The exogenous application of naringenin and rosmarinic acid modulates functional traits in Lepidium sativum.

BACKGROUND: Phenolic modulators have attracted attention for their potential in shaping functional traits in plants. This work investigated the impact of naringenin (Nar) and rosmarinic acid (RA) on the functional properties of Lepidium sativum leaves and roots. RESULTS: Untargeted metabolomics identified a diverse phenolic profile, including flavonoids, phenolic acids, low molecular weight phenolics, lignans, and stilbenes. Cluster, analysis of variance multiblock orthogonal partial least squares (AMOPLS), and orthogonal projection to latent structures discriminant analysis (OPLS-DA) multivariate analyses confirmed tissue-specific modulation of bioactive compounds. The tissue was the hierarchically most influential factor, explaining 27% of observed variability, while the treatment and their interaction were statistically insignificant. Thereafter, various in vitro assays were employed to assess antioxidant capacity, including 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2,2'-azino-bis(3-ethylbenzothiazoline)-6-sulfonic acid (ABTS) radical scavenging activity, cupric ion reducing antioxidant capacity (CUPRAC), and ferric ion reducing antioxidant power (FRAP), metal chelating ability, and phosphomolybdenum (PMD) assays. Extracts were also tested for inhibitory effects on cholinesterase, amylase, glucosidase, and tyrosinase enzymes. RA application positively impacted antioxidant and enzyme inhibitory activities, holding valuable implications in shaping the health-promoting properties of L. sativum. CONCLUSION: The untargeted metabolomics analysis showed a significant tissue-dependent modulation of bioactive compounds, determining no synergistic effect between applying phenolic compounds in combination. Specifically, the sole application of RA increased anthocyanins and hydroxyphenyl propanoic acid content on leaves, which was strictly related to enhancing the biological activities. © 2023 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Foliar applications of a Malvaceae-derived protein hydrolysate and its fractions differentially modulate yield and functional traits of tomato under optimal and suboptimal nitrogen application.

BACKGROUND: Protein hydrolysates (PHs) can enhance plant nitrogen nutrition and improve the quality of vegetables, depending on their bioactive compounds. A tomato greenhouse experiment was conducted under both optimal (14 mM) and suboptimal (2 mM) nitrogen (N-NO3) conditions. Tomatoes were treated with a new Malvaceae-derived PH (MDPH) and its molecular fractions (MDPH1, >10 kDa; MDPH2, 1-10 kDa and MDPH3, <1 kDa). RESULTS: Under optimal N conditions, the plants increased biomass and fruit yield, and showed a higher photosynthetic pigment content in leaves in comparison with suboptimal N, whereas under N-limiting conditions, an increase in dry matter, soluble solid content (SSC) and lycopene, a reduction in firmness, and changes in organic acid and phenolic compounds were observed. With 14 mM N-NO3, MDPH3 stimulated an increase in dry weight and increased yield components and lycopene in the fruit. The MDPH2 fraction also resulted in increased lycopene accumulation in fruit under 14 mM N-NO3. At a low N level, the PH fractions showed distinct effects compared with the whole MDPH and the control, with an increase in biomass for MDPH1 and MDPH2 and a higher pigment content for MDPH3. Regardless of N availability, all the fractions affected fruit quality by increasing SSC, whereas MDPH2 and MDPH3 modified organic acid content and showed a higher concentration of flavonols, lignans, and stilbenes. CONCLUSION: The molecular weight of the peptides modifies the effect of PHs on plant performance, with different behavior depending on the level of N fertilization, confirming the effectiveness of fractioning processes. © 2024 Society of Chemical Industry.

Comprehensive Analysis of the Chemical and Bioactivity Profiles of Endemic Crataegus turcicus Dönmez in Comparison with Other Crataegus Species.

