Search Results (919)

A derivatisation-free HPLC–MS/MS method was developed and validated for the simultaneous quantification of selenium- and sulphur-containing amino acids in soybean leaves, and applied to a 3 × 3 factorial hydroponic experiment probing selenium–sulphur metabolic interactions. The method resolves five biologically informative analytes (Cys₂, SeCys₂, MeSeCys, Met, SeMet) within 1.5 min through multiple reaction monitoring (MRM). Ultrasound-assisted extraction (UAE) of the free fraction was jointly optimised for both analyte classes by the response-surface methodology; enzymatic hydrolysis of the extraction residue recovered the protein-bound fraction on the same platform. Limits of detection ranged from 0.036 to 0.556 µg L⁻¹, intra-day relative standard deviations were below 5%, and spike recoveries fell between 92.3 and 117.4%. Free SeAA and SAA pools were negatively correlated across the nine treatments (R² = 0.83), consistent with competitive Se–S assimilation, whereas bound pools were positively correlated (R² = 0.89), reflecting proportional protein-level incorporation. A regime of 1–5 mM of sulphate with 20 µM of selenite yielded the highest bound organo-Se with near-normal growth, providing leaf-level evidence that may inform future seed-focused studies aimed at Se-enriched soy-protein ingredient development. Full article

Rosehip (the “fruit” of Rosa canina L, commonly known as dog rose) is an emerging functional food, yet native Greek populations remain under-explored. This study screened 76 wild genotypes from Northern Greece for radical scavenging activity (% RSA), total phenolic content (TPC), and ascorbic acid (AsA). The results showed remarkable antioxidant potential (RSA > 70%), with TPC ranging from 1.02 to 35.96 mg g⁻¹ DW, and AsA between 0.72 and 3.57 mg g⁻¹ FW. Stepwise multiple regression analysis identified altitude as the primary predictor for RSA (adjusted R² = 0.139, p = 0.001) and latitude as a significant modulator for TPC (p = 0.034), reflecting subtle environmental adaptations over a robust genetic baseline. HPLC-PDA-MS characterization revealed a complex profile dominated by procyanidins, catechin derivatives, flavanonols (eriodictyol conjugates), and flavonol 3-O-glycosides (mainly quercetin hexosides and pentosides). Exploratory multivariate analysis (PCA) visualized high phenotypic plasticity and identified elite chemotypes (e.g., RPK-5, RCZ-12). Notably, the Rhodopi population exhibited the most extensive multidimensional dispersion despite its geographically restricted collection radius, suggesting a diverse local genetic reservoir. These findings establish a phytochemical map of Greek dog rose germplasm, providing essential criteria for selecting high-quality genotypes for future domestication and exploitation in the nutraceutical and pharmaceutical sectors. Full article

Myrtus communis L. (common myrtle) is an economically valuable Mediterranean shrub with diverse applications in food, pharmaceutical, and ornamental sectors. However, the biochemical diversity of myrtle genotypes from Mediterranean environments remains insufficiently characterized, particularly regarding the relationship between primary and secondary metabolism and stress adaptation. This study investigated the biochemical and aroma profiles of six myrtle genotypes selected from natural populations in Antalya, Turkey, to identify chemotypic diversity and elucidate metabolic diversity observed in Mediterranean genotypes. Volatile compounds were analyzed using HS-SPME/GC-MS, while sugars and organic acids were quantified by HPLC. Multivariate statistical analyses (PCA, hierarchical clustering) were employed to evaluate metabolic relationships and genotype classification. Descriptive analysis suggested three potential chemotypic patterns: (i) 1,8-cineole-type (G34, G36) with G29 showing a transitional profile, (ii) α-Pinene-type (G15, G37), and (iii) Ester-aldehyde type (G9). These groupings are based on single volatile measurements and should be considered preliminary patterns pending validation through replicate analyses. Significant genotypic variation was observed for primary metabolites (sugars and organic acids) (p < 0.001, η² > 0.90), as evaluated by ANOVA with triplicate biological replicates. Volatile compound differences were evaluated as descriptive exploratory patterns only. Hierarchical clustering revealed three metabolic strategies: balanced metabolism integrating diverse volatile and primary metabolite profiles (Cluster 1: G9, G15, G37), terpene-rich volatile defense with enhanced organic acid metabolism (Cluster 2: G29, G36), and specialized 1,8-cineole-dominant biosynthesis (Cluster 3: G34). These findings highlight substantial metabolic diversity and provide a basis for germplasm evaluation and selection and potential applications. Full article

