Search Results (44)

Plant materials rich in proanthocyanidins are fractionated to determine the structure of these compounds and relate it to bioactivity. The aim of this study was to fractionate a procyanidin-rich hawthorn bark extract using low-pressure liquid chromatography and to determine the compound profile and antioxidant activity of the obtained fractions. We identified and quantified the phenolics of four fractions (I–IV) separated on a Toyopearl HW-40S column with methanol as the mobile phase, using HPLC-DAD and LC-ESI-MS techniques. The antioxidant activity was determined to comprise ABTS^•+ and DPPH^• scavenging activity, ferric-reducing antioxidant power (FRAP), and inhibition of β-carotene-linoleic acid emulsion oxidation. Characteristic data were subjected to principal component analysis (PCA). Fraction I contained mainly (−)-epicatechin (741.3 mg/g) and a lower amount of flavones and quercetin derivatives (100.7 mg/g). Fraction II was almost pure procyanidin B2, which accounted for 88.8% of the total phenolics. The subsequent fractions were rich in B-type procyanidin dimers, trimers, and tetramers. FRAP and antiradical activity against ABTS^•+ and DPPH^• of the fraction containing low-molecular weight phenolics was lower than those of the fractions with procyanidin oligomers. The antioxidant activity of fractions II–IV ranged from 8.95 to 9.28 and from 6.45 to 6.71 mmol TE/g in the ABTS and DPPH assays, respectively. Their FRAP was in the range of 17.67–21.06 mmol Fe²⁺/g. According to PCA, the procyanidin dimers of fractions II and III were associated with antioxidant activity in these assays. In turn, the procyanidins with the highest degree of polymerization (trimers and tetramers) present in fraction IV were related to the antioxidant activity measured in the β-carotene-linoleic acid emulsion system. Overall, the separation of purified hawthorn bark extract using low-pressure Toyopearl HW-40S column chromatography resulted in a fraction rich in procyanidin B2, as well as fractions containing procyanidins with an increasing degree of polymerization, all with high levels of antioxidant activity under various conditions and the potential for future applications in food, pharmaceuticals, and cosmetics products. Full article

Rhododendron tomentosum Harmaja is a marsh plant known for its high content of bioactive components, including essential oil, flavonoids, and phenolic acids. In the current work, the effects of cultivation mode (agar, liquid stationary, shake flask, and temporary immersion) and experiment duration (30, 60, and 90 days) on the growth and contents of non-volatile phenolics in Rhododendron tomentosum microshoots were investigated. Agar and liquid stationary cultures provided the highest dry biomass yield per liter, but their dry weight productivities per day were the lowest among the tested systems. Agitated and temporary immersion cultures, on the other hand, were the most productive in terms of fresh and dry biomass yield per day. LC-DAD-ESI-MS analysis of extracts from microshoots and wild-grown plants revealed similarities in phenolic composition: in both cases, the presence of catechin, chlorogenic acid, and flavonoids of flavonol type (mainly glycosides of quercetin and myricetin) was confirmed. The qualitative composition of the phenolic fraction was not affected by experiment duration and cultivation mode. As determined by HPLC analysis, shake flask and temporary immersion cultures were characterized by the highest phenolic contents: up to 37.5 and 26 mg/g dry weight, respectively. The maximum productivities of the above systems were equal to 18 and 13.5 mg/L/d, respectively. Full article

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

In Argentina and globally, pistachio (Pistacia vera) production has significantly grown, driven by its high nutritional value and food industry demand. Its harvesting and processing generate about 40% of pistachio waste (PW), including leaves, twigs, seed coats, green, and empty kernels. Underutilized PW has led to environmental problems, including soil and water contamination by landfill accumulation. However, it could be a potential source of undiscovered bioactive compounds. This study aimed to characterize the chemical profile and to evaluate the bioactive properties of PW. The dried pistachio waste (dPW) was used to prepare the pistachio waste decoction (PWD) (10% w/v). The total phenolic content (TPC) and flavonoid content (FC) were quantified, and the chemical profile was analyzed using UPLC-DAD-ESI-MS/MS. Nematicidal activity against Meloidogyne incognita (J2), acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) inhibition, antioxidant capacity (ABTS, DPPH, FRAP), and phytotoxicity on Allium cepa, Lactuca sativa, and Raphanus sativus were evaluated. The UPLC-DAD-ESI-MS/MS analysis identified 26 compounds, including phenolics, flavonoids, and unsaturated fatty acids. The main compounds were gallic acid, anacardic acid, and quercetin derivatives. The TPC and FC were 212.65 mg GAE/g dPW and 0.022 mg QE/g dPW, respectively, displaying strong antioxidant activity across the assays DPPH, ABTS, and FRAP. PWD exhibited nematicidal activity against M. incognita (J2) (LC₅₀ = 0.12% at 24 h). Alterations in the cuticle were observed, including structural disorganization and detachment from internal tissues. Additionally, a remarkable cholinesterase inhibitory effect was detected at 2.0% PWD (42.65% for AChE and 58.90% for BuChE). PWD showed low phytotoxic effects across the tested species, and the germination percentage (GP) and the mean germination time (MGT) were not significantly affected (GP > 79%). These findings highlight the potential of PW as a sustainable alternative for M. incognita control, the remarkable nematicidal, anticholinesterase, and antioxidant properties, and the low phytotoxicity, supporting its use in sustainable agricultural practices. Full article

