Search Results (112)

Various metabolic modulators have been widely used in recent years to increase the accumulation of desired secondary metabolites in medicinal plants, although most studies to date have focused on in vitro systems. Although simpler and cheaper, their potential application in vivo is still limited. Therefore, the aim of this study was to compare the effect of three chemically different elicitors (150 mg/L chitosan lactate—ChL; 10 mg/L selenium as selenite—Se; 100 mg/L salicylic acid—SA) applied to the soil substrate on some aspects of the secondary metabolism and physiological responses of Chelidonium majus L. Using HPLC-DAD, six isoquinoline alkaloids were identified and quantified in shoot extracts. LC-ESI-TOF-MS analysis confirmed the molecular identity of all target alkaloids, supporting the identification. The strongest stimulatory effect on the accumulation of protopine, berberine, and allocryptopine was observed with the Se and SA treatment, whereas ChL was less effective. In turn, the dominant alkaloids (coptisine and chelidonine) remained unaffected. There was also an increase in total phenolic compounds, but not in soluble flavonols. The elicitor treatments caused an increase in the antioxidant activity of the plant extracts obtained. Regardless of the metabolic modulator type, the strongest effect was generally observed on days 7 and 10 after application. No visual signs of toxicity and no effect on shoot biomass were found, although some elicitor-induced changes in the oxidative status (increased H₂O₂ accumulation and enhanced lipid peroxidation) and free proline levels in leaves were observed. We suggest that Se or SA can be applied to C. majus grown in a controlled pot culture to obtain high-quality raw material and extracts with increased contents of valuable specialized metabolites and enhanced antioxidant capacity. Full article

Eleutherococcus senticosus (ES) has been used in traditional medicine for immune-boosting, stress-reducing, and endurance-enhancing properties. In this study, the chemical composition and biological activity of polar and non-polar fractions obtained from 75% methanol E. senticosus roots extract were evaluated. Spectrophotometric methods were used to assess the antioxidant (DPPH, ABTS, FRAP, CUPRAC, O₂^•−) and anti-enzymatic (hyaluronidase, acetylcholinesterase, butyrylcholinesterase, α-amylase, and tyrosinase) activities. Metabolic profiling was carried out using HPLC-DAD and UHPLC-DAD/ESI-TOF-MS. The ethyl acetate fraction (EtOAc) showed the highest antioxidant activity with IC₅₀ values of 82.73 ± 0.065 µg/mL (DPPH) and 9.92 ± 0.17 µg/mL (ABTS). The EtOAc fraction also exhibited strong anti-enzymatic effects against hyaluronidase and α-amylase (125.24 ± 12.29 and 97.34 ± 9.18 µg/mL, resp.). In turn, the hexane fraction exhibited the most potent anti-AChE activity with IC₅₀ equal 245.72 ± 11.82 µg/mL. The HPLC-DAD analysis revealed the presence of caffeic acid derivatives. These results suggest that the ethyl acetate fraction may have therapeutic relevance in inflammation- and metabolic-related diseases. We perceive the potential of this fraction as a rich source of compounds with an anti-inflammatory activity; however, more advanced research in in vivo model is required. Full article

