Search Results (387)

Given NMN’s (Nicotinamide mononucleotide, NMN) potential pet health benefits and wide use in pet foods, the lack of standardized detection methods hinders quality control. This study developed and validated a simple, rapid HPLC (High-Performance Liquid Chromatography, HPLC) method for NMN determination in pet foods. The method showed good linearity (5–500 μg/mL), LOD (1.0 mg/kg), LOQ (2.0 mg/kg), precision, stability, reproducibility, and spiked recoveries (97.3–109%, RSD < 6.0%). And most tested commercial samples met the standards. This method is simple, efficient, and accurate, supporting pet food NMN detection, quality control, regulation, and pet health protection. Full article

(1) Background: Medicinal plants are widely used in folk medicine. Hypericum perforatum L. (St. John’s wort) is a medicinal plant that is used domestically and exported to other countries. This study addresses the need to develop methods for determining the composition and content of St. John’s wort to determine its biological activity. (2) Methods: High-performance liquid chromatography (HPLC) equipped with an Electrochemical Detector (ECD) and a Photodiode Array Detector (PDA) was employed to identify and quantify major phenolic compounds—gallic acid, catechin, epicatechin, hyperoside, quercetin, and hyperforin—in extracted and lyophilized St. John’s wort flower; stem; and leaf samples. Key analytes exhibited linear responses across both detection systems, within a quantification range of 0.5–10 µg/mL. (3) Results: The PDA method, validated according to ICH Q2(R1) guidelines, demonstrated specificity, linearity, precision, and accuracy, with limits of detection (LOD) ranging from 0.24 to 0.61 µg/mL and limits of quantification (LOQ) between 0.26 and 0.62 µg/mL. PDA effectively identified gallic acid, epicatechin, hyperoside, quercetin, and hyperforin, although catechin was not detected. ECD yielded comparable compound levels across the samples. (4) Conclusions: The novelty of this study lies in identifying the influence of climatic factors associated with the altitude at which St. John’s wort is grown on the content and ratio of biologically active components. Overall, the chemometric approach demonstrates the utility of raw chromatographic data in distinguishing samples by plant part and geographic origin; even when traditional compound-based comparisons may be limited. Full article

Functional foods are gaining heightened popularity in diet modifications. Green bananas contain a significant quantity of resistant starch, dietary fibre, and phytochemicals that demonstrate strong antioxidant properties, particularly due to the high concentration of polyphenols. The community demand for incorporating these essential components into food products, such as bread, has increased. Therefore, the aim of this study was to evaluate the differences in the content and bio-accessibility of phenolic compounds in bread enriched with 5, 10, and 15% of Australian green banana powder (GBF) from (Cavendish “Musa acuminata”, Ladyfinger “Musa paradisiaca L.”, and Ducasse “Musa balbisiana”), as well as their antioxidant capabilities and the generation of short-chain fatty acids (SCFAs) after in vitro gastrointestinal digestion and colonic fermentation. The 15% Cavendish bread exhibited significant TPC and TFC at 1.31 mg GAE/g and 0.05 mg QE/g, respectively, along with substantial antioxidant activity (DPPH, 0.40 mg TE/g), observed following stomach and intestinal phases. However, the 15% Ladyfinger bread exhibited the highest TTC following the stomach digestion, with 17.4 mg CE/g. The bio-accessibility of most phenolic components from 10% GBF-bread was elevated following the gastric and intestinal phases. Nonetheless, a substantial total phenolic content (50.3% in Ladyfinger bread) was still observable in the residue during colonic fermentation. The highest SCFAs production occurred in Cavendish and Ducasse bread after 24 h of fermentation. Overall, the consumption of GBF bread can positively influence intestinal health and provide antioxidant properties, facilitating future advancements in the creation of nutrient-dense and health-enhancing bakery products. Full article

Small ruminant production under grazing conditions plays a crucial role in the global primary sector economy. However, these animals are highly susceptible to gastrointestinal nematodes (GINs), which significantly impact their health and welfare. Given the increasing resistance to conventional anthelmintics, there is a pressing need to explore sustainable alternatives, such as plant secondary metabolites. This study aimed to identify phenolic compounds with anthelmintic activity from Tithonia diversifolia aerial parts, using Haemonchus contortus as a biological model. Egg hatching inhibition (EHI) and larval mortality assays were used to evaluate the anthelmintic activity of a hydroalcoholic extract (HA-E), an aqueous (Aq-F) and ethyl acetate fraction (EtOAc-F), and eight bioactive subfractions (TdR1-TdR8) obtained from EtOAc-F. The identification of major compounds was performed using HPLC-PDA. The E-HA and EtOAc-F achieved 100% EHI at 40 and 4 mg/mL, respectively. The subfractions TdR2 (EC₉₀ = 0.55 mg/mL), TdR3 (EC₉₀ = 0.12 mg/mL), and TdR4 (EC₉₀ = 0.26 mg/mL) exhibited the highest ovicidal activity. In the larval mortality test, EtOAc-F showed an LC₈₅ of 56.74 mg/mL. The major identified compounds included cinnamates, hydroxycinnamic acids (e.g., caffeic acid and chlorogenic acid), gallates, flavonoids (flavones and flavanones), and coumarins. These findings support the potential of T. diversifolia as a promising natural source for the control of GINs in small ruminants. Full article

