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The lipophilization of polyphenols (phenolipids) may increase their affinity for membranes, leading to better antioxidant protection. Cholesteryl esters of caffeic, dihydrocaffeic, homoprotocatechuic and protocatechuic acids were synthetized in a one-step procedure with good to excellent yields of ~50-95%. After evaluation of their radical scavenging capacity by the DPPH method and establishing the anodic peak potential by cyclic voltammetry, their antioxidant capacity against AAPH-induced oxidative stress in soybean PC liposomes was determined. Their interaction with the liposomal membrane was studied with the aid of three fluorescence probes located at different depths in the membrane. The cholesteryl esters showed a better or similar radical scavenging capacity to that of α-tocopherol and a lower anodic peak potential than the corresponding parental phenolic acids. Cholesteryl esters were able to protect liposomes to a similar or greater extent than α-tocopherol. However, despite their antiradical capacity and being able to penetrate and orientate in the membrane in a parallel position to phospholipids, the antioxidant efficiency of cholesteryl esters was deeply dependent on the phenolipid polyphenolic moiety structure. When incorporated during liposome preparation, cholesteryl protocatechuate and caffeate showed more than double the activity of α-tocopherol. Thus, cholesteryl phenolipids may protect biomembranes against oxidative stress to a greater extent than α-tocopherol.

This review explores the potential health benefits and applications of phenolic secoiridoids derived from olive oil by-products in the prevention of Alzheimer's disease (AD). As reviewed herein, polyphenols, such as epigallocatechin-3-gallate, epicatechin, and resveratrol, show in vitro and in vivo antioxidant, anti-inflammatory, and neuroprotective properties, and are particularly relevant in the context of AD, a leading cause of dementia globally. The olive oil industry, particularly in the Mediterranean region, produces significant amounts of waste, including leaves, pomace, and wastewater, which pose environmental challenges but also offer an untapped source of bioactive compounds. Despite promising in vitro and in vivo studies indicating that olive-derived polyphenols, such as oleuropein and hydroxytyrosol, may mitigate AD pathology, human clinical trials remain limited. The variability in extraction methods and the complex nature of AD further complicate research. Future studies should focus on standardizing the protocols and conducting robust clinical trials to fully assess the therapeutic potential of these compounds. This approach not only supports the development of new treatments for AD but also promotes environmental sustainability by valorizing olive oil industry waste.

Background: Sulphur containing natural compounds are among the most biologically relevant metabolites in vivo . Naringenin 8-sulphonate from Parinari excelsa Sabine was evaluated in a previous work, demonstrating ability to act as a natural anti-inflammatory. Although the interference of this molecule against different inflammatory mediators was described, there is no information regarding its potential toxicity and pharmacokinetics, which are essential for its capacity to reach its therapeutic targets. In fact, despite the existence of reports on naringenin ADMET properties, the influence of sulphation patterns on them remains unknown., Objectives: This work aims to assess the in vitro pharmacokinetic and toxicological behavior of naringenin 8-sulphonate, as well as to understand the importance of the presence and position of the sulphur containing group for that., Methods: Naringenin 8-sulphonate physicochemical and ADMET properties were investigated using in silico tools and cell-based in vitro models. At the same time, naringenin and naringenin 4'- O -sulphate were investigated to evaluate the impact of the sulphonate group on the results. ADMETlab 2.0 in silico tool was used to predict the compounds' physicochemical descriptors. Pharmacokinetic properties were determined experimentally in vitro . While MRC-5 lung fibroblasts and HaCaT keratinocytes were used to evaluate the cytotoxicity of samples through MTT and LDH assays, Caco-2 human intestinal epithelial cells were used for the determination of genotoxicity, through alkaline comet assay, and as a permeability model to assess the ability of compounds to cross biological barriers., Results: Experimental determinations showed that none of the compounds was cytotoxic. In terms of genotoxicity, naringenin 8-sulphonate and naringenin caused significant DNA fragmentation, whereas naringenin 4'- O -sulphate did not. When it comes to permeability, the two sulphur-containing compounds with a sulphur containing group were clearly less capable to cross the Caco-2 cell barrier than naringenin., Conclusion: In this study, we conclude that the sulphur containing group from naringenin 8-sulphonate is disadvantageous for the molecule in terms of ADMET properties, being particularly impactful in the permeability in intestinal barrier models. Thus, this work provides important insights regarding the role of flavonoids sulphation and sulphonation upon pharmacokinetics and toxicity., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The author(s) declared that they were an editorial board member of Frontiers, at the time of submission. This had no impact on the peer review process and the final decision., (Copyright © 2024 Macedo, Paiva-Martins, Valentão and Pereira.)

In this work, we aim to find physical evidence demonstrating the crucial role that the effective concentration of antioxidants (AOs) present at the interfacial region of emulsions has in controlling the inhibition of the lipid oxidation reaction. We prepared a series of antioxidants of different hydrophobicities derived from chlorogenic and protocatechuic acids. We first monitored, in intact emulsions, the (sigmoidal) production of conjugated dienes and determined the corresponding induction times, t ind . Independently, we determined the effective concentrations of the antioxidants in the same intact emulsions. Results show that both the length of the induction periods and the antioxidant interfacial concentrations parallel each other, with a maximum at the octyl-dodecyl derivatives. The ratio between the interfacial antioxidant concentrations and the induction periods remains constant for all AOs in the same series, so that the rates of initiation of lipid oxidation are the same regardless of the hydrophobicity of the antioxidant employed. The constancy in the rate of initiation provides strong experimental evidence for a direct relationship between interfacial concentrations and antioxidant efficiencies. Results suggest new possibilities to investigate lipid peroxidation under non-forced conditions and are of interest to formulators interested in preparing emulsions with antimicrobial properties.

The chemistry of the phenolic compounds found in virgin olive oil (VOO) is very complex due, not only to the different classes of polyphenols that can be found in it, but, above all, due to the existence of a very specific phenol class found only in oleaceae plants: the secoiridoids. Searching in the Scopus data base the keywords flavonoid, phenolic acid, lignin and secoiridoid, we can find a number of 148174, 79435, 11326 and 1392 research articles respectively, showing how little is devote to the latter class of compounds. Moreover, in contrast with other classes, that include only phenolic compounds, secoiridoids may include phenolic and non-phenolic compounds, being the articles concerning phenolic secoiridoids much less than the half of the abovementioned articles. Therefore, it is important to clarify the structures of these compounds and their chemistry, as this knowledge will help understand their bioactivity and metabolism studies, usually performed by researchers with a more health science's related background. In this review, all the structures found in many research articles concerning VOO phenolic compounds chemistry and metabolism was gathered, with a special attention devoted to the secoiridoids, the main phenolic compound class found in olives, VOO and olive leaf.

