Flavonol Glycosides Research Articles

The SlDOG1 Affect Biosynthesis of Steroidal Glycoalkaloids by Regulating GAME Expression in Tomato.

Steroidal alkaloids (SAs) and steroidal glycoalkaloids (SGAs) are common constituents of plant species belonging to the Solanaceae family. However, the molecular mechanism regulating the formation of SAs and SGAs remains unknown. Here, genome-wide association mapping was used to elucidate SA and SGA regulation in tomatoes: a SlGAME5-like glycosyltransferase (Solyc10g085240) and the transcription factor SlDOG1 (Solyc10g085210) were significantly associated with steroidal alkaloid composition. In this study, it was found that rSlGAME5-like can catalyze a variety of substrates for glycosidation and can catalyze SA and flavonol pathways to form O-glucoside and O-galactoside in vitro. The overexpression of SlGAME5-like promoted α-tomatine, hydroxytomatine, and flavonol glycoside accumulation in tomatoes. Furthermore, assessments of natural variation combined with functional analyses identified SlDOG1 as a major determinant of tomato SGA content, which also promoted SA and SGA accumulation via the regulation of GAME gene expression. This study provides new insights into the regulatory mechanisms underlying SGA production in tomatoes.

Transformation of Strawberry Plants’ Phenolic Profile after Treatment with a Mechanocomposite Based on Silicon Chelates in the Course of Development under In Vitro, Ex Vitro, and In Vivo Conditions

The positive effect of silicon on plants is thought to be mediated by a modification of phenolic metabolism. The purpose of the study was to evaluate the effect of a silicon-based mechanocomposite (MC) on alterations of the phenolic profile of strawberry plants in the course of development under in vitro, ex vitro, and in vivo conditions. Aqueous ethanol extracts of aboveground parts of in vitro–derived plants (Fragaria × ananassa cv. ‘Solnechnaya polyanka’) were subjected to HPLC. Nineteen individual phenolic compounds (hydroxybenzoic and hydroxycinnamic acids, catechins, ellagic acid derivatives, and flavonol glycosides) were quantified. The results revealed phenolic profiles specific to each studied stage and significant transformations of the profiles by the MC. It induced strong upregulation of hydroxycinnamic acid during in vitro rooting and of catechins and hydroxybenzoic acids during ex vitro acclimation. At ex vitro and in vivo stages, the emergence of quercetin glycosides and ellagitannins was registered, and the MC elevated their levels during ex vitro acclimation and field growth. Principal component analysis confirmed the significant effect of the MC on the phenolic profile at all stages, and this effect was the strongest during ex vitro acclimation. The results are consistent with previous reports on the modification of phenolic profiles of plants by silicon-derived biostimulants.

Biochemical Characterization of Black and Green Mutant Elderberry during Fruit Ripening.

The content of sugars, organic acids, phenolic compounds and selected enzyme activities in the anthocyanin pathway were analyzed in NIGRA (Sambucus nigra var. nigra-black fruits) and VIRIDIS (S. nigra var. viridis-green fruits) fruits over four stages of ripening. The share of glucose and fructose in green fruits was higher than in colored fruits, and the sugar content increased significantly until the third developmental stage. Ripe NIGRA berries had 47% flavonol glycosides, 34% anthocyanins, 3% hydroxycinnamic acids and 14% flavanols, whereas the major phenolic group in the VIRIDIS fruits, making up 88% of the total analyzed polyphenols, was flavonols. NIGRA fruits were rich in anthocyanins (6020 µg g-1 FW), showing strong activation of the late anthocyanin pathway (dihydroflavonol 4-reductase, anthocyanidin synthase). In both color types, phenylalanine ammonia lyase and chalcone synthase/chalcone isomerase activities were highest in the first stage and decreased during ripening. In VIRIDIS fruit, no anthocyanins and only one flavanol (procyanidin dimer) were found. This was most likely caused by a lack of induction of the late anthocyanin pathway in the last period of fruit ripening. The VIRIDIS genotype may be useful in studying the regulatory structures of anthocyanin biosynthesis and the contribution of distinct flavonoid classes to the health benefits of elderberries.

Branched montbretin A mimics allow derivatisation and potent amylase inhibition.

