Search Results (188)

Background/Objectives: Dietary supplements are sources of nutrients or other substances that added to a healthy lifestyle help to preserve human homeostasis. Since inflammation is one of the major contributors to the alteration of homeostasis, this work investigated the effects of a multi-ingredient dietary supplement on human macrophages, cells involved in the inflammatory response. Methods: THP-1 cells were differentiated into macrophage-like cells and polarized in M₁ or M₂ phenotypes. Cell migration was evaluated by Boyden chamber assay; phenotypic markers by qRT-PCR; cytokine release by ELISA and LPS/ATP-induced pyroptosis by LDH assay. The antioxidant properties of the supplement were evaluated in human and mouse fibroblasts by DCF-DA assay. After supplement treatment, cell extracts were analyzed by HPLC-PDA-MS/MS and GC-MS to evaluate the presence of the ingredients. Results: Our results showed that the dietary supplement promoted M₂ migration and polarization and significantly reduced migration of M₁. In a model of LPS-induced inflammation in M₀, it significantly reduced NF-κB activation, COX-2 expression, and cytokine release. The supplement was not a specific inhibitor of NLRP-3, but it was able to modulate LPS priming. In addition, the supplement decreased granulocyte adhesion to HUVEC and reduced the oxidative stress in fibroblasts. The analysis of cell extracts showed the presence of the following ingredients of the formulation inside the cells: CoQ10, spermidine, resveratrol, 5-hydroxytryptophan from Griffonia simplicifolia (Vahl ex DC.) Baill., bacosides from Bacopa monnieri (L.) Wettst, vit B₂, B₅, E acetate. Conclusions: Our results demonstrate how a combination of natural active ingredients may contribute to the maintenance of homeostasis in human cells. Full article

The olive oil industry, a key component of Southern Europe’s agricultural sector, generates large amounts of by-products during processing, including olive leaves, branches, stones, and seeds. In the context of growing environmental concerns and limited natural resources—particularly in the Mediterranean regions—there is increasing interest in circular economy approaches that promote the valorization of agricultural residues. These by-products are rich in bioactive compounds, particularly phenolics such as oleuropein and hydroxytyrosol, which are well known for their antioxidant and anti-inflammatory activities. This study aimed to evaluate the phenolic content and antioxidant capacity of by-products from three olive cultivars using high-performance liquid chromatography with photodiode array detection (HPLC–PDA) and mass spectrometry (MS). The leaves and seeds, particularly from the “Cobrança” and a non-identified variety, presented the highest antioxidant activity, as well as the highest concentration of phenolic compounds, demonstrating once again the direct relationship between these two parameters. The identification of the compounds present demonstrated that the leaves and branches have a high diversity of phenolic compounds, particularly secoiridoids, flavonoids, phenylpropanoids, phenylethanoids, and lignans. An inverse relationship was observed between the chlorophyll and carotenoid content and the antioxidant activity, suggesting that phenolic compounds, rather than pigments, are the major contributors to antioxidant properties. Therefore, the by-products of the olive oil industry are a valuable source of sustainable bioactive compounds for distinct industrial sectors, such as the food, nutraceutical, and pharmaceutical industries, aligning with the European strategies for resource efficiency and waste reduction in the agri-food industries. Full article

In this study, water residue obtained from Salvia desoleana Atzei et Picci steam distillation was evaluated for its antioxidant activity in vitro using different experimental models. In particular, the study evaluated the antiradical and antioxidant activity of Salvia desoleana extracts using CUPRAC, FRAP, DPPH^•, and ABTS^•+ assays; and tested ROS scavenging activity in Caco-2 cell cultures. Phenolic compounds were identified by (HR) LC-ESI-QTOF MS/MS and quantified with HPLC-PDA. Furthermore, Keap1-Nrf2, iNOS, and NOX enzymes involved in oxidative stress and antioxidant defences were the targets of molecular docking on key polyphenols. Hydroxycinnamic acids and flavonoids are the most important classes of compounds detected in the extracts. Among these compounds, the most significant was rosmarinic acid, followed by caffeic acid, luteolin glucuronide, and methyl rosmarinate. Although all extracts have shown encouraging results, the ethanolic extract solubilised with water (SEtOHA) was the one with the highest hydroxycinnamic acid content and total phenol content (518.64 ± 5.82 mg/g dw and 106.02 ± 6.02 mg GAE/g dw), as well as the highest antioxidant and antiradical activity. The extracts have shown anti-inflammatory activity by inhibiting NO release in LPS-stimulated Caco-2 cells. Finally, the in silico evaluation against the three selected enzymes showed interesting results for both numerical affinity ranking and predicted ligand binding models. The outcome of this study suggests this by-product as a possible ally in counteracting oxidative stress, as established by its favourable antioxidant compound profile, thus suggesting an interesting future application as a nutraceutical. Full article