The inflammatory response is a vital mechanism for repairing damage induced by aberrant health states or external insults; however, persistent activation can be linked to numerous chronic diseases. The nuclear factor kappa β (NF-κB) inflammatory pathway and its associated mediators have emerged as critical targets for therapeutic interventions aimed at modulating inflammation, necessitating ongoing drug development. Previous studies have reported the inhibitory effect of a hydroethanol extract derived from Parinari excelsa Sabine (Chrysobalanaceae) on tumour necrosis factor-alpha (TNF-α), but the phytoconstituents and mechanisms of action remained elusive. The primary objective of this study was to elucidate the phytochemical composition of P. excelsa stem bark and its role in the mechanisms underpinning its biological activity. Two compounds were detected via HPLC-DAD-ESI(Ion Trap)-MS 2 analysis. The predominant compound was isolated and identified as naringenin-8-sulphonate (1), while the identity of the second compound (compound 2) could not be determined. Both compound 1 and the extract were assessed for anti-inflammatory properties using a cell-based inflammation model, in which THP-1-derived macrophages were stimulated with LPS to examine the treatments' effects on various stages of the NF-κB pathway. Compound 1, whose biological activity is reported here for the first time, demonstrated inhibition of NF-κB activity, reduction in interleukin 6 (IL-6), TNF-α, and interleukin 1 beta (IL-1β) production, as well as a decrease in p65 nuclear translocation in THP-1 cells, thus highlighting the potential role of sulphur substituents in the activity of naringenin (3). To explore the influence of sulphation on the anti-inflammatory properties of naringenin derivatives, we synthesized naringenin-4'-O-sulphate (4) and naringenin-7-O-sulphate (5) and evaluated their anti-inflammatory effects. Naringenin derivatives 4 and 5 did not display potent anti-inflammatory activities; however, compound 4 reduced IL-1β production, and compound 5 diminished p65 translocation, with both exhibiting the capacity to inhibit TNF-α and IL-6 production. Collectively, the findings demonstrated that the P. excelsa extract was more efficacious than all tested compounds, while providing insights into the role of sulphation in the anti-inflammatory activity of naringenin derivatives., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Inc. All rights reserved.)

In recent years, partitioning of antioxidants in oil-water two-phase systems has received great interest because of their potential in the downstream processing of biomolecules, their benefits in health, and because partition constant values between water and model organic solvents are closely related to important biological and pharmaceutical properties such as bioavailability, passive transport, membrane permeability, and metabolism. Partitioning is also of general interest in the oil industry. Edible oils such as olive oil contain a variety of bioactive components that, depending on their partition constants, end up in an aqueous phase when extracted from olive fruits. Frequently, waste waters are subsequently discarded, but their recovery would allow for obtaining extracts with antioxidant and/or biological activities, adding commercial value to the wastes and, at the same time, would allow for minimizing environmental risks. Thus, given the importance of partitioning antioxidants, in this manuscript, we review the background theory necessary to derive the relevant equations necessary to describe, quantitatively, the partitioning of antioxidants (and, in general, other drugs) and the common methods for determining their partition constants in both binary ( P W OIL ) and multiphasic systems composed with edible oils. We also include some discussion on the usefulness (or not) of extrapolating the widely employed octanol-water partition constant ( P W OCT ) values to predict P W OIL values as well as on the effects of acidity and temperature on their distributions. Finally, there is a brief section discussing the importance of partitioning in lipidic oil-in-water emulsions, where two partition constants, that between the oil-interfacial, P O I , and that between aqueous-interfacial, P w I , regions, which are needed to describe the partitioning of antioxidants, and whose values cannot be predicted from the P W OIL or the P W OCT ones.

During the last years, the formalism of the pseudophase kinetic model (PKM) has been successfully applied to determine the distributions of antioxidants and their effective interfacial concentrations, and to assess the relative importance of emulsion and antioxidant properties (oil and surfactant nature, temperature, acidity, chemical structure, hydrophilic-liphophilic balance (HLB), etc.) on their efficiency in intact lipid-based emulsions. The PKM permits separating the contributions of the medium and of the concentration to the overall rate of the reaction. In this paper, we report the results of a specifically designed experiment to further test the suitability of the PKM to evaluate the distributions of antioxidants among the various regions of intact lipid-based emulsions and provide insights into their chemical reactivity in multiphasic systems. For this purpose, we employed the antioxidants α- and δ-TOCopherol (α- and δ-TOC, respectively) and determined, at different acidities well below their p K a, the interfacial rate constants k I for the reaction between 16-ArN 2 + and α- and δ-TOC, and the antioxidant distributions in intact emulsions prepared with olive and soybean oils. Results show that the effective interfacial concentration of δ-TOC is higher than that of α-TOC in 1:9 ( v / v ) soybean and 1:9 olive oil emulsions. The effective interfacial concentrations of tocopherols are much higher (15-96-fold) than the stoichiometric concentrations, as the effective interfacial concentrations of both δ-TOC and α-TOC in soybean oil emulsions are higher (2-fold) than those in olive oil emulsions. Overall, the results demonstrate that the PKM grants an effective separation of the medium and concentration effects, demonstrating that the PKM constitutes a powerful non-destructive tool to determine antioxidant concentrations in intact emulsions and to assess the effects of various factors affecting them.

