Search Results (320)

The European Regulation (UE) 432/2012 includes a specific health claim for olive-oil-associated with its phenolic content, which is based on its protective role against lipid oxidation in the blood. To make use of the health claim, olive oil must have a minimum concentration in phenolic compounds of 250 mg/kg. Reviewing the health claim, the phenolic compounds referred to are the secoiridoid derivatives of hydroxytyrosol and tyrosol. A method based on absolute quantification of phenolic compounds in olive oil is proposed for the recognition of the health claim. The method involves liquid–liquid extraction with a 1:8 (v/v) oil:extract ratio to avoid saturation of the extract in oils with a higher phenolic content and its subsequent determination through LC–MS/MS in multiple reaction monitoring (MRM) mode, the gold standard technique in many application fields because of its analytical features. The optimized method was applied to a set of 100 extra virgin olive oils (EVOOs), and the results obtained were compared with the classic Folin–Ciocalteu method. The comparison between the two methods showed that the classic method is a non-selective method that can be affected by many interferences and that the Folin method underestimates the real phenolic content. Full article

This study evaluated the sustainability of removing phenolic compounds from olive mill effluents using a nanobiochar synthesized from olive pomace. Catechol, tyrosol, hydroxytyrosol, and homovanillic alcohol were chosen as model pollutants due to their presence in agro-industrial wastewater. The surface morphology, elemental composition, crystallographic structure, functional groups, porosity, and thermal stability of the nanobiochar were investigated by SEM, EDX, XRD, FTIR, BET analysis, and TGA/DTA. The developed nanobiochar exhibited a predominantly amorphous carbon structure, enriched in carbon (85.6%), with localized graphitic domains. Its mesoporous architecture (S_BET = 15.478 m² g⁻¹; Dp = 2.14 nm) promotes accessibility to active sites, while its thermal stability confirmed its suitability for adsorption applications. In this batch adsorption study, the technological aspect considered is the influence of operating parameters on adsorption efficiency, using kinetic and equilibrium models. Pseudo-first-order and pseudo-second-order kinetic models, as well as Freundlich and Langmuir isotherms, were used to analyze the experimental data. The pseudo-second-order model proved to be the most suitable for describing adsorption, suggesting that the process is primarily dominated by chemisorption. Similarly, the Langmuir model gave the least satisfactory results regarding equilibrium data, indicating monolayer adsorption on homogeneous active sites. The adsorption capacity of phenolic compounds was variable. The highest adsorption capacities were observed for catechol (250 mg g⁻¹), tyrosol (19.23 mg g⁻¹), homovanillic alcohol (15.38 mg g⁻¹), and hydroxytyrosol (13.16 mg g⁻¹). The results of this research indicate that adsorption affinity depends on molecular structure and electronic properties. Furthermore, computer modeling based on molecular simulations and electronic descriptors was performed to explain the adsorption mechanism. Linear regression, principal component analysis, and elastic regression revealed strong correlations between adsorption parameters and molecular descriptors. These results demonstrate that olive pomace-based nanobiochar is an environmentally friendly adsorbent for the treatment of phenolic effluents, with adsorption primarily controlled by surface interactions. Full article

