Olive Oil Phenols Research Articles

Metrological aspects of a gas-phase DFT/B3LYP quantum-chemical approach to prioritize radical scavenging activity among a group of olive oil phenols

Metrological aspects of a gas-phase DFT/B3LYP quantum-chemical approach to prioritize radical scavenging activity among a group of olive oil phenols

Storage effect on olive oil phenols: cultivar-specific responses.

Olive oil is a widely recognized and appreciated food commodity, its quality and health benefits can be compromised when the oil goes through oxidative processes that may occur during production and storage. This study aimed to investigate the effects of the olive genotype on polar phenolic content after seven months of storage. Oil produced from eight different olive cultivars (Leccino, Leccio del Corno, Moraiolo, Frantoio, Bianchera, Pendolino, Maurino, and Caninese) grown in southern Tuscany, Italy, were subjected to chemical analysis such as free fatty acids, peroxide value, K232 and K268, phenolics and UPLC-DAD at the beginning of the trial (Control) and seven months later (Stored). Free fatty acids, peroxide values, K232 and K268, significantly increased, suggesting heightened hydrolysis and oxidation after storage. A cultivar effect was observed, with Leccino, Moraiolo, and Pendolino showing less susceptibility to oxidation (low differences between Control and Stored). In contrast, others (Bianchera and Caninese) are more affected (higher differences between Control and Stored). Phenolics analysis supports this observation, revealing that samples with higher resistance to oxidation exhibit elevated levels of hydroxytyrosol, tyrosol, vanillic acid, caffeic acid, p-coumaric acid, and ferulic acid. Principal Component Analysis highlights that Bianchera and Caninese cultivars correlate with rutin, tyrosol, and pinoresinol. As this research delves into the intricate relationship between genotype diversity, phenolic composition, and oxidative stability, a nuanced understanding emerges, shedding light on how different cultivars may present varying compositions and concentrations of phenols, ultimately influencing the oil's resistance to the oxidation that occurred during storage.

Proton-Coupled Electron Transfer and Hydrogen Tunneling in Olive Oil Phenol Reactions.

Olive oil phenols are recognized as molecules with numerous positive health effects, many of which rely on their antioxidative activity, i.e., the ability to transfer hydrogen to radicals. Proton-coupled electron transfer reactions and hydrogen tunneling are ubiquitous in biological systems. Reactions of olive oil phenols, hydroxytyrosol, tyrosol, oleuropein, oleacein, oleocanthal, homovanillyl alcohol, vanillin, and a few phenolic acids with a DPPH• (2,2-diphenyl-1-picrylhydrazyl) radical in a 1,4-dioxane:water = 95:5 or 99:1 v/v solvent mixture were studied through an experimental kinetic analysis and computational chemistry calculations. The highest rate constants corresponding to the highest antioxidative activity are obtained for the ortho-diphenols hydroxytyrosol, oleuropein, and oleacein. The experimentally determined kinetic isotope effects (KIEs) for hydroxytyrosol, homovanillyl alcohol, and caffeic acid reactions are 16.0, 15.4, and 16.7, respectively. Based on these KIEs, thermodynamic activation parameters, and an intrinsic bond orbital (IBO) analysis along the IRC path calculations, we propose a proton-coupled electron transfer mechanism. The average local ionization energy and electron donor Fukui function obtained for the phenolic compounds show that the most reactive electron-donating sites are associated with π electrons above and below the aromatic ring, in support of the IBO analysis and proposed PCET reaction mechanism. Large KIEs and isotopic values of Arrhenius pre-exponential factor AH/AD determined for the hydroxytyrosol, homovanillyl alcohol, and caffeic acid reactions of 0.6, 1.3, and 0.3, respectively, reveal the involvement of hydrogen tunneling in the process.

Hydroxytyrosol, a Promising Supplement in the Management of Human Stroke: An Exploratory Study.

