Unexpected Antioxidant Efficiency of Chlorogenic Acid Phenolipids in Fish Oil-in-Water Nanoemulsions: An Example of How Relatively Low Interfacial Concentrations Can Make Antioxidants to Be Inefficient.

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, ΦI, 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.

Effect of new antioxidants: phenolipids on quality of fried French fries and rapeseed oil.

The purpose of this work was to evaluate the effect of five novel synthetic antioxidants: octyl sinapate (OSA), octyl ferulate (OFA), octyl caffeate (OCA), cetyl sinapate (CSA) and cetyl ferulate (CFA) added to refined rapeseed oil for the first time on antioxidant activity (AA) and total phenolic content (TPC) in French fries. French fries after frying in enriched oils had higher the AA determined by three assays: 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS = 2907-20,029 µmol TE/100 g), 2,2-diphenyl-1-picrylhydrazyl (DPPH = 657-6886 µmol TE/100 g), ferric reducing antioxidant power (FRAP = 332-2659 µmol TE/100 g) and total phenolic content (TPC = 44-378 mg SA/100 g) than those prepared in refined rapeseed oil (ABTS = 2146 µmol TE/100 g, DPPH = 403 µmol TE/100 g, FRAP = 218 µmol TE/100 g, TPC = 14 mg SA/100 g). Moreover, the presence of phenolipids in rapeseed oil decreased the oil content in fried French fries by 22-45%. However, the AA and TPC in the fortified oils significantly increased, whereas slower changes in oxidation parameters of supplemented oils after frying were observed. A new lipophilic antioxidants can be useful for the development of management system for the preparation of French fries and the prolongation of frying oil shelf life.

Phenolic compounds in germinated cereal and pulse seeds: Classification, transformation, and metabolic process.

Natural phenolic compounds are rich in cereal and pulse seeds and their dietary functions tend to improve dramatically during germination. This article reviews recent research on the transformation of phenolic compounds during seed germination. In particular, it highlights the classification of crude phenolic compounds that can be divided into extractable and non-extractable phenolic compounds based on the biosynthesis process and extraction method. It also recommends grouping resorcinol lipids in the category of extractable phenolic compounds as non-polar solvent extractable phenolic compounds. Moreover, it discusses the variation of the different form of phenolic compounds and proposes a possible metabolic model of these phenolic compounds for seeds germination. This article is crucial for phenolic compounds research, cereal and pulse seeds germination, and food ingredients industry.

Antioxidant activity of hydroxytyrosyl esters studied in liposome models.

The properties and the antioxidant activity of a series of hydroxytyrosyl esters having different carbon chain lengths (C4, C8, C12 and C18) have been measured in phosphatidylcholine model membrane (liposomes) using specific probes for the bilayer and liposome lumen microenvironment, i.e., 1,6-diphenyl-1,3,5-hexatriene (DPH) and 2',7'-dichlorodihydrofluorescein (H2DCF), respectively. Antioxidants self-assembly and their interaction with liposomes has been evaluated by light scattering, fluorescence, turbidimetry, gel filtration chromatography and microfiltration measurements, allowing the determination of critical aggregation concentration, bound fraction, capacity of crossing the lipid bilayer. The distribution of hydroxytyrosyl long chain esters has been proved to depend quite specifically on their lipophilic chain length, and this turns to have deep effects on their antioxidant behaviour. Shedding new light on the cut off effect and antioxidant behaviour of phenolipids, this study also put forward the relevance of cell-free liposome-based cellular models, like giant liposomes, for further characterization of analogous systems.

Synthesis of novel octyl sinapate to enhance antioxidant capacity of rapeseed-linseed oil mixture.