This study critically examines the limitations of the official Italian methodology used for detecting bovine adulteration milk in Protected Designation of Origin (PDO) Mozzarella di Bufala Campana (MdBC). This method focuses on the whey fraction of cheese samples, which comprises about 1% of total MdBC proteins, and is based on a high-performance liquid chromatography (HPLC) quantification of the bovine β-lactoglobulin A (β-Lg A) as a marker. Here, we have demonstrated that this official methodology suffers from measurement inconsistencies due to its reliance on raw bovine whey standards, which fail to account for β-Lg genetic polymorphisms in real MdBC samples and protein thermal modifications during cheesemaking. To overcome these limitations, we propose a dual proteomics-based approach using matrix-assisted laser desorption ionization (MALDI-TOF) mass spectrometry (MS) and nano-HPLC-electrospray (ESI)−tandem mass spectrometry (MS/MS) analysis of MdBC extracted whey. MALDI-TOF-MS focused on identifying proteotypic peptides specific to bovine and buffalo β-Lg and α-lactalbumin (α-La), enabling high specificity for distinguishing the two animal species at adulteration levels as low as 1%. Complementing this, nano-HPLC-ESI-MS/MS provided a comprehensive profile by identifying over 100 bovine-specific peptide markers from β-Lg, α-La, albumin, lactoferrin, and osteopontin. Both methods ensured precise detection and quantification of bovine milk adulteration in complex matrices like pasta filata cheeses, achieving high sensitivity even at minimal adulteration levels. Accordingly, the proposed dual proteomics-based approach overcomes challenges associated with whey protein polymorphism, heat treatment, and processing variability, and complements casein-based methodologies already validated under European standards. This integrated framework of analyses focused on whey and casein fraction enhances the reliability of adulteration detection and safeguards the authenticity of PDO buffalo mozzarella, upholding its unique quality and integrity. Full article

Calix[4]resorcinarenes are polyhydroxylated macrocyclic compounds with four units of resorcinol. These compounds can be derivatized through modifications at the upper rim, allowing reactivity with secondary amines to produce Mannich base derivatives via Mannich-type aminomethylation reactions. In this paper, we report the reaction of C-tetra(propyl)calix[4]resorcinarene with piperidine in acetonitrile. The aminomethylated compound C-2,8,14,20-tetra(propyl)-5,11,17,23-tetrakis(N–(piperidine)methyl)calix[4]resorcinarene was obtained with a 52% yield, with an exact mass of 1044.6994 u and a mass error of 7.6 ppm. The reaction progress and product formation were monitored by RP-HPLC, and the compound was characterized using LC ESI-TOF/MS, one- and two-dimensional ¹H and ¹³C NMR, and FTIR spectroscopy. Chromatographic and spectroscopy data are presented and discussed. Full article

The olive leaf is one of the main by-products from the olive oil industry. This by-product is a rich source of phenolic compounds that have been shown to possess beneficial health activities, which are due in part to their antioxidant activities. Therefore, the revaluation of this by-product would be of great importance for the food industry. For this reason, this study focuses on the pretreatment of olive leaves with a technology based on the use of pulsed electric fields (PEF) and their following extraction by ultrasounds in order to obtain an extract enriched in phenolic compounds. A Box-Behnken design of 15 experiments with three independent factors has been carried out: electric field strength (kV/cm), frequency (Hz) and total treatment time (s). The response variables were the sum of phenolic compounds, hydroxytyrosol and oleuropein measured by HPLC-MS-ESI-TOF and the antioxidant activity measured by DPPH. The validity of the experimental design was confirmed by ANOVA and the optimal conditions were established by using the response surface methodology in combination with a desirability function. The PEF optimal conditions were 0.6 kV/cm at 90 Hz for 11 s, which allowed for obtaining an olive leaf extract with 26.8, 21.7 and 15.6% higher contents of hydroxytyrosol, oleuropein and total phenolic compounds, respectively, compared to the non-treated sample with PEF. The antioxidant activity measured by DPPH was increased significantly by 32.3%. The data confirmed that the pre-treatment with PEF under these optimal conditions has proven to be effective in improving the extraction of phenolic compounds in olive leaves. Full article