Hydroxytyrosol (HT) is a bioactive olive oil phenol with beneficial effects in a number of pathological situations. We have previously demonstrated that an HT-enriched diet could serve as a beneficial therapeutic approach to attenuate ischemic-stroke-associated damage in mice. Our exploratory pilot study examined this effect in humans. Particularly, a nutritional supplement containing 15 mg of HT/day was administered to patients 24 h after the onset of stroke, for 45 days. Biochemical and oxidative-stress-related parameters, blood pressure levels, serum proteome, and neurological and functional outcomes were evaluated at 45 and 90 days and compared to a control group. The main findings were that the daily administration of HT after stroke could: (i) favor the decrease in the percentage of glycated hemoglobin and diastolic blood pressure, (ii) control the increase in nitric oxide and exert a plausible protective effect in oxidative stress, (iii) modulate the evolution of the serum proteome and, particularly, the expression of apolipoproteins, and (iv) be beneficial for certain neurological and functional outcomes. Although a larger trial is necessary, this study suggests that HT could be a beneficial nutritional complement in the management of human stroke.

Dilation of Pregnant Rat Uterine Arteries with Phenols from Extra Virgin Olive Oil Is Endothelium-Dependent and Involves Calcium and Potassium Channels.

During pregnancy, uterine vasculature undergoes significant circumferential growth to increase uterine blood flow, vital for the growing feto-placental unit. However, this process is often compromised in conditions like maternal high blood pressure, particularly in preeclampsia (PE), leading to fetal growth impairment. Currently, there is no cure for PE, partly due to the adverse effects of anti-hypertensive drugs on maternal and fetal health. This study aimed to investigate the vasodilator effect of extra virgin olive oil (EVOO) phenols on the reproductive vasculature, potentially benefiting both mother and fetus. Isolated uterine arteries (UAs) from pregnant rats were tested with EVOO phenols in a pressurized myograph. To elucidate the underlying mechanisms, additional experiments were conducted with specific inhibitors: L-NAME/L-NNA (10-4 M) for nitric oxide synthases, ODQ (10-5 M) for guanylate cyclase, Verapamil (10-5 M) for the L-type calcium channel, Ryanodine (10-5 M) + 2-APB (3 × 10-5 M) for ryanodine and the inositol triphosphate receptors, respectively, and Paxilline (10-5 M) for the large-conductance calcium-activated potassium channel. The results indicated that EVOO-phenols activate Ca2+ signaling pathways, generating nitric oxide, inducing vasodilation via cGMP and BKCa2+ signals in smooth muscle cells. This study suggests the potential use of EVOO phenols to prevent utero-placental blood flow restriction, offering a promising avenue for managing PE.

Qualitative and quantitative determination of phenols and their metabolites in urine by in-syringe solid-phase extraction and LC–MS/MS analysis for evaluation of virgin olive oil metabolism

To know the bioavailability of virgin olive oil (VOO) phenols and its impact on health, it is necessary to determine the levels of phenols excreted in urine. We present here a novel strategy for in-syringe solid-phase extraction and analysis of the extract by liquid chromatography–tandem mass spectrometry (LC–MS/MS), using ammonium fluoride as ionization agent to enhance sensitivity. This approach allows avoiding additional steps such as solvent evaporation or analytes derivatization. The method can be used with a previous acid hydrolysis for quantitative determination of tyrosol and hydroxytyrosol to estimate metabolized phenols. We tested this application by analysis of a cohort of volunteers (n = 20) after a standardized intake of VOO. Additionally, the method can be used as such for metabolite profiling of phenolic derivatives in urine using LC–MS/MS in high-resolution data-independent acquisition (DIA). Information about the phenolic profile of the consumed VOO and the human metabolism is thus obtained. The proposed approach represents a simple and versatile tool for qualitative and quantitative characterization of VOO phenolic metabolism.

A Systematic Ex-Vivo Study of the Anti-Proliferative/Cytotoxic Bioactivity of Major Olive Secoiridoids' Double Combinations and of Total Olive Oil Phenolic Extracts on Multiple Cell-Culture Based Cancer Models Highlights Synergistic Effects.