BACKGROUND: Lipophilisation allows the formation of new functionalised antioxidants having beneficial properties compared to natural hydrophilic phenolic acids. Therefore, this work focused on the synthesis of lipophilic antioxidants, such as a new octyl sinapate, octyl caffeate and octyl ferulate using the modified Fischer esterification of selected hydroxycinnamic acids with 1-octanol. RESULTS: The lipophilic octyl sinapate was obtained for the first time with satisfactory yield (83%) after purification by column chromatography. The identity of the synthesised phenolipids was confirmed by chromatographic and spectroscopic analyses. Antioxidant capacity of phenolipids was determined by DPPH (IC50 = 35.87-52.24 µg mL-1 ) and ABTS (IC50 = 39.45-48.72 µg mL-1 ) methods and compared with IC50 values (7.37-35.30 µg mL-1 and 7.55-41.67 µg mL-1 , respectively) for well known antioxidants. The antioxidant capacity of rapeseed-linseed oil enriched with the purified esters was about two to 30 times higher in comparison with a non-supplemented oil. CONCLUSION: The novel octyl sinapate as well as octyl caffeate and octyl ferulate have antioxidant properties and lipophilic character, therefore they may be added to vegetable oils as potential antioxidants for tackling oxidative processes. © 2017 Society of Chemical Industry.

Effect of Alkylresorcinols Isolated from Wheat Bran on the Oxidative Stability of Minced-Meat Models as Related to Storage.

Due to their antioxidant activity, alkylresorcinols (ARs) extracted from by-products could represent promising natural and innovative antioxidants for the food industry. This study tested the ability of ARs isolated from wheat bran to increase the shelf-life of minced-meat models stored at 4 °C for 9 days. Fifteen alk(en)ylresorcinols (C17-C25) were recognized by GC/MS, showing good radical-scavenging (200.70 ± 1.33 µmolTE/g extract) and metal-chelating (1.38 ± 0.30 mgEDTAE/g extract) activities. Two ARs concentrations (0.01% and 0.02%) were compared to sodium ascorbate (0.01% and 0.10%) on color (CIELAB values L*, a*, b*, chroma, and hue) and oxidative stability (lipid hydroperoxides, thiobarbituric acid reactive substances (TBARS), and volatile organic compounds (VOCs)) of minced-beef samples. ARs-treated samples were oxidatively more stable than those formulated with sodium ascorbate and the negative control, with significantly lower contents of hydroperoxides and VOCs (hexanal, 1-hexanol, and 1-octen-3-ol) throughout the experiment (p < 0.001). However, no effect on color stability was observed (p > 0.05). Since 0.01% of ARs was equally or more effective than 0.10% sodium ascorbate, those results carry important implications for the food industry, which could reduce antioxidant amounts by ten times and replace synthetic antioxidants with natural ones.

Ascorbic acid potentiates photodynamic inactivation mediated by octyl gallate and blue light for rapid eradication of planktonic bacteria and biofilms.

This study reported for the first time that Ascorbic acid (AA) could appreciably boost the efficiency of Octyl gallate (OG)-mediated photodynamic inactivation (PDI) on Escherichia coli and Staphylococcus aureus in planktonic and biofilm states. The combination of OG (0.075 mM) and AA (200 mM) with 420 nm blue light (212 mW/cm2) led to a >6 Log killing within only 5 min for E. coli and S. aureus and rapid eradication of biofilms. The mechanism of action appears to be the generation of highly toxic hydroxyl radicals (•OH) via photochemical pathways. OG was exposed to BL irradiation to generate various reactive oxygen radicals (ROS) and the addition of AA could transform singlet oxygen (1O2) into hydrogen peroxide (H2O2), which could further react with AA to generate enormous •OH. These ROS jeopardized bacteria and biofilms by nonspecifically attacking various biomacromolecules. Overall, this PDI strategy provides a powerful microbiological decontamination modality to guarantee safe food products.

New anti-α-Glucosidase and Antioxidant Ingredients from Winery Byproducts: Contribution of Alkyl Gallates.