The effect of bovine back fat replacement by oleogels containing pork skin and olive oil on the oxidative stability, physicochemical, technological, nutritional, and sensory parameters of burgers was evaluated. Four different hamburger (H) were manufactured: with 90 % of lean beef and 10 % of bovine back fat (control, HC), or with 10 % of pork skin/water/virgin olive oil (HVOO), stripped olive oil added of an olive leaf extract (HESOO) or stripped olive oil (HSOO) oleogels, at 20:60:20 ratio. Physical-chemical stability was assessed after storage for 7 days at 4 °C and for 90 days at -20 °C, under non-vacuum and vacuum packaging. A reduction in the fat content by 80 % and in the energy content by 35 %, an increase in the protein content by 15 % and a better fatty acid profile were achieved in the oleogel containing burgers. After processing at 180 °C (grill), hardness, chewiness, sensory parameters and overall acceptability were high and comparable to control. All burgers were oxidative stable during 7 days at 4 °C. After storage for 90 days at -20 °C, only HSOO samples stored under non-vacuum packaging were oxidized. The antioxidant content in samples HVOO and HESOO efficiently prevented the oxidation of these samples., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Natural antioxidants from fruits and vegetables, meats, eggs and fish protect cells from the damage caused by free radicals. They are widely used to reduce food loss and waste, minimizing lipid oxidation, as well as for their effects on health through pharmaceutical preparations. In fact, the use of natural antioxidants is among the main efforts made to relieve the pressure on natural resources and to move towards more sustainable food and pharmaceutical systems. Alternative food waste management approaches include the valorization of by-products as a source of phenolic compounds for functional food formulations. In this review, we will deal with the chemistry of antioxidants, including their molecular structures and reaction mechanisms. The biochemical aspects will also be reviewed, including the effects of acidity and temperature on their partitioning in binary and multiphasic systems. The poor bioavailability of antioxidants remains a huge constraint for clinical applications, and we will briefly describe some delivery systems that provide for enhanced pharmacological action of antioxidants via drug targeting and increased bioavailability. The pharmacological activity of antioxidants can be improved by designing nanotechnology-based formulations, and recent nanoformulations include nanoparticles, polymeric micelles, liposomes/proliposomes, phytosomes and solid lipid nanoparticles, all showing promising outcomes in improving the efficiency and bioavailability of antioxidants. Finally, an overview of the pharmacological effects, therapeutic properties and future choice of antioxidants will be incorporated.

The anti-inflammatory potential of oleacein, the main polyphenolic compound found in olive oil, and its main metabolites were characterized by their effects on RAW 264.7 macrophages challenged with lipopolysaccharide (LPS), and by their ability to inhibit enzymes of the arachidonic acid metabolism with a key role in the synthesis of pro-inflammatory lipid mediators. Oleacein at 12.5 µM significantly decreased the amount of L-citrulline and ● NO generated by LPS-stimulated macrophages. Hydroxytyrosol, hydroxytyrosol acetate and hydroxytyrosol acetate sulfate were also able to reduce the cellular amount of ● NO, although to a lesser extent. In contrast, hydroxytyrosol glucuronide and sulfate did not show detectable effects. Oleacein was also able to inhibit the coupled PLA 2 + 5-LOX enzyme system (IC 50 = 16.11 µM), as well as the 5-LOX enzyme (IC 50 = 45.02 µM). Although with lower activity, both hydroxytyrosol and hydroxytyrosol acetate were also capable of inhibiting these enzymes at a concentration of 100 µM. None of the other tested metabolites showed a capacity to inhibit these enzymes. In contrast, all compounds, including glucuronides and sulfate metabolites, showed a remarkable capacity to inhibit both cyclooxygenase isoforms, COX-1 and COX-2, with IC 50 values lower than 3 µM. Therefore, oleacein and its metabolites have the ability to modulate ● NO- and arachidonic acid-dependent inflammatory cascades, contributing to the anti-inflammatory activity associated with olive oil polyphenols.

Selecting effective antioxidants is challenging since their efficiency in inhibiting lipid oxidation depends on the rate constants of the chemical reactions involved and their concentration at the reaction site, i.e., at the interfacial region. Accumulation of antioxidants at the interface of emulsions is key to modulate their efficiency in inhibiting lipid oxidation but its control was not well understood, especially in emulsions. It can be optimized by modifying the physicochemical properties of antioxidants or the environmental conditions. In this work, we analyze the effects of surfactant concentration, droplet size, and oil to water ratio on the effective interfacial concentration of a set of chlorogenic acid (CGA) esters in fish oil-in-water (O/W) emulsions and nanoemulsions and on their antioxidant efficiency. A well-established pseudophase kinetic model is used to determine in the intact emulsified systems the effective concentrations of the antioxidants (AOs). The relative oxidative stability of the emulsions is assessed by monitoring the formation of primary oxidation products with time. Results show that the concentration of all AOs at the interfacial region is much higher (20-90 fold) than the stoichiometric one but is much lower than those of other phenolipid series such as caffeic or hydroxytyrosol derivatives. The main parameter controlling the interfacial concentration of antioxidants is the surfactant volume fraction, Φ I , followed by the O/W ratio. Changes in the droplet sizes (emulsions and nanoemulsions) have no influence on the interfacial concentrations. Despite the high radical scavenging capacity of CGA derivatives and their being concentrated at the interfacial region, the investigated AOs do not show a significant effect in inhibiting lipid oxidation in contrast with what is observed using other series of homologous antioxidants with similar reactivity. Results are tentatively interpreted in terms of the relatively low interfacial concentrations of the antioxidants, which may not be high enough to make the rate of the inhibition reaction faster than the rate of radical propagation.

Hypothesis: A detailed quantitative description of the effects of antioxidants in inhibiting lipid peroxidation in oil-in-water emulsions can be achieved by determining the relationships between the rates of initiation of the lipid peroxidation reaction, the length of the induction period preceding the propagation step of the radical oxidation process and the effective antioxidant interfacial concentrations., Experiments: We successfully prepared and characterized a series of olive oil-in-water nanoemulsions and allowed them to spontaneously oxidize. Their oxidative stability was evaluated by carrying out in the presence, and absence, of antioxidants derived from gallic acid, by monitoring the formation of primary oxidation products with time, by determining the corresponding induction periods, and by determining the effective interfacial concentrations of the antioxidants in the intact emulsions., Findings: Results show that both, the length of the induction periods and the antioxidant interfacial concentrations change concomitantly, increasing with the hydrophobicity of the antioxidant up to a maximum at the octyl derivative; longer aliphatic chains decrease their efficiency. The ratio between the interfacial antioxidant concentration and the induction period remains constant independently of the antioxidant, demonstrating that the effective concentrations of antioxidant at the interface control their efficiencies in emulsions., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 The Author(s). Published by Elsevier Inc. All rights reserved.)

Toxicity caused by the exposure to human-made chemicals and environmental conditions has become a major health concern because they may significantly increase the formation of reactive oxygen species (ROS), negatively affecting the endogenous antioxidant defense. Living systems have evolved complex antioxidant mechanisms to protect cells from oxidative conditions. Although oxidative stress contributes to various pathologies, the intake of molecules such as polyphenols, obtained from natural sources, may limit their effects because of their antioxidant and antimicrobial properties against lipid peroxidation and against a broad range of foodborne pathogens. Ingestion of polyphenol-rich foods, such as fruits and vegetables, help to reduce the harmful effects of ROS, but the use of supramolecular and nanomaterials as delivery systems has emerged as an efficient method to improve their pharmacological and therapeutic effects. Suitable exogenous polyphenolic antioxidants should be readily absorbed and delivered to sites where pathological oxidative damage may take place, for instance, intracellular locations. Many potential antioxidants have a poor bioavailability, but they can be encapsulated to improve their ideal solubility and permeability profile. Development of effective antioxidant strategies requires the creation of new nanoscale drug delivery systems to significantly reduce oxidative stress. In this review we provide an overview of the oxidative stress process, highlight some properties of ROS, and discuss the role of natural polyphenols as bioactives in controlling the overproduction of ROS and bacterial and fungal growth, paying special attention to their encapsulation in suitable delivery systems and to their location in colloidal systems where interfaces play a crucial role.