This study investigates the transformation of Acmella oleracea with the Agrobacterium rhizogenes rolC gene and evaluates its impact on phytochemical composition and biological activity. A total of 480 plant nodes were subjected to Agrobacterium−mediated transformation, leading to the regeneration of 35 putative transgenic plants. Molecular analysis confirmed the presence of the rolC transgene in 17 clones, of which four (C123, C127, C129, and C132) exhibited rolC mRNA expression. Phytochemical profiling of hydroalcoholic extracts of aerial parts (AP) and roots (R) revealed significant differences (p ≤ 0.05) between transgenic and non-transgenic plants (CTR). Compared to non−transgenic plants, transgenic AP exhibited lower total phenolic content but retained or increased flavonoid concentrations, particularly flavan−3−ols, whereas R extracts consistently showed reduced secondary metabolite levels. LC−DAD−ESI−MS analysis identified a diverse metabolite profile, with AP being notably rich in flavonoids (48.65%) and alkylamides (32.43%), including spilanthol. Functional assessments across antioxidant and anti−inflammatory assays demonstrated that R extracts exhibited stronger bioactivity compared to AP extracts, as indicated by lower IC₅₀ values (0.004–2.18 mg/mL for R vs. 0.007–7.24 mg/mL for AP). However, iron−chelating capacity was higher in AP extracts, correlating with flavonoid concentration. Hierarchical clustering confirmed that transgenic lines C123 and C127 most closely resembled the control, while C129 and C132 displayed distinct metabolic profiles. These findings highlight rolC’s role in modulating secondary metabolite synthesis, influencing both the phytochemical composition and functional properties of A. oleracea extracts. Full article

This study investigates the potential reuse of Prunus spinosa (blackthorn) seeds, a food industry by-product. Traditionally discarded, these seeds are now being explored for their bioactive compounds. In this work, seeds were used as raw material for supercritical CO₂ extraction. Two distinct extracts were obtained at low and high pressure (SFE90 and SFE200) and both extracts presented an aqueous phase (WE90 and WE200). SFE90 analysis by GC/MS allowed us to identify benzaldehyde and fatty acids (mainly oleic and linoleic acids). The fatty acid profile of SFE200, determined by HPLC-DAD/ELSD, showed that oleic and linoleic acids were predominant in supercritical oil. The phytochemical composition of the water extracts, analyzed via LC-DAD-ESI-MS, revealed that higher pressure enhanced the recovery of specific flavonols and anthocyanins, while lower pressure preserved various polyphenolic subclasses. WE90 was rich in 3-feruloylquinic acid and cyanidin-3-O-rutinoside, whereas WE200 was rich in caffeic acid hexoside 2 and dihydro-o-coumaric acid glucoside. Benzaldehyde was individuated in WE90 and WE200 by HPLC-DAD analysis. Cytotoxicity assays demonstrated that WE90, WE200 and SFE200 had anticancer effects on SH-SY5Y neuroblastoma cells, while all extracts did not remarkably affect the viability and morphology of human skin keratinocytes (HaCaT cells). These results suggest that P. spinosa seed extracts have potential nutraceutical and pharmaceutical applications. Full article

