Characterization Of Polyphenols Research Articles

Extraction and Analytical Methods for the Characterization of Polyphenols in Marine Microalgae: A Review.

Marine microalgae are emerging as promising sources of polyphenols, renowned for their health-promoting benefits. Recovering polyphenols from microalgae requires suitable treatment and extraction techniques to ensure their release from the biomass and analytical methodologies to assess their efficiency. This review provides a comprehensive comparison of traditional and cutting-edge extraction and analytical procedures applied for polyphenolic characterization in marine microalgae over the past 26 years, with a unique perspective on optimizing their recovery and identification. It addresses (I) cell disruption techniques, including bead milling, high-speed homogenization, pulsed electric field, ultrasonication, microwave, freeze-thawing, and enzymatic/chemical hydrolysis; (II) extraction techniques, such as solid-liquid extraction, ultrasound and microwave-assisted extraction, pressurized-liquid extraction, and supercritical CO2; (III) analytical methods, including total phenolic and flavonoid content assays and advanced chromatographic techniques like GC-MS, HPLC-DAD, and HPLC-MS. Key findings showed bead milling and chemical hydrolysis as effective cell disruption techniques, pressurized-liquid extraction and microwave-assisted extraction as promising efficient extraction methods, and HPLC-MS as the finest alternative for precise phenolic characterization. Unlike previous reviews, this study uniquely integrates both extractive and analytical approaches in one work, focusing exclusively on marine microalgae, a relatively underexplored area compared to freshwater species, offering actionable insights to guide future research and industrial applications.

Bioactivity-guided purification and characterization of antioxidant, anti-gout and anti-diabetic polyphenols from Panchkuta: A traditional food combination of (semi) arid regions

Desert ecosystems have thrived in part because of the contribution of local traditions, along with conventional knowledge passed down through generations. Panchkuta, a combination of five plants, including Prosopis cineraria, Cordia dichotoma, Capparis decidua, Acacia senegal, and Mangifera indica, is a classic example of ethnic desert food and how communities utilize existing food resources to ensure nutritional security. In this study, we determined the total phenolic content, total flavonoid content, antioxidant, anti-gout, and anti-diabetic activities of crude and purified (using Amberlite XAD7HP and Sephadex LH-20) fractions of Panchkuta. Further, the phenolic compounds (quercetin, tiliroside, myricetin-3-O-galactoside, and others) responsible for these activities were identified by analytical techniques, including TLC, LC-MS, and HR-MS. The purified fraction showing maximum antioxidant activity in DPPH assay (Fraction 16, IC50–156.65 ± 12.6 μg/ml) was validated for its cytoprotective and anti-lipid peroxidative effects in the in vitro cell culture system (murine fibroblast cells). Fraction 9 showed the maximum anti-gout activity in xanthine oxidase assay (IC50- 80.59 ± 9.89 μg/ml) and fraction 11 showed the maximum anti-diabetic activity in α-amylase assay (IC50 of 101.54 ± 14.7 μg/ml). The study reinforces the understanding of traditional knowledge, relevance to One Health, and authenticates the potential medicinal value of Panchkuta through scientific validation.

Polyphenolic characterization, nutritional and microbiological assessment of newly formulated semolina fresh pasta fortified with grape pomace

The application of an innovative patented technology on grape pomace allowed to obtain a food ingredient no matter the seasonality and with a great shelf-life thanks to its high polyphenolic content. Then, fresh pasta samples fortified with 5 and 10 g/100 g of this innovative ingredient were prepared and their polyphenolic content, nutritional and microbiological assessments were investigated. The two levels of fortification led to an increase of polyphenols content before and after pasta cooking, which resulted in an increased antibacterial activity. Only the fortification with 5 g/100 g of grape pomace powder determined a high level of Slowly Digestible Starch and a low predicted Glycaemic Index, highlighting a possible ‘limit effect’ for the fortification of semolina fresh pasta. Overall, results confirmed that the use of this concentration obtained with the employment of the new technology allowed the production of fresh pasta with better polyphenolic profile and nutritional value even after cooking.

