Search Results (5,359)

Wolffia globosa, the world’s smallest flowering plant, is a sustainable protein- and phytochemical-rich food source with emerging neuroprotective potential. This study presents the first comprehensive characterization that integrates extraction optimization, multitarget bioactivity profiling, drug synergy assessment, high-resolution metabolomics, molecular docking, and in vivo validation in Drosophila melanogaster. Ethanol-based extraction was optimized using a Box–Behnken design and response surface methodology (RSM), yielding optimal conditions of 32% ethanol, 1:40 (g/mL) solid-to-solvent ratio, 60 °C, and 28 min. The optimized extract exhibited a total phenolic content (TPC) of 15.17 ± 0.27 mg GAE/g DW, a total flavonoid content (TFC) of 22.34 ± 1.71 mg QE/g DW, and an exceptional ORAC antioxidant activity of 4374.33 ± 395.65 µmol TE/g DW. Potent and selective BACE-1 inhibition was observed (IC₅₀: 1.59 ± 0.11 mg/mL), alongside moderate AChE (IC₅₀: 7.42 mg/mL) and BChE (IC₅₀: 6.55 mg/mL) inhibition. Synergistic interactions with donepezil were confirmed by the Chou–Talalay combination index method, with combined AChE, BChE, and BACE-1 inhibitions reaching 60.3%, 77.0%, and 82.2%, respectively—substantially exceeding theoretical additive values. Multi-platform metabolite profiling by HPLC-QTOF-MS/MS and LC-ESI-MS/MS identified luteolin, apigenin, caffeic acid, isovitexin, schaftoside, and pinolenic acid as the principal bioactives. Molecular docking predicted favorable binding of luteolin (delta-G: −10.24 kcal/mol, AChE) and naringenin (delta-G: −8.76 kcal/mol, BACE-1) against crystal structures of human AChE (PDB: 7E3H), BChE (PDB: 4TPK), and BACE-1 (PDB: 6EQM). In vivo, the W. globosa extract significantly improved locomotor performance and suppressed brain BACE-1 activity in an Drosophila AD model over 28 days. These findings establish W. globosa as a multifunctional nutraceutical candidate for neurodegenerative disease prevention. Full article

Ocimum gratissimum L. (OGL) is a medicinal plant known for various therapeutic properties, yet its potential in genetic-level skin anti-aging remains underexplored. This study aimed to evaluate the phytochemical profiles, antioxidant activities, and anti-aging potential of OGL leaf extracts obtained via aqueous boiling (OGL-B) and ethanolic maceration (OGL-M). Total phenolic and flavonoid contents were quantified using colorimetric assays, while rutin, catechin, and quercetin were analyzed using HPLC. Biological activities were assessed using antioxidant models, tyrosinase inhibition, and B₁₆F₁₀ anti-melanogenesis assays, while skin regenerative potential was evaluated through collagen biosynthesis and expression of longevity-related genes (SIRT1, FOXO1, TERT, and KLOTHO) in human skin fibroblasts. OGL-B provided a significantly higher extraction yield (14.70%) and superior multifunctional antioxidant capacity than OGL-M. OGL-M showed higher phenolic content and cell-free tyrosinase inhibition, whereas OGL-B demonstrated greater cellular anti-melanogenic efficacy and more than twofold higher collagen stimulatory effects than L-ascorbic acid. OGL-B significantly upregulated mRNA expression (271.25 ± 0.21%), exceeding the reference activator resveratrol, and enhanced fibroblast viability to 121.53 ± 3.85% at 1 mg/mL, indicating high biocompatibility. These findings suggest that OGL-B is a multifunctional extract capable of modulating longevity-related pathways, with potential for advanced anti-aging food supplement and cosmeceutical applications. Full article