Despite the widespread use of COVID-19 vaccines, there is still a global need to find effective therapeutics to deal with the variants of SARS-CoV-2. Moslae herba (MH) is a herbal medicine credited with antiviral effects. This study aims to investigate the antiviral effects and the underlying mechanism of aqueous extract of Moslae herba (AEMH) for treating SARS-CoV-2. The in vitro anti-SARS-CoV-2 activity of AEMH was evaluated using cell viability and viral load. Component analysis was performed by HPLC-ESI-Q-TOF/MS. The connection between COVID-19 and AEMH was constructed by integrating network pharmacology and transcriptome profiles to seek the core targets. The components with antiviral activities were analyzed by molecular docking and in vitro pharmacological verification. AEMH exerted anti-SARS-CoV-2 effects by inhibiting viral replication and reducing cell death caused by infection (IC₅₀ is 170 μg/mL for omicron strain). A total of 27 components were identified from AEMH. Through matching 119 intersection targets of ‘disease and drug’ with 1082 differentially expressed genes of COVID-19 patients, nine genes were screened. Of the nine, the PNP and TPI1 were identified as core targets as AEMH treatment significantly regulated the mRNA expression level of the two genes on infected cells. Three components, caffeic acid, luteolin, and rosmarinic acid, displayed antiviral activities in verification. Molecular docking also demonstrated they could form stable bonds with the core targets. This study explored the antiviral activity and possible mechanism of AEMH for treating SARS-CoV-2, which could provide basic data and reference for the clinical application of MH. Full article

The separation and purification of molecular compounds and their functionalized derivatives is a common challenge in organic synthesis. In particular, calix[4]resorcinarenes present a high potential for chemical derivatization at their upper edge by aminomethylation reactions, and these compounds and their derivatives require appropriate analytical methodologies for their analysis, separation, and purification. In this study, C-tetra(propyl)calix[4]resorcinarene was synthesized and functionalized with maleimide groups by optimized aminomethylation reactions, obtaining a mixture of mono-, di-, tri-, and tetrasubstituted compounds. Initial separation by RP-HPLC with a core-shell column showed poorly resolved peaks, indicating a loss of separation efficiency. Therefore, a monolithic C18 column was used, which significantly improved the separation, thanks to its larger pore volume and continuous structure facilitating the diffusion of these bulky molecules, notably improving efficiency. Finally, the six compounds functionalized with maleimide groups were efficiently separated and enriched by RP-SPE by analytical method transfer, and the two peptides of six and the thirteen residues derived from LfcinB (20–25): RRWQWR were synthesized by SPPS-Fmoc/tBu and purified. These were modularly linked by the Michael thiol-maleimide addition reaction obtaining six (peptide)_n-resorcinarene conjugates. The analytical method by RP-HPLC with a monolithic C18 column, the separation and purification by RP-SPE were used transversally in all the steps to obtain compounds with adequate purities and quantities. Finally, the antibacterial activities of the six conjugates were evaluated against E. coli and E. faecalis strains, and it was determined that three aminomethylated compounds and one monosubstituted conjugate showed activity against E. faecalis. Our work established a new modular conjugation strategy between calix[4]resorcinarenes and peptides by thiol-maleimide click chemistry, and a methodology of separation, purification, and enrichment for these products by RP-HPLC and RP-SPE, which permitted us to obtain quantities with purities appropriate for their characterization by NMR, LC-MS and antibacterial activity assays. Full article

Dendrobium fimbriatum is a perennial herb, and its stems are high-grade tea and nourishing medicinal materials. Various solvent extracts of D. fimbriatum were evaluated for their anti-inflammatory, anti-acetylcholinesterase (AChE), antioxidant, and anti-α-glucosidase properties. Acetone and EtOAc extracts showed significant antioxidant effects. Acetone, n-hexane, and EtOAc extracts revealed potent inhibition against α-glucosidase. EtOAc, n-hexane, and dichloromethane extracts displayed significant anti-AChE activity. Among the isolated constituents, gigantol, moscatin, and dendrophenol showed potent antioxidant activities in FRAP, DPPH, and ABTS radical scavenging tests. Moscatin (IC₅₀ = 161.86 ± 16.45 μM) and dendrophenol (IC₅₀ = 165.19 ± 13.25 μM) displayed more potent anti-AChE activity than chlorogenic acid (IC₅₀ = 236.24 ± 15.85 μM, positive control). Dendrophenol (IC₅₀ = 14.31 ± 3.17 μM) revealed more efficient anti-NO activity than quercetin (positive control, IC₅₀ = 23.09 ± 1.43 μM). Analysis of AChE and iNOS inhibitory components was performed using molecular docking and/or the bioaffinity ultrafiltration method. In bioaffinity ultrafiltration, the binding affinity of compounds to the enzyme (acetylcholinesterase and inducible nitric oxide synthase) was determined using the enrichment factor (EF). Among the main components of the EtOAc extract from D. fimbriatum stem, moscatin, dendrophenol, gigantol, and batatasin III with acetylcholinesterase exhibited the highest binding affinities, with affinity values of 66.31%, 59.48%, 54.60%, and 31.87%, respectively. Moreover, the affinity capacity of the identified compounds with inducible nitric oxide synthase can be ranked as moscatin (88.99%) > dendrophenol (65.11%) > gigantol (44.84%) > batatasin III (27.18%). This research suggests that the bioactive extracts and components of D. fimbriatum stem could be studied further as hopeful candidates for the prevention or treatment of hyperglycemia, oxidative stress-related diseases, and nervous disorders. Full article