This study evaluated olive leaves from three cultivars (Hojiblanca, Picual, and Arbequina) and olive pomace as complementary sources of bioactive compounds, comparing ultrasound-assisted extraction using organic solvents (UAE) with supercritical CO₂ extraction (SFE). The aim was to determine how the plant matrix and extraction method influence phytochemical composition and functional properties, including antioxidant and antimicrobial activity. The results showed that both factors strongly affected extract composition and bioactivity. UAE favored the recovery of phenolic compounds associated with antioxidant activity, particularly in leaf extracts, while SFE promoted a distinct compositional profile enriched in flavonoids and lipophilic constituents, especially in olive pomace. Multivariate analysis confirmed a clear differentiation between matrices and extraction methods. Leaf extracts from Picual and Arbequina were mainly associated with phenolic compounds linked to antioxidant activity, including luteolin, ethyl vanillin, tyrosol, and isorhamnetin-3-O-glucoside. In contrast, olive pomace extracts were more strongly associated with flavonoids and lipophilic metabolites, such as triterpenes (oleanolic, maslinic, and ursolic acids) and lipid derivatives (oleic acid and lauric isopropanolamide). These compositional differences were reflected in biological activity: UAE extracts showed higher antioxidant activity, whereas SFE extracts, enriched in lipophilic and triterpenic compounds, exhibited stronger antimicrobial effects against Pseudomonas savastanoi and Hanseniaspora sp. Overall, these findings demonstrate that extraction-driven selectivity enables the production of olive-derived extracts with targeted functionalities, with UAE favoring antioxidant-oriented extracts and SFE promoting extracts enriched in lipophilic compounds with antimicrobial potential, particularly from olive pomace. Full article

Extra virgin olive oil (EVOO) is a key component of the Mediterranean diet, valued for its bioactive constituents and associated health benefits. This study evaluated the influence of four agronomic and processing factors—harvest month, destoning, fruit washing, and bottling delay—on the chemical composition of Kolovi EVOOs from the PGI Lesvos region. A total of 34 oils were produced under standardized conditions and analyzed for phenolic compounds, tocopherols, pigments, and squalene using UPLC-QTOF-MS and HPLC-DAD. The oils were characterized by consistently high nutritional quality, with most samples fulfilling EFSA health claim thresholds for hydroxytyrosol, tyrosol and its derivatives, and α-tocopherol. Harvest month was the most influential parameter: early harvested oils (October) contained significantly higher levels of phenolics, α-tocopherol, and lutein, whereas later harvests (November) were richer in squalene. Destoning produced modest changes, with slightly higher phenolics in non-destoned oils and reduced lipophilic antioxidants in destoned samples. Fruit washing selectively decreased hydrophilic phenolics, while lipophilic compounds were largely unaffected. Bottling delays of up to 48 h under protective conditions had negligible effects on composition, aside from minor increases in specific phenolic derivatives. These findings suggest that early harvesting and careful consideration of destoning are the most effective strategies for supporting the antioxidant profile of Kolovi EVOOs, while other practices can be adjusted with limited impact on quality. Full article

Background/Objectives: Salidroside, a bioactive phenylethanol glycoside primarily derived from Rhodiola rosea, and its major in vivo metabolite tyrosol exhibit diverse pharmacological activities. However, their direct molecular targets remain poorly defined. Methods: In the present study, an integrated strategy combining transcriptomic profiling, Connectivity Map (CMap) analysis, and multi-level experimental validation was employed. Transcriptomic signatures derived from A549 cells treated with salidroside or tyrosol were queried against the CMap database. Molecular docking, surface plasmon resonance (SPR), and cellular thermal shift assays (CETSA) were performed to predict and validate binding interactions. Functional validation was performed in SH-SY5Y cells. The phosphorylation level of extracellular signal-regulated kinase (ERK), a downstream signaling event of dopamine D2 receptor (DRD2), was detected after salidroside and tyrosol treatment. DRD2 antagonist sulpiride pre-intervention and small interfering RNA (siRNA)-mediated DRD2 knockdown were conducted to verify the receptor dependence of the compounds’ effects. Results: CMap analysis revealed that the transcriptomic signatures of salidroside and tyrosol showed significant similarity to known DRD2 modulators. Molecular docking predicted potential binding interactions between the two compounds and DRD2, which was confirmed by SPR and CETSA to be direct physical binding. Functional studies showed that both compounds rapidly induced DRD2 downstream ERK phosphorylation in SH-SY5Y cells; this effect was abrogated by sulpiride or DRD2 knockdown, indicating DRD2-dependent signaling activation. Conclusions: These findings identify DRD2 as a direct molecular target of salidroside and tyrosol and provide mechanistic insight into their dopaminergic regulatory effects. This study highlights the utility of CMap-guided target discovery combined with rigorous experimental validation for elucidating the molecular mechanisms of natural products. Full article