Caffeic acid (CA) has demonstrated a strong intracellular antioxidant ability by scavenging ROS, contributing to the maintenance of cell membrane structural integrity and to reduce oxidative injuries in other cell components. Nevertheless, caffeic acid has limited usage, due to its hydrophilic character. In this work, the introduction of alkyl chains in the caffeic acid molecule by esterification (methyl - C1, ethyl - C2, butyl - C4, hexyl - C6, octyl - C8 and hexadecyl - C16), significantly increased its lipophilicity. All caffeates tested showed a much higher protective activity than caffeic acid against red blood cells (RBCs) AAPH-induced oxidative stress; this protection was heavily dependent on the length of the alkyl chain of the esters, and on their concentration. At 2.5 and 5 μM, the more lipophilic compounds (C8 and C16) showed a remarkable antioxidant activity, inhibiting hemolysis; probably, their better location within the membrane leads to a better antioxidative protection; however, at 50 μM, the more hydrophilic compounds (C1-C4) showed a better activity against hemolysis than the more lipophilic ones (C8-C16). At this higher concentration, the better interaction of the more lipophilic compounds with the membrane seems to cause changes in RBC membrane fluidity, disturbing membrane integrity. Our data show that the antioxidant activity of these compounds could play an important role for the protection of different tissues and organs, by protecting cell membranes from oxidative injuries., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Reversible acetylcholinesterase (AChE) inhibitors are key therapeutic tools to modulate the cholinergic connectivity compromised in several degenerative pathologies. In this work, four alkyl esters of homarine were synthesized and screened by using Electrophorus electricus AChE and rat brain AChE-rich fraction. Results showed that all homarine alkyl esters are able to inhibit AChE by a competitive inhibition mode. The effectiveness of AChE inhibition increases with the alkyl side chain length of the homarine esters, being HO-C 16 (IC 50 =7.57±3.32 μM and K i =18.96±2.28 μM) the most potent inhibitor. The fluorescence quenching studies confirmed that HO-C 16 is the compound with higher selectivity and affinity for the tryptophan residues in the catalytic active site of AChE. Preliminary cell viability studies showed that homarine esters display no toxicity for human neuronal SH-SY5Y cells. Thus, the long-chain homarine esters emerge as new anti-cholinesterase agents, with potential to be considered for therapeutic applications development., (© 2021 Wiley-VCH GmbH.)

Surfactants have been used for decades in the food industry for the preparation of lipid-based emulsified food stuffs. They play two main roles in the emulsification processes: first they decrease the interfacial tension between the oil and water, facilitating droplet deformation and rupture; second, they reduce droplet coalescence by forming steric barriers. However, addition of surfactants to binary oil-water mixtures also brings up the formation of three-dimensional interfacial layers, surrounding each emulsion droplet, that significantly alter chemical reactivity. This is the case, for instance, in the inhibition reaction between antioxidants and the lipid radicals formed in the course of the spontaneous oxidation reaction of unsaturated lipids, which are commonly employed in the preparation of food-grade emulsions. The rate of the inhibition reaction depends on the effective concentrations of antioxidants, which are mostly controlled by the amount of surfactant employed in the preparation of the emulsion. In this work, we analyze the effects of the surfactant Tween 20 on the oxidative stability and on the effective concentrations of two model antioxidants derived from cinnamic acid, determining their interfacial concentrations in the intact emulsions to avoid disrupting the existing equilibria and biasing results. For this purpose, a recently developed methodology was employed, and experimental results were interpreted on the grounds of a pseudophase kinetic model.

Bulk phase chemistry is hardly ever a reasonable approximation to interpret chemical reactivity in compartmentalized systems, because multiphasic systems may alter the course of chemical reactions by modifying the local concentrations and orientations of reactants and by modifying their physical properties (acid-base equilibria, redox potentials, etc.), making them-or inducing them-to react in a selective manner. Exploiting multiphasic systems as beneficial reaction media requires an understanding of their effects on chemical reactivity. Chemical reactions in multiphasic systems follow the same laws as in bulk solution, and the measured or observed rate constant of bimolecular reactions can be expressed, under dynamic equilibrium conditions, in terms of the product of the rate constant and of the concentrations of reactants. In emulsions, reactants distribute between the oil, water, and interfacial regions according to their polarity. However, determining the distributions of reactive components in intact emulsions is arduous because it is physically impossible to separate the interfacial region from the oil and aqueous ones without disrupting the existing equilibria and, therefore, need to be determined in the intact emulsions. The challenge is, thus, to develop models to correctly interpret chemical reactivity. Here, we will review the application of the pseudophase kinetic model to emulsions, which allows us to model chemical reactivity under a variety of experimental conditions and, by carrying out an appropriate kinetic analysis, will provide important kineticparameters.

Our previous research was focused on the effects of hydrophobicity on the antioxidant (AO) efficiency of series of homologous antioxidants with the same reactive moieties. In this work we evaluate the antioxidant efficiency of hydrophobic phenolipids in 4:6 olive oil-in-water emulsions, with different phenolic moieties (derived from caffeic, 4-hydroxycinnamic, dihydrocaffeic acids, tyrosol and hydroxytyrosol), with alkyl chains of 8 and 16 carbons, and compare the antioxidant efficiency with that of the parent compounds. All catecholic phenolipids, in particular the C8 derivatives, have proven to be better antioxidants for the oxidative protection of emulsions than their parental compounds with octyl dihydrocafffeate being the most efficient (16-fold increase in relation to the control). To understand the importance of some factors on the antioxidant efficiency of compounds in emulsions, Pearson's correlation analysis was carried out between antioxidant activity and the first anodic potential ( E pa ), reducing capacity (FRAP value), DPPH radical scavenging activity (EC 50 ) and the concentration of antioxidants in each region of the emulsified system. Results confirm the importance of the effective concentration of AOs in the interfacial region (AO I ) (ρ = 0.820) and of the E pa (ρ = -0.677) in predicting their antioxidant efficiency in olive oil-in-water emulsions.