This study focuses on the extraction of betalain compounds from Opuntia stricta as a natural alternative to synthetic colorants and sustainable environmentally friendly technology solutions. Non-conventional extraction technologies including microwave (MW) and ultrasound (US) were used alone or in combination. The extraction process was conducted for both undried Opuntia stricta (OS) and dried Opuntia stricta (DOS) plant material at two distinct drying temperatures, 40 °C and 60 °C, to assess the stability of betalain molecules. The colorant’s potential was evaluated by determining the betalain content, total phenolic content, and antioxidant activity. The MW (2 min) and MW (2 min) + US (10 min) extraction processes yielded the greatest betalain content in OS fresh weight (FW), with 48.54 ± 0.29 mg/100 g FW and 51.01 ± 0.16 mg/100 g FW, respectively. Furthermore, the results showed a considerable drop in betalain content when the plant material was dried at 40 °C and 60 °C, with reduction rates of 53.75% and 24.82%, respectively, compared to the betalain content before the drying process. The LC-DAD-ESI-MS analysis supported this result, revealing the presence of 17-decarboxy betanin, 17-decarboxy neobetanin, and Cyclo-dopa5-O-βglucoside in DOS at 40 °C. This study highlights the potential future in the sustainable green extraction of betalain compounds with less heat degradation to offer a stable natural colorant. Full article

The dried leaves of Lannea velutina A.Rich. and Sorindeia juglandifolia Planch. ex Oliv. are commonly used in traditional medicine throughout West Africa to treat inflammatory diseases. The aim of the present study was to evaluate the anti-inflammatory activity of the standardized hydroethanolic (70%) extracts of these plants and to investigate the underlying mechanisms, with a focus on their antioxidant properties. The anti-inflammatory effects were evaluated using a rat model of induced paw edema, while the antioxidant activity was evaluated by DPPH^• radical scavenging and iron-reducing antioxidant power assays. Chemical fingerprint was achieved by LC-UV/DAD-ESI/MS and the main classes of secondary metabolites were quantified by colorimetric analysis. The results showed that Sorindeia juglandifolia extract significantly inhibited the increase in paw edema volume, with the maximum effect observed at doses of 100 and 200 mg/kg (20.51 ± 1.07% and 35.50 ± 6.90%, respectively). For L. velutina, the strongest inhibition was observed at 200 and 400 mg/kg (47.48 ± 11.37% and 35.40 ± 1.70%, respectively). Both extracts also showed remarkable antioxidant activity. Phenol derivatives were identified as the main classes of secondary metabolites, with L. velutina containing 350.1 ± 20.6 mg GAE/g DE and S. juglandifolia containing 463.4 ± 29.4 mg GAE/g DE. Ten phenolic markers were identified in L. velutina and six in S. juglandifolia extracts. The main components of L. velutina include myricetin-3-O-glucuronide, quercetin-3-O-glucuronide, catechin, and gallic acid, while S. juglandifolia contains gallic acid, isoquercitrin, and ethyl gallate. These results confirm the anti-inflammatory potential of L. velutina and S. juglandifolia and highlight their prospects as candidates for the development of standardized anti-inflammatory herbal medicines based on their chemical and biological properties. Full article

Plumeria rubra L. is an ornamental Caribbean plant widely known for its ethnobotanical uses and pharmacological activities. The ‘Tonda Palermitana’ cultivar, on which no data are to date available, is commonly cultivated in Sicily. The aim of our study was to characterize the micro-morphological features of leaves and flowers of this cultivar by light and Scanning Electron Microscopy and to investigate the phytochemical profile and the biological properties of their food-grade extracts (LE and FE, respectively) by LC-DAD-ESI-MS analysis and different in vitro assays. Numerous branched laticifers were observed, and their secretion contained alkaloids and lipophilic compounds as confirmed by histological analyses. Phytochemical analyses showed the presence of alkaloids (9%), terpenoids (13%) and fatty acids (6%), together with a very abundant presence of iridoids (28%) and polyphenols (39%). The most notable biological activity of both extracts appears to be the antioxidant one, showing half-inhibitory concentrations (IC₅₀) about 5 times lower than those detected in anti-inflammatory assays (383.74 ± 5.65 and 232.05 ± 2.87 vs. 1981.23 ± 12.82 and 1215.13 ± 10.15, for FE and LE, respectively), with LE showing the best, and statistically significant (p < 0.001), biological activity. These results allow us to speculate promising nutraceutical and cosmeceutical applications for this old Sicilian cultivar. Full article