Cannabis sativa (C. sativa) is a psychoactive plant that has been used for millennia for medicinal, recreational, and industrial purposes. The main constituents of cannabis are the cannabinoids, with other constituents including terpenes and flavonoids that contribute to its bioactivity. Among the flavonoid class, there is a subclass, specific to cannabis, namely the cannflavins (A, B, and C), which are biologically active. This study is directed to the analysis of these constituents in various cannabis chemovars. In this study, an HPLC-PDA method was validated and applied to determine the content of cannflavins, namely, cannflavin A (CF-A), cannflavin B (CF-B), and cannflavin C (CF-C), in six different cannabis chemovars. The HPLC separation was achieved using a Luna^® C18 (150 × 4.6 mm × 3 μm) with isocratic elution using a mobile phase consisting of acetonitrile and water (65:35, v/v), both containing 0.1% formic acid at a flow rate of 1 mL/min, with the detector set at 342.4 nm. The method was validated according to the ICH guidelines and exhibited a linear relationship in the 5–500 ppm range with R² > 0.99. The method showed good recovery, ranging from 82% to 98%. The intra-day and inter-day relative standard deviations (% RSDs) were ≤5.29%. Consequently, the method was applied for the determination of all these cannflavins in the different cannabis chemovars. CF-A was the most abundant cannflavin in the examined samples (15.2–478.38 ppm). The method was shown to be simple, accurate, and selective. Full article

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

Dialium guineense (velvet tamarind), Parkia biglobosa Jacq. (African locust bean) and Adanosonia digitata L. (baobab) are fruits from African plants whose nutritional potential remains poorly characterised. As such, their pulps and seeds were analysed for colour (CIELab system), moisture, ash, protein, fat, soluble and insoluble dietary fibre, free sugars (HPLC-RI), organic acids (HPLC-PDA), macro and microelements (XRF analyser) and amygdalin (HPLC-PDA). The colours of their pulps differed considerable (ΔE > 38 between the velvet tamarind and African locust bean) and the moisture content was lower in seeds (about 7%) compared to pulps (9–13%). Seeds were more concentrated in protein (20–28%) and fat (5–22%), whereas pulps were richer in sugar (1–12%). African locust bean pulp was the sweetest (39% total sugar), while baobab pulp contained the highest soluble fibre (>30%) and citric acid (3.2%), and velvet tamarind pulp was distinguished by its tartaric acid content (3.4%). Seeds of the African fruits presented higher Ca, P, S and Fe contents, whereas pulps had higher K content. No amygdalin (<6.34 mg per 100 g of dry weight) or toxic heavy metal contents were detected. The PCA segregated samples by pulp and seed and the PC1 explains the sugar and moisture of the pulps, while protein, fat and minerals are associated with the seeds. These data confirm that African fruit pulps and seeds have distinct functional profiles, are safe for food use and can be consumed, which is important for efforts to promote the conservation of these tropical plant species. Full article