Mimics of the complex flavonol glycoside montbretin A in which a flavonol moiety is coupled to a caffeic acid via partially peptidic linkers have proved to be potent inhibitors of human pancreatic alpha-amylase with potential as therapeutics for control of blood glucose levels. After exploring optimal linker length, a synthetic route to a version with a branched linker was devised based on the structure of the enzyme/inhibitor complex. The resultant branched inhibitors were shown to retain nanomolar potency even when decorated with polymers as a means of modifying solubility. Similar improvements, along with nanomolar affinity, could also be achieved through conjugation to cyclodextrins which have the potential to bind to starch binding sites found on the surface of human amylase. Incorporation of a conjugatable branch into this unusual pharmacophore thereby affords considerable flexibility for further modifications to improve pharmacokinetic behaviour or as a site for attachment of capture tags or fluorophores.

Chemical constituents from the fruits of Ferula ovina (Boiss.) Boiss. and their chemotaxonomic significance

Phytochemical analysis of the fruits of Ferula ovina (Boiss.) Boiss. resulted in isolation and identification of eleven compounds, including two monoterpene-phenyl esters (1 and 4), four sesquiterpene-phenyl esters (2, 3, 5 and 6), three caffeoylquinic acid derivatives (7, 9 and 10) and two flavonol glycosides (8 and 11). The structures of isolated compounds were elucidated on the basis of NMR spectral data. Compounds 8–10 were isolated for the first time in the genus Ferula and this is the first report of the presence of the compounds 3 and 5–10 in Ferula ovina. Chemotaxonomic significance of the isolated compounds was also discussed.

Distribution and Stability of Polyphenols in Juices Made from Traditional Apple Cultivars Grown in Bosnia and Herzegovina

The present research was undertaken to investigate polyphenolic profiles of peel, pulp and juices made from two standard commercial and five traditional apple cultivars from Bosnia and Herzegovina. The main goal of the study was to monitor the distribution and changes of polyphenolic profiles through different phases of apples' processing into cloudy and clear juices, with regard to L-ascorbic acid pretreatment. Quantitative determination of phenolic compounds was carried out by using high-performance liquid chromatography with diode-array detection. The obtained results showed that traditional cultivars, namely 'Paradija' and 'Prijedorska zelenika', displayed significantly higher content of these compounds compared to commercial ones. Flavan 3-ols and flavonol glycosides were mostly found in peels of all cultivars (21.2-44.1 and 5.40-33.3%, respectively), while phenolic acids along with flavan 3-ols were predominant in the pulp (8.20-30.8 and 5.10-13.9%, respectively). Apples' processing into juices caused decrease (more than 90%) in the content of all polyphenols and the distribution of these compounds from fruits to final products had a negative trend, particularly evident in clear juices. The most drastic loss occurred in the flavonol glycosides and dihydrochalcones content, while chlorogenic acid displayed quite stable distribution from apples to final products due to its good solubility. Apple mash pretreatment with L-ascorbic acid had a positive impact on the preservation and retention of polyphenols.

Encapsulating Calendula arvensis (Vaill.) L. Florets: UHPLC-HRMS Insights into Bioactive Compounds Preservation and Oral Bioaccessibility.

Wild edible plants, once consumed in times of famine or for health purposes, today represent an interesting dietary supplement, aimed at enriching local dishes and/or formulating healthy nutraceutical products. In fact, the broad content of different, and diversely bioactive, specialized metabolites therein suggests new scenarios of use which, in order to be as functional as possible, must maximize the bioactivity of these compounds while preserving their chemistry. In this context, based on a recent investigation on the metabolic profile of the organs of Calendula arvensis that highlighted that florets are abundant in flavonol glycosides and triterpene saponins, the freeze-drying encapsulation of their alcoholic extract (FE) into maltodextrin (MD) was investigated. FE-MD chemical composition was evaluated using Fourier Transform InfraRed spectroscopy (FTIR), while ultra-high performance liquid chromatography coupled with high-resolution tandem mass spectrometry (UHPLC-HRMS/MS) techniques were employed to unravel FE compound preservation also during in vitro simulated digestion. The establishment of H-bonds between FE compounds and MD hydroxyl groups was in line with FE-MD biocompatibility in Caco-2 cells, while in vitro digestion mostly affected structural integrity and/or diversity. Flavonol compounds underwent deglycosylation and demethylation, while deacylation, beyond oxidation, involved triterpene saponins, which massively preserve their aglycone core.