Several individual olive oil phenols (OOPs) and their secoiridoid derivatives have been shown to exert anti-proliferative and pro-apoptotic activity in treatments of human cancer cell lines originating from several tissues. This study evaluated the synergistic anti-proliferative/cytotoxic effects of five olive secoiridoid derivatives (oleocanthal, oleacein, oleuropein aglycone, ligstroside aglycone and oleomissional) in all possible double combinations and of total phenolic extracts (TPEs) on eleven human cancer cell lines representing eight cell-culture-based cancer models. Individual OOPs were used to treat cells for 72 h in half of their EC50 values for each cell line and their synergistic, additive or antagonistic interactions were evaluated by calculating the coefficient for drug interactions (CDI) for each double combination of OOPs. Olive oil TPEs of determined OOPs' content, originating from three different harvests of autochthonous olive cultivars in Greece, were evaluated as an attempt to investigate the efficacy of OOPs to reduce cancer cell numbers as part of olive oil consumption. Most combinations of OOPs showed strong synergistic effect (CDIs < 0.9) in their efficacy, whereas TPEs strongly impaired cancer cell viability, better than most individual OOPs tested herein, including the most resistant cancer cell lines evaluated.

Cp2TiCl/H2O as a Sustainable System for the Reduction of Organic Functional Groups: Potential Application of Cp2TiCl/D2O to the Analysis of Bioactive Phenols in Olive Oil

Significant efforts have been made toward developing sustainable reduction reactions of organic and bioorganic compounds. In these studies, the selection of reagents and solvents has played a very important role in the development of environment-friendly methodologies. In this context, the reducing agent Cp2TiCl/H2O has been introduced as a safe, efficient, selective, and low-cost reagent, and thus as a sustainable alternative for the reduction of organic compounds. To facilitate understanding of the reductions mediated by this system, in this study we focus on describing the intermediates, mechanisms, and representative examples. Finally, a reflection is made on the future perspectives of this reducing agent, including its analog Cp2TiCl/D2O as a powerful tool for the preparation of deuterated phenols, which can be successfully used as an internal standard for analyzing bioactive phenols in olive oil.

Virgin Olive Oil By-Product Valorization: An Insight into the Phenolic Composition of Olive Seed Extracts from Three Cultivars as Sources of Bioactive Molecules.

Olives are very rich in phenolic compounds with important health-promoting properties. The profile and content of phenols in olive pulp and virgin olive oil are strongly influenced by the fruit ripening degree, but little is known concerning the evolution of phenolic compounds in the seed. In this work, the phenolic composition of seed from Tuscan cultivars (Frantoio, Moraiolo, Leccino) was studied over maturation. Starting from each seed sample, a phenolic extract was prepared and analyzed by HPLC-DAD-MS. Nüzhenide and nüzhenide 11-methyl oleoside were by far the most abundant phenolic compounds; their content reached up to 46 g/kg in dry seeds, although this diminished in the final stage of fruit maturation. At the same time, the phenolic composition of the pulp was also characterized over the course of maturation, showing that oleuropein was by far the most abundant compound, with concentrations comparable to those of nüzhenide and nüzhenide 11-methyl oleoside in the seeds. Overall, the total amount of phenols in seed dry extracts was significant, reaching approx. 100 g/kg. The chemically characterized dry phenolic extracts from seeds could be used for future biological assays aimed at evaluating the potential bioactivities of these phytocomplexes.

Effect of olive oil phenols on oxidative stress biomarkers: A systematic review and dose-response meta-analysis of randomized clinical trials.