Wine-making activity entails the production of solid and semisolid byproducts (grape stems and pomace and wine lees) that negatively impact the environment and industrial sustainability. Their features as sources of bioactive compounds support valorization procedures for functional and healthy ingredients. This work uncovers the quantitative alkyl gallates (gallic acid esters, C1-C12) profile of fresh (freeze-dried) materials and the effect of oven-drying on their stability by UHPLC-ESI-QqQ-MS/MS. The functionality was established concerning DPPH• scavenging and antihyperglycemic power. Wine lees exerted the highest high-free concentration of galloyl derivatives, ethyl gallate being the most abundant ester (3472.62 ng/g dw, on average). About the impact of the stabilization process, although as a general trend, the thermal treatment reduced the concentration, the reduction dimensions depended on the compound/matrix, remaining in valuable concentrations. Concerning radical scavenging, ze-dried stems and pomace displayed the highest capacity (24.11 and 18.46 mg TE/g dw, respectively), being correlated with propyl gallate (r2 = 0.690), butyl gallate (r2 = 0.686), and octyl gallate (r2 = 0.514). These two matrices exerted α-glucosidase inhibitory activity (1.58 and 1.46 units/L) equivalent to that of acarbose (a recognized α-glucosidase inhibitor). The newly described bioactive phytochemicals in winery residues (galloyl esters) and their correlation with functional traits allow for envisioning valorization alternatives.

Interfacial effects of gallate alkyl esters on physical and oxidative stability of high fat fish oil-in-water emulsions stabilized with sodium caseinate and OSA-modified starch.

Effects of sodium caseinate (SC) and its combination with OSA-modified starch (SC-OS; 1:1) alone and with n-alkyl gallates (C0-C18) on the physical and oxidative stability of high-fat fish oil-in-water emulsion were evaluated. SC emulsion contained the smallest droplets and highest viscosity due to the fast adsorption at droplet surfaces. Both emulsions had non-Newtonian and shear-thinning behavior. A lower accumulation of lipid hydroperoxides and volatile compounds was found in SC emulsion due to its better Fe2+ chelating activity. The incorporated short-chain gallates (G1 > G0 âˆ¼ G3) in SC emulsion had a strong synergistic effect against lipid oxidation compared to that of SC-OS emulsion. The better antioxidant efficiency of G1 can be related to its higher partition at the oil-water interface, while G0 and G3 had a higher partition into the aqueous phase. In contrast, G8, G12, and G16 added emulsions indicated higher lipid oxidation due to their internalization inside the oil droplets.

Targeting Interfacial Location of Phenolic Antioxidants in Emulsions: Strategies and Benefits.

It is important to have larger proportions of health-beneficial polyunsaturated lipids in foods, but these nutrients are particularly sensitive to oxidation, and dedicated strategies must be developed to prevent this deleterious reaction. In food oil-in-water emulsions, the oil-water interface is a crucial area when it comes to the initiation of lipid oxidation. Unfortunately, most available natural antioxidants, such as phenolic antioxidants, do not spontaneously position at this specific locus. Achieving such a strategic positioning has therefore been an active research area, and various routes have been proposed: lipophilizing phenolic acids to confer them with an amphiphilic character; functionalizing biopolymer emulsifiers through covalent or noncovalent interactions with phenolics; or loading Pickering particles with natural phenolic compounds to yield interfacial antioxidant reservoirs. We herein review the principles and efficiency of these approaches to counteract lipid oxidation in emulsions as well as their advantages and limitations.

Enzymatic Modification of Pomace Olive Oil with Natural Antioxidants: Effect on Oxidative Stability.