The variation of total flavonoids, anthocyanins and total phenols in Vaccinium uliginosum fruits in Changbai Mountain of China is closely related to spatial distribution

BACKGROUND: The fruit of Vaccinium uliginosum is a natural berry resource that is rich in polyphenols, flavonol glycosides, anthocyanins, and other active substances, indicating its high developmental potential. However, research on V. uliginosum is limited, with no literature available to clarify the germplasm resources suitable for breeding. OBJECTIVE: This study aimed to evaluate the contents of total flavonoids (TF), total anthocyanins (TA), and total phenols (TP) in 10 different populations of V. uliginosum from the Changbai Mountains, China, and investigate the correlation between these functional components and spatial distribution. METHODS: The components and contents of TA, TF, and TP were determined using mass spectrometry, high-performance liquid chromatography, and the Folin–Ciocalteu method, respectively. RESULTS: A total of 15 anthocyanins were detected, and the content of Mal-glu, Pet-glu, and Del-glu was the highest among these anthocyanins. The TF and TA and TP contents were highest in the DFHI and LJII populations, respectively, which can be reasonably developed as excellent populations. The TF content of sample DFHI-8, TA content of LJIII-1 and TP content of LJIII-4 were higher than other samples, which can be used as important breeding germplasm. The content of TF is positively correlated with altitude, while the content of TA and TP is bidirectional, which is positively correlated at 740–838 m and negatively correlated at >838 m. CONCLUSIONS: Significant differences in the contents of TF, TA, and TP in V. uliginosum fruit were found among and within populations, and there was a certain correlation between these contents and their spatial distribution.

A Novel 3-O-rhamnoside: 2″-O-xylosyltransferase Responsible for Terminal Modification of Prenylflavonol Glycosides in Epimedium pubescens Maxim.

Prenylated flavonol glycosides in Epimedium plants, as key medicinal components, are known to have great pharmaceutical activities for human health. Among the main prenylated flavonol glycosides, the modification mechanism of different sugar moieties is still not well understood. In the current study, a novel prenylated flavonol rhamnoside xylosyltransferase gene (EpF3R2″XylT) was cloned from E. pubescens, and the enzymatic activity of its decoding proteins was examined in vitro with different prenylated flavonol rhamnoside substrates and different 3-O-monosaccharide moieties. Furthermore, the functional and structural domains of EpF3R2″XylT were analyzed by bioinformatic approaches and 3-D protein structure remodeling. In summary, EpF3R2″XylT was shown to cluster with GGT (glycosyltransferase that glycosylates sugar moieties of glycosides) through phylogenetic analysis. In enzymatic analysis, EpF3R2″XylT was proven to transfer xylose moiety from UDP-xylose to prenylated flavonol rhamnoside at the 2″-OH position of rhamnose. The analysis of enzymatic kinetics showed that EpF3R2″XylT had the highest substrate affinity toward icariin with the lowest Km value of 75.96 ± 11.91 mM. Transient expression of EpF3R2″XylT in tobacco leaf showed functional production of EpF3R2″XylT proteins in planta. EpF3R2″XylT was preferably expressed in the leaves of E. pubescens, which is consistent with the accumulation levels of major prenylflavonol 3-O-triglycoside. The discovery of EpF3R2″XylT will provide an economical and efficient alternative way to produce prenylated flavonol trisaccharides through the biosynthetic approach.

Functional Analysis of a Methyltransferase Involved in Anthocyanin Biosynthesis from Blueberries (Vaccinium corymbosum)

Anthocyanins are natural water-soluble pigments that widely exist in plants, with various biological activities, including antioxidant, anti-obesity, and anti-diabetic activities. Currently, monomeric anthocyanins are mainly obtained through natural sources, which limits their availability. In the biosynthesis of anthocyanins, anthocyanin methyltransferases are recognized to play important roles in the water solubility and structural stability of anthocyanins. Blueberries are a rich source of anthocyanins with more than 30 chemical structures. However, the enzymes that were responsible for the methylation of anthocyanidin cores in blueberries had not been reported. Here, blueberries (Vaccinium corymbosum) have been selected as the candidate for characterization of the key enzyme. Phylogenic analysis, enzymatic activity assay, homology modeling, molecular simulation, protein expression and purification assay, site-directed mutation, isothermal titration calorimetry assay, and enzyme kinetic assay were used to identify the enzymatic function and molecular mechanism of VcOMT, which was responsible for the methylation of anthocyanidin cores. VcOMT could use delphinidin as a substrate but not cyanidin, petunidin, anthocyanins, flavonols, and flavonol glycosides. Ile191 and Glu198 were both identified as important amino acid residues for the binding interactions of anthocyanidins with VcOMT.