Valorization of Date Fruit (Phoenix dactylifera L.) as a Potential Functional Food and Ingredient: Characterization of Fiber, Oligosaccharides, and Antioxidant Polyphenols.

The fruit of the date tree (Phoenix dactylifera L.) is increasingly recognized for its nutritional and functional value. This exotic fruit shows variable composition, influenced by factors such as variety, ripening stage, and climatic conditions. In this context, this study aimed to investigate the nutritional profile and the bioactive components, including phenolic compounds and oligosaccharides, in different varieties of dates from Saudi Arabia collected at the Tamr ripening stage. The HPLC-ESI-MS analysis identified a total of 15 phenolic compounds, principally phenolic acids and flavonoids. Among the varieties tested, Safawi exhibited the highest phenolic concentration (1132 µg/100 g dw). To the best of our knowledge, the oligosaccharide composition is described for the first time among different varieties, with Sukari showing the highest concentration (3.37 g/100 g dw). Moreover, the antioxidant capacity (DPPH, ABTS, and FRAP assays) was assessed following a solid-phase extraction (SPE) clean-up to remove interferents, especially sugars. These results provide valuable insights into the health-promoting properties of date fruit as a functional food and provide a foundation for further research into their industrial applications as functional ingredients.

Metabolic characterization and transcriptional profiling of polyphenols in Cannabis sativa L. inflorescences with different chemical phenotypes.

After the most comprehensive analysis of the phenolic composition in Cannabis reported to date, a total of 211 compounds were identified, phenolic profiles were able to discriminate cannabis varieties and a complex regulatory network for phenolics accumulation in Cannabis chemovars was highlighted. Female inflorescences of Cannabis sativa L. are plenty of secondary metabolites, of which flavonoids and phenolic acids have been investigated by far less than phytocannabinoids and terpenoids. Understanding the biochemical composition in phenylpropanoids of Cannabis inflorescences, the molecular basis of flavonoid synthesis and how their content can be modulated by specific transcription factors will shed light on the variability of this trait in the germplasm, allowing the identification of biologically active metabolites that can be of interest to diverse industries. In this work, an untargeted metabolomic approach via UHPLC-HRMS was adopted to investigate the composition and variability of phenylpropanoids in thirteen Cannabis genotypes differentiated for their profile in phytocannabinoids, highlighting that phenolic profiles can discriminate varieties, with characteristic, unique genotype-related patterns. Moreover, the transcription profile of candidate phenolics regulatory MYB and bHLH transcription factors, analyzed by RT-qPCR, appeared strongly genotype-related, and specific patterns were found to be correlated between biochemical and transcriptional levels. Results highlight a complex regulatory network for phenolic accumulation in Cannabis chemovars that will need further insights from the functional side.

Antioxidant Activity and UHPLC-MS/MS Characterization of Polyphenol and Nicotine Content in Nicotiana Glauca Leaf Extracts: A Comparative Study of Conventional and Deep Eutectic Solvent Extraction Methods.

The leaves of Nicotiana glauca (N. glauca; Solanaceae) plant are a known, major human health concern. This study investigated the antioxidant activity and polyphenols composition of aerial parts of N. glauca collected from its wild habitat in Jordan, using Methanol-Conventional (MC) and deep eutectic solvents (DES) extraction methods in addition to nicotine content determination using UHPLC. Our results showed that the MC extract contains fewer total phenols and flavonoid content than the 90% DES extract, (0.1194 ± 0.009 and 0.311 ± 0.020 mg/mL equivalent to gallic acid) and (0.01084 ± 0.005 and 0.928 ± 0.09 mg/mL equivalent to rutin), respectively. Moreover, this study showed that the prepared MC extract contain 635.07 ppm nicotine, while the 90% DES extract contain 1194.91 ppm nicotine. Extracts prepared using the MC and the DES methods exhibited weak antioxidant activities; the highest was a 33% inhibition rate (equivalent to ascorbic acid), obtained by the 90% DES extract,. The performed UHPLC-MS/MS analysis in this study also revealed the presence of variations in the detected compounds between the two extraction methods. Furthermore, this study found that environmentally friendly DES extraction of N. glauca produced higher phenol and flavonoid content than the MC method; this highlights the superior efficiency and environmental benefits of sustainable chemistry methods for extracting valuable phytoconstituents.