Crataegus turcicus is a plant endemic to Türkiye. For the first time, this study aimed to comparatively assess its flower-bearing branches, leaves, and fruits with other well-known Crataegus species (C. monogyna, C. pentagyna, and C. orientalis) in terms of chemical composition and bioactivity studies to evaluate its potential use as a food supplement. Firstly, the contents of total phenolics (TPC), flavonoids (TFC), proanthocyanidin (TPAC), and anthocyanin (TAC) in different plant parts of Crataegus species were evaluated. The highest TPAC was found in the hydroalcoholic extract of C. turcicus flower-bearing branches. Moreover, all plant parts had comparatively higher amounts of TPC, TFC, and TAC compared to other Crataegus species. The chemical screening by high-performance thin-layer chromatography (HPTLC) resulted that C. turcicus parts were rich with chlorogenic acid, neochlorogenic acid, quercetin and vitexin derivatives, epicatechin, procyanidin, etc., and their quantities were evaluated by high-performance liquid chromatography (HPLC). In terms of several in vitro antioxidant activity outcomes, the flower-bearing branches of C. turcicus showed the highest antioxidant activity by a 2,2-diphenyl-1-picrylhydrazyl (DPPH) test among the assessed antioxidant assays. Additionally, hydroalcoholic extracts of C. turcicus significantly decreased LPS-induced nitric oxide, tumor necrosis factor-alpha, and interleukin-6 production more potently than indomethacin (positive control). In addition to its remarkable anti-inflammatory activity, C. turcicus showed analgesic activity by reducing prostaglandin E2 levels.

Rewired phenolic metabolism and improved saccharification efficiency of a Zea mays cinnamyl alcohol dehydrogenase 2 (zmcad2) mutant.

Lignocellulosic biomass is an abundant byproduct from cereal crops that can potentially be valorized as a feedstock to produce biomaterials. Zea mays CINNAMYL ALCOHOL DEHYDROGENASE 2 (ZmCAD2) is involved in lignification, and is a promising target to improve the cellulose-to-glucose conversion of maize stover. Here, we analyzed a field-grown zmcad2 Mutator transposon insertional mutant. Zmcad2 mutant plants had an 18% lower Klason lignin content, whereas their cellulose content was similar to that of control lines. The lignin in zmcad2 mutants contained increased levels of hydroxycinnamaldehydes, i.e. the substrates of ZmCAD2, ferulic acid and tricin. Ferulates decorating hemicelluloses were not altered. Phenolic profiling further revealed that hydroxycinnamaldehydes are partly converted into (dihydro)ferulic acid and sinapic acid and their derivatives in zmcad2 mutants. Syringyl lactic acid hexoside, a metabolic sink in CAD-deficient dicot trees, appeared not to be a sink in zmcad2 maize. The enzymatic cellulose-to-glucose conversion efficiency was determined after 10 different thermochemical pre-treatments. Zmcad2 yielded significantly higher conversions compared with controls for almost every pre-treatment. However, the relative increase in glucose yields after alkaline pre-treatment was not higher than the relative increase when no pre-treatment was applied, suggesting that the positive effect of the incorporation of hydroxycinnamaldehydes was leveled off by the negative effect of reduced p-coumarate levels in the cell wall. Taken together, our results reveal how phenolic metabolism is affected in CAD-deficient maize, and further support mutating CAD genes in cereal crops as a promising strategy to improve lignocellulosic biomass for sugar-platform biorefineries.

Transcriptional and metabolic changes associated with internode development and reduced cinnamyl alcohol dehydrogenase activity in sorghum.

The molecular mechanisms associated with secondary cell wall (SCW) deposition in sorghum remain largely uncharacterized. Here, we employed untargeted metabolomics and large-scale transcriptomics to correlate changes in SCW deposition with variation in global gene expression profiles and metabolite abundance along an elongating internode of sorghum, with a major focus on lignin and phenolic metabolism. To gain deeper insight into the metabolic and transcriptional changes associated with pathway perturbations, a bmr6 mutant [with reduced cinnamyl alcohol dehydrogenase (CAD) activity] was analyzed. In the wild type, internode development was accompanied by an increase in the content of oligolignols, p-hydroxybenzaldehyde, hydroxycinnamate esters, and flavonoid glucosides, including tricin derivatives. We further identified modules of genes whose expression pattern correlated with SCW deposition and the accumulation of these target metabolites. Reduced CAD activity resulted in the accumulation of hexosylated forms of hydroxycinnamates (and their derivatives), hydroxycinnamaldehydes, and benzenoids. The expression of genes belonging to one specific module in our co-expression analysis correlated with the differential accumulation of these compounds and contributed to explaining this metabolic phenotype. Metabolomics and transcriptomics data further suggested that CAD perturbation activates distinct detoxification routes in sorghum internodes. Our systems biology approach provides a landscape of the metabolic and transcriptional changes associated with internode development and with reduced CAD activity in sorghum.