This study aimed to evaluate genotype-dependent variation in biochemical composition, antioxidant capacity, and aroma profiles of dried Chinese mulberry (Morus spp.) genotypes. Three cultivars, Lvmeiren (green), Zhenzhubai (white), and Yunsang No 2 (red), were analyzed. Organic acids and sugars were determined using HPLC, while total phenolic content, antioxidant capacity (DPPH and FRAP), and total anthocyanins were quantified using spectrophotometric methods. Volatile compounds were analyzed by HS-SPME/GC–MS. Significant differences were observed among genotypes for all measured parameters. Among the studied genotypes, Yunsang No 2 exhibited the highest total phenolic content (379.59 mg GAE g⁻¹ DW), FRAP value (21.51 μmol g⁻¹ DW), and anthocyanin content (37.1 mg L⁻¹). In contrast, Lvmeiren was characterized by markedly higher sucrose (22.57%) and succinic acid (3.69%) contents. Zhenzhubai exhibited the highest glucose (25.82%) and fructose (32.65%) contents, together with elevated citric (2.58%) and malic acid (2.93%) levels. Yunsang No 2 showed markedly higher total phenolics, anthocyanins, and antioxidant capacity, indicating superior nutraceutical potential. Volatile compound analysis revealed aldehydes and alcohols as dominant groups in Lvmeiren and Zhenzhubai, while acids were predominant in Yunsang No 2. Multivariate analyses (PCA and hierarchical clustering) clearly separated genotypes based on biochemical and antioxidant traits. These findings demonstrate that genotype plays a critical role in determining the nutritional quality and aroma profile of dried mulberries and provide valuable insights for breeding, cultivar selection, and functional food applications. Full article

The genus Leonurus L. is renowned for its diverse secondary metabolites with significant pharmacological value; however, the chemical biodiversity and biological potential of its indigenous members in Türkiye remain largely unexplored. This study investigates four species (L. cardiaca, L. quinquelobatus, L. glaucescens, and L. persicus) to elucidate their phytochemical architecture and therapeutic capacities. Characterization of the ethanol, methanol, and aqueous (5% infusion) extracts via HPLC-MS/MS identified verbascoside, genkwanin, and caffeoylquinic acids as the major representative bioactive constituents across the studied Leonurus species. The extracts exhibited measurable biological activity, with L. cardiaca displaying the highest antioxidant profile (EC₅₀ 0.117 ± 0.01 mg/mL for DPPH, 2.731 ± 0.01 mM/Trolox for ABTS), correlating with its phenolic content. Notably, the extracts demonstrated notable anticandidal activity (MIC 0.1–1 mg/mL) and negligible to moderate antibacterial effects, alongside varying levels of susceptibility against breast (MCF7) and glioma (C6) cancer cell lines. These effects showed differentiation in toxicity compared to lung (A549) cells. This investigation provides scientific evidence supporting the traditional medicinal use of Leonurus species while highlighting their potential as standardized sources for the pharmaceutical and nutraceutical sectors. Our results lay a robust foundation for future bioactivity-guided isolation studies to further elucidate the molecular mechanisms behind their differential biological effects. Full article