Characterization of wine polyphenols with a carbon/nanoparticle TiO2 electrode

The positive effects of polyphenolic compounds on the sensory characteristics of wine and human health indicate a great need for a simple, fast and easily accessible method to determine the content of polyphenols in wine. The aim of this study is the electrochemical characterization of polyphenolic compounds in natural wine samples using a modified carbon paste electrode with TiO2 nanoparticles (MCPE/npTiO2) and improved voltammo­gram processing to obtain indicative data for polyphenols. The most marked influence of the modification of CPE with TiO2 nanoparticles was an increased sensitivity to electrooxi­dation, which is reflected in an increase of the anodic peak current. Of the five tested poly­phenols, i.e., gallic acid (GA), caffeic acid (CA), catechin (CT), quercetin (QV) and resveratrol (RE), the current maximum of the first two oxidation peaks increased only for GA by a factor of about 2 and for CT by a factor of about 1.5. Of the three red wines, Vranac (VR), Merlot (ME), Cabernet Sauvignon (CS); three white wines, Graševina (GR), Temjanika (TE), Char­donnay (CS) and one rose wine, Belrose Mediterranée Rosé (RO) tested, an increase by a factor of about 2.5 was observed for two red wines (VR, CS) and by a factor of 1.5 for one red wine (ME), one white wine (GR), and the rose wine (RO), while no increase in the current signal was observed for one of the white wines (CS). The most significant increase in the voltam­metric signal of GA at the MCPE/npTiO2 compared to other studied polyphenols can be explained by its higher affinity for adsorption on TiO2 nanoparticles. The investigated modified electrode provides an improved, linear and reproducible voltammetric response in wine samples. Consequently, MCPE/npTiO2 represents a good basis for the further deve­lopment of an integrated sensor/data system with the possibility of a broader application for the detection of polyphenols, especially GA, as an aroma and visually relevant parameter in winemaking.

Characterization of Polyphenols in Baru (Dipteryx alata) and Their Effects on Palmitic Acid-Induced Oxidative Stress in Rat Aortic Vascular Smooth Muscle Cells

Characterization of Polyphenols in Baru (Dipteryx alata) and Their Effects on Palmitic Acid-Induced Oxidative Stress in Rat Aortic Vascular Smooth Muscle Cells

Purification, Characterization, and Anti-Inflammatory Potential of Free and Bound Polyphenols Extracted from Rosa roxburghii Tratt Pomace.

Rosa roxburghii Tratt pomace (RRTP), an underutilized byproduct, is rich in polyphenol compounds. This study aimed to further explore the purification, characterization, anti-inflammatory activities, and underlying molecular mechanisms of free polyphenols (RRTP-FP) and bound polyphenols (RRTP-BP) from RRTP. The results indicated that AB-8 macroporous resin emerged as the preferred choice for subsequent separation and purification. The purities of purified RRTP-FP (P-RRTP-FP) and purified RRTP-BP (P-RRTP-BP) increased by 103.34% and 66.01%, respectively. Quantitative analysis identified epigallocatechin, epicatechin, and ellagic acid as the main phenolic compounds in P-RRTP-FP. In P-RRTP-BP, the primary phenolic compounds were ellagic acid, epicatechin, and gallic acid. In vitro antioxidant assays demonstrated the superior DPPH and ABTS radical scavenging activities of P-RRTP-FP and P-RRTP-BP compared to vitamin C. Treatment with P-RRTP-FP and P-RRTP-BP reduced nitric oxide (NO) and reactive oxygen species (ROS) production, mitigated the decline in cellular membrane potential, and significantly downregulated the mRNA expression of pro-inflammatory cytokines and inducible nitric oxide synthase (iNOS) in lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages. Additionally, P-RRTP-FP and P-RRTP-BP inhibited the phosphorylation of pertinent proteins in the nuclear factor kappa-B (NF-κB) and mitogen-activated protein kinase (MAPK) signaling pathways. This finding suggests potential utility of RRTP-derived polyphenols as anti-inflammatory agents for managing severe inflammatory conditions.