Coupling the high-resolution LC-MS characterisation of the phenolic compounds with the antimicrobial and antibiofilm properties of helencha (Enydra fluctuans Lour.).

This study reports the polyphenol profile of helencha (Enydra fluctuans Lour.), an underutilised, aquatic leafy vegetable, based on high resolution accurate mass analysis. The methanolic extract of helencha leaves was screened by ultra-high performance liquid chromatography with quadrupole time of flight mass spectrometry (LC-QToF-MS). An in-house developed database of phytochemical metabolites was referred for compound identifications. Based on the detection of the pseudomolecular ion and at least one molecule-specific fragment ion (each with < 5 ppm of mass error), 25 potentially-bioactive phenolic compounds were putatively identified. These included 6 flavonols, 4 phenolic acids, 3 lignans, 3 flavones and 1 each of flavanol, flavanone, dihydroflavonol, tetramethoxyflavone, isoflavonoid and methylated flavonol. In addition, 3 unclassified compounds are also reported. The helencha extract showed antibiofilm properties with a potent bacteriostatic activity against the clinical isolates of Pseudomonas aeruginosa, a human pathogenic bacteria. The complementary molecular docking studies indicated strong binding interactions of the identified compounds with the active site of LasR protein of P. aeruginosa. The in vitro and in silico study results would be useful to develop novel neutraceutical products based on helencha-extract and design new lead compounds to control the biofilm producing pathogenic microorganisms. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at (10.1007/s13197-021-04968-y).

Bioactive compounds and total antioxidant capacity of cane residues from different grape varieties.

BACKGROUND: Every year, the viticulture activity generates considerable amounts of underused lignocellulosic residues as grape cane, which are generally composted or burned despite their potential value as a source of bioactive compounds. Determination of their phytochemical composition and total antioxidant capacity (TAC) may be a useful way of exploiting different high-added value applications. RESULTS: Twenty-one phenolic compounds (PC) and two carotenoids (Car) were quantified by high performance-liquid chromatography-diode array detection in eight grape varieties from different locations in Mendoza, Argentina. The maximum concentrations corresponded to the stilbene ϵ-viniferin [10 552 µg g-1 dry weight (DW)], followed by the flavanols (+)-catechin (3718 µg g-1 DW) and (-)-epicatechin (2486 µg g-1 DW). In addition, lutein and ß-carotene were quantified at levels ranging between 350 and 2400 ng g-1 DW. The TAC of the extracts was assessed by oxygen radical absorbance capacity, 2,20-azino-bis-3-ethylbenzthiazoline-6-sulphonic acid and 1,1-diphenyl-2-picrylhydrazyl assays, with a good correlation between TAC and total PC for each sample (r ≥ 0.82). CONCLUSION: Samples of cv. Malbec, the most representative variety of Argentina's winemaking industry, presented high contents of PC, particularly ϵ-viniferin, (+)-catechin and (-)-epicatechin. Quercetin-3-galactoside, OH-tyrosol and Car were reported for the first time in grape canes of the eight varieties. The results add to the existing knowledge related to this inexpensive source of high-value bioactive compounds, which could be used as functional ingredients. © 2019 Society of Chemical Industry.

Profiling of polyphenols in phalsa (Grewia asiatica L) fruits based on liquid chromatography high resolution mass spectrometry.