This study investigated the natural bioactive compounds in Polygonum cuspidatum Sieb. et Zucc. (P. cuspidatum) by fractionating a 70% ethanol extract using n-hexane, chloroform, ethyl acetate, n-butanol, and water. The total polyphenol and flavonoid contents of each fraction were determined, and their antioxidant activities were evaluated using DPPH, ABTS, and FRAP assays. Additionally, the anti-diabetic potential was assessed via α-glucosidase inhibitory activity, while anti-obesity activity was evaluated using lipase inhibitory activity. The fractions were also tested for tyrosinase and elastase inhibitory activities to assess their skin-whitening and anti-wrinkle potential, and their antibacterial activity against Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa was determined using the agar diffusion method. Finally, bioactive compounds were identified and quantified using HPLC and GC–MSD. The results showed that the ethyl acetate fraction possessed the highest total polyphenol content (0.53 ± 0.01 g GAE/g) and total flavonoid content (0.19 ± 0.02 g QE/g). It also exhibited strong antioxidant activity, with the lowest DPPH radical scavenging IC₅₀ (0.01 ± 0.00 mg/mL), ABTS radical scavenging IC₅₀ (0.06 ± 0.00 mg/mL), and the highest FRAP value (6.02 ± 0.30 mM Fe²⁺/mg). Moreover, it demonstrated potent enzyme inhibitory activities, including tyrosinase inhibitory activity (67.78 ± 2.50%), elastase inhibitory activity (83.84 ± 1.64%), α-glucosidase inhibitory activity (65.14 ± 10.29%), and lipase inhibitory activity (85.79 ± 1.04%). In the antibacterial activity, the ethyl acetate fraction produced a clear inhibitory zone of 19.50 mm against Staphylococcus aureus, indicating notable antibacterial activity. HPLC-PDA and GC–MSD analyses identified tannic acid and emodin as the major bioactive constituents. These findings suggest that the ethyl acetate fraction of P. cuspidatum extract, rich in polyphenol and flavonoid compounds, is a promising natural source of bioactive ingredients for applications in the food, pharmaceutical, and cosmetic industries. Further research is needed to explore its mechanisms and therapeutic applications. Full article

Increased market demand for plant herbs has prompted growers to ensure a continuous and assured supply of superior nutritional quality over the years. Apart from the nutritional value, culinary herbs contain phytochemical benefits that can improve human health. However, a significant amount of research has focused on enhancing yield, frequently overlooking the impact of production practices on the antioxidant and phytonutritional content of the produce. Thus, the study aimed to evaluate the yield, phytonutrients, and essential mineral profiling in selected aromatic herbs and their intricate role in nutritional quality when grown under different production systems. Five selected aromatic herbs (coriander, rocket, fennel, basil, and moss-curled parsley) were evaluated at harvest when grown under three production systems: in a gravel-film technique (GFT) hydroponic system and in soil, both under the 40% white shade-net structure, as well as in a soilless medium using sawdust under a non-temperature-controlled plastic tunnel (NTC). The phytonutritional quality properties (total phenolic, flavonoids, β-carotene-linoleic acid, and condensed tannins contents) as well as 1,1-diphenyl-2-picrylhydrazyl (DPPH) were assessed using spectrophotometry, while vitamin C and β-carotene were analyzed using HPLC-PDA, and leaf mineral content was evaluated using ICP-OES (Inductively Coupled Plasma Optical Emission Spectrometry). The results show that the health benefits vary greatly owing to the particular culinary herb. The fresh leaf mass (yield) of coriander, parsley, and rocket was not significantly affected by the production system, whereas basil was high in soil cultivation, followed by GFT. Fennel had a high yield in the GFT system compared to in-soil and in-soilless cultivation. The highest levels of vitamin C were found in basil leaves grown in GFT and in soil compared to the soilless medium. The amount of total phenolic and flavonoid compounds, β-carotene, β-carotene-linoleic acid, and DPPH, were considerably high in soil cultivation, except on condensed tannins compared to the GFT and soilless medium, which could be a result of Photosynthetic Active Radiation (PAR) values (683 μmol/m²/s) and not favoring the accumulation of tannins. Overall, the mineral content was greatly influenced by the production system. Leaf calcium and magnesium contents were highly accumulated in rockets grown in the soilless medium and the GFT hydroponic system. The results have highlighted that growing environmental conditions significantly impact the accumulation of health-promoting phytonutrients in aromatic herbs. Some have positive ramifications, while others have negative ramifications. As a result, growers should prioritize in-soil production systems over GFT (under the shade-net) and soilless cultivation (under NTC) to produce aromatic herbs to improve the functional benefits and customer health. 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