Olive oil is a food of great importance in the Mediterranean diet and culture. However, during its production, the olive oil industry generates a large amount of waste by-products that can be an important source of bioactive compounds, such as phenolic compounds and terpenes, revalorizing them in the context of the circular economy. Therefore, it is of great interest to study the distribution and abundance of these bioactive compounds in the different by-products. This research is a screening focused on phytochemical analysis, with particular emphasis on the identification and quantification of the phenolic and terpenic fractions. Both the main products of the olive industry (olives, olive paste and produced oil) and the by-products generated throughout the oil production process (leaf, “alpeorujo”, liquid and solid residues generated during decanting commonly named “borras” and washing water) were analyzed. For this purpose, different optimized extraction procedures were performed for each matrix, followed by high-performance liquid chromatography coupled with electrospray time-of-flight mass spectrometry (HPLC-ESI-TOF/MS) analysis. Although no phenolic alcohols were quantified in the leaf and the presence of secoiridoids was low, this by-product was notable for its flavonoid (720 ± 20 µg/g) and terpene (5000 ± 300 µg/g) contents. “Alpeorujo” presented a complete profile of compounds of interest, being abundant in phenolic alcohols (900 ± 100 µg/g), secoiridoids (4500 ± 500 µg/g) and terpenes (1200 ± 100 µg/g), among others. On the other hand, while the solid residue of the borras was the most abundant in phenolic alcohols (3700 ± 200 µg/g) and secoiridoids (680 ± 20 µg/g), the liquid fraction of this waste was notable for its content of elenolic acid derivatives (1700 ± 100 µg/mL) and phenolic alcohols (3000 ± 300 µg/mL). Furthermore, to our knowledge, this is the first time that the terpene content of this by-product has been monitored, demonstrating that it is an important source of these compounds, especially maslinic acid (120 ± 20 µg/g). Finally, the phytochemical content in wash water was lower than expected, and only elenolic acid derivatives were detected (6 ± 1 µg/mL). The results highlighted the potential of the olive by-products as possible alternative sources of a wide variety of olive bioactive compounds for their revalorization into value-added products. Full article

Orange processing generates peel by-products rich in phenolic compounds, particularly flavanones like hesperidin and narirutin, offering potential health benefits. Utilizing these by-products is of significant interest in supporting Spain’s circular bioeconomy. Therefore, the aim of this study was to investigate the fermentation of orange peels by different lactic acid bacteria (LAB) strains and its impact on phenolic composition and antioxidant activity. Three different LAB strains, two Lactiplantibacillus plantarum, and one Levilactobacillus brevis were utilized. The phenolic compounds were measured by HPLC-ESI-TOF-MS, and antioxidant activity was assessed using DPPH and ABTS methods. The growth of the LAB strains varied, showing initial increases followed by gradual declines, with strain-specific patterns observed. Medium acidification occurred during fermentation. A phenolic analysis revealed an 11% increase in phenolic acids in peels fermented by La. plantarum CECT 9567-C4 after 24 h, attributed to glycosylation by LAB enzymes. The flavonoid content exhibited diverse trends, with Le. brevis showing an 8% increase. The antioxidant assays demonstrated strain- and time-dependent variations. Positive correlations were found between antioxidant activity and total phenolic compounds. The results underscore the importance of bacterial selection and fermentation time for tailored phenolic composition and antioxidant activity in orange peel extracts. LAB fermentation, particularly with La. plantarum CECT 9567 and Le. brevis, holds promise for enhancing the recovery of phenolic compounds and augmenting antioxidant activity in orange peels, suggesting potential applications in food and beverage processing. Full article