Cenostigma pluviosum var. peltophoroides, known as “sibipiruna,” is a plant rich in polyphenols used in traditional medicine for gastrointestinal disorders. The study aimed to investigate the chemical composition of the crude ethanolic extract of the stem bark (CEECP), evaluating its in vivo toxicity, genotoxicity, mutagenicity and anti-inflammatory activity. The plant material was macerated in 95% ethanol for 72 h, and the solvent was removed by rotary evaporation to obtain CEECP. Chemical characterization was performed by HPLC-ESI-MS/MS in negative mode. In vivo approaches were performed using male/female Swiss albino mice. Acute toxicity was assessed at a single high dose of 2000 mg/kg. Mutagenicity was investigated by the micronucleus test and genotoxicity by the comet assay, both at a dose of 2000 mg/kg. Anti-inflammatory activity was evaluated in carrageenan-induced paw edema and peritonitis models, at doses of 50, 100, and 200 mg/kg. HPLC-ESI-MS/MS analysis showed the presence of hydrolyzable tannins, phenolic acid heterosides, and biflavonoids. The safety profile of the CEECP was demonstrated for the first time, with no evidence of acute toxicity, mortality, mutagenicity, or genotoxicity at the tested doses. The extract significantly reduced paw edema in a dose-dependent manner at doses of 100 and 200 mg/kg, with inhibition rates of 65.78% and 73.12%, respectively, and also decreased leukocyte migration in the peritonitis model by 61.81% and 72.79% at the same doses. These findings indicate the CEECP as a source of pharmacologically relevant phytocompounds and, most notably, demonstrate its pronounced anti-inflammatory activity. Furthermore, the extract exhibited a favorable safety profile in the toxicological evaluations, highlighting the extract as a promising anti-inflammatory agent. Full article

Polygonum capitatum (PC) is an ethnomedicine with reported antibacterial and anti-inflammatory activities and has been clinically used in urinary tract infection (UTI)-related disorders. However, its in vivo exposure characteristics and metabolically associated therapeutic mechanisms in acute pyelonephritis (AP) remain insufficiently understood. To address this issue, this study aimed to evaluate the therapeutic effects of PC in an Escherichia coli (E. coli)-induced rat model of AP and to explore constituents and metabolic pathways associated with its activity. Different PC extracts were screened for antibacterial and anti-inflammatory activities, and the 70% ethanol extract was selected for further study. Seven major compounds were quantified by HPLC. In AP rats, the pharmacokinetic profiles of these compounds in plasma and the renal cortex were analyzed by microdialysis-coupled HPLC-MS/MS. Pharmacodynamic evaluation included urinary bacterial load, urinalysis, renal function, inflammatory cytokines, and renal histopathology. Exploratory PK–PD analysis, untargeted renal metabolomics, and targeted metabolomics of the tryptophan–kynurenine (Trp–Kyn) pathway were also performed. The 70% ethanol extract of PC exhibited the strongest antibacterial and anti-inflammatory activities. The total content of seven active compounds was 3.85%, with gallic acid being the predominant compound (3.42%). Pharmacokinetic analysis revealed that gallic acid, protocatechuic acid, methyl gallate, and quercitrin achieved relatively high systemic exposure and renal distribution. In AP rats, the pharmacokinetic profiles of several compounds were altered, with increased plasma exposure of protocatechuic acid, vanillic acid, ethyl gallate, and syringic acid, while protocatechuic acid also showed higher exposure in the renal cortex. PC treatment reduced urinary bacterial load, improved renal function and urinalysis parameters, alleviated histopathological injury, and decreased inflammatory mediator levels, particularly in renal tissue. Exploratory PK–PD correlations were observed between several compounds and selected pharmacodynamic indicators. Metabolomic analysis suggested disturbances in glycerophospholipid metabolism and the Trp–Kyn pathway in AP rats, some of which were partially reversed after PC treatment. PC showed antibacterial and anti-inflammatory effects in AP rats. Gallic acid, protocatechuic acid, methyl gallate, and quercitrin may be candidate constituents associated with the therapeutic effects of PC, while modulation of glycerophospholipid metabolism and the Trp–Kyn pathway may be involved in its action against AP. These findings provide preclinical pharmacological evidence supporting the therapeutic potential of PC in AP. Full article