The phenolic content of olive oil has a role in cardiovascular protection. Some clinical trial studies demonstrated that phenolic compounds of olive oil have antioxidant activity which can protect macronutrients from oxidative damages. The aim of this study was to summarize the results of clinical trials which assessed the effects of high- versus low-phenol olive oil on oxidative stress biomarkers levels. We searched Scopus, PubMed, Web of Science, Google Scholar, ProQuest, and Embase up to July 2021. Eight clinical trials which evaluated the effect of the phenolic content of olive oil on oxidized-LDL (ox-LDL), malondialdehyde (MDA), or ferric-reducing ability of plasma (FRAP) were included the meta analysis. A significant decrease was observed in ox-LDL level (WMD: -0.29 U/L; 95% CI: -0.51, -0.07) and MDA (WMD: -1.82 μmoL/L; 95% CI: -3.13, -0.50). However, after subgroup analysis for MDA, the result was not significant for not serious limitation (SMD: -0.05, 95% CI: -0.35 to 0.24), but significant for serious limitation (SMD: -3.64, 95% CI: -4.29 to -2.99). Also, no significant change was found in FRAP (WMD: 0.0 mmoL/L; 95% CI: -0.03, 0.04) level. Dose-response analysis indicated a significant linear relationship between the phenolic content of olive oil and ox-LDL. The present study showed some beneficial effects of high-phenol compared with low-phenol olive oil on ox-LDL and MDA levels. According to the meta-regression analysis along with the increasing phenolic content of olive oil, a reduction in oxidative stress biomarkers was observed.

Maximising olive oil by‐products

Maximising olive oil by‐products

Effect of Controlled Oxygen Supply during Crushing on Volatile and Phenol Compounds and Sensory Characteristics in Coratina and Ogliarola Virgin Olive Oils.

In virgin olive oil industries, the technological choices of the production plant affect the biochemical activities that take place in the olives being processed throughout the entire process, thereby affecting the quality of the final product. The lipoxygenase pool enzymes that operated their activity during the first phases of the process need the best conditions to work, especially concerning temperature and oxygen availability. In this study, a system was equipped to supply oxygen in the crusher at a controllable concentration in an industrial olive oil mill at pilot plant scale, and four oxygen concentrations and two cultivars, Coratina and Ogliarola, were tested. The best concentration for oxygen supply was 0.2 L/min at the working capacity of 0.64 Ton/h. Further, using this addition of oxygen, it was possible to increase the compound's concentration, which is responsible for the green, fruity aroma. The effect on volatile compounds was also confirmed by the sensory analyses. However, at the same time, it was possible to maintain the concentration of phenols in a good quality olive oil while also preserving all the antioxidant properties of the product due to the presence of phenols. This study corroborates the importance of controlling oxygen supply in the first step of the process for process management and quality improvement in virgin olive oil production.

S-(-)-Oleocanthal Ex Vivo Modulatory Effects on Gut Microbiota.

Compelling evidence points to the critical role of bioactive extra-virgin olive oil (EVOO) phenolics and gut microbiota (GM) interplay, but reliable models for studying the consequences thereof remain to be developed. Herein, we report an optimized ex vivo fecal anaerobic fermentation model to study the modulation of GM by the most bioactive EVOO phenolic S-(-)-oleocanthal (OC), and impacts therefrom, focusing on OC biotransformation in the gut. This model will also be applicable for characterization of GM interactions with other EVOO phenolics, and moreover, for a broadly diverse range of bioactive natural products. The fecal fermentation media and time, and mouse type and gender, were the major factors varied and optimized to provide better understanding of GM-OC interplay. A novel resin entrapment technique (solid-phase extraction) served to selectively entrap OC metabolites, degradation products, and any remaining fraction of OC while excluding interfering complex fecal medium constituents. The effects of OC on GM compositions were investigated via shallow shotgun DNA sequencing. Robust metabolome analyses identified GM bacterial species selectively altered (population numbers/fraction) by OC. Finally, the topmost OC-affected gut bacterial species of the studied mice were compared with those known to be extant in humans and distributions of these bacteria at different human body sites. OC intake caused significant quantitative and qualitative changes to mice GM, which was also comparable with human GM. Results clearly highlight the potential positive health outcomes of OC as a prospective nutraceutical.

New Affordable Methods for Large-Scale Isolation of Major Olive Secoiridoids and Systematic Comparative Study of Their Antiproliferative/Cytotoxic Effect on Multiple Cancer Cell Lines of Different Cancer Origins.