Cynometra iripa Kostel. is a Fabaceae species of mangrove used in traditional Ayurvedic medicine for treating inflammatory conditions. The present study aims to establish monographic botanical and chemical quality criteria for C. iripa leaf and bark as herbal substances and to evaluate their in vitro antioxidant potential. Macroscopic and microscopic qualitative and quantitative analyses, chemical LC-UV/DAD-ESI/MS profiling, and the quantification of key chemical classes were performed. Antioxidant activity was evaluated by DPPH and FRAP assays. Macroscopically, the leaf is asymmetrical with an emarginated apex and cuneate base. Microscopically, it shows features such as two-layered adaxial palisade parenchyma, vascular bundles surrounded by 3–6 layers of sclerenchyma, prismatic calcium oxalate crystals (5.89 ± 1.32 μm) along the fibers, paracytic stomata only on the abaxial epidermis (stomatal index–20.15), and non-glandular trichomes only on petiolules. The microscopic features of the bark include a broad cortex with large lignified sclereids, prismatic calcium oxalate crystals (8.24 ± 1.57 μm), and secondary phloem with distinct 2–5 seriated medullary rays without crystals. Chemical profile analysis revealed that phenolic derivatives, mainly condensed tannins and flavonoids, are the main classes identified. A total of 22 marker compounds were tentatively identified in both plant parts. The major compounds identified in the leaf were quercetin-3-O-glucoside and taxifolin pentoside and in the bark were B-type dimeric proanthocyanidins and taxifolin 3-O-rhamnoside. The total phenolics content was higher in the leaf (1521 ± 4.71 mg GAE/g dry weight), while the total flavonoids and condensed tannins content were higher in the bark (82 ± 0.58 mg CE/g and 1021 ± 5.51 mg CCE/g dry weight, respectively). A total of 70% of the hydroethanolic extracts of leaf and bark showed higher antioxidant activity than the ascorbic acid and concentration-dependent scavenging activity in the DPPH assay (IC₅₀ 23.95 ± 0.93 and 23.63 ± 1.37 µg/mL, respectively). A positive and statistically significant (p < 0.05) correlation between the phenol content and antioxidant activity was found. The results obtained will provide important clues for the quality control criteria of C. iripa leaf and bark, as well as for the knowledge of their pharmacological potential as possible anti-inflammatory agents with antioxidant activity. Full article

This study investigates the chemical composition, nutritional, and biological properties of extracts obtained from A. melanocarpa berries using different extraction methods and solvents. Hydrodistillation and supercritical fluid extraction with CO₂ allowed us to isolate fruit essential oil (HD_EX) and fixed oil (SFE_EX), respectively. A phenol-enriched extract was obtained using a mild ultrasound-assisted maceration with methanol (UAM_M). The HD_EX most abundant component, using gas chromatography-mass spectrometry (GC/MS), was italicene epoxide (17.2%), followed by hexadecanoic acid (12.4%), khusinol (10.5%), limonene (9.7%), dodecanoic acid (9.7%), and (E)-anethole (6.1%). Linoleic (348.9 mg/g of extract, 70.5%), oleic (88.9 mg/g, 17.9%), and palmitic (40.8 mg/g, 8.2%) acids, followed by α-linolenic and stearic acids, were the main fatty acids in SFE_EX determined using high-performance liquid chromatography coupled with a photodiode array detector and an evaporative light scattering detector (HPLC-DAD/ELSD). HPLC-DAD analyses of SFE_EX identified β-carotene as the main carotenoid (1.7 mg/g), while HPLC with fluorescence detection (FLU) evidenced α-tocopherol (1.2 mg/g) as the most abundant tocopherol isoform in SFE_EX. Liquid chromatography-electrospray ionization-MS (LC-ESI-MS) analysis of UAM_M showed the presence of quercetin-sulfate (15.6%, major component), malvidin 3-O-(6-O-p-coumaroyl) glucoside-4-vinylphenol adduct (pigment B) (9.3%), di-caffeoyl coumaroyl spermidine (7.6%), methyl-epigallocatechin (5.68%), and phloretin (4.1%), while flavonoids (70.5%) and phenolic acids (23.9%) emerged as the most abundant polyphenol classes. UAM_M exerted a complete inhibition of the cholesterol oxidative degradation at 140 °C from 75 μg of extract, showing 50% protection at 30.6 μg (IA₅₀). Furthermore, UAM_M significantly reduced viability (31–48%) in A375 melanoma cells in the range of 500–2000 μg/mL after 96 h of incubation (MTT assay), with a low toxic effect in normal HaCaT keratinocytes. The results of this research extend the knowledge of the nutritional and biological properties of A. melanocarpa berries, providing useful information on specific extracts for potential food, cosmetic, and pharmaceutical applications. Full article