Industrial growth led to the expansion of existing environmental problems, where different kinds of pollutants can enter the environment by many known routes, particularly through wastewater. Among other contaminants, pharmaceuticals, such as diazepam, once released, pose a significant challenge related to their removal from complex environmental matrices due to their persistence and potential toxicity. For this reason, it is a great challenge to find suitable methods for the treatment of wastewater. The aim of this paper was to investigate the stability of diazepam, subjecting it to various degradation processes (hydrolysis and photolysis), focusing on photocatalysis, an advanced oxidation process commonly used for the purification of industrial wastewater. The photocatalytic system consisted of UV-A and simulated solar irradiation with titanium dioxide (TiO₂) immobilized on a glass mesh as a photocatalyst, with an additional reaction performed in the presence of an oxidizing agent, i.e., hydrogen peroxide, to improve diazepam removal from water matrices. The kinetic rate of diazepam degradation was monitored with a high-performance liquid chromatograph coupled with a photodiode array detector (HPLC-PDA). The target compound was characterized as a hydrolytically and photolytically stable compound with t_1/2 = 25 h. The presence of an immobilized TiO₂ catalyst contributed significantly to the degradation of diazepam under the influence of UV-A and simulated solar radiation, with t_1/2 in the range of 1.61–2.56 h. Five degradation products of diazepam were identified at the laboratory scale by MS analysis (m/z = 267, m/z = 273, m/z = 301, m/z = 271, and m/z = 303), while the toxicity assessment revealed that diazepam exhibited developmental toxicity and a low bioaccumulation factor. The pilot-scale process resulted in significant improvements in diazepam degradation with the fastest degradation kinetics (0.6888 h⁻¹). These results obtained at the pilot scale highlight the potential for industrial-scale implementation, offering a promising and innovative solution for pharmaceutical removal from wastewater. Full article

Chinese herbal medicines have played a significant role in the development of new and effective drugs, but how to identify the active ingredients from complex extracts of traditional Chinese herbal medicines was a research difficulty. In recent years, few studies have focused on high-efficiency identification of small-molecule inhibitors of Programmed Death Ligand 1 with lower antigenicity and flexible structure tunability. In order to identify small molecule inhibitors of PD-L1 from complex Chinese herbal extracts, this study established a protein–ligand trapping system based on high-performance liquid chromatography coupled with a photo-diode array detector, ion trap/quadrupole time-of-flight tandem mass spectrometry, and a Programmed Death Ligand 1 affinity chromatography unit (ACPD-L1-HPLC-PDA-IT-TOF (Q-TOF)-MS) to rapidly screen and identify small-molecule inhibitors of Programmed Death Ligand 1 from Toddalia asiatica (L.) Lam. Fourteen components were then identified as PD-L1 binders, and surface plasmon resonance (SPR) validation results showed that six of them—magnoflorine (6), nitidine (22), chelerythrine (24), jatrorrhizine (13), toddaculin (68), and toddanol (45)—displayed PD-L1 binding activity. Laser scanning confocal microscopy results demonstrated that these compounds effectively inhibited the binding of PD-1 to PD-L1 in a dose-dependent manner. Additionally, flow cytometry analysis indicated they could promote human lung cancer cell line (A549) apoptosis when co-cultured with Peripheral Blood Mononuclear Cells (PBMCs). The system’s innovation lies in its first integration of dynamic protein–ligand trapping with multi-dimensional validation, coupled with high-throughput screening capacity for structurally diverse natural products. This workflow overcomes traditional phytochemical screening bottlenecks by preserving native protein conformations during affinity capture while maintaining chromatographic resolution, offering a transformative template for accelerating natural product-derived immunotherapeutics through the PD-1/PD-L1 pathway. Full article