This study involved 45 finishing Limousin young bulls (initial BW 350 ± 15 kg; aged 250 ± 20 days) reared under commercial conditions and randomly assigned for 250 days to three dietary treatments: a control diet without olive cake (CTR), and diets including partially destoned dried olive cake at 10% (OC10) or 15% (OC15) of dry matter. The effects of the dietary inclusion of partially destoned dried olive cake (DOC) on the physicochemical properties, intramuscular fatty acid composition, nutritional lipid indices, and antioxidant profile of the Longissimus dorsi muscle were evaluated. Meat pH was measured at 45 min, 24 h, and 7 days of aging. Instrumental color, proximate composition, fatty acid profile, lipid nutritional indices, total phenolic content (TPC), individual polyphenols, and antioxidant capacity were determined. Data were analyzed by ANOVA using PROC GLIMMIX in SAS, with orthogonal contrasts to evaluate linear and quadratic responses to olive cake inclusion. Treatments did not affect post mortem pH, and color differences observed at 24 h were not present after 7 days of aging. Crude protein content was lower in OC-fed groups, while total lipid content tended to increase; sodium chloride concentration was higher in OC15 meat. Olive cake supplementation reduced hypercholesterolemic saturated fatty acids and increased stearic, oleic, and polyunsaturated fatty acids, including α-linolenic acid, EPA, and DPA, improving PUFA/SFA and UFA/SFA ratios and reducing atherogenic and thrombogenic indices. Despite a higher peroxidability index, OC groups showed greater muscle TPC and antioxidant capacity, the presence of detectable hydroxytyrosol and tyrosol suggests a possible contribution of dietary olive phenolics or their metabolites, although the exact mechanisms underlying their appearance in muscle tissue remain to be fully elucidated. Overall, partially destoned DOC can be included up to 15% of the finishing diet to improve beef nutritional and functional quality within sustainable circular systems. Full article

Olive pomace (OP), a by-product of olive oil production, is a sustainable resource rich in bioactive compounds with potential applications in cosmetics and pharmaceuticals. This study investigates the protective effects of olive pomace juice (OPJ) against H₂O₂-induced neuronal damage and LPS-induced inflammatory responses in HT22 and BV2 cells, respectively. OPJ suppressed H₂O₂-induced cell death and exerted anti-apoptotic effects by reducing the BAX/BCL2 ratio and caspase-3 cleavage. OPJ also mitigated neurodegenerative hallmarks by decreasing amyloid fibrils formation and inhibiting β-secretase and acetylcholinesterase (AChE) activity. Mechanistically, OPJ enhanced antioxidant response by upregulating Nrf2 and its downstream molecule HO-1, along with increasing mRNA levels of antioxidant enzymes, including catalase, SOD1, and GPx. OPJ further activated AMPKα–SIRT1–PGC1α signaling and CREB–BDNF–TrkB signaling, suggesting modulation of key antioxidant, anti-apoptotic, and neurotrophic pathways. In BV2 cells, OPJ downregulated pro-inflammatory cytokines (IL-6 and IL-1β) and decreased iNOS and COX-2 expression through suppression of NF-κB and MAPK signaling pathways. HPLC analysis identified hydroxytyrosol (10.92%) as the major active compound in OPJ, which compared with tyrosol (2.18%), and hydroxytyrosol exhibited greater neuroprotective and anti-inflammatory effects than tyrosol. This study highlights the potential of OPJ and its major compound, hydroxytyrosol, as functional agents for mitigating neurodegeneration-related cellular response, supporting its application in the food and pharmaceutical industries. Full article