Olive oil phenols (OOPs) are associated with the prevention of many human cancers. Some of these have been shown to inhibit cell proliferation and induce apoptosis. However, no systematic comparative study exists for all the investigated compounds under the same conditions, due to difficulties in their isolation or synthesis. Herein are presented innovative methods for large-scale selective extraction of six major secoiridoids from olive oil or leaves enabling their detailed investigation. The cytotoxic/antiproliferative bioactivity of these six compounds was evaluated on sixteen human cancer cell lines originating from eight different tissues. Cell viability with half-maximal effective concentrations (EC50) was evaluated after 72 h treatments. Antiproliferative and pro-apoptotic effects were also assessed for the most bioactive compounds (EC50 ≤ 50 μM). Oleocanthal (1) showed the strongest antiproliferative/cytotoxic activity in most cancer cell lines (EC50: 9-20 μM). The relative effectiveness of the six OOPs was: oleocanthal (1) &gt; oleuropein aglycone (3a,b) &gt; ligstroside aglycone (4a,b) &gt; oleacein (2) &gt; oleomissional (6a,b,c) &gt; oleocanthalic acid (7). This is the first detailed study comparing the bioactivity of six OOPs in such a wide array of cancer cell lines, providing a reference for their relative antiproliferative/cytotoxic effect in the investigated cancers.

Olive and echium oil gelled emulsions: Simulated effect of processing temperature, gelling agent and in vitro gastrointestinal digestion on oxidation and bioactive compounds

The impact and relative relevance of in vitro gastrointestinal digestion, processing temperature (room temperature or 180 °C), and gelling agent (GA) (carrageenan and alginate) on the bioactive compounds and oxidation status of olive and echium oils gelled formulations with 40% lipid incorporation was assessed. In vitro digestion was not affected by the GA, with >90% lipolysis in all formulations, but was the most relevant variable, promoting oxidation (MDA) regardless of the oil type, GA or temperature applied. Tocopherols and phenolic decreased with digestion, which could be interpreted as a protective response to pro-oxidative conditions during digestion. Temperature decreased olive oil phenolics. Gelification of echium oil using alginate reduced secondary oxidation products formation in comparison with carrageenan, with oxidation degrees after digestion equivalent to those shown with olive oil. The use of alginate with olive oil resulted in the most stable formulations, although not protecting its minor bioactive compounds from thermal degradation.

The influence of olive tree fertilization on the phenols in virgin olive oils. A review

The total phenols in virgin olive oil are highly dependent on cultivar, but also on ripening stage and other agronomic factors. The focus of most studies on agronomic factors has been irrigation, while fertilization has received less attention. Most of the fertilization works find that nitrogen over-fertilization leads to a decrease in phenol contents in virgin olive oil (VOO) and extra virgin olive oil (EVOO), under rain-fed or irrigation management. Ortho-diphenols also decrease with high doses of nitrogen, with no effect on secoiridoids. Phosphorous has a minor effect on irrigated trees; while the role of potassium is controversial, with a lack of trials with calcium and micro-nutrients. Due to the great impact of the fertilization on the phenol content and quality of VOO, new research is necessary with focus aimed at different cultivars and agronomic factors.

Characterization and Quantification of Capsaicinoids and Phenolic Compounds in Two Types of Chili Olive Oils, Using HPLC/MS.

Chili olive oil is a popular addition to various foods in many countries. In a detailed study, the content of phenols and capsaicinoids in chili olive oil was determined using chili flakes and whole chilies. A total of 99.8% of the phenols in chili olive oil were secoiridoids, with elenolic acid, oleuropein aglycones, and ligostride aglycones being the most abundant. Chili olive oil with chili flakes contained higher levels of capsaicinoids (+21.6%) compared to whole chili olive oil. Capsaicin and dihydrocapsaicin accounted for about 95% of all capsaicinoids in the chili olive oil. The extraction rate of dry “Cayenne” chili was 7.1% in whole chili olive oil and 9% in chili olive oil with flakes, confirming that chili flakes are better extracted in olive oil. With the determination of 29 individual phenols and five individual capsaicinoids, the study provided a detailed insight into the secondary metabolites of chili olive oil and confirmed that it is a health source.

Olive Oil Phenolics and Platelets-From Molecular Mechanisms to Human Studies.