(1) Background: Variations in bean temperature before grinding are a little-studied factor, but they can potentially influence the characteristics of the resulting powder and the chemical and physical properties of the espresso beverage. This study investigated the effect of two heat treatments, heating and cooling, applied to coffee beans immediately before grinding. (2) Methods: The analyses focused on powder particle size distribution (laser diffraction), impact on the operation of the coffee grinder (noise and electrical absorption), chemical-physical properties of the beverage, caffeine and chlorogenic acid content (HPLC-DAD) and profile of volatile organic compounds (HS-SPME-GC-MS). (3) Results: Heating induced a decrease in the content of caffeine and chlorogenic acids and a change in the aromatic profile consistent with phenomena like accelerated aging (increase in hexanal). Cooling treatment had similar, but less pronounced, effects, although it reduced caffeine extraction and some aromatic compounds. (4) Conclusions: The study demonstrated that the temperature of the coffee beans prior to grinding is a key factor to consider in terms of the particle size distribution of the resulting coffee grounds, as well as the content of bioactive compounds and volatile organic compounds, which can significantly influence various aspects of the final espresso’s quality. Full article

Green seeds of Coffea arabica represent a rich source of bioactive compounds. This study aimed to compare the butanol-soluble (CA-BU) and the ethyl acetate-soluble (CA-EtAc) fractions in terms of their phytochemical composition and biological activity. As a first step, the fractions were analyzed by Fourier-transform infrared spectroscopy (FT-IR) and high-performance liquid chromatography coupled with mass spectrometry (HPLC–MS) in order to investigate the major constituents. Subsequently, CA-BU and CA-EtAc were evaluated for antioxidant effect, antimicrobial activity, antiproliferative properties, effects on the mitochondrial function, and on the chorioallantoic membrane. The CA-EtAc fraction was enriched in chlorogenic acids and catechins and showed superior antioxidant activity, whereas CA-BU displayed a broader profile of semi-polar polyphenols, conferring moderate antimicrobial effects and stronger antiproliferative activity in MCF-7 human breast adenocarcinoma cells, although with limited selectivity compared with HaCaT non-tumorigenic cells. Respirometric analysis demonstrated that CA-BU selectively inhibited mitochondrial oxidative phosphorylation Complex I (OXPHOS CI), without affecting Complex II (CII) or basal respiration, indicating a specific mitochondria-targeted mechanism. Both fractions were non-irritant and well tolerated in the chorioallantoic membrane (CAM) assay; CA-BU reduced vascular density. These findings demonstrate a clear mechanistic differentiation between the fractions, highlighting the decisive role of solvent polarity in obtaining extracts with distinct and targeted biological effects. Full article

This article is the first report highlighting Rhodope avens as a natural source of bioactive compounds with potential pharmaceutical applications. Dry tinctures obtained from herba (HDT) and rhizoma (RDT) from the Balkan Peninsula endemic plant Geum rhodopaeum were analyzed and tested for antioxidant activity (DPPH, ABTS, FRAP, CUPRAC assays) and DNA nicking protection potential. The tinctures were characterized by a high total polyphenol content (total the polyphenols in RDT were more than twice that of HDT, i.e., 516.6 mg GAE/g dt; p < 0.01). HPLC analysis revealed the presence of 14 phenolics, with the main one being (over 3 mg/g dt) (+)-catechin (28.21 mg/g dt for RDT; 9.67 mg/g dt HDT), followed by protocatechuic acid, salicylic acid, and rutin. Rosmarinic acid content is herein first reported for the genus Geum. GC/MS analysis identified 41 compounds, among which monosaccharides predominated (with the highest fructose content), followed by gluconic acid, glucose-1-phosphate, mannitol, sorbitol, and glycerol in HDT, along with xylitol in RDT. The tinctures exhibited strong antioxidant potential as evaluated by the methods used, with the most pronounced effect found in the CUPRAC assay (5 458.87 μM TE/g dt for HDT; 10 073.99 μM TE/g dt for RDT; p < 0.01), as confirmed in comparison to proven antioxidants such as BHT and vitamin C. DNA-protective activity against oxidative damage was established and microscopic identification of the herbal substances was also performed. Full article