This study addresses the ongoing challenge of accurately classifying beers by fermentation type and product category, an issue of growing importance for quality control, authenticity assessment, and product differentiation in the brewing sector. We applied a multiblock chemometric framework that integrates phenolic profiling obtained via GC–MS, antioxidant and antiradical activity derived from in vitro assays, and complementary colorimetric and physicochemical measurements. Principal Component Analysis (PCA) revealed clear compositional structuring within the dataset, with p-coumaric, gallic, syringic, and malic acids emerging as major contributors to variance. Supervised machine-learning classification demonstrated robust performance, achieving approximately 93% accuracy in discriminating top- from bottom-fermented beers, supported by a well-balanced confusion matrix (25 classified and 2 misclassified samples per group). When applied to ale–lager categorization, the model retained strong predictive ability, reaching 90% accuracy, largely driven by the C* chroma value and the concentrations of tyrosol, acetic acid, homovanillic acid, and syringic acid. The integration of multiple analytical blocks significantly enhanced class separation and minimized ambiguity between beer categories. Overall, these findings underscore the value of multi-block chemometrics as a powerful strategy for beer characterization, supporting brewers, researchers, and regulatory bodies in developing more reliable quality-assurance frameworks. Full article

Game meats are particularly prone to oxidation and microbial spoilage due to their specific characteristics and the procedures required to obtain them. Various sustainable bioactive molecules derived from food industry by-products, such as olive mill wastewater, have the potential to enhance the stability and safety of game meats. The use of different levels of polyphenolic extracts from olive mill vegetation water, encapsulated through a freeze-drying process, was tested on wild boar meat patties as an antioxidant and antimicrobial. Two separate trials were performed. Trial 1 was carried out by adding different concentrations of polyphenolic extract (0, 1, and 2%) during the production of wild boar patties, and trial 2 by adding 1.5% salt and adding or not adding 2% polyphenolic extract. The first trial revealed antioxidant effects on the raw patties during storage time, both on colour (increasing in saturation index) and thiobarbituric acid-reactive substances (0.306, 0.268, and 0.254 mg MDA/kg after 5 days of storage in the control with 1% and 2% polyphenolic extract groups, respectively). Oxidation was also reduced during cold storage of cooked patties. Trial 1 also revealed a dose-dependent antimicrobial effect, mainly on Enterobacteriaceae and Pseudomonas spp. Trial 2 confirmed that salt plus extract addition had an overall higher antimicrobial effect than when singularly added, but with a moderate increase in the hardness of the products. Full article

Atherosclerosis is a chronic inflammatory cardiovascular disease that may result from the interaction between oxidative stress, immune dysregulation, and metabolic disorders. Recent studies indicate that the well-known phenolic compounds, hydroxytyrosol (HTyr) and tyrosol (Tyr) present in extra virgin olive oil, confer cardioprotection through various mechanisms of action that include antioxidant, anti-inflammatory, and metabolic regulatory properties. The gut microbiota modulates the structure, bioavailability, and bioactivity of these phenolic compounds, thereby influencing their therapeutic potential. This review explores the intricate interactions between Tyr, HTyr, and gut microbiota within the context of atherosclerosis prevention and management. We explore how gut microbial metabolism can magnify or alter the biological effects of the Tyr and HTyr, and how interindividual differences in microbiota composition may influence their efficacy. A deeper understanding of these mechanisms could support the development of precision nutrition strategies aimed at reducing the risk of atherosclerosis. Full article

Kinetic evolution of quality parameters in 21 extra-virgin olive oils (EVOOs) from Coratina cultivar was evaluated during 18 months of dark storage at room temperature (RT) and 4 °C (LT). The aim was to identify the most discriminating variables—fatty acids, peroxide value, spectrophotometric indices, and phenolic compounds—using kinetic analysis and multivariate statistics. Fatty acids remained stable, while peroxide value and ultraviolet absorbance indices increased significantly at RT, following zero-order kinetics. Polyphenols declined markedly after 6 months, especially at RT, with degradation rates influenced by initial concentrations. Hydroxytyrosol and tyrosol followed pseudo-zero-order kinetics, whereas secoiridoids and lignans followed second-order kinetics. Discriminant analysis achieved 90% accuracy (p = 0.000012) in classifying oils by storage condition. The most relevant discriminants were associated with phenolic degradation and oxidative changes. These findings support the importance of low-temperature storage in preserving the biochemical quality and shelf life of EVOOs. Full article