Characterization of polyphenols and carbohydrates exuded by Phaeodactylum tricornutum diatom grown under Cu stress

This study is focused on analysing polyphenols and carbohydrates released by Phaeodactylum tricornutum (P. tricornutum) diatoms cultured in natural seawater enriched with sublethal and lethal Cu doses. Cu concentrations of 0.31, 0.79 and 1.57 µM reduced cell densities by 37, 82 and 91%, respectively, compared to the control. The total sum of all identified polyphenols and total carbohydrates released by cells grown under lethal Cu levels increased up to 18.8 and 107.4 times, respectively, compared to data from a control experiment. Four different in vitro assays were used to estimate the antioxidant activities of the extracellular compounds: 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical inhibition, cupric ion reducing antioxidant capacity (CUPRAC), ferric reducing antioxidant power and Cu complexing ability (CCA). The highest antioxidant activities were observed in the Cu lethal treatments, where the CCA assay exhibited a greater increase (up to 32.2 times higher than that found in the control experiment) to reduce the concentration of free Cu in the medium and its toxicity. The presence of Cu stimulated the release of polyphenols and carbohydrates to the medium as a detoxification mechanism to survive under lethal levels of Cu regulating its speciation.

Assessing the stability of polyphenol content in red rooibos herbal tea using traditional methods and high-resolution mass spectrometry: Implications for studying dietary interventions in preclinical rodent studies

Preclinical rodent models are used to examine the relationship between tea consumption and bone health, where tea is available for rodents and typically replaced weekly. However, the extent to which the tea polyphenols change over time remains uncertain, despite its importance in preparing tea during preclinical rodent trials. Using an untargeted molecular approach, we applied a liquid chromatography quadrupole-time-of-flight mass spectrometry (LC-QTOFMS) system to assess the molecular profile of red rooibos teas throughout a 6-day aging period. We found a significant, 3-fold decrease of polyphenols involved in bone metabolism, including m-coumaric acid, catechin derivatives and courmaroyl tartaric acid over 6 days, likely due to photochemical decomposition and autooxidation within tea extracts. Using a novel untargeted workflow for polyphenol characterization, our findings revealed the complexity of polyphenols in red rooibos teas that can inform the evidence-based decisions of how often to change teas during in vivo rodent trials.

Polyphenolic characterization and biological assessment of Acacia nilotica (L.) wild. Ex delilie: An In vitro and In vivo appraisal of wound healing potential