Background/Objectives: This study explores the roles of kynurenine and 5-hydroxytryptophan (5-HTP) in modulating gut microbiota and their potential implications for exudative age-related macular degeneration (AMD). By examining the interplay between these metabolites and the microbiome, we aim to uncover novel pathways that may influence the pathogenesis of AMD. Understanding these associations could lead to innovative therapeutic approaches for managing this leading cause of vision loss in the elderly. To investigate the roles of kynurenine and 5-HTP, alongside the composition of the nasopharyngeal microbiota, in patients with exudative AMD. Methods: Blood metabolite profiling was performed using LC–MS–based metabolomics. Metabolites were extracted with cold methanol/water containing internal standards, filtered through a 10 kDa cutoff filter, separated on a ZIC-HILIC HPLC column, and detected using an Orbitrap mass spectrometer. Metabolites were identified using MZmine 2 software. Results: Patients with exudative AMD exhibited a profound systemic shift in tryptophan metabolism, characterized by significantly lower plasma levels of 5-HTP and higher levels of kynurenine compared to control subjects (p < 0.001 for both). Logistic regression analysis confirmed that both metabolites were independent predictors of AMD status; higher kynurenine levels were associated with increased disease probability, while higher 5-HTP levels demonstrated a protective effect. The kynurenine/5-HTP ratio emerged as a robust biomarker, achieving an area under the curve (AUC) of 0.85 with an optimal threshold of 3.43 (74.1% sensitivity, 84.4% specificity). When integrated with age and gender, the diagnostic performance of the model reached an excellent AUC of 0.92. After adjusting for demographic factors, the kynurenine/5-HTP ratio remained a potent independent risk factor, with each unit increase associated with a 6.30-fold increase in the odds of exudative AMD. Conclusions: Exudative AMD is characterized by a shift in tryptophan metabolism toward the kynurenine pathway, with decreased 5-HTP, increased kynurenine, and an elevated kynurenine/5-HTP ratio. This ratio showed a strong independent association with AMD and excellent diagnostic performance, highlighting its potential as a biomarker and its role in disease pathogenesis. Full article

Due to its abundant content of biologically active compounds, Althaea officinalis L. (marshmallow) has been extensively researched and applied in both the food and pharmaceutical industries. This study aimed to evaluate hydroethanolic and natural deep eutectic solvent (NADES) extracts from leaves, flowers, and roots in terms of their chemical composition and biological activities. Extracts were characterized using chromatographic and spectrophotometric methods. Phytochemical profiling by HPLC-MS revealed a diverse composition, with 35 secondary metabolites identified or tentatively assigned, mainly hydroxycinnamic acid derivatives and flavonoid glycosides. GC-MS analysis of the ethanol extracts identified ten free amino acids, seven organic acids, several mono- and disaccharides, and one oligosaccharide. Their concentrations varied across different parts of the plant depending on the specific metabolism of the respective organ. Ethanolic extracts showed the highest total phenolic content (up to 176 mg GAE/L), while flower extracts exhibited the strongest antioxidant activity (DPPH up to 89 µmol TE/L). The antimicrobial potential of the extracts was assessed by the agar well diffusion method. NADES1 extracts demonstrated significant antibacterial activity, with inhibition zones reaching up to ~34 mm, whereas NADES2 extracts were largely inactive. In contrast, antifungal activity was negligible or absent across all samples. Full article

Globba bicolor Gagnep., an ornamental ginger of cultural importance in Thailand’s “Tak Bat Dok Mai” festival, faces conservation challenges due to climate change and slow natural propagation. Limited understanding of its cultivation and chemical composition further constrains sustainable utilization. This study provides the first integrated investigation of micropropagation using rhizome-derived explants under various combinations of exogenous hormones, acclimatization strategies, and comparative phytochemical profiling between wild and in vitro-propagated plants. An optimized clonal regeneration system was established from plantlets, with Murashige and Skoog (MS) medium containing 2.0 mg/L 6-benzylaminopurine (BA) and 0.5 mg/L 1-naphthaleneacetic acid (NAA), yielding the highest multiplication (9.10 shoots/explant and 12.40 roots/explant) after eight weeks of cultivation. During acclimatization, sand substrate proved superior, facilitating a 90% survival rate and enhanced physiological vigor. Comparative analysis revealed that while wild plants possessed significantly higher total phenolic (TPC) and total flavonoid (TFC) contents and antioxidant activities (DPPH, ABTS, and FRAP) than their in vitro counterparts, both sources maintained a rich diversity of chemical constituents. HPLC analysis identified cinnamic acid, rutin, and quercetin as major metabolites, while GC–MS detected 90 volatile compounds, with β-caryophyllene and β-pinene as predominant constituents. Notably, rhizomes of wild plants exhibited particularly high-value detections. To provide a rapid and non-destructive approach for linking chemical composition with antioxidant activity, FTIR-based chemometric models were applied, demonstrating high predictive accuracy (R²–cv = 0.9712–0.9862). These results provide a scientific foundation for the conservation and sustainable commercial utilization of G. bicolor as a potential source of bioactive natural products. Full article