The aim of the study was to screen the metabolite profile of phalsa (Grewia asiatica), an underutilized fruit crop, using liquid chromatography-high resolution mass spectrometric analysis. A total of 50 compounds were tentatively identified based on their molecular mass and characteristic fragment ions, each with less than 5 ppm of mass error. These compounds included 21 flavonols, 2 dihydroflavonols, 7 flavones, 3 flavanols, 6 anthocyanins, 3 isoflavonoids, 2 phenolic acids, 2 flavanones, and 4 other phenolics. Flavonols were the predominant group of compounds, representing around 52.6% of the total phenolics. The paper has also discussed the potentiality of phalsa as an emerging functional food for the management of various human diseases in relation to the existing literature.

Phenolic Profiling of Duchesnea indica Combining Macroporous Resin Chromatography (MRC) with HPLC-ESI-MS/MS and ESI-IT-MS.

Duchesnea indica (D. indica) is an important traditional Chinese medicine, and has long been clinically used to treat cancer in Asian countries. It has been described previously as a rich source of phenolic compounds with a broad array of diversified structures, which are the major active ingredients. However, an accurate and complete phenolic profiling has not been determined yet. In the present work, the total phenolic compounds in crude extracts from D. indica were enriched and fractionated over a macroporous resin column, then identified by HPLC-ESI-MS/MS and ESI-IT-MS (ion trap MS). A total of 27 phenolic compounds were identified in D. indica, of which 21 compounds were identified for the first time. These 27 phenolic compounds encompassing four phenolic groups, including ellagitannins, ellagic acid and ellagic acid glycosides, hydroxybenzoic acid and hydroxycinnamic acid derivatives, and flavonols, were then successfully quantified using peak areas against those of the corresponding standards with good linearity (R² > 0.998) in the range of the tested concentrations. As a result, the contents of individual phenolic compounds varied from 6.69 mg per 100 g dry weight (DW) for ellagic acid to 71.36 mg per 100 g DW for brevifolin carboxylate. Not only did this study provide the first phenolic profiling of D. indica, but both the qualitative identification and the subsequent quantitative analysis of 27 phenolic compounds from D. indica should provide a good basis for future exploration of this valuable medicinal plant.

Bio-functional properties of Jilungin (Terminalia canescens).

Jilungin (Terminalia canescens) is a native Australian plant and the Indigenous "Nyul Nyul" people of the  Kimberley region of Western Australia use its leaves to make herbal tea. Due to the rise in the popularity of drinking Jilungin tea among the consumers in Australia and internationally, it is important to study the nutritional and health-beneficial properties as well as safety of Jilungin leaves. This study aims to determine the nutritional composition, anti-nutritional factors, antimicrobial and antidiabetic properties of Jilungin leaves. Also, the phytochemical profiling using UHPLC-MS/MS (Ultra-performance liquid chromatographymass spectrometry) and antioxidant activity of Jilungin methanolic extracts and herbal infusion were investigated. The safety of the leaves and infusion was also investigated by using in vitro mammalian cell lines (Caco2, HT29, and HepG2) through cell viability assays. The leaves are rich in dietary fiber (43.9%) and linoleic acid (30.4% of total fatty acids). Phytochemical profiling revealed ellagic acid, geraniin, pedunculagin, and punicalagin as the major bioactive compounds. The results also demonstrated that Jilungin has strong antioxidant and antidiabetic activities. A significant (p < 0.01) strong positive correlation was observed between the high antioxidant activity of Jilungin infusion with the major bioactive compounds. Jilungin extracts (50 mg/mL) exhibited strong antimicrobial activity against Staphylococcus aureus and Bacillus cereus. Its infusion and methanolic extract were safe on the studied cell lines (Caco-2, HT29, and HepG2) at higher concentrations of 66.6 and 98 mg/mL, respectively. Therefore, Jilungin teas or infusions could be a safe and effective way to promote health and well-being. PRACTICAL APPLICATION: Jilungin tea is very popular among consumers in Australia and is gaining popularity worldwide. The current study will increase knowledge on the nutritional aspects and safety of the Jilungin use.