Ethnopharmacological relevanceAcacia nilotica (L.) Wild. Ex Delilie is a shrub with significant ethnomedicinal stature. Therefore, in the undertaken study, its wound healing attributes are determined. Aim of the studyThe current study provided evidence of the traditional use of A. nilotica species and conferred A. nilotica bark extract as a potent candidate for wound healing agents. Materials & methodsA. nilotica leaves extract (ANL-E); A. nilotica bark extract (ANB-E), and A. nilotica stem extract (ANS-E) were prepared using methanol-chloroform (1:1). Phytochemical analysis was performed using gallic acid equivalent (GAE) total phenolic content (TPC), quercetin equivalent (QE) total flavonoid content (TFC) assays and High-performance liquid chromatography (HPLC). In vitro antioxidant potential (free radical scavenging activity (FRSA), total antioxidant capacity (TAC), and ferric reducing antioxidant power (FRAP) assay), antibacterial activity (broth microdilution method) and hemolytic analysis was carried out. Wound healing proficiency of ANB-E was determined by wound excision model followed by estimating hydroxyproline content and endogenous antioxidant markers. ResultsMaximum phenolic and flavonoid content were depicted by ANB-E i.e., 50.9 ± 0.34 μg gallic acid equivalent/mg extract and 28.7 ± 0.13 μg quercetin equivalent/mg extract, respectively. HPLC analysis unraveled the presence of a significant amount of catechin in ANL-E, ANB-E and ANS-E (54.66 ± 0.02, 44.9 ± 0.004 and 31.36 ± 0.02 μg/mg extract) respectively. Highest percent free radical scavenging activity, total antioxidant capacity, and ferric reducing action power (i.e., 93.3 ± 0.42 %, 222.10 ± 0.76, and 222.86 ± 0.54 μg ascorbic acid equivalent/mg extract) were exhibited by ANB-E. Maximum antibacterial potential against Staphylococcus aureus was exhibited by ANB-E (MIC 12.5 μg/ml). Two of the extracts i.e., ANL-E and ANB-E were found biocompatible with less than 5 % hemolytic potential. Based upon findings of in vitro analysis, ANB-E (10, 5, and 2.5 % w/w, C1, C2, and C3, respectively) was selected for evaluating its in vivo wound healing potential. Maximum contraction of wound area and fastest epithelization i.e., 98 ± 0.05 % and 11.2 ± 1.00 (day) was exhibited by C1. Maximum hydroxyproline content, glutathione, catalase, and peroxidase were demonstrated by C1 i.e., 15.9 ± 0.52 μg/mg, 9.3 ± 0.17 mmol/mg, 7.2 ± 0.17 and 6.2 ± 0.14 U/mg, respectively. Maximal curbed lipid peroxidation i.e., 0.7 ± 0.15 mmol/mg was also depicted by C1. ConclusionsIn a nutshell, the current investigation endorsed the wound healing potential of ANB-E suggesting it to be an excellent candidate for future studies.

Polyphenolic Characterization and Anti-Inflammatory Effect of In Vitro Digested Extracts of Echinacea purpurea L. Plant Parts in an Inflammatory Model of Human Colon Cells.

Echinacea purpurea L. (EP) preparations are globally popular herbal supplements known for their medicinal benefits, including anti-inflammatory activities, partly related to their phenolic composition. However, regarding their use for the management of inflammation-related intestinal diseases, the knowledge about the fate of orally ingested constituents throughout the human gastrointestinal tract and the exposition of in vitro digested extracts in relevant inflammatory models are unknown. This study investigated for the first time the impact of in vitro gastrointestinal digestion (INFOGEST) on the phenolic composition and anti-inflammatory properties of EP extracts from flowers (EF), leaves (EL), and roots (ER) on IL-1β-treated human colon-derived CCD-18Co cells. Among the seven hydroxycinnamic acids identified using HPLC-UV-MS/MS, chicoric and caftaric acids showed the highest concentrations in EL, followed by EF and ER, and all extracts exerted significant reductions in IL-6, IL-8, and PGE2 levels. After digestion, despite reducing the bioaccessibility of their phenolics, the anti-inflammatory effects were preserved for digested EL and, to a lesser extent, for EF, but not for digested ER. The lower phenolic content in digested EF and ER could explain these findings. Overall, this study emphasizes the potential of EP in alleviating intestinal inflammatory conditions and related disorders.

Characterization and stability assessment of polyphenols bound to Lycium barbarum polysaccharide: Insights from gastrointestinal digestion and colon fermentation