The autoxidation of lipids in complex systems such as emulsions or biological membranes, although known to occur readily and to be associated with important pathological events, is lacking in quantitative data in spite of the huge efforts that have been made in attempting to unravel the complex mechanisms of lipid oxidation and its inhibition by antioxidants. Lipids are present as oil-in-water emulsions in many foods and pharmaceutical formulations, and the prevalent role of the interfacial region is critical to understand the antioxidant behavior and to correctly interpret antioxidant efficiencies. The aim of this review is to summarize the current knowledge on the chemical fate of antioxidants before they react with peroxyl radicals. Many researchers highlighted the predominant role of interfaces, and although some attempts have been made to understand their role, in most instances, they were essentially qualitative and based on putative hypotheses. It is only recently that quantitative reports have been published. Indeed, knowledge on the effects of relevant experimental variables on the effective concentrations of antioxidants is necessary for a successful design of alternate, effective antioxidative solutions.

Hydroxytyrosol (HT) is a well-known olive oil polyphenol for its high antioxidant capacity and important cardio and neuroprotective effects. However, its use in lipidic systems is limited, due to its hydrophilic character. In this study, we approach the particular structure of xanthophylls and synthetize HT esters specially designed for the protection of liposomal systems. These HT esters contain two polyphenolic moieties separated by a lipophilic alkyl spacer of different length (12, 16 or 22 carbons). To evaluate the antioxidant activity of these compounds against the 2,2'-azobis(2-amidinopropane) hydrochloride induced oxidation, soybean phospholipid liposomes were used. Fluorescence quenching studies were used to assess the insertion of the compounds in the liposomes. The synthetized HT derivatives were able to protect liposomes from induced oxidation when added to the suspensions. The rank of activity was severely influenced by the alkyl chain length of the spacer molecule, being the C12 derivative the most active antioxidant, with an increase in the oxidative stability of liposomes of 2.2 times when compared with the control. The incorporation of compounds during liposome preparation improved the antioxidant capacity of all HT derivatives by about 2.8 times, when compared to the control. This is probably due to a similar transmembrane position with both polyphenolic rings located at the phospholipid polar heads. The synthesis of bis-ester derivatives seems to be a promising strategy to fine-tune antioxidant molecules at biomembranes, thus increasing the oxidative stability of liposomal systems by improving the antioxidant activity of hydrophilic phenolic compounds with high free radical scavenging activity., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Reports on the effect of droplet size on the oxidative stability of emulsions and nanoemulsions are scarce in the literature and frequently contradictory. Here, we have employed a set of hydroxytyrosol (HT) esters of different hydrophobicity and fish oil-in-water emulsified systems containing droplets of different sizes to evaluate the effect of the droplet size, surfactant, (Φ I ) and oil (Φ O ) volume fractions on their oxidative stability. To quantitatively unravel the observed findings, we employed a well-established pseudophase kinetic model to determine the distribution and interfacial concentrations of the antioxidants (AOs) in the intact emulsions and nanoemulsions. Results show that there is a direct correlation between antioxidant efficiency and the concentration of the AOs in the interfacial region, which is much higher (20-200 fold) than the stoichiometric one. In both emulsified systems, the highest interfacial concentration and the highest antioxidant efficiency was found for hydroxytyrosol octanoate. Results clearly show that the principal parameter controlling the partitioning of antioxidants is the surfactant volume fraction, Φ I , followed by the O/W ratio; meanwhile, the droplet size has no influence on their interfacial concentrations and, therefore, on their antioxidant efficiency. Moreover, no correlation was seen between droplet size and oxidative stability of both emulsions and nanoemulsions.

The consumption of extra virgin olive oil (EVOO) has been associated with a lower incidence of cardiovascular diseases partly due to its polyphenol content. The metabolites hydroxytyrosol sulfate and hydroxytyrosol acetate sulfate were shown to be the most concentrated polyphenol metabolites found in plasma after EVOO consumption. Therefore, the capacity of hydroxytyrosol, hydroxytyrosol acetate, homovanillyl alcohol, homovanillyl alcohol acetate and tyrosol sulfate metabolites, to protect red blood cells (RBCs) from oxidative injury induced by the radical initiator 2,2'-azo-bis(2-amidinopropane) dihydrochloride (AAPH) was evaluated. In the presence of AAPH, all non-sulfated compounds and the hydroxytyrosol and hydroxytyrosol acetate monosulfate metabolites showed a significant protective activity against RBCs induced oxidative hemolysis. Moreover, even at 5 μM, the protection was highly significant for hydroxytyrosol acetate, hydroxytyrosol and hydroxytyrosol acetate 3' and 4' monosulfates. The morphological changes of RBC and the nature of their hemoglobin were in accordance with the hemolysis assay. Results showed that a free phenolic hydroxyl group is needed for the antioxidant protection given by compounds. Hydroxytyrosol metabolites present as phase II sulfate conjugates are actually able to protect RBC from oxidative injury by a non-transcriptional mechanism and are likely to contribute for the anti-atherosclerosis properties of regular EVOO consumption.

The distribution of a homologous series of polyphenol derivatives of increasing lipophilicity has been determined in fish oil-in-water emulsions and nanoemulsions by the pseudophase model. One of the hypotheses on which the pseudophase model is based, is that its application is independent of the size of emulsion droplets. In agreement with our hypothesis, results showed that the smaller droplet size found in nanoemulsions does not affect partition constants of gallic acid (GA) and its esters. The antioxidant efficiency of GA and gallates in the emulsified systems used, correlated positively with the concentration of antioxidant at the interfacial region. The increase in the oil/water ratio increased the overall oxidative stability of emulsions but decreased the antioxidant efficiency of the more lipophilic derivatives. This can be assigned to the fact that, increasing the oil phase volume, the interfacial concentration decreased for the more lipophilic antioxidants., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Hypothesis: One fundamental and unsolved question in colloid chemistry, and also in the food industry, is whether molecular distributions, specifically the interfacial concentrations of antioxidants (AO I ), are independent of the droplet sizes. Pseudophase kinetic models, widely employed to interpret chemical reactivity in colloidal systems and to determine antioxidant distributions, assume that they are independent., Experiments: To prove, or discard, the above hypothesis, we prepared and characterized a series of olive and fish oil-in-water nanoemulsions with different droplet sizes, carried out a kinetic study to evaluate their oxidative stability, both in the presence and absence of gallic acid (GA), and determined its interfacial concentrations., Findings: Results indicate that a change in the droplet size (80-1300 nm) does not alter the oxidative stability of the nanoemulsions in the absence of GA. Addition of GA increases their oxidative shelf-life and, at constant surfactant volume fraction, Φ I , the oxidative stability and the antioxidant distribution do not depend on the droplet size. Overall, results suggest that the droplet size does not affect the ratio between the rates of radical production and of inhibition by antioxidants, ratio that defines an "efficient" (or inefficient) antioxidant, providing experimental evidence supporting the operative assumption of pseudophase kinetic models., (Copyright © 2019 Elsevier Inc. All rights reserved.)