Artichoke by-products (ABP) represent valuable sources of bioactive compounds with relevant health benefits. In this study, a green extraction strategy based on pressurized liquid extraction (PLE) was optimized to enhance the recovery of phenolic and flavonoid compounds from ABP using a response surface methodology. Extraction temperature and solvent composition were identified as the key factors driving extraction performance. Optimal conditions using a mixture of ethyl acetate and ethanol (90/10, v/v) at 180 °C significantly enhanced extraction yield, total phenolic and flavonoid content, and antioxidant activities, as measured by ORAC and DPPH assays. Chemical characterization via HPLC-C18-Q-TOF-MS/MS revealed a diverse profile of phenolic and flavonoid compounds, including caffeoylquinic acid derivatives and related transformation products. The neuroprotective potential of the optimized extract was further evaluated through in vitro inhibition assays targeting acetylcholinesterase (AChE), butyrylcholinesterase (BChE) and lipoxygenase (LOX), alongside a permeability assessment using an in vitro blood–brain barrier (BBB) model. Molecular docking simulations were performed to explore the interactions of apigenin—the most representative flavonoid in the optimal extract—with the three target enzymes. Overall, these findings support the valorization of ABP as a source of bioactive compounds and highlight the potential of PLE as an efficient and sustainable extraction approach. Full article

Pyrolysis liquid (PL) derived from biomass pyrolysis exhibits biopesticidal properties and represents a promising value-added product within the sustainable circular economy framework. However, knowledge about the antimicrobial potential of PLs produced from walnut residue at different pyrolysis temperatures remains limited. We investigated the chemical composition and antimicrobial activity of PLs obtained from agricultural walnut residue (Juglans regia L.) against selected plant pathogenic bacteria and fungi. PLs were produced at four temperature ranges: 200–300 °C (W-1), 300–400 °C (W-2), 400–500 °C (W-3), and 500–600 °C (W-4). Chemical characterization was performed using Gas chromatography–mass spectrometry (GC-MS), High-performance liquid chromatography (HPLC), and Inductively coupled plasma optical emission spectrometry (ICP-OES), with determination of total phenolic and flavonoid contents. Pyrolysis temperature significantly influenced the chemical profile and bioactive compound content of the PLs, with W-4 showing the highest total phenolic and flavonoid levels. Heavy metal analysis indicated minimal contamination in all samples. Antibacterial activity was observed in stock solutions, whereas diluted applications showed limited effects. The W-4 fraction showed the strongest antibacterial activity and exhibited MIC values of 12.50 µL/mL against Clavibacter michiganensis subsp. michiganensis, Xanthomonas euvesicatoria, and Pseudomonas syringae pv. syringae, and 25.00 µL/mL against Erwinia amylovora. Antifungal activity differed markedly across temperature ranges, with W-3 and W-4 displaying superior activity against Fusarium oxysporum and Verticillium dahliae, achieving complete mycelial growth inhibition at 5%, compared to 10% for W-2 and 20% for W-1. Positive controls confirmed assay validity (ciprofloxacin for antibacterial assays and cycloheximide for antifungal assays), whereas negative controls showed no inhibitory effect. Overall, higher pyrolysis temperatures, particularly 400–600 °C, enhanced the antimicrobial potential of walnut residue-derived PLs, supporting their possible use as bio-based antifungal agents for sustainable crop protection. Full article