Polysaccharides and polyphenols are biologically active components that coexist in Lycium barbarum fruit, and there may be interactions between them that affect the release of each other. In this study, polyphenols bound to L. barbarum polysaccharide (LBP) were characterized, and the stability of bound phenolics (BP) was assessed by gastrointestinal digestion and colon fermentation. The results showed that a total of 65 phytochemicals such as flavonoids, phenolic acids, and coumarins were identified by UPLC-MS/MS. Quantitative analysis revealed that the major phenolic constituents were rutin, p-coumaric acid, catechin, ferulic acid, protocatechuic acid, and gallic acid, and their contents were 58.72, 24.03, 14.24, 13.28, 10.39, and 6.7 mg GAE/100 g DW, respectively. The release of BP by gastric digestion and gastrointestinal digestion was 9.67 % and 19.39 %, respectively. Most polyphenols were greatly affected by gastric digestion, while rutin was released in small intestine. The BP were fully released (49.77 %) and metabolized by gut microorganisms, and a considerable number of intermediates and end-products were detected, such as phloroglucinol, phenylacetic acid, and phenyllactic acid. Microbiomics data emphasized the positive impact of LBP on gut bacteria of Bacteroides, Parabacteroides, and Clostridioides. These findings could deepen our understanding of the bioavailability and biological fate of BP and also provide reference data for nutrient release and utilization of L. barbarum as a whole.

Phenological Variations in the Content of Polyphenols and Triterpenoids in Epilobium angustifolium Herb Originating from Ukraine.

The composition of secondary metabolites undergoes significant changes in plants depending on the growth phase and the influence of environmental factors. Therefore, it is important to determine the harvesting time of plant material for the optimum secondary metabolite profile and therapeutic activity of the primary material. The shoots of Epilobium angustifolium are used as a healing tea due to the presence of polyphenolic compounds. The aim of this study was to assess the composition of phenolic compounds and triterpenoid saponins in E. angustifolium leaves and flowers and to estimate the dynamics of their content depending on the flowering phase. Qualitative and quantitative characterisation of polyphenols and triterpenoids in E. angustifolium samples from Ukraine of three flowering phases were performed using the high-performance liquid chromatography photo diode array (HPLC-PDA) method. During the present study, 13 polyphenolic compounds and seven triterpenoids were identified in the plant material. It was noted that the largest content and the best polyphenol profile was in late flowering. The most important polyphenolic compounds in the plant material were chlorogenic acid, hyperoside, isoquercitin, and oenothein B. The triterpenoid profile was at its maximum during mass flowering, with corosolic and ursolic acids being the dominant metabolites. The results of the analysis revealed that the quantity of many of the tested metabolites in the raw material of E. angustifolium is dependent on the plant organ and flowering phase. The largest content of most metabolites in the leaves was in late flowering. In the flowers, the quantity of the metabolites studied was more variable, but decreased during mass flowering and increased significantly again in late flowering. The results show that E. angustifolium raw material is a potential source of oenothein B and triterpenoids.

Comprehensive characterization of the overlooked residue generated during roselle calyxes brewing with potential use as functional ingredient

A significant quantity Hibiscus sabdariffa L. calyxes is generated as a by-product during the decoction process commonly used for roselle beverage preparation. We conducted an extensive characterization of polyphenols, organic acids, and antioxidant potential in roselle calyxes, decoction, and their by-product. Roselle calyxes were found to be a rich source of diverse polyphenols, including delphinidin sambubioside, caffeoylquinic acids, hibiscus acid, and citric acid as major components. Importantly, we extracted a significant proportion of these bioactive compounds during the decoction process, resulting in a polyphenol-rich beverage. The used calyces (decoction by-product) retained from 23 % to 140 % of the extractable polyphenols and organic acids found in roselle calyxes. Additionally, due to the leaching of hydrophobic components like soluble dietary fiber and extractable polyphenols and organic acids, the roselle by-product was enriched with non-extractable constituents attached to dietary fiber (126 % - 272 %). Therefore, roselle calyxes and their decoction by-products emerge as promising sources of polyphenols with the potential for use in dietary supplements, alongside the commonly consumed roselle decoction.