Chronically activated, dysfunctional platelets mediate the progression of the majority of non-communicable diseases in a pleiotropic fashion. Antiplatelet therapy remains an attractive therapeutic means which however hasn't reached the expected targets according to the promising preclinical studies. It is therefore obvious that the consumption of foods demonstrating antiplatelet activity may be a less drastic but on the other hand a more sustainable way of achieving daily antiplatelet therapy, either alone or in combination with antiplatelet drugs. Olive oil is probably the main cardioprotective component of the Mediterranean Diet according to the results of observational and dietary intervention studies. Among all phytochemicals of olive oil, its unique phenolics seems to be responsible for the majority of its cardioprotective properties. This review article aims to highlight the platelet modulating roles of olive oil polyphenols, trying to critically assess whether those properties could partially explain the cardioprotective role of olive oil. The cellular and animal studies clearly show that extra virging olive oil (EVOO) phenolics, mainly hydroxytyrosol, are able to inhibit the activation of platelets induced by several endogenous agonists and pathologies. However, the outcomes of the pre-clinical studies are difficult to be translated to humans mainly because the dosages and the chemical forms of the phenolics used to these studies are much higher and different to that found in human circulation. Despite the heterogeneity of the few human trials on the field so far, the results are promising showing that EVOO can exert antiplatelet actions in real, acute or long-term, trials and at least part of this antiplatelet action can be attributed to the phenolic content of EVOOs. Although we clearly need better, well-powered studies to give certain answers on this field the antiplatelet properties of olive oil phenolics is a promising, emerging mechanism which may explain some of the health properties of EVOO and the Mediterranean Diet.

The secoiridoid profile of virgin olive oil conditions phenolic metabolism

The European Food Safety Authority highlights the beneficial effects of olive oil phenols, mainly, secoiridoids. Nevertheless, the metabolism of secoiridoids in humans has not been fully elucidated. This research evaluated the metabolism of secoiridoids in humans after intake of olive oils with diverse phenolic profiles. For this purpose, three extra virgin olive oils (EVOOs) were ingested by six volunteers at scheduled meals, and urine samples were collected the following morning for subsequent LC–MS/MS analysis. Using untargeted analysis, urinary metabolites revealed representative patterns associated with the various olive oil phenolic contents in absolute and relative terms. We were able to identify metabolites obtained through phase I, phase II, and microbial metabolism with discrimination between tyrosol and hydroxytyrosol derivatives. Metabolism of phenols is differentially activated as a function of the olive oil secoiridoids content, and this proof-of-concept study shows how urinary metabolites represent olive oil phenolic content.

Olive Oil Phenols Prevent Mercury-Induced Phosphatidylserine Exposure and Morphological Changes in Human Erythrocytes Regardless of Their Different Scavenging Activity.

Phosphatidylserine (PS) translocation to the external membrane leaflet represents a key mechanism in the pathophysiology of human erythrocytes (RBC) acting as an “eat me” signal for the removal of aged/stressed cells. Loss of physiological membrane asymmetry, however, can lead to adverse effects on the cardiovascular system, activating a prothrombotic activity. The data presented indicate that structurally related olive oil phenols prevent cell alterations induced in intact human RBC exposed to HgCl2 (5–40 µM) or Ca2+ ionophore (5 µM), as measured by hallmarks including PS exposure, reactive oxygen species generation, glutathione depletion and microvesicles formation. The protective effect is observed in a concentration range of 1–30 µM, hydroxytyrosol being the most effective; its in vivo metabolite homovanillic alcohol still retains the biological activity of its dietary precursor. Significant protection is also exerted by tyrosol, in spite of its weak scavenging activity, indicating that additional mechanisms are involved in the protective effect. When RBC alterations are mediated by an increase in intracellular calcium, the protective effect is observed at higher concentrations, indicating that the selected phenols mainly act on Ca2+-independent mechanisms, identified as protection of glutathione depletion. Our findings strengthen the nutritional relevance of olive oil bioactive compounds in the claimed health-promoting effects of the Mediterranean Diet.