A New Method for Fractionation and Characterization of Polyphenols and Tannins from Grapevine Leaf Tissue.

Plants accumulate different types of phenolic material in their tissue as a response to biotic as well as abiotic stress. Monomeric polyphenols and smaller oligomers can serve as protection against ultraviolet radiation or prevent oxidative tissue damage, while larger molecules such as tannins can be the plant's reaction to an infection or physical damage. Therefore, characterization, profiling, and quantification of diverse phenolics can provide valuable information about the plant and the stress status at any given time. A method was developed that allows the extraction of polyphenols and tannins from leaf tissue, followed by fractionation and quantification. Extraction was performed with liquid nitrogen and 30% acetate-buffered ethanol. The method was tested with four cultivars under varying extraction conditions (solvent strength and temperature) and showed great improvements of the chromatography that would otherwise be impacted by tannins. The separation of tannins from smaller polyphenols was achieved by bovine serum albumin precipitation and resuspension in a urea-triethanolamine buffer. Tannins were reacted with ferric chloride and analyzed spectrophotometrically. Monomeric non-protein-precipitable polyphenols were then analyzed via HPLC-DAD from the supernatant of the precipitation sample. This way, a more complete spectrum of compounds can be analyzed from the same plant tissue extract. With the fractionation suggested here, hydroxycinnamic acids and flavan-3-ols can be separated and quantified with good accuracy and precision. Possible applications include the assessment of plant stress and response monitoring using the total concentrations of polyphenols and tannins, as well as the ratios between those compound classes.

Optimization and Characterization of Phenolic Extraction Conditions and Antioxidant Activity Evaluation of Adenanthera pavonina L. Bark.

The presence of high levels of secondary metabolites in medicinal plants can significantly influence the progress of drug development. Here, we aimed to maximize phenolic extraction from Adenanthera pavonina L. stem bark using various solvents such as ethyl acetate, methanol, petroleum ether, and chloroform. A response surface method (RSM) with a central composite design (CCD) statistical technique was applied to optimize the extraction process, employing three important extracting parameters such as extraction time (h), temperature (°C), and solvent composition (% v/v of methanol/water) to obtain the highest phenolic content. Total phenolic content (TPC) and antioxidant activity (IC50 of extract's DPPH radical scavenging activity) were used as response variables to find the influence of these extracting parameters. Among the various solvents used, methanol extract showed the highest contents of phenolics and the maximum level of antioxidant activity with a lower IC50 value. The notable TPC and IC50 value of the extract's DPPH radical scavenging capacity were found to be 181.69 ± 0.20 mg GAE/g dry tissue and 60.13 ± 0.11 mg/mL, respectively, under the optimal conditions with a solvent composition of 71.61% (v/v) of methanol/water, extraction temperature of 42.52 °C, and extraction time of 24 h. The optimized extract of A. pavonina stem bark was further subjected to HPLC analysis, where six phenolic compounds, including coumarin, p-coumaric acid, chlorogenic acid, sinapic acid, gallic acid, and caffeic acid, were identified along with their respective quantities. Overall, the findings of this study uncover a low-cost analytical model for maximizing phenolic extraction from A. pavonina bark with enhanced antioxidant activity.

Recovery of phenolic compounds from wine lees using green processing: Identifying target molecules and assessing membrane ultrafiltration performance.

Winery wastes are rich in polyphenols with high added value to be used in cosmetics, pharmaceuticals, and food products. This work aims at recovering and purifying the polyphenolic fraction occurring in the malolactic fermentation lees generated during the production of Albariño wines. Phenolic acids, flavonoids, and related compounds were recovered from this oenological waste by green liquid extraction using water as the solvent. The resulting extract solution was microfiltered to remove microparticles and further treated by ultrafiltration (UF) using membranes of 30 kDa and 5 kDa molecular weight cut-offs (MWCOs). The feed sample and the filtrate and retentate solutions from each membrane system were analyzed by reversed-phase liquid chromatography (HPLC) with UV and mass spectrometric (MS) detection. The most abundant polyphenols in the extracts were identified and quantified, namely: caftaric acid with a concentration of 200 µg g-1 and trans-coutaric acid, cis-coutaric acid, gallic acid, and astilbin with concentrations between 15 and 40 µg g-1. Other minor phenolic acids and flavanols were also found. The UF process using the 30 kDa membrane did not modify the extract composition, but filtration through the 5 kDa poly-acrylonitrile membrane elicited a decrease in polyphenolic content. Hence, the 30 kDa membrane was recommended to further pre-process the extracts. The combined extraction and purification process presented here is environmentally friendly and demonstrates that malolactic fermentation lees of Albariño wines are a valuable source of phenolic compounds, especially phenolic acids.