Supercritical CO2 and ultrasound extraction and characterization of lipids and polyphenols from Phacelia tanacetifolia Benth. pollen

The phytochemical profile of Phacelia tanacetifolia Benth. pollen was characterized in terms of phenolic compounds and fatty acids. Fatty acids were extracted from pollen using supercritical CO2 and characterized by GC-FID. The main constituents found in the extract were palmitic and oleic acid accounting for 50% of all fatty acids. The polyphenols were extracted from exhausted material after CO2 extraction using ultrasound-assisted extraction (UAE) and aqueous methanol solution as solvent. The major compounds identified in the UAE extract by UHPLC-DAD-QqTOF-MS belonged to flavonoid glycosides (isorhamnetin O-rutinoside-rhamnoside) and phenolamides (di-p-coumaroylputrescine, N′,N''-di-p-coumaroyl-N'''-caffeoylspermidine, and N′,N'',N'''-tri-p-coumaroylspermidine isomers). The UAE extraction of flavonoids and phenolamides was optimized considering time, temperature, methanol percentage and solvent-solid ratio. The determined optimized conditions were: temperature of 66.1 °C, time of 44.9 min, solvent-solid ratio of 49.9 mL/g and 70.1% of aqueous methanol solution as a solvent. The statistically significant impact on the extraction of the analyzed compounds had the solvent-solid ratio and methanol percentage as well as interactions between those two parameters. The optimized conditions may be useful for efficient extraction of fractions rich in bioactive compounds e.g. N′,N'',N'''-tri-p-coumaroylspermidine with proven biological potential.

Characterization of Polyphenols and Volatile Compounds from Understudied Algerian Pallenis spinosa by HS-SPME-GC-MS, NMR and HPLC-MSn Approaches

Pallenis spinosa (L.) Cass. is a widespread plant in the Mediterranean region. Traditionally, it is used as a medicinal species to treat several ailments, from inflammation to skin injuries. Although the phytochemical content of this plant has already been investigated, there is currently limited data on Algerian P. spinosa. In this work, we focused on volatile compounds and non-volatile secondary metabolites extracted using HS-SPME and methanol from the aerial parts of P. spinosa collected from Northeast Algeria. Volatile constituents were analyzed by GC-MS, while non-volatile compounds were analyzed by NMR and HPLC-MSn. In total, 48 volatile compounds were identified, including sesquiterpene hydrocarbons (65.8%), monoterpene hydrocarbons (16.9%), and oxygenated monoterpenes and sesquiterpenes (8.3% and 6.5%, respectively). β-Chamigrene (16.2%), α-selinene (12.8%), β-pinene (10.6%), and β-caryophyllene (9.2%) were assessed as the main constituents. Concerning non-volatile metabolites, 23 polyphenols were identified (7.26 mg/g DW), and phenolic acids were predominant (5.83 mg/g DW). Tricaffeoylhexaric acid (1.76 mg/g DW), tetracaffeoylhexaric acid (1.41 mg/g DW), 3,5-dicaffeoylquinic acid (1.04 mg/g DW), caffeoyl dihexoside (0.35 mg/g DW), and chlorogenic acid (0.29 mg/g DW) were the most abundant ones. Several known flavonoids, such as tricin and patuletin glycosides, kaempferol, and apigenin, were also identified, and myricetin hexoside was detected in P. spinosa for the first time. Overall, our work is the first to report an exhaustive characterization of volatile and non-volatile secondary metabolites from Algerian P. spinosa. The results represent a step forward in revealing the chemistry of this widespread plant species. Furthermore, they may contribute to rationalizing its traditional medicinal applications and preserve the biodiversity of Algerian flora.

Extraction, characterization of polyphenols from certain medicinal plants and evaluation of their antioxidant, antitumor, antidiabetic, antimicrobial properties, and potential use in human nutrition.