Lentil is a leguminous crop with a high content of health-beneficial polyphenols. Lentil sprouts are popularly consumed in fresh vegetable markets, although their phytochemical qualities are not well understood. In this study, we investigated the accumulation of phenolics in lentil sprouts in response to photosynthetic and stress light qualities, including fluorescent light (FL), red LED (RL), blue LED (BL), ultraviolet A (UV-A), and ultraviolet B (UV-B). Three lentil cultivars, Lentil Green (LG), French Green (FG), and Lentil Red (LR), were used to evaluate sprouts grown under each light condition. The adequate light intensities for enhancing the antioxidant activity of lentil sprouts were found to be 11 W/m² under photosynthetic lights (FL, RL, BL), and 1 W/m² under stress lights (UV-A, UV-B). Subsequently, HPLC-ESI/Q-TOF MS analysis was conducted for the quantitative analysis of the individual phenolics that were accumulated in response to light quality. Four main phenolic compounds were identified: ferulic acid, tricetin, luteolin, and kaempferol. Notably, tricetin accumulation was significantly enhanced under BL across all three lentil cultivars examined. Furthermore, the study revealed that the other phenolic compounds were highly dependent on FL, BL, or UV-B exposure, exhibiting cultivar-specific variations. Additionally, the antioxidant activities of lentil extracts indicated that BL was most effective for LG and FG cultivars, whereas FL was most effective for enhancing antioxidant activity of LR cultivars as the sprouts grew. Full article

Cherry stems, prized in traditional medicine for their potent antioxidant and anti-inflammatory properties, derive their efficacy from abundant polyphenols and anthocyanins. This makes them an ideal option for addressing skin aging and diseases. This study aimed to assess the antioxidant and anti-inflammatory effects of cherry stem extract for potential skincare use. To this end, the extract was first comprehensively characterized by HPLC-ESI-qTOF-MS. The extract’s total phenolic content (TPC), antioxidant capacity, radical scavenging efficiency, and its ability to inhibit enzymes related to skin aging were determined. A total of 146 compounds were annotated in the cherry stem extract. The extract effectively fought against NO^· and HOCl radicals with IC₅₀ values of 2.32 and 5.4 mg/L. Additionally, it inhibited HYALase, collagenase, and XOD enzymes with IC₅₀ values of 7.39, 111.92, and 10 mg/L, respectively. Based on the promising results that were obtained, the extract was subsequently gently integrated into a cosmetic gel at different concentrations and subjected to further stability evaluations. The accelerated stability was assessed through temperature ramping, heating-cooling cycles, and centrifugation, while the long-term stability was evaluated by storing the formulations under light and dark conditions for three months. The gel formulation enriched with cherry stem extract exhibited good stability and compatibility for topical application. Cherry stem extract may be a valuable ingredient for creating beneficial skincare cosmeceuticals. Full article