Chemical constituents of Pteris excelsa and their antioxidant activity

Pteris excelsa Gaud. is a perennial evergreen herbaceous plant belonging to the Pteridaceae family. In the present study, the chemical constituents of this species were obtained by utilizing MCI, C18 and silica gel and Sephadex LH-20 gel column chromatography. A new flavonol glycoside was identified, kaempferol-4′,7-dimethyl-3-O-β-D-glucosyl-(1 → 4)-α-L-rhamnoside (1), together with fourteen known compounds (2–15), kaempferol-4′-methyl-3-O-β-D-glucoside (2), kaempferol-4′,7-dimethyl-3-O-glucoside (3), kaempferol-3-O-β-D-glucoside (4), kaempferol- 4′-methyl-3,7-α-O-L-dirhamnoside (5), kaempferol-3,7-di-O-α-L-rhamnoside (6), pterosin B (7), 3β-hydroxycinnamolide (8), N-(3-carboxypropyl)-2-acetylpyrrole (9), vanillic acid (10), sauropunol C/D (11), 6-anhydro-2-deoxy-β-O-furanosides (12), aquilegiolide (13), menisdaurilide (14), 2-(D-glycerol-1,2-dihydroxyethyl) furan (15). The structures of these compounds were established based on their high-resolution electrospray ionization mass spectrometry results and nuclear magnetic resonance spectroscopy data and by comparing their spectroscopic data with those reported in the literature. Importantly, we found that the antioxidant activity of PE, EtOAc and n-BuOH fractions of 98% methanol extract of P. excelsa (mediated by scavenging free radicals) was 33.57 ± 0.67%, 52.98 ± 1.55% and 45.25 ± 1.10% at a concentration of 400 μg/mL, respectively. Besides, compounds 1–6 displayed an antioxidant effect against the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical.

Biological Investigation and Chemical Study of Brassica villosa subsp. drepanensis (Brassicaeae) Leaves

Brassica villosa subsp. drepanensis (Caruel) Raimondo & Mazzola, belonging to the Brassica oleracea complex, is a wild edible plant endemic to western Sicily and a relative of modern cultivated Brassica crops. In this study, the antioxidant properties, anti-inflammatory activities, enzymatic inhibition, and cytotoxicity in cancer cells of B. villosa subsp. drepanensis leaf ethanolic extract were analysed for the first time. In addition, its chemical profile was investigated partitioning the total 70% ethanol extract among ethyl acetate, n-butanol, and water to obtain three residues that were subjected to chromatographic separation. Two flavonol glycosides, a phenol glucoside, two amino acids, and purine/pyrimidine bases were obtained. The presence of the glucosinolate glucoiberin was detected in the water extract by UHPLC-MS analysis. The total polyphenol and flavonoid content of the 70% ethanol extract showed good antioxidant capacities and anti-inflammatory properties by reducing nitric oxide release and reactive oxygen species levels and increasing glutathione in lipopolysaccharide-stimulated RAW 264.7 cells. The extract inhibited the enzymatic activity of α-amylase, α-glucosidase, and, significantly, of lipase. The MTT assay showed that the extract did not affect the viability of normal HFF-1 and RAW 264.7 cells. Among the cancer cell lines tested, an antiproliferative action was only observed in CaCo-2. The cytotoxicity of the extract was further confirmed by LDH release assay and by the destabilization of the oxidative balance. Results confirmed the antioxidant properties of the crude extract responsible for the anti-inflammatory effect on healthy cells and cytotoxicity in cancer cells.

Antioxidants in Animal Nutrition: UHPLC-ESI-QqTOF Analysis and Effects on In Vitro Rumen Fermentation of Oak Leaf Extracts.