Extra virgin olive oil (EVOO) polyphenols are recognized for their beneficial effects on human health, yet how their concentration shapes biological outcomes remains insufficiently explored. While a daily intake of 25 mL EVOO is generally regarded as beneficial for cardiovascular protection, the high-phenolic EVOO examined in this study contains markedly higher levels of polyphenols than most EVOOs reported previously. This suggests that oils richer in polyphenols may exert distinct biological effects. To investigate this, we compared extracts from a standard EVOO and a naturally high-phenolic EVOO, along with their key phenolic compounds, hydroxytyrosol (HT) and tyrosol (Tyr). Antioxidant effects were assessed by quantifying intracellular reactive oxygen species (ROS) and lipid peroxidation. Anti-inflammatory activity was evaluated in THP-1-derived macrophages stimulated with LPS by analyzing inflammatory surface markers’ expression, cytokines’ production, and the NLRP3-inflammasome pathway. Atheroprotective potential was investigated by measuring cholesterol efflux in J774 macrophages. Both EVOO polyphenols extracts and (HT and Tyr) significantly reduced ROS and lipid peroxidation. High phenolic EVOO extract (EVOOPE+) displayed superior antioxidant activity at lower concentrations, while standard EVOO phenolic extract (EVOOPE) showed more consistent effects across doses. Both extracts favored an anti-inflammatory macrophage phenotype, as indicated by increased CD163 and IL-10 expression and reduced CD86, IFN-α, and NLRP3. Moreover, all treatments enhanced cholesterol efflux in a dose-dependent manner, with EVOOPE+ and HT producing the strongest effects. Collectively, these results highlight the capacity of EVOO polyphenols to modulate, through key bioactivity mechanisms, cardioprotective effects and emphasize the importance of polyphenols concentration in their biological efficacy. Full article

Olive mill wastewater (OMW), a by-product of olive oil extraction, poses significant environmental challenges due to its toxicity and heterogeneity. This study evaluates the phenolic and mineral composition of OMW and alpechin sludges from abandoned ponds in Spain, and establishes a standardized conventional method to recover phenolic fractions and promote their safe valorization as bioactive ingredients. Phenolic compounds were identified by triple-TOF-LC-MS/MS, and minerals and heavy metals were quantified by ICP-MS. Across thirteen ponds analyzed, samples from Cordoba, Tarragona, Alicante and Toledo showed higher phenolic levels, ranging from 7.2 g GAE/kg to 18.9 g GAE/kg, with methanolic extracts reaching 10.98–15.67 mg GAE/mL. Thirty-one phenolic compounds were identified, predominantly luteolin, apigenin, quercetin, and secoiridoid derivatives, notably hydroxytyrosol and tyrosol, supporting their functional potential as natural antioxidants. The mineral profile was dominated by K and Ca and showed negligible carryover to the phenolic organic fraction (<1%). Heavy metal concentrations in fresh OMW were 0.32–1.06 µg/kg for Cd and Hg and 9–43.9 µg/kg for As and Pb. In OMW sludge, they ranged between 0.033 and 0.19 mg/kg for Cd, 0.01 and 0.12 mg/kg for Hg, 5.45 and 8.06 mg/kg for As, and 4.45 and 23.70 mg/kg for Pb, whereas phenolic extracts contained only 0.15–21.50 µg/kg, remaining below EU food safety limits. This work presents one of the first integrated approaches to risk-benefit mapping of abandoned ponds in Spanish soils and advances extraction standardization by jointly considering functional potential, contaminant profiles, and matrix location. Full article