Watermelon (Citrullus lanatus) is a widely consumed fruit rich in carotenoids, phenolic compounds, vitamins, and other bioactive constituents associated with antioxidant and health-promoting properties. The present study aimed to develop novel low-sugar watermelon jelly formulations enriched with pectin and single-cell protein (SCP), and to evaluate their bioactive and functional properties, including the antioxidant and enzyme inhibitory activities of their 70% ethanol (70% EtOH) extracts. Six jelly formulations (W1–W6) differing in pectin and SCP supplementation were prepared using concentrated watermelon pulp processed under low-temperature vacuum conditions to preserve thermolabile bioactive compounds. The total phenolic content (TPC) and total flavonoid content (TFC) were determined in the 70% EtOH extracts using spectrophotometric methods. TPC remained relatively stable among the investigated formulations, with the highest value recorded for sample W6, which contained 0.58 g pectin and 0.215 g SCP (1.37 mg gallic acid equivalents (GAE)/mL extract). The highest TFC was observed for sample W5, corresponding to the formulation supplemented with 0.5 g pectin and 0.300 g SCP (5.87 mg quercetin equivalents (QE)/mL extract). Antioxidant activity, evaluated using ABTS, DPPH, and FRAP assays, demonstrated considerable antioxidant potential in all 70% EtOH extracts. The strongest antioxidant activity was observed in pectin-rich samples, with ABTS and FRAP values reaching 89.06 mg Trolox equivalents (TE)/mL extract and 8.99 mg ascorbic acid equivalents (AAE)/mL extract, respectively. The potential carbohydrate-hydrolyzing enzyme inhibitory activity of the 70% EtOH extracts was assessed through α-amylase and α-glucosidase inhibition assays. The formulation containing both pectin and SCP exhibited the strongest α-amylase inhibitory activity (58.93%), whereas the highest α-glucosidase inhibition (60.54%) was observed in the pectin-containing formulation without SCP supplementation. These findings indicate that both pectin and SCP significantly influenced the nutritional and bioactive properties of the developed watermelon jelly formulations. Overall, the results demonstrate that enrichment with pectin and SCP significantly enhances the nutritional and bioactive properties of watermelon jelly formulations. These findings highlight indicate that pectin and SCP were associated with differences in the bioactive and functional properties of the developed watermelon jelly formulations. Overall, the results demonstrate the potential of pectin- and SCP-enriched watermelon jellies as sustainable functional food products with promising antioxidant and enzyme inhibitory properties. Full article

Despite previous phytochemical investigations of Medicago marina L., comparative information on the phenolic composition and multifunctional bioactivities of extracts obtained with different solvents remains limited. This study compared water and methanol extracts prepared by ultrasound-assisted extraction using targeted liquid chromatography–tandem mass spectrometry (LC-MS/MS), antioxidant assays, and enzyme inhibitory assays. The extracts had comparable total phenolic contents, whereas the methanol extract was richer in flavonoids. LC-MS/MS profiling showed distinct phenolic compositions between the investigated extraction batches: the water extract contained higher levels of ferulic acid, apigenin, and several hydroxybenzoic acid derivatives, whereas the methanol extract was enriched in hyperoside, hesperidin, chlorogenic acid, and other flavonoid derivatives. On an extract-mass basis, the water extract showed higher 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical-scavenging activity, ferric reducing antioxidant power (FRAP), and phosphomolybdenum antioxidant capacity, whereas the methanol extract showed higher ferrous ion-chelating and enzyme inhibitory activity values. Yield normalization to the initial dry plant material altered the relative performance of the two extraction batches in several assays because of the higher water extraction yield. These findings show that extract-level enrichment and yield-normalized activity provide complementary perspectives on the phytochemical and biological characteristics of the specific M. marina extraction batches investigated. Full article

To optimize the varietal structure of fruit crops in arid environments and screen apricot-plum varieties suitable for cultivation in the Aksu region, a systematic agronomic evaluation was conducted to comprehensively analyze the phenotypic diversity and fruit quality traits of seven apricot-plum cultivars in Aksu, Xinjiang. In 2025, seven-year-old trees cultivated under identical and standardized agronomic conditions were selected as experimental materials. Key agronomic parameters, including flowering and fruiting phenology, floral and foliar morphological traits, fruit appearance properties, and internal nutritional profiles (soluble solids, soluble sugars, titratable acids, vitamin C, flavonoids, and total phenolics) were systematically measured. Correlation analysis and principal component analysis (PCA) were applied to quantitatively integrate these multi-dimensional datasets, identify key factors driving agronomic variation, and evaluate varietal adaptability to the arid climate. The results revealed a high degree of phenotypic diversity and significant variations in agronomic performance among the seven cultivars. Specifically, ‘Weihou’ recorded the highest single-fruit weight (111.27 g), indicating superior physical development, whereas ‘Konglongdan’ achieved the maximum soluble solid content (21.43%) and vitamin C level (96.41 mg/100 g), exhibiting exceptional nutritional quality. Despite these outstanding individual traits in specific cultivars, evaluating overall adaptability requires a holistic approach. PCA effectively captured 93.84% of the total variance through five extracted principal components, successfully modeling the interrelationships among vegetative vigor, reproductive morphology, and fruit chemical profiles. Based on the comprehensive agronomic scores, the varieties ranked as follows: ‘Fengweihuanghou’ > ‘Weihou’ > ‘Hongtianerong’ > ‘Konglongdan’ > ‘Weidi’ > ‘WeiWang’ > ‘Fengweimeigui’. Our findings identify ‘Fengweihuanghou’ as the most superior cultivar, with optimal agronomic adaptability and fruit quality balance under arid conditions. This study suggests that ‘Fengweihuanghou’ is a highly promising cultivar for large-scale cultivation in the Aksu region, providing a valuable reference for optimizing the local crop varietal structure, although further multi-year and multi-location validations are warranted. Full article