The genus Quercus supplies a large amount of residual material (e.g., bark, acorns, leaves, wood), the valorization of which can favor a supply of antioxidant polyphenols to be used in the pharmaceutical, nutraceutical, or cosmeceutical sector. The recovery of specialized metabolites could also benefit livestock feeding, so much so that polyphenols have gained attention as rumen fermentation modifiers and for mitigating the oxidative imbalance to which farm animals are subject. In this context, leaves of Quercus robur L. from Northern Germany were of interest and the alcoholic extract obtained underwent an untargeted profiling by means of ultra-high-performance liquid chromatography/high-resolution tandem mass spectrometry (UHPLC-HRMS/MS) techniques. As triterpenes and fatty acids occurred, the alcoholic extract fractionation pointed out the obtainment of a polyphenol fraction, broadly constituted by coumaroyl flavonol glycosides and condensed tannins. Total phenol, flavonoid and condensed tannins content assays, as well as antiradical (DPPH● and ABTS+●) and reducing activity (PFRAP) were carried out on the alcoholic extract and its fractions. When the effects on rumen liquor was evaluated in vitro in terms of changes in fermentation characteristics, it was observed that oak leaf extract and its fractions promoted an increase in total volatile fatty acids and differently modulated the relative content of each fatty acid.

Quality evaluation of ginkgo biloba leaves based on non-targeted metabolomics and representative ingredient quantification

Ultra-performance liquid chromatography coupled with time-of-flight mass spectrometry (UPLC-Q-TOF-MS/MS) combined with multivariate statistical analysis was applied to the study of plant metabolomics to reveal the factors affecting the content of ginkgo leaf compounds. As a follow-up analysis, the terpene lactones and ginkgolic acids were quantified simultaneously using ultra-performance liquid chromatography coupled with triple quadrupole mass spectrometry (UPLC-QqQ-MS/MS), and subsequently total flavonol glycosides were quantified by high-performance liquid chromatography (HPLC). The results revealed that a total of 52 compounds were potentially identified by establishing a database, and 10 compounds were verified by reference standards; terpene lactones, ginkgolic acids, and flavonoids were the differential compounds; and ginkgolide A was identified as an important indicator compound for tree age. In addition, quantitative analysis showed that the contents of total flavonol glycosides and terpene lactones were highest during April and August in young ginkgo leaves, and differed based on origin. In summary, numerous compounds were rapidly detected by liquid chromatography coupled with MS, the ginkgo leaf samples were compared, and the differential metabolites were screened out. The content changing rules of the target compounds in ginkgo leaves from different regions with different tree ages and harvesting periods were clarified.

Rutin alleviates bisphenol A-glycidyl methacrylate-induced generation of proinflammatory mediators through the MAPK and NF-κB pathways in macrophages.

Bisphenol A-glycidyl methacrylate (BisGMA) is a methacrylate monomer that is mainly used in three-dimensional structures to reconstruct dental and bony defects. BisGMA has toxic and proinflammatory effects on macrophages. Rutin is a natural flavonol glycoside that is present in various plants and has useful biological effects, such as anti-inflammatory, anticancer, and antioxidative effects. The aim of this study was to investigate the anti-inflammation of rutin in macrophages after exposure to BisGMA. Pretreatment of the RAW264.7 macrophage with rutin at 0, 10, 30, and 100 μM for 30 min before being incubated with BisGMA at 0 or 3μM. Proinflammatory cytokines and prostaglandin (PG) E2 were detected by enzyme-linked immunosorbent assay (ELISA). Nitric oxide (NO) was detected by the Griess assay. Expression and phosphorylation of proteins were measured by Western blot assay. Pretreatment with rutin inhibited the BisGMA-induced generation of proinflammatory cytokines, including tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-6, and PGE2, in macrophages. Rutin also suppressed the BisGMA-induced secretion of NO and expression of inducible nitric oxide synthase (iNOS) in a concentration-dependent manner. Furthermore, rutin suppressed the mitogen-activated protein kinase (MAPK) phosphorylation in a concentration-dependent manner. Finally, rutin suppressed the BisGMA-induced phosphorylation of nuclear factor (NF)-κB p65 and degradation of inhibitor of κB (IκB). These results indicate that the concentration of rutin has an inhibitory effect on proinflammatory mediator generation, MAPK phosphorylation, NF-κB p65 phosphorylation, and IκB degradation. In conclusion, rutin is a potential anti-inflammatory agent for BisGMA-stimulated macrophages through NF-κB p65 phosphorylation and IκB degradation resulting from MAPK phosphorylation.