The valorization of olive oil production residues represents an innovative and sustainable strategy aligned with circular economy principles and the United Nations Sustainable Development Goals. In this study, we aimed to explore the phytochemical composition and neuroprotective potential of organic extracts obtained from olive pomace of the Arbequina and Arbosana cultivars. Extracts were prepared through solid–liquid extraction and analyzed by high-performance liquid chromatography coupled with mass spectrometry (HPLC-MS), enabling the comprehensive identification of bioactive metabolites. The analysis revealed a diverse profile of phenolic compounds, including hydroxytyrosol, tyrosol, and multiple oleuropein derivatives, as well as triterpenic acids such as oleanolic and maslinic acids. These compounds are widely recognized for their antioxidant, anti-inflammatory, and neuroprotective activities. The antioxidant potential of the extracts was evaluated in vitro using DPPH and ABTS radical scavenging assays, showing significant activity comparable to that of standard antioxidants. Moreover, cholinesterase inhibitory assays demonstrated moderate to strong inhibition of acetylcholinesterase, an enzyme implicated in neurodegenerative diseases such as Alzheimer’s disease. To further elucidate the molecular basis of these effects, in silico molecular docking studies were performed on the most abundant compounds, revealing favorable binding affinities and interactions with key active site residues of acetylcholinesterase. Overall, these findings highlight olive pomace as a promising, underutilized source of bioactive compounds with potential applications in the development of functional foods, nutraceuticals, and neuroprotective therapeutic agents. The integration of in vitro and in silico approaches strengthens the evidence supporting the use of these extracts in future biomedical and industrial applications. Full article

Background/Objectives: Lavandula stoechas has reported bioactivities, but its selective anticancer potential in human models remains insufficiently defined. This study aimed to compare cytotoxicity and selectivity of ethanol and methanol extracts prepared from fresh and dried L. stoechas and to profile candidate bioactive metabolites. Methods: Aerial parts Lavandula stoechas L. subsp. stoechas (L. stoechas L.) were extracted with ethanol or methanol from fresh (LsFE, LsFM) and dried (LsDE, LsDM) material. Cytotoxicity was assessed in cancer (MDA-MB-231, T98G, RT4) and non-malignant (hGF, ARPE-19) cells using Hoechst 33342-stained nuclear counts and MTS viability at 24–48 h. Metabolite identification was performed using LC–QTOF–MS in both positive and negative ESI modes, supported by database search results. Results: All extracts reduced viability in a dose- and time-dependent manner. Among them, the ethanol extract from fresh material (LsFE) displayed the highest cytotoxic potency and the most favorable selectivity profile, markedly reducing viability in breast (MDA-MB-231) and glioblastoma (T98G) cells while exerting only mild effects on non-malignant fibroblast (hGF) and retinal epithelial (ARPE-19) cells. In contrast, extracts from dried material, particularly LsDE, showed broader cytotoxicity across both cancerous and non-cancerous lines. LC–MS highlighted sesquiterpenoids (Kikkanol A; 3(4→5)-Abeo-4,11:4,12-diepoxy-3-eudesmanol), phenolics (tyrosol; 3,4-dihydroxybenzoic acid), flavonoid/ionone derivatives (luteolin 5,3′-dimethyl ether; 3-hydroxy-β-ionone), oxidized fatty acids (9(S)-HpODE, α-EpODE, 5,12-dihydroxy-eicosatetraenoic acid), and jasmonates (12-hydroxyjasmonic acid; dihydrojasmonic acid methyl ester), especially enriched in LsFE. Conclusions: Ethanol extracts of L. stoechas L., especially LsFE, demonstrated selective cytotoxicity against cancer cells while exerting relatively mild effects on non-malignant cells. The metabolite profile of L. stoechas L. extracts revealed a diverse composition, including phenolics, terpenoids, flavonoids, and oxidized lipids, which are commonly associated with biological activity. These results suggest that LsFE is a promising candidate for further studies focusing on compound isolation and mechanistic analysis. Full article