The development of sustainable and efficient sample-preparation strategies remains a key challenge in analytical workflows for plant-derived bioactive compounds. In this study, choline chloride (ChCl)-based natural deep eutectic solvents (NADES) were systematically investigated as green extraction media and combined with chemical and enzyme assays to identify the bioactive effect of phenolic compounds from Bellevalia dubia, an understudied Mediterranean plant. A design of experiments approach was applied to evaluate the influence of hydrogen bond donors (lactic, malic, and citric acid) and their molar ratios on extraction performance. Among the tested systems, the ChCl–malic acid (MA) mixture at a molar ratio of 1:2.5 (precisely 1:2.47) was identified as the best overall solution according to the desirability approach, which integrated both phytochemical and enzyme-inhibitory responses. This system also showed strong enzyme-inhibitory effects compared to conventional extractants (water and 80% methanol). To identify the extract composition, a suspect screening workflow was applied based on ultra-high-performance liquid chromatography hybrid quadrupole Orbitrap mass spectrometry (UHPLC–q-Orbitrap-MS). Twenty-one compounds were detected and annotated, mainly belonging to the flavonoid class. Principal component analysis (PCA) showed that ChCl-MA extracts were discriminated, particularly at molar ratio 1:2.5, from the rest, indicating a distinct chemical and bioactivity profile. PCA further suggested a possible association of the ChCl-MA 1:2.5 system with vitexin and AChE inhibition, which requires further investigation. Overall, the proposed NADES-based approach offers a tunable, promising, and potentially more sustainable alternative for the extraction of bioactive compounds from plant-based sources. Full article

Agricultural and agro-industrial byproducts from tropical crops represent abundant sources of structurally bound and freely extractable bioactive compounds. However, variability in feedstock composition, the incomplete characterization of microbial transformations, and limited translation beyond chemical antioxidant assays constrain their development as standardized ingredients. This review critically evaluates metabolomics-guided fermentation as a strategy for converting tropical agricultural byproducts into functional and circular products. It covers fruit-processing residues, coffee and cocoa byproducts, cereal bran, cassava-processing residues, coconut and oilseed materials, and selected medicinal-crop tissues. Solid-state, submerged, lactic-acid-bacterial, yeast, filamentous fungal, Bacillus-based, and mixed-culture fermentations are compared. Particular attention is given to cell-wall deconstruction, phenolic release, flavonoid deglycosylation, phenolic-acid conversion, carotenoid stability, peptide generation, fiber modification, organic-acid production, antinutrient reduction, and undesirable metabolite formation. Analytical strategies based on untargeted and targeted LC-MS, GC-MS, NMR, volatilomics, peptidomics, and multiomics integration are assessed, together with experimental design, quality control, annotation confidence, and statistical validation. A substrate–microorganism–process–metabolite–function framework and a six-level evidence-grading system are proposed to distinguish exploratory chemical findings from mechanistically validated and industrially translatable outcomes. Applications in functional foods, nutraceutical ingredients, natural preservatives, animal feed, active packaging, agricultural inputs, and cascade biorefineries are evaluated alongside microbial safety, mycotoxins, biogenic amines, contaminants, regulatory classification, life-cycle assessment, and techno-economic feasibility. The central conclusion is that fermentation can become a programmable platform for tropical byproduct valorization only when metabolite-resolved characterization, causal validation, process standardization, safety, and sustainability are developed as an integrated pipeline. Full article