Glochidion chodoense, a rare and endangered plant endemic to Republic of Korea, is known for containing a wide variety of phytochemicals, including triterpenoid saponins and flavonoids. This study sought to profile the phytochemical composition of the leaves and branches of G. chodoense harvested during three different periods (May, July, and October 2023) using liquid chromatography–electrospray ionization/mass spectrometry (LC-ESI/MS) and high-performance liquid chromatography–photodiode array detection (HPLC/PDA). Plant materials were harvested, authenticated, and subjected to ethanol extraction prior to chemical analysis. LC-ESI/MS and quantitative HPLC/PDA analyses were conducted to identify and quantify nine key phytochemicals: norbergenin (1), bergenin (2), epigallocatechin (3), ethyl gallate (4), orientin (5), epicatechin gallate (6), isovitexin 2″-O-arabinoside (7), ellagic acid (8), and cynaroside (9). Our findings revealed significant seasonal variations in major phytochemicals, with leaves containing higher concentrations than branches. Notably, bergenin (2) showed the highest content in May leaves (43.42 mg/g extract), followed by October (17.60 mg/g extract) and July branches (8.56 mg/g extract). Ethyl gallate (4), which was absent or present in trace amounts in branches, was abundant in leaves, with concentrations of 22.24 mg/g extract in October, 21.75 mg/g extract in May, and 17.48 mg/g extract in July. A similar trend was observed for norbergenin (1). These findings provide valuable insights into the phytochemical composition of G. chodoense, emphasizing its potential applications in pharmaceuticals, cosmetics, and functional foods, while highlighting the critical importance of conserving this endangered species. Full article

Barley sprouts, rich in bioactive compounds, have gained attention as functional food ingredients because of their antioxidant potential. This study evaluated their bioactive composition and antioxidant capacity, focusing on the saponarin, chlorophyll, policosanol, total polyphenol (TP), and total flavonoid (TF) contents. The antioxidant capacity was assessed using 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), and ferric-reducing antioxidant power (FRAP) assays. The results showed that barley sprouts contained 8.14 ± 0.02 mg/g of saponarin, 15.36 ± 0.18 mg/g of total chlorophyll, 396.99 mg/100 g of policosanols, 12.64 ± 0.04 mg of gallic acid equivalent (GAE)/g, and 5.99 ± 0.09 mg of rutin equivalent (RE)/g. The half-maximal inhibitory concentration (IC₅₀) values exhibited the trend FRAP > DPPH > ABTS. Significant correlations (R > 0.891, p < 0.05) were observed between the antioxidant assays and TP and TF contents, indicating their substantial role in the antioxidant properties of barley sprout extracts. These findings suggest that barley sprouts are a valuable natural source of antioxidants for functional food applications. Nevertheless, additional in vivo and clinical research is necessary to improve their bioavailability and expand their potential use in food formulations. Full article

Phenolic compounds are strong antioxidant and antibacterial agents with great pharmacological, medicinal, nutritional, and industrial value. The potential of Morus nigra in stem node culture was investigated for the production of phenolic compounds and their elicitation with CuSO₄. Individual phenolic compounds in the samples were identified and quantified by using HPLC-PDA and HPLC-MS methods, while the content of total phenolic compounds, the content of total flavonoids, and the antioxidant activity of methanolic extracts were evaluated spectrophotometrically. The highest fresh and dry weights were obtained in plantlets treated with 0.5 mM CuSO₄ for 42 days. The highest total phenolic content, total flavonoid content, and antioxidant activity of the extracts were determined in stem node cultures treated with 3 mM CuSO₄ for 42 days. Under the latter conditions, the predominant representatives of the caffeoylquinic acids, p-coumaric acid derivatives, kaempferol derivatives, and quercetin derivatives also achieved the highest content. The most abundant phenolic compound in all samples was the chlorogenic acid. The nodal culture of M. nigra elicited with CuSO₄ could potentially be used for the industrial production of phenolic compounds, especially caffeoylquinic acids. Moreover, considering the biochemical response to CuSO₄ treatment and the ability to tolerate and accumulate copper, the potential application of M. nigra in phytoremediation is also highlighted. Full article

This article represents the first consideration of the peculiarities of the fatty acid (FAs) composition and structure of storage triacylglycerols (TAGs) of the relict plant Lunaria rediviva L. The composition of storage TAGs was found to comprise 21 individual FAs, with an unsaturated FA content of 96.8%. Additionally, monounsaturated acids with a very long chain (VLCFAs), specifically C20:1–C24:1, constituted over 60% of the total FAs. The ethylene bond position isomers of unsaturated FAs were accurately identified and the presence of unusual isomers, including 20:1$\Delta$13, 22:1$\Delta$15, and 24:1$\Delta$17 acids. Furthermore, the unusual minor 24:2$\Delta$15,18 acid was identified and characterised for the first time. The pathways of the mentioned VLCFA’s biosynthesis have been proposed. The distribution of FA acyls between the sn positions of triacylglycerols was found to be highly specific. Thus, VLCFAs exclusively acylate the $\alpha$ positions of the carbon atoms of the glycerol residue of the TAG molecule (sn-1 and sn-3 positions), while unsaturated C18 acids exclusively acylate the $\beta$-carbon atom (sn-2 position). The composition of the molecular species of TAGs was analysed using a calculation method based on the Vander Wal model and by RP-HPLC-ESI-MS. A significant discrepancy from the statistical model was observed, indicating a preference for the formation of symmetrical TAGs, such as sn-1,3-dierucoyl-2-oleoyl-glycerol and related molecular species. This observation led to the formulation of a hypothesis regarding the potential existence of at least two specialised enzyme isoforms involved in the biosynthesis of such TAGs via the Kennedy pathway, exhibiting unusual substrate specificity. Consequently, this plant can be regarded not only as a producer of unusual molecular types of triacylglycerols but also as a source of genetic material for the search of genes encoding the aforementioned enzymes with unusual substrate specificity. Full article