HPLC-DAD-MS Identification and Quantification of Phenolic Components in Japanese Knotweed and American Pokeweed Extracts and Their Phytotoxic Effect on Seed Germination.

We performed a detailed HPLC-MSn analysis of the phenolic compounds from the extracts of two invasive alien plant species (IAPS): Japanese knotweed (Fallopia japonica (Houtt.) Ronse Decr.) and American pokeweed (Phytolacca americana L.). The major phenolic groups were hydroxycinnamic acids and flavanols in Japanese knotweed (J. knotweed) and flavonols, hydroxycinnamic acids, and stilbenes in American pokeweed (A. pokeweed). We investigated the influence of solvent type and extraction time on the extraction efficiency of the phenolic compounds. The solvent 80% methanol had a higher polyphenolic extraction efficiency than water, since 14.5 times more flavonols and 2.3 times more stilbenes were extracted from J. knotweed and 5.2 times more flavonols and 2.6 times more stilbenes were extracted from A. pokeweed. In contrast, with water, we obtained a 52% higher hydroxycinnamic acids (HCA) content from J. knotweed. Hydroxycinnamic acids were best extracted in water after 24 h, flavanols after 12 h, stilbenes between 12 and 24 h, and flavonol glycosides after 48 h of extraction. We also tested the allelopathic effect of the aqueous extract of A. pokeweed and J. knotweed on seed germination and shoot and root growth of perennial ryegrass. The results showed that the water extract of J. knotweed resulted in 38 to 48% lower seed germination of perennial ryegrass, and the extract of A. pokeweed resulted in 83 to 90% lower seed germination. The phytotoxic effect of the extract of J. knotweed and A. pokeweed was also reflected in a characteristic reduced growth of shoots and roots of perennial ryegrass. The phytotoxic action of IAPS could also be applied for beneficial purposes, since this would be an effective strategy for their control and a reduction of their spread in the environment.

Ultrasensitive Determination of Natural Flavonoid Rutin Using an Electrochemical Sensor Based on Metal-Organic Framework CAU-1/Acidified Carbon Nanotubes Composites.

Rutin, a natural flavonol glycoside, is widely present in plants and foods, such as black tea and wheat tea. The antioxidant and anti-inflammatory effects of flavonoids are well known. In this study, a new electrochemical rutin sensor was developed using multiwalled carbon nanotubes/aluminum-based metal-organic frameworks (MWCNT/CAU-1) (CAU-1, a type of Al-MOF) as the electrode modification material. The suspension of multiwalled carbon tubes was dropped on the surface of the GCE electrode to make MWCNT/GCEs, and CAU-1 was then attached to the electrode surface by electrodeposition. MWCNTs and CAU-1 were characterized using scanning electron microscopy (SEM) and X-ray photoelectron spectroscopy (XPS). Due to the synergistic effect of CAU-1 and MWCNT-COOH, the prepared sensor showed an ultrasensitive electrochemical response to rutin. Under optimized conditions, the sensor showed a linear relationship between 1.0 × 10-9~3.0 × 10-6 M with a detection limit of 6.7 × 10-10 M (S/N = 3). The sensor also showed satisfactory stability and accuracy in the detection of real samples.

Medicinal Values and Potential Risks Evaluation of Ginkgo biloba Leaf Extract (GBE) Drinks Made from the Leaves in Autumn as Dietary Supplements.

