocratic elution in reversed phase mode, provides the separation of polydatin and resveratrol in less than 10.0 min. Chromatographic analysis was performed on a C18, 10 cm x 3.0 mm, 2.7 µm stationary phase, by using triethanolamine phosphate solution (0.1 M, pH = 3.7) and ACN 85:15 (v/v) as mobile phase at a flow rate of 0.5 mL/min. The UV detector was set at 306 nm for the analysis of both polydatin and resveratrol. The limit of detection (LoD) and the limit of quantification (LoQ) for polydatin in plasma samples were found to be 7.82 ± 0.38 nM and 26.06 ± 1.28 nM respectively. The method was found to be accurate and precise with a coefficient for intra- and inter-day variation below 5 %. All the reported data demonstrate how the developed method is rapid and sensitive. Moreover, results of the analysis of plasma samples, obtained from orally treated volunteers with nutritional supplements containing polydatin, have shown the method to be suitable for the pharmacokinetic characterization of polydatin and resveratrol, as metabolite, in humans.