Alkanna tinctoria L. Tausch (“alkanet” or “alkanna”) is a medicinal plant. Extracts from alkanet roots have applications as natural food coloring agents. In addition, they have proven antioxidant effects. Three classical solvents (ethanol and acidified ethanol/water) and four natural eutectic solvents (NADES)—choline chloride/urea; choline chloride/citric acid; choline chloride/lactic acid; and sodium acetate/formic acid—were compared for their effectiveness as “green” solvents for the extraction of the alkanet pigment. Notably, this study is the first to apply choline chloride-based NADESs for alkanet extraction, providing a comprehensive profile of key bioactive compounds and their contributions to antioxidant activity using UV/Vis and FT-IR spectrometry, GC-MS, and HPLC-PDA-MS. Among the classical solvents, 50% acidified ethanol showed the highest extraction capacity, as indicated by its total flavonoid (708 ± 32 mgCE/L) and total phenolic (1318 ± 63 mgGAE/L) content. However, this extract exhibited the highest total alkaloid content (256 ± 15 µg/L) compared to the other classical extraction solvents. Consequently, absolute ethanol was identified as a more suitable alternative. Among the NADES, the sodium acetate/formic acid (1:2 mol/mol, NADES4) extract was the only one to show the presence of alkannins. This extract also contained high levels of phenols (355 ± 21 mg GAE/L) and tannins (163 ± 10 mg CE/L), exhibited strong antioxidant potential (DPPH: 131 µmol TE/g dw, FRAP: 7.49 mg Fe(II)/mg dw), and contained significantly lower alkaloid levels (7.0 ± 0.5 µg/L). Comparative analyses indicated that the sodium acetate/formic acid extract outperformed those prepared with ethanolic solvents and other NADES. Full article

The environmental impact of invasive species necessitates creating a strategy for managing their spread by utilising them as a source of potentially high-value raw materials. Amelanchier × spicata (Lam.) K. Koch (dwarf serviceberry) is a shrub species in the Rosaceae Juss. family. The evaluation of different populations of plants that accumulate great amounts of biologically active compounds is requisite for the quality determination of plant materials and medicinal and nutritional products. The assessment of natural resources from a phytogeographic point of view is relevant. Phytochemical analysis of A. spicata leaf samples was carried out using spectrophotometric methods, HPLC-PDA, and HPLC-MS techniques, while antioxidant activity was determined using ABTS, FRAP, and CUPRAC assays. A significant diversification of phenolic compounds and antioxidant activity was determined in the A. spicata leaf samples collected in different habitats. Due to their characteristic chemical heterogeneity, natural habitats lead to the diversity of indicators characterising the quality of plant raw materials. Chlorogenic acid and neochlorogenic acid, as well as quercitrin, rutin, and hyperoside, were found to be predominant among the phenolic compounds. Thus, these compounds can be considered phytochemical markers, characteristic of the A. spicata leaf material from northern Europe. Full article

In order to compare the grape juice sensory properties of four common seedless grape varieties (Kyoho, Summer Black, Moldovan, and Sweet Sapphire) in China, a thorough comparison of these grape varieties was conducted. Physicochemical indicators, aroma, and color characteristics were analyzed and compared to a commercially available not-from-concentrate Concord grape juice. The contents of fructose, glucose, and seven organic acids were evaluated. Kyoho and Sweet Sapphire possessed optimal Brix–acid ratio in the range around 35–40. In terms of aroma, 60 volatiles were identified by GC-MS, including 16 alcohols, 9 terpenes, 6 aldehydes, and 4 ketones. Kyoho demonstrated the highest aroma intensity with superior floral and fruity notes, while Sweet Sapphire showed the lowest aroma intensity with a grassy scent. Additionally, grape pigment profiles were investigated by HPLC-PDA-MS. Summer Black grapes featured a vibrant color, and 52.5% of their anthocyanins were acylated, which helps provide good stability in follow-up processing. Concord juice showed the best overall properties, with the most saturated color and delightful aroma. It was suggested that blending Summer Black with Kyoho could be a promising way to achieve balanced color, taste, and aroma of grape juice. This study provides a feasible and promising combination of Chinese local grapes for making all-round high-quality juice products. Full article