Ginkgo tea and ginkgo wine are two familiar Ginkgo biloba leaf extract (GBE) drinks in the form of dietary supplements (DS) used for healthcare in east Asia. Nevertheless, a comprehensive evaluation of their safety and efficacy is still lacking. In this study, GBE drinks were prepared from naturally newly senescent yellow leaves (YL) and green leaves (GL) in autumn. Their total flavonoids, antioxidant capacity and prescribed ingredients were investigated. In brief, the proportions of total flavonoids, total flavonol glycosides (TFs), total terpene trilactones (TTLs) and ginkgolic acids in the GBE drinks all did not meet the standards of worldwide pharmacopoeias. Specifically, the levels of TFs in the ginkgo tea prepared from YL were significantly higher than that prepared from GL. Further analyses revealed a substandard ratio of isorhamnetin/quercetin and an accumulation of leaf-age-related compounds, which were both unqualified. The proportions of specific TTLs varied between the ginkgo tea and ginkgo wine, although no significant differences were detected in terms of the total levels of TTLs. Noticeably, numerous biflavones and thousands of times over the limiting concentration of ginkgolic acids, including newly identified types, were only detected in ginkgo wine. Finally, the use of the GBE drinks as DSs was comprehensively evaluated according to the acceptable daily intake. This study showed the limited healthcare effects of GBE drinks despite their powerful antioxidant capacity.

Glycine max (L.) Merr. (Soybean) metabolome responses to potassium availability

Potassium (K+) has vital physiological and metabolic functions in plants and its availability can impact tolerance to biotic and abiotic stress conditions. Limited studies have investigated the effect of K+ fertilization on soybean metabolism. Using integrated omics, ionomics and metabolomics, we investigated the field-grown Glycine max (soybean) response, after four K+ soil fertilization rates. Soybean leaf and pod tissue (valves and immature seeds) extracts were analysed by ultra-performance liquid chromatography coupled to high-resolution mass spectrometry (UPLC-HRMS) and inductively coupled plasma optical emission spectroscopy (ICP-OES). Multivariate analyses (PCA-X&Y e O2PLS-DA) showed that 51 compounds of 19 metabolic pathways were regulated in response to K+ availability. Under very low potassium availability, soybean plants accumulated of Ca2+, Mg2+, Fe2+, Cu2+, and B in young and old leaves. Potassium fertilization upregulated carbohydrate, galactolipid, and flavonol glycoside biosynthesis in leaves and pod valves, while K+ deficient pod tissues showed increasing amino acids, oligosaccharides, benzoic acid derivatives, and isoflavones contents. Severely K+ deficient soils elicited isoflavones, coumestans, pterocarpans, and soyasaponins in trifoliate leaves, likely associated to oxidative and photodynamic stress status. Additionally, results demonstrate that L-asparagine content is higher in potassium deficient tissues, suggesting this compound as a biomarker of K+ deficiency in soybean plants. These results demonstrate that potassium soil fertilization did not linearly contribute to changes in specialised constitutive metabolites of soybean. Altogether, this work provides a reference for improving the understanding of soybean metabolism as dependent on K+ availability.

Revitalizing Unfermented Cabernet Sauvignon Pomace Using an Eco-Friendly, Two-Stage Countercurrent Process: Role of pH on the Extractability of Bioactive Phenolics

As the major byproduct of the winemaking industry, grape pomace remains an untapped source of valuable bioactive phenolic compounds. This study elucidated the optimal aqueous extraction parameters for maximizing phenolic extractability, while avoiding the use of harsh conventional solvents and limiting water usage, from Cabernet Sauvignon grape pomace in which the red grape was processed for white wine. In the single-stage aqueous extraction process (AEP), the concurrent impact of pH (2.64–9.36), solids-to-liquid ratio (SLR, g pomace/mL water) (1:50–1:5), and temperature (41.6–58.4 °C) on the total phenolic content (TPC) of Cabernet Sauvignon pomace was evaluated alongside a kinetic study (15–90 min). Optimal single-stage extraction conditions (pH 9.36, 1:50 SLR, 50 °C, 75 min) guided the development of a two-stage countercurrent extraction process (pH 9.36, 1:10 SLR, 50 °C, 75 min) to further reduce water consumption without compromising overall extractability. The countercurrent process reduced fresh water usage by 80%, increased the TPC of the extracts by 18%, and improved the in vitro antioxidant activities (ABTS and ORAC) of the extracts. Untargeted metabolomics enabled the identification of a diverse pool of phenolics, especially flavonol glycosides, associated with grape pomace, while further phenolic quantitation detected improvements in the release of commonly bound phenolics such as ferulic acid, p-coumaric acid, syringic acid, and protocatechuic acid in alkaline extracts compared to the ethanolic extract. This investigation provides an efficient, eco-friendly extraction strategy suitable for applications in functional food, beverage, nutraceutical, and cosmetic industries.