Breast cancer remains the most prevalent malignancy among middle-aged and elderly women with underlying health conditions, with triple-negative breast cancer (TNBC) representing a notably aggressive subtype associated with poorer clinical outcomes. Prunus africana (P. africana) has been reported to contain bioactive compounds with notable antioxidant and anticancer properties. Bark extracts obtained using water, as well as single (100%) and binary (50% v/v) solvents of methanol and ethanol (MeOH-100, EtOH-100, MeOH-50, and EtOH-50), were evaluated for their antioxidant activity, apoptosis-inducing effects, and modulation of signaling pathways in selected breast cancer cell lines. Antioxidant activity was measured via FRAP, DPPH radical scavenging, and TEAC assays; cytotoxic activity and pathway modulation were assessed using MTS, human apoptosis, and phospho-RTK array assays. While the EtOH-100 extract presented the lowest extraction yield of 3.22%, it showed the highest total phenolic (505.96 mg GAE/g) and total flavonoid (2.53 mg RU/g) contents and antioxidant activities (2.25 mM Fe²⁺/g and 311.68 mM TE/g), and the lowest EC₅₀ (0.21 mg/mL), comparable to ascorbic acid (0.18 mg/mL). EtOH-100 also notably induced apoptosis in HCC1806 as shown by increased cleaved caspase-3, HSP60, and HSP27 expression, and sustained catalase levels. Phospho-RTK array analysis revealed early silencing of HGF and transient upregulation of EGF and Insulin R following EtOH-100 extract treatment, suggesting modulation of oncogenic signaling. These findings highlight the therapeutic potential of P. africana, particularly the EtOH-100 extract, as a viable treatment for TNBC. Full article

Bee products, including propolis, royal jelly, honey, bee pollen, and bee bread, are natural functional foods recognized for their nutritional value and health-promoting properties. In the present study, propolis, royal jelly, honey, bee pollen, and bee bread collected from Mersin, one of Türkiye’s major beekeeping regions, were comprehensively characterized in terms of their phytochemical composition, antioxidant potential, enzyme inhibition effects, and physicochemical properties. The antioxidant capacities of the samples were evaluated by determining total phenolic content (TPC), total flavonoid content (TFC), and their radical scavenging and reducing abilities using the DPPH, ABTS, and DMPD radical scavenging assays and CUPRAC, FRAP, and Fe³⁺-reducing assays. Enzyme inhibition effects were assessed against human carbonic anhydrase I and II (hCA I and hCA II), acetylcholinesterase (AChE), butyrylcholinesterase (BChE), α-glycosidase, and α-amylase. Furthermore, the phytochemical profiles of all bee products were determined by LC–MS/MS through the targeted analysis of 53 phenolic compounds and related metabolites. Furthermore, the honey sample was evaluated for its principal physicochemical quality parameters, including hydroxymethylfurfural (HMF), sugar composition (glucose, fructose, sucrose, and maltose), naphthalene residue, diastase activity, proline content, free acidity, pH, moisture content, and electrical conductivity. Consistent with its phytochemical composition, propolis showed the strongest antioxidant activity among the investigated bee products under the same experimental conditions. These findings highlight the antioxidant potential of Mersin bee products and indicate their potential as natural sources of bioactive compounds, with possible applications in functional food and nutraceutical research. Further comprehensive studies are warranted to better characterize their bioactive constituents and explore their potential applications. Full article

Background: Plectranthus scutellarioides is an ornamental and medicinal species of the Lamiaceae family characterized by substantial phytochemical diversity and variability. This review critically synthesizes current evidence on its bioactive compounds, factors determining phytochemical composition, extraction efficiency, analytical characterization, and biological activity, with particular emphasis on relationships between plant material, extraction conditions, metabolite profiles, and experimentally determined bioactivity. Methods: A narrative review of the available scientific literature was conducted using studies addressing the phytochemical composition of P. scutellarioides, cultivar and cultivation-related variability, extraction procedures, analytical characterization, and biological activities of extracts and isolated metabolites. Particular attention was given to quantitative phytochemical and biological data and to methodological factors limiting direct comparison among studies. Results: P. scutellarioides contains phenolic acids, flavonoids, anthocyanins, abietane-type diterpenoids, and volatile constituents, with rosmarinic acid representing a major phytochemical marker. Substantial quantitative variability is associated with cultivar, geographical origin, cultivation conditions, plant material, and extraction procedure. Optimized ultrasound-assisted extraction yielded 110.8 ± 4.5 mg g⁻¹ DW of rosmarinic acid after 45 min, approximately twice the recovery obtained by conventional heat-reflux extraction over the same period. Quantitative profiling also demonstrated marked cultivation-dependent changes in volatile metabolites. Biological studies revealed assay- and compound-dependent antioxidant, antimicrobial, anti-inflammatory, and cytotoxic/antiproliferative activities. In particular, isolated diterpenoids inhibited NF-κB activity with IC₅₀ values of 4.5–11.2 µM, providing compound-specific quantitative evidence of anti-inflammatory potential. However, differences in extract composition, experimental models, and assay conditions, together with limited standardization and the predominance of in vitro studies, substantially restrict cross-study comparisons and assessment of translational relevance. Conclusions: The available evidence indicates that the biological potential of P. scutellarioides should be interpreted through an integrated extraction–composition–activity framework rather than from the simple presence of individual metabolites. Future studies should prioritize standardized and botanically characterized plant material, clearly defined harvesting conditions, quantitative phytochemical characterization, comparable biological endpoints, bioactivity-guided identification of active constituents, and in vivo validation. Establishing reproducible relationships between plant material, extraction conditions, chemical composition, and biological effects will be essential for assessing the pharmaceutical, cosmetic, and functional potential of this species. Full article