Optimized Ultrasound-Assisted Enzymatic Extraction of Phenolic Compounds from Rosa canina L. Pseudo-Fruits (Rosehip) and Their Biological Activity.

Two techniques, namely, optimized ultrasound-assisted extraction (UAE) and enzyme-assisted extraction (EAE), were used to promote the extraction of phenolic compounds from the pseudo-fruits of Rosa canina L. (RC). For UAE, an optimization process based on the design of experiment (DoE) principles was used for determining the dependence between three variables (i.e., time of extraction, ultrasound amplitude, and the material-to-water ratio) and the total phenolic content of the samples. For EAE, a 2:1:1 pectinase, cellulase, and hemicellulase enzymatic blend was used as pre-treatment for optimized UAE, inducing a higher total phenolic content. The untargeted phenolic profiling approach revealed a great abundance of lower molecular weight phenolics (1.64 mg Eq./g) in UAE-RC extracts, whilst gallic acid (belonging to hydroxybenzoic acid derivatives) was the most abundant individual compound of both extracts. The unsupervised multivariate statistics clearly discriminated the impact of enzymatic pre-treatment on the phenolic profile of RC pseudo-fruits. Finally, Pearson's correlation coefficients showed that anthocyanins, phenolic acids, and tyrosol derivatives were those compounds mostly correlated to the in vitro antioxidant potential of the extracts, whilst negative and significant (p < 0.05) correlation coefficients were recorded when considering the enzymatic inhibition activities. The highest enzyme-inhibitory activity has been identified against α-glucosidase, which indicates an antidiabetic effect.

Phenolic-rich feijoa extracts from flesh, peel and whole fruit activate apoptosis pathways in the LNCaP cell line.

This study aimed to explore the potential anticancer activity of phenolic-rich feijoa extracts from the flesh, peel, and whole fruit on the human prostate cancer cell line (LNCaP). Results showed that feijoa extracts had cancer-specific anti-proliferative activity on the LNCaP cell line. The anticancer activity of feijoa extracts was shown through activation of the caspase-dependent apoptosis pathway based on the increase of sub-G1 phase in the cell cycle, the decrease of mitochondrial membrane potential, as well as the elevated caspase 3, 8, and 9 activity in the treated LNCaP cells. The anti-cancer activity of feijoa extracts could be attributed to the high total phenolic contents (0.14-0.37 mg GAE/mg dw) and, in particular, the high ellagic acid content (2.662-9.119 µg/mg dw). The successful activation of the caspase-dependent apoptosis pathway indicates that phenolic-rich feijoa extracts have a good potential to be utilized as a functional ingredient in foods and nutraceuticals.

Chemical characterization and 5α-reductase inhibitory activity of phenolic compounds in goji berries.