A simple and efficient procedure for the synthesis of N-acyl 4-hydroxy, 4-hydroxy-3-methoxy and 3,4-dihydroxy phenylglycine amides by a strategy based on the multicomponent Ugi reaction is proposed. Hydroxybenzaldehyde derivatives were reacted with 4-methoxybenzylamine, cyclohexyl isocyanide and benzoic acid or 2-naphthylacetic acid to give Ugi adducts that were treated with trifluoroacetic acid yielding N-acyl hydroxyphenylglycine amides in good yields. The same procedure using as acid component protocatechuic acid or hydrocaffeic acid gave N-catechoyl 3,4-dihydroxyphenylglycine amides. The use of N-benzyloxycarbonylglycine as acid component allowed the preparation of a 3,4-dihydroxyphenylglycyl dipeptide derivative. Radical-scavenging activity studies of the polyphenolic amino acid derivatives showed a sharp increase in activity with the increase in number of hydroxyl or catechol groups present. Cyclic voltammetry experiments established a correlation between oxidation peak potentials and the radical-scavenging activity., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Background: Controlling the interfacial concentrations of antioxidants (AOs) in oil-in-water emulsions can be regarded as a unique approach for increasing the efficiency of AOs in inhibiting the oxidation of lipids. Classical methods to determine the AO distribution in binary systems cannot be employed and their distribution needs to be assessed in the intact emulsion., Results: We have employed a well-established kinetic method to determine the distribution of a homologous series of AOs derived of chlorogenic acid in olive oil-in-water emulsions and analyse the effects of AO hydrophobicity on their distributions and their efficiencies. Results indicate that variations in the efficiency of chlorogenates in emulsions are due to differences in their interfacial concentrations. Their interfacial concentrations AO I were much higher (20- to 150-fold) than their stoichiometric concentrations. On the other hand, their concentrations in the oil region were 1.5- to 0.1-fold. Results also show the complex effect of the oil-to-water ratio employed in the preparation of the emulsions on the (AO I ) values., Conclusion: Results highlight the key role of the interfacial region and of its composition (interfacial AO molarity, emulsifier concentration, oil-to-water ratio) in interpreting the efficiency of AOs in inhibiting lipid oxidation in emulsions. Thus, a careful modulation of these parameters is necessary to ensure optimum AO efficiency. © 2019 Society of Chemical Industry., (© 2019 Society of Chemical Industry.)

The oxidation of lipid-based emulsions and nanoemulsions strongly affects their overall quality and safety. Moreover, introduction of oxidatively unstable emulsions into biological systems either as an energy source in parenteral nutrition or as delivery systems of bioactives may promote oxidation "in situ" leading to the overproduction of reactive oxygen species, initiating new harmful oxidative reactions and increasing the oxidative damage. Addition of antioxidants, AOs, may help to prevent the oxidative degradation of unsaturated lipids. Nevertheless, prediction of the optimal antioxidant or set of antioxidants and their efficiency is still far from being completely understood because the site of reaction is often uncertain and because the effective concentrations of reactants in the different regions of the emulsion have been frequently overlooked. Furthermore, the absence of quantitative relationships between the hydrophobicity of the antioxidants and their partitioning among the oil, water, and interfacial regions hampers their optimal use. Here we investigated the effects of gallic acid and some of its alkyl derivatives on the oxidative stability of soybean oil-in-water emulsions and determined their effective concentrations in the different regions of the emulsion (aqueous, oil, and interface). The results provide physical evidence for the crucial role played by the interfacial region in the reaction between antioxidants and lipid radicals: a direct relationship between interfacial concentrations and the oxidative stability could be established. The results indicate that AOs accumulate in the interfacial region, where the effective concentration is 20-180 times higher than the stoichiometric concentrations. Control of the hydrophobicity of the AOs and of the surfactant concentration allows control of interfacial concentrations: the lower the concentration of surfactant employed, the higher the effective interfacial concentration.

Distributions of a homologous series of gallic acid (GA) derivatives of increasing lipophilicity were determined in stripped fish oil-in-water emulsions. The distribution was determined by employing a kinetic method based on the reaction between the hydrophobic 4-hexadecylbenzenediazonium ions (16-ArN 2 + ) and the antioxidants (AOs), that allows to determine the partition constants between the oil-interfacial, P O I , and water-interfacial, P W I , regions of the emulsion. Results showed that the antioxidant activity of gallic acid and its derivatives was positively correlated with the percentage of AO at the interface. The percentage of butyl, octyl and dodecyl gallates in the interfacial region is similar but much higher than that of the other derivatives, showing the highest AO efficiency. When increasing the surfactant fraction, the percentage of all compounds at the interface increased but the emulsion stability decreased due to the dilution of AO at the interface region. Both butyl and octyl gallates were found the most suitable AOs to carry out fish oil-in-water emulsions protection. For maximum effectiveness, butyl and octyl gallate should be used with the lowest emulsifier volume fraction necessary to physically stabilize emulsions., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Phenolipids are a class of phenolic compounds with a lipidic moiety that have been receiving increasing attention due to their promising biological activities; however data regarding their toxicity and mechanism of action are scarce. A series of 11 phenolipids consisting of alkyl esters derivatives of the natural molecule protocatechuic acid was synthesized and evaluated against a panel of cancer and non-cancer cell lines. The macrophage cell line RAW 264.7, widely used as a tool for screening anti-inflammatory drugs, was more susceptible to the toxicity of these molecules than human cancer cells, reason for which mechanist studies were conducted. The parent molecule was not toxic up to 100μM, however structural modifications by inclusion of carbon side chains resulted in increased toxicity, compounds bearing 8-14 carbons being the most toxic and displaying IC 50 in the nanomolar range. Mechanistic studies showed that phenolipids elicit chromatin condensation, loss of cell viability and disruption of mitochondrial membrane potential (ΔΨm), increased reactive oxygen species (ROS) and activation of caspase-9/3, thus pointing to the involvement of mitochondria in the programmed cell death process taking place. This is the first study addressing the toxicity and mechanism of action of protocatechuic acid derivatives, which is relevant in light of the recent interest in these molecules., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Marine invertebrates have been attracting the attention of researchers for their application in nutrition, agriculture, and the pharmaceutical industry, among others. Concerning sea anemones (Cnidaria), little is known regarding their metabolic profiles and potential value as a source of pharmacologically-active agents. In this work, the chemical profiles of two species of sea anemones Actinia equina and Anemonia sulcata , were studied by high-performance liquid chromatography with diode-array detection (HPLC-DAD) and its impact upon immune and gastric cells was evaluated. In both species, the methylpyridinium alkaloid homarine was the major compound in aqueous extracts. The extracts were effective in reducing lipopolysaccharide (LPS)-induced levels of nitric oxide (NO) and intracellular reactive oxygen species (ROS) in a macrophage model of inflammation. Both the extracts and the alkaloid homarine were effective in inhibiting phospholipase A₂ (PLA₂), a pivotal enzyme in the initial steps of the inflammatory cascade. In order to mimic the oral consumption of these extracts; their effect upon human gastric cells was evaluated. While no caspase-9 activation was detected, the fact that the endoplasmic reticulum-resident caspase-4, and also caspase-3, were activated points to a non-classical mechanism of apoptosis in human gastric cells. This work provides new insights on the toxicity and biological potential of sea anemones increasingly present in human nutrition.