Enzymatic lipophilization has been proposed as a cost-effective strategy to produce new liposoluble antioxidant compounds. In this study, modified oils rich in structured phenolipids were prepared via one-pot enzymatic acylation of hydroxytyrosol (HTYR), vanillyl alcohol (VA) and homovanillyl alcohol (HVA) with pomace olive oil (POO) in solvent-free conditions using immobilized lipase on biogenic nanoparticles. The effect of temperature (30-70 °C) and enzyme concentration (0.1-1%, w/w) on the efficiency of the bioprocess as well as the reusability of the nanobiocatalyst were thoroughly investigated. The modified oils exhibited increased antioxidant activity compared to the control oil according to DPPH and CUPRAC assays (p < 0.05). The oxidative stability of pomace olive oil was also significantly enhanced after modification, as depicted by the K232 values and TBARS contents under accelerated oxidation at 60 °C (p < 0.05). Moreover, a fortified mayonnaise containing modified oil with HTYR was prepared that was noticeably stable compared to the control mayonnaise at 28 °C for 5 months (p < 0.05). Enzymatically modified oils have great potential for application in the nutraceutical and food industry, encouraging the exploitation of immobilized lipases as effective and green catalytic tools.

Phenolipids, Amphipilic Phenolic Antioxidants with Modified Properties and Their Spectrum of Applications in Development: A Review.

Polyphenols, as secondary metabolites from plants, possess a natural antioxidant capacity and biological activities attributed to their chemical and structural characteristics. Due to their mostly polar character, polyphenols present a low solubility in less polar environments or hydrophobic matrices. However, in order to make polyphenols able to incorporate in oils and fats, a transformation strategy is necessary. For the above, the functionalization of polyphenols through chemical or enzymatic lipophilization has allowed the synthesis of phenolipids. These are amphipilic molecules that preserve the natural phenolic core to which an aliphatic motif is attached by esterification or transesterification reactions. The length of the aliphatic chain in phenolipids allows them to interact with different systems (such as emulsions, oily molecules, micelles and cellular membranes), which would favor their use in processed foods, as vehicles for drugs, antimicrobial agents, antioxidants in the cosmetic industry and even in the treatment of degenerative diseases related to oxidative stress.

Toxicity of phenolipids: Protocatechuic acid alkyl esters trigger disruption of mitochondrial membrane potential and caspase activation in macrophages.

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 IC50 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.

Synthesis and in vitro antioxidant and antimicrobial studies of novel structured phosphatidylcholines with phenolic acids.

Novel phenoylated phosphatidylcholines were synthesized from 1,2-dipalmitoyl phosphatidylcholine/egg 1,2-diacyl phosphatidylcholine and phenolic acids such as ferulic, sinapic, vanillic and syringic acids. The structures of phenoylated phosphatidylcholines were confirmed by spectral analysis. 2-acyl-1-lyso phosphatidylcholine was synthesized from phosphatidylcholine via regioselective enzymatic hydrolysis and was reacted with hydroxyl protected phenolic acids to produce corresponding phenoylated phosphatidylcholines in 48-56% yields. Deprotection of protected phenoylated phosphatidylcholines resulted in phenoylated phosphatidylcholines in 87-94% yields. The prepared compounds were evaluated for their preliminary in vitro antimicrobial and antioxidant activities. Among the active derivatives, compound 1-(4-hydroxy-3,5-dimethoxy) cinnamoyl-2-acyl-sn-glycero-3-phosphocholine exhibited excellent antioxidant activity with EC50 value of 16.43µg/mL. Compounds 1-(4-hydroxy-3-methoxy) cinnamoyl-2-acyl-sn-glycero-3-phosphocholine and 1-(4-hydroxy-3,5-dimethoxy) cinnamoyl-2-palmitoyl-sn-glycero-3-phosphocholine exhibited good antioxidant activity with EC50 values of 36.05 and 33.35µg/mL respectively. Compound 1-(4-hydroxy-3-methoxy) cinnamoyl-2-palmitoyl-sn-glycero-3-phosphocholine exhibited good antibacterial activity against Klebsiella planticola with MIC of 15.6µg/mL and compound 1-(4-hydroxy-3-methoxy) benzoyl-2-acyl-sn-glycero-3-phosphocholine exhibited good antifungal activity against Candida albicans with MIC of 15.6µg/mL.

Effects of Different Lipophilized Ferulate Esters in Fish Oil-Enriched Milk: Partitioning, Interaction, Protein, and Lipid Oxidation.