Lycium fruits have a high content of phenolics as bioactive constituents with various pharmacological effects, but there is a lack of comparative analysis and chemical profiling of phenolics in Lycium fruit varieties. An ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MSE) combined with chemometrics was developed to characterize the phenolics in fruits from four Lycium species, including Lycium barbarum L. (LBL), L. chinense Mill. (LCM), L. barbarum var. auranticarpum (LBA) and L. ruthenicun Murr. (LRM). 63 phenolics were identified according to reported tandem mass fragmentation patterns and the UNIFI scientific informatics platform. Nine phenolics (5, 18, 20, 29, 31, 37, 41, 43, 60) were common and predominant components among four types of Lycium fruit. The partial least squares discriminant analysis (PLS-DA) and the orthogonal partial least squares discriminant analysis (OPLS-DA) were analyzed on the basis of a matrix created from 653 sets of data, and 20 Lycium fruits were classified into four groups. Further analysis identified that phenolics profiles were characteristic for each Lycium species, and five markers (13, 29, 31, 35, 99) could be utilized for fruit identification. Subsequently, inhibitory activity against 5α-reductase of phenolic extracts of Lycium fruits showed that LBL extract was the relative better effective, followed by LCM, whereas LBA and LRM showed no activity, which might be associated with the high contents of marker compounds (29, 31, 35, 43, 71, 99) in LBL. These findings will provide guidance for the development of Lycium phenolics with beneficial properties for the prevention and treatment of Benign prostatic hyperplasia (BPH).

Feasibility of using integrated fingerprinting, profiling and chemometrics approach to understand (bio) chemical changes throughout commercial red winemaking: A case study on Merlot.

This study assessed the feasibility of using a multiplatform approach; integrating untargeted fingerprinting of volatiles and targeted profiling of phenolic and oenological attributes (soluble solids, pH, titratable acidity and colour properties) coupled with chemometrics to understand complex (bio) chemical reactions occurring during Merlot red winemaking. The changes were investigated at three winemaking stages, starting from pre-maceration (PM), maceration-alcoholic fermentation (MAF) up to completion of malolactic fermentation (MLF). Merlot musts at PM were characterised by lighter colour and higher amount of green aroma-related volatiles. Completion of MAF led to increased extraction of anthocyanins, flavonols, and stilbenes, resulting in a more intense and darker fermenting juice. Furthermore, development of yeast-fermentation associated volatiles such as esters and alcohols was observed at this stage. The final wine, when MLF was completed, was rich in phenolic acids, esters, alcohols, and terpenes. The multiplatform analytical approach was effective to unravel the complex reactions throughout Merlot winemaking process and find relevant markers, which could help to predict expected quality attributes in the finished wine.

Phytochemical Profile and Antioxidant Activity of Aerial and Underground Parts of Salvia bulleyana Diels. Plants.

Plants have been used for medical purposes since ancient times. However, a detailed analysis of their biological properties and their associated active compounds is needed to justify their therapeutic use in modern medicine. The aim of the study was to identify and quantify the phenolics present in hydromethanolic extracts of the roots and shoots of the Chinese Salvia species, Salvia bulleyana. The qualitative and quantitative analyses were carried out by ultrahigh-performance liquid chromatography with electrospray ionization mass spectrometry detection (UHPLC-PDA-ESI-MS), and high-performance liquid chromatography with photodiode array (HPLC-PDA) detection. The extracts of S. bulleyana were also screened for their antioxidant activity using ferric ion (Fe3+) reducing antioxidant power (FRAP), 1,1-diphenyl-2-picrylhydrazyl (DPPH), diammonium 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonate) cation (ABTS), superoxide radical anion (O2•-), and inhibition of lipid peroxidation assays. The S. bulleyana extracts were found to contain 38 substances, of which 36 were phenols, with a total level of 14.4 mg/g DW (dry weight) in shoots, and 23.1 mg/g DW in roots. Twenty-eight phenols were polyphenolic acids or their derivatives, the most abundant in shoots being rosmarinic acid, and in roots, salvianolic acid K followed by rosmarinic acid. The other major phenolic acids were caffeic acid, caffeoyl-threonic acids, isomers of lithospermic acid, salvianolic acid F, salvianolic acid B, and yunnaneic acid E. In addition to polyphenolic acids, nine flavonoids were detected in the shoot extract. While both extracts showed significant antioxidant activity, the shoot extract, containing both polyphenolic acids and flavonoids, demonstrated a slightly greater antioxidant potential in some of the anti-radical tests than the roots. However, the root extract proved to be slightly more effective in the lipid peroxidation inhibition test. Thus, S. bulleyana was demonstrated as a promising source of antioxidants, and worthy of further more detailed studies.