Background/Objectives: Rubus crataegifolius Bunge (Rosaceae, RC), a wild raspberry native to East Asia, has been investigated for the biological properties of its leaves and roots, while the dermatological potential of its fruit remains largely unexplored. This research evaluated the dermatological effects of RC fruit extracts, focusing on their potential for skin regeneration and anti-aging applications. Methods: RC fruit extracts were prepared by ultrasound-assisted extraction (UAE) and maceration extraction (ME) with distilled water or 70% ethanol as solvents. Phytochemical profiling was performed by liquid chromatography–electrospray ionization–time-of-flight mass spectrometry (LC-ESI-TOF-MS). Dermatological activities were evaluated using fibroblast and keratinocyte proliferation, collagen synthesis, wound healing, and gene expression assays. Safety was assessed by cytotoxicity and inflammation assays. Skin hydration was evaluated in a human application test. Results: Various phytochemicals, including flavonoids, phenolic acids, organic acids, and terpenoids, were putatively annotated by LC-ESI-TOF-MS in the RC fruit extracts. Ethanol extracts exhibited higher flavonoid content and antioxidant activity; however, dermatological activities were not correlated with antioxidant activity. UAE-derived extracts enhanced fibroblast proliferation. All extracts promoted collagen synthesis, with ME-EtOH showing the highest increase. In wound-healing assays, ME-DW extracts promoted fibroblast wound closure, while UAE-DW enhanced keratinocyte migration. In keratinocytes, UAE-DW specifically upregulated HAS1 and HAS2 mRNA expression. Clinically, UAE-DW extract produced a significant increase in skin hydration and moisture distribution in healthy volunteers. Conclusions: RC fruit extracts exhibit biological activities, including enhanced fibroblast proliferation, collagen synthesis, wound healing, and skin hydration. Full article

Acne vulgaris is a multifactorial inflammatory disorder involving bacterial colonization, oxidative stress, and lipid-mediated inflammation, necessitating a comprehensive multitarget approach. This study evaluated the anti-acne potential of five morphological parts of Limonia acidissima L. using an integrated in vitro and in silico approach. Ultrasound-assisted extracts were evaluated for antibacterial activity against Cutibacterium acnes and Staphylococcus epidermidis, antioxidant activity using DPPH and ABTS assays, and total phenolic and flavonoid contents (TPC and TFC). TOPSIS was applied to minimize selection bias and identified the leaf extract as the optimal candidate (closeness coefficient = 0.88325). LC-MS/MS tentatively identified major metabolites, including vitexin, quercetin, cyanidin-3-O-galactoside, caffeate, methyleugenol, indole, and linolenic acid. Molecular docking predicted favorable interactions of linolenic acid with C. acnes KAS III (Rerank Score = −78.470) and 15-lipoxygenase (15-LOX) (Rerank Score = −101.025), as well as cyanidin-3-O-galactoside with S. epidermidis (Rerank Score = −109.588) and KEAP1 (Rerank Score = −132.253). SwissADME analysis further suggested favorable drug-likeness and skin-permeation profiles. Collectively, these findings support the potential of L. acidissima leaf extract as a promising multitarget anti-acne candidate. Full article