Background: The relationships between the hydrophilic-lipophilic balance (HLB) of antioxidants (AOs) and their distributions and efficiencies in emulsions are not fully understood. Recent reports indicate that, for series of homologous antioxidants of different hydrophobicity, the variation of their efficiency with the HLB of the AO increases with the alkyl chain length up to a maximum (C 3 -C 8 ester) followed by a decrease (cut-off effect)., Results: We determined the distributions of a series of caffeic acid derivatives in intact soybean emulsions by employing a specifically designed chemical probe located in the interfacial region of the emulsion. We also determined the AO efficiencies in the very same emulsions. We demonstrate that the variation of the percentage of AO in the interfacial region of soybean oil-in-water emulsions with the AO HLB parallels that of their antioxidant efficiency., Conclusion: The results provide physical evidence that the variations in the efficiency of homologous series of antioxidants in emulsions are the result of differences in their distribution. The results confirm that, with other things being equal, there is a direct relationship between the percentage of AO in the interfacial region of the emulsions and their efficiency, providing a natural explanation, based on molecular properties, of the cut-off effect. © 2016 Society of Chemical Industry., (© 2016 Society of Chemical Industry.)

We determined the interfacial molarities of the antioxidants, AOs, hydroxytyrosol (HT), and HT fatty acid esters with chain lengths of 1 to 16 carbons in intact olive oil/water/Tween 20 emulsions. The results were compared with chain length effects on the oxidative stability of the same emulsions, and a direct correlation was established. Both (AOI) molarities (varying 50-250 times greater than the stoichiometric 3.5 × 10(-3) M AO concentration) and antioxidant efficiencies show similar parabola-like dependences on AO chain length with a maximum at C8, consistent with the "cut-off" effect often observed at longer chain lengths. Results should aid in understanding the complex structure-reactivity relationships between AO efficiencies in emulsified systems and their hydrophobilic-hydrophobic balance.

The growing interest in the bioactivity of natural polyphenols and of their metabolites requires metabolites to be used in bioassays and as standards in research protocols. We report here on the synthesis of several hydroxytyrosol metabolite monosulfates achieved using a simplified protocol with improved yields. A synthetic solution based on avoidance of high temperature conditions during the synthesis and of low pressure conditions during purification has been established. Monosulfates of several phenolic compounds, namely, hydroxytyrosol, hydroxytyrosol acetate, homovanillyl alcohol, homovanillyl alcohol acetate, homovanillic acid, ferulic acid, and 3,4-dihydroxyphenylethanoic acid, were efficiently synthesized in 1-2 steps in good yield and isolated using simple procedures. The proposed protocol was shown to be relatively rapid, efficient, cheap, and widely applicable to a number of catechol scaffolds.

Red blood cells (RBCs), as anucleated cells, have poor repair and biosynthetic mechanisms, suffering and accumulating oxidative lesions whenever oxidative stress develops. RBCs are particularly exposed to endogenous oxidative damage because of their specific role as oxygen carriers. However, as the most abundant blood cells, RBCs also play an important role in the oxidative status of the whole blood constituents. In previous studies by our group, the most important polyphenolic compounds found in virgin olive oil, 3,4-dihydroxyphenylethanol-elenolic acid (3,4-DHPEA-EA) and 3,4-dihydroxyphenylethanol-elenolic acid dialdehyde (3,4-DHPEA-EDA), were shown to significantly protect RBCs from oxidative damage initiated by AAPH and H2O2, with the most active compound being 3,4-DHPEA-EDA. However, the in vivo protective effects of these phenols are dependent on their bioavailability. It has been demonstrated that 3,4-DHPEA-EDA is absorbed by intestinal cells and is then metabolized, yielding a reduced metabolite, 3,4-DHPEA-EDAH2. In order to assess the importance of VOO phenolic compound metabolites for the overall in vivo protective activity, the capacity of this phase I metabolite to protect RBCs in the presence of the radical initiators AAPH or H2O2 was evaluated in the presence and absence of the naturally occurring antioxidant, ascorbic acid. The metabolite was shown to protect RBCs from haemolysis induced by both initiators, in a dose dependent way, after 2 h and 4 h of incubation. The protective effect was however lower than that of the parental compound. The analysis of the membrane proteins of erythrocytes showed that the metabolite can interact with these biological structures.