The olive oil industry generates large volumes of by-products, creating notable environmental and economic concerns. Among these, olive cake (OC)—a primary by-product of olive oil extraction—stands out due to its high content of bioactive compounds and potential for value-added recycling. This study focused on characterizing six OC samples from the Trás-os-Montes and Alto Douro regions, collected at different processing times and mills. The samples included two derived from pressing (COC), two from two-phase centrifugation (TPOC; one partially pitted and one dehydrated), and two exhausted OC (EOC) samples. Fundamental analyses assessed total phenols, ortho-diphenols, flavonoids, antioxidant capacity, and tannin content. Results revealed significant variation (p < 0.05) in phenolic composition, namely ortho-diphenols and flavonoid levels among the samples. EOC 2 exhibited the highest concentrations (19.61, 21.82, and 20.12 mg CAT/g, respectively), while COC 2 had the lowest (5.08, 5.08, and 2.76 mg GA/g, respectively). This correlated with elevated antioxidant activity in EOC 2, as measured by FRAP, DPPH, and ABTS assays (129.98, 78.00, and 56.65 μmol Trolox/g). In contrast, COC 1 and COC 2 displayed the lowest antioxidant activities (32.61 μmol Trolox/g in FRAP and 17.24 and 18.98 μmol Trolox/g in DPPH). Tannin analysis showed the highest total tannin content in the dehydrated and pitted OC samples (250.31 and 240.89 mg CAT/100 g), with COC 2 showing the lowest (88.17 mg CAT/100 g). Condensed tannin content varied significantly, with EOC 2 presenting the highest level (328.17 mg CAT/100 g) and COC 2 the lowest one (20.56 mg CAT/100 g). Through HPLC-PDA-MS, 22 compounds were identified, with luteolin and verbascoside being particularly prevalent. This in-depth characterization supports the potential valorization of olive by-products, advancing sustainability and promoting a circular economy in the olive oil sector. Full article

Background/Objectives: Horseradish (Armoracia rusticana L.) roots—largely used in traditional medicine for their multiple therapeutic effects—are a rich source of health-promoting phytochemicals. However, their efficacy can be compromised by low chemical stability and poor bioavailability. Incorporation into phospholipid vesicles is often proposed to tackle this problem. Methods: In this study, a hydroalcoholic extract was produced from horseradish roots. The extract was characterized by UPLC-MS and HPLC-PDA and formulated in conventional liposomes and Penetration Enhancer-containing Vesicles (PEVs) for skin application. Results: The obtained nanovesicles were small in size (<100 nm), negatively charged, uni/bilamellar, and with high values of entrapment efficiency (>85%) for the flavonoids identified in the extract. Both the free and the nanoformulated extract showed optimal biocompatibility, measured as the absence of hemolysis of erythrocytes and absence of cytotoxicity in skin cell lines. Furthermore, the nanoformulations displayed antioxidant activity in vitro. Conclusions: The proposed nananoformulations could be exploited to counteract oxidative stress involved in the pathogenesis and progression of numerous skin disorders. Full article

Triticum aestivum (wheat) is one of the most significant crops worldwide. This study compares the chemical composition and radical-scavenging activities of two cultivars of T. aestivum, Saekeumkang wheat (SW) and Baekkang wheat (BW). Sprouted wheatgrass extracts of SW and BW were analyzed using assessments of total polyphenol and flavonoid contents, liquid chromatography–electrospray ionization/mass spectrometry (LC-ESI/MS), and high-performance liquid chromatography with a photodiode array (HPLC-PDA). Radical-scavenging activities were evaluated using 2,2-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS·⁺) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) assays. The results indicated that SW had a higher total polyphenol content than BW, while no significant differences were observed regarding total flavonoid content. HPLC-PDA analysis, guided by LC-ESI/MS, identified four compounds—saponarin, schaftoside, isoorientin, and isovitexin—with isoorientin (3.02 mg/g extract) and schaftoside (4.23 mg/g extract) present in higher concentrations in SW compared to BW. In the ABTS·⁺ assay, the two samples did not show noticeable differences, with SW displaying a scavenging ability with an IC₅₀ of 3.36 mg/mL, and BW with an IC₅₀ of 3.19 mg/mL. Contrarily, the DPPH assay results showed an inverse trend, suggesting that the radical-scavenging behavior may be influenced by the synergistic and antagonistic interactions of the compounds in SW and BW extracts. Full article