Thymus zygis subsp. zygis an Endemic Portuguese Plant: Phytochemical Profiling, Antioxidant, Anti-Proliferative and Anti-Inflammatory Activities.

Thymus zygis subsp. zygis is an endemic Portuguese plant belonging to the Thymus zygis species. Although T. zygis is commonly used as a condiment and as a medicinal herb, a detailed description of the polyphenol composition of hydroethanolic (HE) and aqueous decoction (AD) extracts is not available. In this work, we describe for the first time a detailed phenolic composition of Thymus zygis subsp. zygis HE and AD extracts, together with their antioxidant, anti-proliferative and anti-inflammatory activities. Unlike other Thymus species, T. zygis subsp. zygis extracts contain higher amounts of luteolin-(?)-O-hexoside. However, the major phenolic compound is rosmarinic acid, and high amounts of salvianolic acids K and I were also detected. T. zygis subsp. zygis extracts exhibited significant scavenging activity of ABTS+, hydroxyl (•OH), and nitric oxide (NO) radicals. Regarding the anti-proliferative/cytotoxic effect, tested against Caco-2 and HepG2 cells, the AD extract only slightly reduced cell viability at higher concentrations (IC50 > 600 µg/mL, 48 h exposure), denoting very low toxicity, while the HE extract showed a high anti-proliferative effect, especially at 48 h exposure (IC50 of 85.01 ± 15.10 µg/mL and 82.19 ± 2.46 µg/mL, for Caco-2 and HepG2, respectively). At non-cytotoxic concentrations, both extracts reduced the nitric oxide (NO) release by lipopolysaccharide (LPS)-stimulated RAW 264.7 cells (at 50 µg/mL, HE and AD extracts inhibited NO release in ~89% and 48%, respectively). In conclusion, the results highlight the non-toxic effect of aqueous extracts, both resembling the consumption of antioxidants in foodstuff or in functional food. Furthermore, the HE extract of T. zygis subsp. zygis is a source of promising molecules with antioxidant, anti-inflammatory and anticancer activities, highlighting its potential as a source of bioactive ingredients for nutraceutical and pharmaceutical industries.

In vitro cytotoxic activity of six Syzygium leaf extracts as related to their phenolic profiles: An untargeted UHPLC-QTOF-MS approach.

Untargeted metabolomics was used in this study to discriminate the phenolic fingerprints of six Syzygium species. This approach resulted in the annotation of 441 compounds that belong to different phenolic classes, such as flavonoids, lignans, stilbenes, tyrosols, alkylphenols, and phenolic acids. Multivariate data analysis unraveled the main differences between the studied species. S. paniculatum and S. aqueum were the richest sources in terms of phenolic compounds, cumulatively amounting to 355.3 and 266.4 mg/g dry matter, respectively. Nevertheless, S. jambos showed reduced amounts of phenolics, when compared with other species. The biological activity of Syzygium leaf extracts was assessed on MCF-7 breast adenocarcinoma and MDA-MB-231 breast cancer cell lines. Potent estrogenic activity was detected using the SRB assay on MCF-7. This activity may be ascribable to the presence of phenolic compounds miming phytoestrogens such as lignans, stilbenes, and isoflavonoids in the investigated Syzygium extracts. By examining the biological effect of Syzygium extracts against MDA-MB-231 cell lines, the Syzygium gratum leaf extract exhibited the strongest inhibition, with IC50 = 19.4 µg/mL, followed by S. paniculatum (IC50 = 50.9 µg/mL). However, the Syzygium gratum leaf extract showed a potent cytotoxic effect on normal human skin fibroblasts, HSF (IC50 = 1.24 µg/mL), assuming a nonselective cytotoxic effect. On the other hand, other studied Syzygium leaves proved as safe nutraceuticals (IC50 ≥ 100 µg/mL) on HSF cell lines. Our study suggested a possible implication of Syzygium malaccense and Syzygium aqueum leaves as potential estrogenic candidates in relation to their health-promoting phenolic constituents.