This study defined the physicochemical attributes, composition, and antioxidant capacity of four Polish cultivars of cornelian cherry (CC) at six stages of development and ripening. A total of 52 metabolites were identified by UPLC-ESI-qTOF-MS/MS and quantified by HPLC-PDA. In general, phenolic acids, hydrolyzable tannins, flavonols, iridoids, antioxidant activity, organic acids, and vitamin C decreased, while anthocyanins, malic acid, sugars, and titratable acidity increased. For the first time, we determined the evolution of the CC chemical properties and the metabolic behavior and quantified the individual compounds, and groups of compounds during ripening, in particular gallotannins, ellagitannins, iridoids, and organic acids. The main novelty of our study is that CC is a valuable resource for utilization at different degrees of maturity. We showed that unripe fruits in particular deserve valorization, as they contained the highest content of total bioactive phytocompounds (5589.1–6779.6 mg/100 g dw)—primarily phenolic acids > iridoids > tannins—and the highest antioxidant capacity. The intermediate stages were the most abundant in vitamin C (341.1–495.6 mg/100 g dw), ellagic acid (5.9–31.6 mg/100 g dw), gallotannins (47.8–331.1 mg/100 g dw), and loganic acid (1393.0–2839.4 mg/100 g dw). The ripe fruits contained less bioactive phytocompounds (1403.7–1974.6 mg/100 g dw)—primarily iridoids > phenolic acids > tannins > anthocyanins—and the lowest antioxidant capacity. On the other hand, ripe fruits showed the highest content of anthocyanins (30.8–143.2 mg/100 g dw), sugars (36.4–78.9 g/100 g dw), malic acid (5.5–12.2 g/100 g dw), and, favorably for the nutritional applications, the highest sugar-to-acids ratio (3.0–6.4). Our work illustrates in detail that quality attributes and the content of health-promoting phytocompounds in CC depend on the ripening stage and on the cultivar. These results advance the scientific knowledge about CC. Our findings can be helpful to select the optimal properties of CC for the development of diverse functional foods and phytopharmaceuticals applied in the prevention of civilization diseases. Full article

Plant extracts rich in phenolic compounds have been reported to exert different bioactive properties. Despite the fact that there are plant extracts with completely different phenolic compositions, many of them have been reported to have similar beneficial properties. Thus, the structure–bioactivity relationship mechanisms are not yet known in detail for specific classes of phenolic compounds. In this context, this work aims to demonstrate the relationship of extracts with different phenolic compositions versus different bioactive targets. For this purpose, five plant matrices (Theobroma cacao, Hibiscus sabdariffa, Silybum marianum, Lippia citriodora, and Olea europaea) were selected to cover different phenolic compositions, which were confirmed by the phytochemical characterization analysis performed by HPLC-ESI-qTOF-MS. The bioactive targets evaluated were the antioxidant potential, the free radical scavenging potential, and the inhibitory capacity of different enzymes involved in inflammatory processes, skin aging, and neuroprotection. The results showed that despite the different phenolic compositions of the five matrices, they all showed a bioactive positive effect in most of the evaluated assays. In particular, matrices with very different phenolic contents, such as T. cacao and S. marianum, exerted a similar inhibitory power in enzymes involved in inflammatory processes and skin aging. It should also be noted that H. sabdariffa and T. cacao extracts had a low phenolic content but nevertheless stood out for their bioactive antioxidant and anti-radical capacity. Hence, this research highlights the shared bioactive properties among phenolic compounds found in diverse matrices. The abundance of different phenolic compound families highlights their elevated bioactivity against diverse biological targets. Full article

Paeonia ostii ‘Feng Dan’ is widely cultivated in China for its ornamental, medicinal, and edible properties. The whole plant of tree peony is rich in bioactive substances, while the comprehensive understanding of metabolites in the leaves is limited. In this study, an untargeted metabolomics strategy based on UPLC-ESI-TOF-MS was conducted to analyze the dynamic variations of bioactive metabolites in P. ostii ‘Feng Dan’ leaves during development. A total of 321 metabolites were rapidly annotated based on the GNPS platform, in-house database, and publications. To accurately quantify the selected metabolites, a targeted method of HPLC-ESI-QQQ-MS was used. Albiflorin, paeoniflorin, pentagalloylglucose, luteolin 7-glucoside, and benzoylpaeoniflorin were recognized as the dominant bioactive compounds with significant content variations during leaf development. Metabolite variations during the development of P. ostii ‘Feng Dan’ leaves are greatly attributed to the variations in antioxidant activities. Among all tested bacteria, the leaf extract exhibited exceptional inhibitory effects against Streptococcus hemolytis-β. This research firstly provides new insights into tree peony leaves during development. The stages of S1–S2 may be the most promising harvesting time for potential use in food or pharmaceutical purposes. Full article