Recently published results for a series of homologous antioxidants, AOs, of increasing alkyl chain length show a maximum in AO efficiency followed by a significant decrease for the more hydrophobic AOs, typically called the "cut-off" effect. Here we demonstrate that in olive oil emulsions both antioxidant efficiencies and partition constants for distributions of AOs between the oil and interfacial regions, PO(I), show a maximum at the C8 ester. A reaction between caffeic acid, CA, and its specially synthesised C1-C16 alkyl esters, and a chemical probe is used to estimate partition constants for AO distributions and interfacial rate constants, kI, in intact emulsions based on the pseudophase kinetic model. The model provides a natural interpretation for both the maximum and the "cut-off" effect. More than 70% of the CA esters are in the interfacial region even at low surfactant volume fraction, ΦI=0.005., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Background: In a preliminary study the oxidative stability and tocopherol content of pork meat were shown to be improved by olive leaf (OL) feed supplementation at 50 and 100 g kg(-1) . However, growth performance was affected negatively. Therefore the objective of the present study was to assess the influence of OL supplementation at a lower level on feed digestibility, growth performance and meat quality., Results: Pigs were fed a basal diet (control), a basal diet with 25 g OL kg(-1) (OL2.5) or a basal diet with 50 g OL kg(-1) (OL5). The incorporation of OL significantly decreased growth rates (P = 0.010) and backfat thickness (P = 0.035) and increased feed/gain ratio (P = 0.032) in the OL5 group. Feed/gain ratio increased more for females (P = 0.001). The incorporation of OL decreased the crude fat (P = 0.006) and protein (P = 0.037) digestibility of both OL diets. Nevertheless, OL was effective in increasing the tocopherol content of meat (P = 0.009). However, meat from pigs fed the OL diets showed similar conjugated diene content, pH and colour parameters to that from pigs fed the control diet, even after 6 days of storage at 4 °C., Conclusion: The data indicate that olive leaves may be included in pig diets at 25 g kg(-1) in order to improve the tocopherol content of meat without excessively compromising growth performance., (© 2014 Society of Chemical Industry.)

We investigated the effects of resveratrol (RES) on the oxidative stability of emulsions composed of stripped corn oil, acidic water and Tween 20 and determined its distribution in the intact emulsions by employing a well-established kinetic method. The distribution of RES is described by two partition constants, that between the oil-interfacial region, PO(I), and that between the aqueous and interfacial region, PW(I). The partition constants, PO(I) and PW(I), are obtained in the intact emulsions from the variations of the observed rate constant, kobs, for the reaction between the hydrophobic 4-hexadecylbenzenediazonium ion and RES with the emulsifier volume fraction, ФI. The obtained PO(I) and PW(I) values are quite high, PW(I) = 4374 and PO(I) = 930, indicating that RES is primarily located in the interfacial region of the emulsions, %RESI > 90% at ФI = 0.005, increasing up to 99% at ФI = 0.04. The oxidative stability of the corn oil emulsions was determined by measuring the formation of conjugated dienes at a given time in the absence and in the presence of RES. The addition of RES did not improve their oxidative stability in spite that more than 90% of RES is located in the interfacial region of the emulsion, because of the very low radical scavenging activity of RES.

The capacity of important hydroxytyrosol metabolites (homovanillyl alcohol, hydroxytyrosol acetate, homovanillyl alcohol acetate, hydroxytyrosol 3' and 4'-O-glucuronides, and homovanillyl alcohol 4'-O-glucuronide) to protect red blood cells (RBCs) from oxidative injury induced by the radical initiator 2,2'-azo-bis(2-amidinopropane) dihydrochloride (AAPH) or by the natural radical initiator H2O2 was evaluated. In the presence of AAPH, all compounds showed to protect RBCs from hemolysis in a dose-dependent manner, exccept for the homovanillyl alcohol glucuronide, with the order of activity being at 20 μM hydroxytyrosol > hydroxytyrosol glucuronides = hydroxytyrosol acetate = homovanillyl alcohol = homovanillyl acetate > homovanillyl alcohol glucuronide. At 10 μM, hydroxytyrosol, hydroxytyrosol acetate, and hydroxytyrosol glucuronides still protected hemoglobine from oxidation and from morphological RBC changes. In the presence of H2O2, hydroxytyrosol showed to significantly protect RBCs from oxidative hemolysis in a dose-dependent manner, but the hydroxytyrosol glucuronides showed only a limited protection that was independent of the concentration used.

Antioxidant (AO) efficiencies are reported to go through maxima with increasing chain length (hydrophobicity) in emulsions. The so-called "cutoff" after the maxima, indicating a decrease in efficiency, remains unexplained. This paper shows, for gallic acid (GA) and propyl, octyl, and lauryl gallates (PG, OG, and LG, respectively), that at any given volume fraction of emulsifier, the concentrations of antioxidants in the interfacial region of stripped corn oil emulsions and their efficiency order follow PG > GA > OG > LG. These results provide clear evidence that an AO's efficiency correlates with its fraction in the interfacial region. AO distributions were obtained in intact emulsions by using the pseudophase kinetic model to interpret changes in observed rate constants of the AOs with a chemical probe, and their efficiencies were measured by employing the Schaal oven test. The model provides a natural explanation for the maxima with increasing AO hydrophobicity.

Background: The importance of olive polyphenols as bioactive compounds has grown in recent years as a result of intensive research on their anticancer, antiatherosclerotic and antihypertensive activities. However, there is currently no official method for determining the content of polyphenols in olive oils because of the technical difficulties in their determination. Here a simple method for the analysis of extra virgin olive oil o-diphenols by visible spectrometry is proposed and compared with the traditional method of solid phase extraction followed by colorimetric determination using sodium molybdate or Folin-Ciocalteu reagent or by high-performance liquid chromatography (HPLC) analysis using UV detection. This new approach to determining total o-diphenolic compounds exploits the oxidation of o-diphenols to quinones in a basic medium., Results: Preliminary results showed a better correlation between the total o-diphenol determination by HPLC and by the proposed method (R(2) = 0.9229) than between the total o-diphenol determination by HPLC and by the molybdate colorimetric method (R(2) = 0.8689). A good correlation was also observed between the total phenolic content determined by HPLC and by the proposed method (R(2) = 0.8196), but this correlation was a little lower than the one obtained between the HPLC method and the Folin-Ciocalteu method (R(2) = 0.8752)., Conclusion: The proposed method involves very little sample manipulation, requires inexpensive reagents and can be performed in less than 40 min for several samples at the same time, using olive oil samples of only 1-2 g., (© 2012 Society of Chemical Industry.)

We employed a kinetic method to determine the distributions of the antioxidants hydroxytyrosol (HT) and hydroxytyrosol acetate (HTA) between the oil, aqueous, and interfacial regions of a model food emulsion composed of stripped olive oil, acidic water, and a blend of Tween 80 and Span 80 [hydrophilic–lipophilic balance (HLB) = 8.05] as an emulsifier. HT is oil-insoluble, but HTA is both oil- and water-soluble (partition constant P(O)(W) = 0.61). Results indicate that, at a given emulsifier volume fraction Φ(I), the fraction of HTA in the interfacial region is higher than that of HT. The percentage of both antioxidants increases with an increasing Φ(I), so that % HT > 40% at Φ(I) = 0.005 and % HT > 80% at Φ(I) = 0.04. HTA appears to be a better antioxidant than HT, as shown by an accelerated oxidative test (Schaal oven method). A correlation between their distribution in the emulsion and their efficiency was established.