Guayusa tea is derived from the leaves of the Ilex guayusa Loes. plant, which is native to the Amazon rainforest. Beyond its pleasant sensory properties, Guayusa tea is rich in antioxidants, phenolics, and minerals. In this study, the effects of infusion time, temperature, and solvent conditions on the color, antioxidant capacity, total phenolic content, phenolic profile, and antimicrobial activity of Guayusa (Ilex guayusa Loes.) tea were investigated. Guayusa tea samples were prepared using two different solvents, ethanol and water, with 4, 6, and 8-h infusions at 60 and 70 °C. Liquid chromatography, diode array detection, and electrospray ionization tandem mass spectrometry (LC-DAD-ESI-MS/MS) were used to determine a comprehensive profile of phenolic compounds and to detect differences due to infusion conditions. Moreover, after the Guayusa tea infusion with the highest bioactive properties was determined, the effects of in vitro gastrointestinal digestion on the total phenolic content, antioxidant capacity, and phenolic compounds of the Guayusa tea infusion were measured. Phenolic profile analysis identified 29 compounds, among which chlorogenic acid and its derivatives were predominant. The increase in infusion time was correlated with an elevation in total phenolic content. Significant differences were observed between water and ethanol infusions of Guayusa in terms of phenolics and antioxidants. The total amount of phenolic compounds in the samples prepared with both solvents was found to increase after oral intake, depending on the digestion stage; meanwhile, the amounts of flavonoid compounds and di-O-caffeoylquinic acid derivatives decreased during digestion. Full article

Saponaria sicula Raf. grows in Sicily, Sardinia, and Algeria on limestone cliffs and volcanic sands 1300–2500 m above sea level. The aim of the present study was to investigate how the pedo-climatic conditions influence the micromorphological, phytochemical, and biological properties of Sicilian S. sicula leaves collected in the Madonie Mountains (SsM) and on Etna Mt (SsE). Micromorphological investigations revealed that leaves from SsM had a higher amount of calcium oxalate druses in the mesophyll and a more intense blue–green staining with Toluidine blue O, indicating a higher content of polyphenols. These data were confirmed by phytochemical analyses carried out on hydroalcoholic extracts, which showed a higher content of total phenols (8.56 ± 0.57 g GAE/100 g DE) and flavonoids (6.09 ± 0.17 g RE/100 g DE) in SsM. Sixty-four compounds were identified by LC-DAD-ESI-MS analysis with propelargonidin dimer as the most abundant compound (10.49% and 10.19% in SsM and SsE, respectively). The higher polyphenol content of SsM leaves matches also with their biological activity, identifying SsM extract as the strongest plant complex (IC₅₀ 2.75–477.30 µg/mL). In conclusion, the present study experimentally demonstrates that not only climatic differences but also soil characteristics affect the micromorphological, phytochemical, and biological features of this plant species. Full article

Cnidoscolus aconitifolius (CA) and Porophyllum ruderale (PR) are representative edible plants that are a traditional food source in Mexico. This research aimed to analyze the phytochemical composition and untargeted metabolomics analysis of CA and PR and evaluate their antiproliferative effect in vitro. The phytochemical composition (UPLC-DAD-QToF/MS-ESI) identified up to 38 polyphenols and selected organic acids that were clustered by the untargeted metabolomics in functional activities linked to indolizidines, pyridines, and organic acids. Compared with PR, CA displayed a higher reduction in the metabolic activity of human SW480 colon adenocarcinoma cells (LC₅₀: 10.65 mg/mL), and both extracts increased the total apoptotic cells and arrested cell cycle at G0/G1 phase. PR increased mRNA Apc gene expression, whereas both extracts reduced mRNA Kras expression. Rutin/epigallocatechin gallate displayed the highest affinity to APC and K-RAS proteins in silico. Further research is needed to experiment on other cell lines. Results suggested that CA and PR are polyphenol-rich plant sources exhibiting antiproliferative effects in vitro. Full article

To improve the nutri-functional quality and, in particular, the antioxidant capacity of corn by-products, fermentation with selected lactic acid bacteria was carried out. To this purpose, white-1 and 2 and yellow corn by-products were fermented and then extracted. In all the samples, the fermentation process shows an improvement in antioxidant activity in comparison to non-fermented by-products. It was observed that the yellow corn by-product extracts have a higher content of total phenols, especially after fermentation with P. pentosaceus, while for white-1 corn by-product extracts, an increment of antioxidant capability was noticed when fermented with L. plantarum. The antioxidant capacity was measured with DPPH and ABTS⁺ assays, showing that yellow corn extracts are more active in comparison with white-1 and white-2 ones. Moreover, L. plantarum and P. pentosaceus provided the best results in increasing the antioxidant activity in all the samples. Analyzing lipid peroxidation in the presence of fermented white-2 corn by-product extracts, we observed an inhibition of the process after treatment with L. citreum compared to the non-fermented control. In all the analyzed samples, through LC-DAD-ESI/MS analysis, the antioxidant dicoumaroyl spermidine (DCS) was detected. The abundance of antioxidant molecules was higher in samples fermented with P. pentosaceus, confirming previous observations. Full article