The major objective was to investigate the protective capabilities of endophytic bacterial strains isolated from a number of medicinal plant species towards Aspergillus spp. secured from the internal tissues of fungi-infected peanuts. Among 32 fungal isolates surveyed for mycotoxin production in various culture media (PDA, RBCA, YES, CA), 10 isolates qualitatively producing AFB₁, besides 10 OTA-producers, were assayed by HPLC for quantitative toxin production. Aspergillus spp. isolate Be 13 produced an extraordinary quantity of 1859.18 μg mL⁻¹ AFB₁, against the lowest toxin level of 280.40 μg mL⁻¹ produced by the fungus isolate IS 4. The estimated amounts of OTA were considerably lower and fell in the range 0.88–6.00 μg mL⁻¹; isolate Sa 1 was superior, while isolate Be 7 seemed inferior. Based on ITS gene sequencing, the highly toxigenic Aspergillus spp. isolates Be 13 and Sa 1 matched the description of A. novoparasiticus and A. ochraceus, respectively, ochraceus, respectively, which are present in GenBank with identity exceeding 99%. According to 16S rRNA gene sequencing, these antagonists labeled Ar6, Ma27 and So34 showed the typical characteristics of Pseudomonas aeruginosa, Bacillus subtilis and Bacillus velezensis, respectively, with similarity percentages of 99–100. The plant growth-promoting activity measurements of the identified endophytes indicated the production of 16.96–80.00 μg/100 mL culture medium of IAA. Phosphate-solubilizing capacity varied among endophytes from 2.50 to 21.38 μg/100 mL. The polysaccharide production pool of bacterial strains ranged between 2.74 and 6.57 mg mL⁻¹. P. aeruginosa Ar6 and B. velezensis successfully produced HCN, but B. subtilis failed. The in vitro mycotoxin biodegradation potential of tested bacterial endophytes indicated the superiority of B. velezensis in degrading both mycotoxins (AFB₁-OTA) with average percentage of 88.7; B. subtilis ranked thereafter (85.6%). The 30-day old peanut (cv. Giza 6) seedlings grown in gnotobiotic system severely injured due to infection with AFB₁/OTA-producing fungi, an effect expressed in significant reductions in shoot and root growth traits. Simultaneous treatment with the endophytic antagonists greatly diminished the harmful impact of the pathogens; B. velezensis was the pioneer, not P. aeruginosa Ar6. In conclusion, these findings proved that several endophytic bacterial species have the potential as alternative tools to chemical fungicides for protecting agricultural commodities against mycotoxin-producing fungi. Full article

Background: The Quantitative Analysis of Multi-components by Single-marker (QAMS) method has been developed as an alternative to the External Standards Method (ESM) for the quality control of medicinal herbs. Objectives: In this study, QAMS was developed to determine saponins in the raw materials of Panax vietnamensis using HPLC-PDA/ELSD. Methods: The method was developed and validated. The relative conversion factors F_x were calculated based on the linear regression for HPLC-PDA and the logarithm equation for HPLC-ELSD. The Standard Method Difference (SMD) was determined to indicate the difference in the results of QAMS and EMS. Results: Relative conversion factors (F_x) were determined for each detector to quantify five saponins (ginsenoside Rb₁, Rd, Rg₁, majnoside R2, and vina-ginsenoside R2) in VG root. The F_x values were calculated based on the ratio of the slopes of the regression equations of a single standard and an external standard. For HPLC-PDA, G-Rb1 was used as a single standard with the F_x values of 1.00 (G-Rb₁), 1.08 (G-Rd), 1.32 (G-Rg₁), and 0.04 (M-R2). For HPLC-ELSD, G-Rb₁ was used for determining the content of G-Rg₁ and G-Rb₁ with the F_x values of 1.00 (G-Rb₁) and 0.95 (G-Rg₁), while M-R2 was used for quantitating M-R2 and V-R2 with F_x of 1.00 (M-R2) and 1.05 (V-R2). An SMD value less than 5.00% confirms the close alignment of the QAMS method with ESM. Conclusions: The QAMS method proved to be a feasible and promising method for the quality control of P. vietnamensis. Full article

Type 2 diabetes mellitus (T2DM) is a chronic metabolic disorder characterized by impaired glucose regulation. This study evaluated the antioxidant and antidiabetic potential of aqueous extracts from four plant species from the southern Algarve: Aristolochia baetica, Chelidonium majus, Dittrichia viscosa, and Lavandula viridis, using non-cellular in vitro assays. HPLC/PDA was used to identify active compounds. Antioxidant activity was assessed by using TAA, FRAP, RP, and DPPH assays; antidiabetic potential through α-glucosidase and α-amylase inhibition; and wound healing relevance through elastase, collagenase, and lipoxygenase inhibition. D. viscosa showed the highest antioxidant activity (FRAP: 1132.99 ± 19.54 mg TE/g dw; DPPH IC₅₀ = 25.85 ± 0.75 μg/mL) and total phenolic/flavonoid content, with a diverse profile including caffeic and chlorogenic acids, isoquercetin, and quercetin. It also exhibited potent α-glucosidase inhibition (IC₅₀ = 0.61 ± 0.06 mg/mL), outperforming acarbose. L. viridis had the highest total phenolic content (39.04 mg/g), while A. baetica demonstrated the strongest anti-elastase, anti-collagenase, and lipoxygenase activity, suggesting wound-healing potential. C. majus showed the weakest effects. A strong correlation was observed between phenolic content and antioxidant/antidiabetic activity. These findings support further in vivo studies on D. viscosa and A. baetica for potential use in T2DM management and diabetic wound healing. 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