Antioxidant effects of ferulic acid and lipophilized ferulate esters were investigated in fish oil-enriched milk. Methyl ferulate (C1) and ethyl ferulate (C2) more efficiently prevented lipid oxidation than dodecyl ferulate (C12) did, followed by ferulic acid (C0). The combination of C1 or C2 with C12 could have a "synergistic" effect indicated by peroxide value, hexanal, and 1-penten-3-ol analysis results. These antioxidants also showed protein oxidation inhibition effects. The most effective antioxidants (C1 and C2) had the highest concentration in the precipitate phase but the lowest concentration in the aqueous phase, which was the opposite of the partitioning of C0. C12 had the highest concentration in the oil and emulsion phase. In particular, the interaction between ferulates esterified with short and medium alkyl chain lengths could lead to their "synergistic" effects in fish oil-enriched milk, which could be caused by the change in their partitioning or localization at the interface.

Antioxidative effect of lipophilized caffeic acid in fish oil enriched mayonnaise and milk.

The antioxidative effect of lipophilized caffeic acid was assessed in two different fish oil enriched food products: mayonnaise and milk. In both emulsion systems, caffeic acid esterified with fatty alcohols of different chain lengths (C1-C20) were better antioxidants than the original phenolic compound. The optimal chain length with respect to protection against oxidation was, however, different for the two food systems. Fish oil enriched mayonnaise with caffeates of medium alkyl chain length (butyl, octyl and dodecyl) added resulted in a better oxidative stability than caffeates with shorter (methyl) or longer (octadecyl) alkyl chains. Whereas in fish oil enriched milk emulsions the most effective caffeates were those with shorter alkyl chains (methyl and butyl) rather than the ones with medium and long chains (octyl, dodecyl, hexadecyl and eicosyl). These results demonstrate that there might be an optimum alkyl chain length for each phenolipid in each type of emulsion systems.

Phosphatidyl derivative of hydroxytyrosol. In vitro intestinal digestion, bioaccessibility, and its effect on antioxidant activity.

Intestinal digestion of phosphatidyl derivatives of HT (PHT) and its bioaccessibility under in vitro conditions was performed. First, an in vitro intestinal digestion model for phospholipids was developed. The impact of digestion in the antioxidant ability of PHT was also assayed. PHT was progressively hydrolyzed to lyso-PHT. However, digestion was slower than the phospholipid control. Nevertheless, most hydrolysis products were found at the micellar phase fraction, meaning a high bioaccessibility. Either PHT or digested PHT showed lower antioxidant activity than HT. However, PHT improved its antioxidant ability after digestion, likely related to lyso-PHT. As a summary, the synthetic phosphatidyl derivative of HT as PHT is recognized by phospholipases during simulation of intestinal digestion, although less efficiently than analogous phospholipids. Nevertheless, taking into account the bioaccessibility and the antioxidant activity of digested PHT, the potential of carriers of HT under the form of phospholipids might be of interest.

Antioxidant properties and efficacies of synthesized alkyl caffeates, ferulates, and coumarates.

Caffeic, ferulic, and coumaric acids were lipophilized with saturated fatty alcohols (C1-C20). The antioxidant properties of these hydroxycinnamic acids and their alkyl esters were evaluated in various assays. Furthermore, the antioxidant efficiency of the compounds was evaluated in a simple o/w microemulsion using the conjugated autoxidizable triene (CAT) assay. All evaluated phenolipids had radical scavenging, reducing power, and metal chelating properties. Only caffeic acid and caffeates were able to form a complex with iron via their catechol group in the phenolic ring. In the o/w emulsion, the medium chain phenolipids of the three homologues series were most efficient. The antioxidant properties and efficacies were dependent upon functional groups substituted to the ring structure and were in the following order: caffeic acid and caffeates > ferulic acid and ferulates > coumaric acid and coumarates. Moreover, the results demonstrated that the test system has an impact on the antioxidative properties measured.