Dietary medicinal plants are among the most sought-after topics in alternative medicine today due to their preventive and healing properties against many diseases. This study aimed to extract and determine the polyphenols from indigenous plants extracts, i.e., Mentha longifolia, M. arvensis, Tinospora cordifolia, Cymbopogon citratus, Foeniculum vulgare, Cassia absus, Camellia sinensis, Trachyspermum ammi, C. sinensis and M. arvensis, then evaluate the antioxidant, cytotoxicity, and antimicrobial properties, besides enzyme inhibition of isolated polyphenols. The antioxidant activity was evaluated by DPPH, Superoxide radical, Hydroxyl radical (OH.), and Nitric oxide (NO.) scavenging activity; the antidiabetic activity was evaluated by enzymatic methods, and anticancer activity using MTT assay, while the antibacterial activity. The results showed that tested medicinal plants' polyphenolic extracts (MPPE) exhibited the most significant antioxidant activity in DPPH, hydroxyl, nitric oxide, and superoxide radical scavenging methods because of the considerable amounts of total polyphenol and flavonoid contents. UHPLC profile showed twenty-five polyphenol complexes in eight medicinal plant extracts, categorized into phenolic acids, flavonoids, and alkaloids. The main polyphenol was 3-Feroylquinic acid (1,302 mg/L), also found in M. longifolia, C. absus, and C. sinensis, has a higher phenolic content, i.e., rosmarinic acid, vanillic acid, chlorogenic acid, p-coumaric acid, ferulic acid, gallic acid, catechin, luteolin, 7-O-neohesperideside, quercetin 3,7-O-glucoside, hesperidin, rutin, quercetin, and caffeine in the range of (560-780 mg/L). At the same time, other compounds are of medium content (99-312 mg/L). The phenolics in C. sinensis were 20-116% more abundant than those in M. longifolia, C. absus, and other medicinal plants. While T. cordifolia is rich in alkaloids, T. ammi has a lower content. The MTT assay against Caco-2 cells showed that polyphenolic extracts of T. ammi and C. citratus had maximum cytotoxicity. While M. arvensis, C. sinensis, and F. vulgare extracts showed significant enzyme inhibition activity, C. sinensis showed minor inhibition activity against α-amylase. Furthermore, F. vulgare and C. sinensis polyphenolic extracts showed considerable antibacterial activity against S. aureus, B. cereus, E. coli, and S. enterica. The principal component analysis demonstrated clear separation among medicinal plants' extracts based on their functional properties. These findings prove the therapeutic effectiveness of indigenous plants and highlight their importance as natural reserves of phytogenic compounds with untapped potential that needs to be discovered through advanced analytical methods.

Characterization of Polyphenols from Chenopodium botrys after Fractionation with Different Solvents and Study of Their In Vitro Biological Activity.

In the present work, we have investigated the polyphenolic composition of Chenopodium botrys from Bulgaria. The polyphenols were fractionated with solvents of varying polarity (n-hexane, chloroform, ethyl acetate, and n-butanol). The fractions were analyzed by HPLC-PDA and UHPLC-MS. The ethyl acetate fraction contained mono- and di-glycosides of quercetin, di-glycosides of kaempferol, and isorhamnetin and monoglycosides of hispidulin and jaceosidine. We found quercetin triglycosides in the butanol fraction. The ethyl acetate and butanol fractions contained 168.82 mg/g Extr and 67.21 mg/g Extr of quercetin glycosides, respectively. The main components of the polyphenolic complex in C. botrys were 6-methoxyflavones (355.47 mg/g Extr), which were found in the chloroform fraction. The flavonoids pectolinarigenin, demethylnobiletin, and isosinensetin, and the glycosides of quercetin (triglycosides, acylglycosides), kaempferol, isorhamnetin, hispidiulin, and jaceosidine, were discovered and reported in Chenopodium botrys for the first time. We used in vitro methods to assess the biological activity against oxidative stress (hydrogen peroxide scavenging activity (HPSA) and hydroxyl radical scavenging activity (HRSA)), nitrosative stress (nitric oxide scavenging activity (NOSA)), anti-inflammatory activity (IAD inhibition), and anti-tryptic activity (ATA). Quercetin mono- and di-glycosides exhibited greater HPSA and HRSA (IC50 = 39.18, 105.03 µg/mL), while 6-methoxyflavones had a greater NOSA (IC50 = 146.59 µg/mL). The same components showed the highest ATA (IC50 ranging from 116.23 to 202.44 µg/mL).