Background/Objectives: Dasiphora parvifolia, a little-studied Central Asian shrub, lacks standardized pharmacognostic data. This study aimed to comprehensively characterize its above-ground organs, establish diagnostic criteria, comparatively evaluate extraction methods, profile phytochemicals, and evaluate antimicrobial activity. Methods: Morphological, morphometric, and anatomical analyses were performed using light microscopy (ANOVA, p ≤ 0.05). Physicochemical parameters were determined gravimetrically. Percolation and ultrasonic-assisted extraction (UAE) were compared using varying ethanol concentrations, ratios, and maceration times. Phenolics were identified by HPLC-UV and confirmed by ESI-MS. Results: Diagnostic features included leaflet size (6.91 × 1.14 mm), stem length (17.39 cm), and flower diameter (6.92 mm). Microscopy revealed anomocytic stomata and multicellular trichomes (120 µm). Quality parameters (moisture 2.61%, total ash 5.55%, extractives 42.36%). UAE with 70% ethanol (1:20, 24 h pre-maceration) gave 7.53% total phenolics in the dry extract and 7.68% for percolation, corresponding to 20.81 and 28.72 mg/g of raw material, respectively. Among the analyzed extracts, the highest concentrations were detected for gallic acid (3.61%), chlorogenic acid (2.94%), cynaroside (2.28%), and hyperoside (1.82%). Total polyphenols, flavonoids, tannins, β-carotene, and α-tocopherol were 3.70%, 1.36%, 2.67%, 330.08 mg/kg, and 138.0 mg%, respectively. DPPE70 showed the highest activity against P. aeruginosa (17.0 mm), whereas DPUE70-24 showed the highest activity against S. aureus (22.0 mm). Conclusions: D. parvifolia can be considered a relevant source of phenolic compounds. The high levels of gallic and chlorogenic acids, together with hyperoside and cynaroside, indicate a substantial phenolic profile and may contribute to the antioxidant and antimicrobial activities of the extracts. Full article

Huai chrysanthemum (Chrysanthemum morifolium Ramat. cv. Huaiju), collected from Jiaozuo, Henan Province, China, is a medicinal and edible plant rich in phenolic compounds. Polyphenols are natural bioactive compounds with potential antibacterial activity. In this study, HPLC-MS/MS was used to identify and quantify the phenolic compounds in huai chrysanthemum polyphenol extract, and its antibacterial activity against representative Gram-negative and Gram-positive foodborne bacteria was evaluated. A total of 22 polyphenolic compounds were identified, including 10 phenolic acids, 10 flavonoids, and two other phenolic compounds. Caffeic acid was the predominant phenolic acid, while luteoloside was the predominant flavonoid. Flavonoids accounted for 94.03% of the total identified polyphenols. The extract inhibited the growth of both Escherichia coli and Staphylococcus aureus, with an MIC₈₀ of 12.5 mg/mL for both strains. Further analysis showed that the extract reduced intracellular protein content and ATP levels, increased extracellular alkaline phosphatase activity, altered cell surface charge, affected superoxide dismutase activity, and caused marked morphological changes in bacterial cells. These results suggested that the antibacterial effect of huai chrysanthemum polyphenol extract was related to cell envelope damage, energy metabolism disturbance, and redox imbalance. This study provided a basis for further application of huai chrysanthemum polyphenol extract as a natural antibacterial ingredient in food preservation. Full article

Citrus aurantium L. by-products are a valuable source of bioactive compounds with potential industrial applications. This study evaluated the use of natural deep eutectic solvents (NADESs), formulated with choline chloride as the hydrogen bond acceptor (HBA) and fructose as the hydrogen bond donor (HBD), combined with ultrasound-assisted extraction to optimize the recovery of flavonoids from Citrus aurantium L. industrial by-products. A central composite design (CCD) was employed to optimize total flavonoid content (TFC). The molar ratio (MR; 1 and 2 mol of HBD per mol of HBA) and the percentage of added water (AW; 40 and 60%) were optimized. In addition to TFC, total phenolic content (TPC), total ascorbic acid (TAA), antioxidant capacity (Ax) and phenolic profiles were also evaluated. The optimal extraction conditions, as obtained by Response Surface Methodology (RSM) for TFC, corresponded to an HBD molar ratio of 1.33 per 1 mol of HBA and 47.48% added water, resulting in an optimal value of 1576.63 ± 4.9 mg quercetin equivalents (QE)/100 g dry mass (DM). Maximum values of 4276.94 ± 25.4 mg gallic acid equivalents (GAE)/100 g DM for TPC, 2077.66 ± 21.3 mg/100 g DM for TAA and 88.09 ± 0.1% inhibition for Ax were also obtained. The optimal NADES was characterized by density, viscosity, pH, Fourier-transform infrared (FTIR) and Raman spectroscopy. Additionally, phenolic profiling revealed high concentrations of hesperidin (2747.1 mg/100 g DM), naringenin (1251.93 mg/100 g DM), and quercetin + luteolin (660.65 mg/100 g DM). Full article