Comparison of antioxidant potency of commonly consumed polyphenol-rich beverages in the United States

A number of different beverage products claim to have antioxidant potency due to their perceived high content of polyphenols. Basic and applied research indicates that pomegranate juice (PJ), produced from the Wonderful variety of Punica granatum fruits, has strong antioxidant activity and related health benefits. Although consumers are familiar with the concept of free radicals and antioxidants, they are often misled by claims of superior antioxidant activity of different beverages, which are usually based only on testing of a limited spectrum of antioxidant activities. There is no available direct comparison of PJ's antioxidant activity to those of other widely available polyphenol-rich beverage products using a comprehensive variety of antioxidant tests. The present study applied (1) four tests of antioxidant potency [Trolox equivalent antioxidant capacity (TEAC), total oxygen radical absorbance capacity (ORAC), free radical scavenging capacity by 2,2-diphenyl-1-picrylhydrazyl (DPPH), and ferric reducing antioxidant power (FRAP)]; (2) a test of antioxidant functionality, that is, inhibition of low-density lipoprotein (LDL) oxidation by peroxides and malondialdehyde methods; and (3) evaluation of the total polyphenol content [by gallic acid equivalents (GAEs)] of polyphenol-rich beverages in the marketplace. The beverages included several different brands as follows: apple juice (3), a?aí juice (3), black cherry juice (3), blueberry juice (3), cranberry juice (3), Concord grape juice (3), orange juice (3), red wines (3), iced tea beverages (10) [black tea (3), green tea (4), white tea (3)], and a major PJ available in the U.S. market. An overall antioxidant potency composite index was calculated by assigning each test equal weight. PJ had the greatest antioxidant potency composite index among the beverages tested and was at least 20% greater than any of the other beverages tested. Antioxidant potency, ability to inhibit LDL oxidation, and total polyphenol content were consistent in classifying the antioxidant capacity of the polyphenol-rich beverages in the following order: PJ>red wine>Concord grape juice>blueberry juice>black cherry juice, a?aí juice, cranberry juice>orange juice, iced tea beverages, apple juice. Although in vitro antioxidant potency does not prove in vivo biological activity, there is also consistent clinical evidence of antioxidant potency for the most potent beverages including both PJ and red wine.     

Assessment of radical scavenging capacity and lipid peroxidation inhibiting capacity of antioxidant

The role of radical scavenging antioxidants against oxidative stress has received much attention, and the antioxidant capacity has been assessed by various methods. Among them, a method that measures the effect of antioxidant on decay of the probe is one of the most widely used methods. The present study was performed to compare the two methods to assess the antioxidant capacity, one to follow the decay of the probe and the other to measure lipid peroxidation products in human plasma. It was shown that the method following probe decay was suitable for assessment of radical scavenging capacity of antioxidant, but not for the capacity to inhibit lipid peroxidation in plasma. This is true whether a hydrophilic or lipophilic probe is used. Such different results arise from the fact that the efficacy of inhibition of lipid peroxidation by antioxidants depends on the fate of antioxidant-derived radical and interaction between antioxidants as well as the capacity of free radical scavenging. Thus, the capacity of antioxidants for inhibition of lipid peroxidation should be assessed from the effect on the extent of oxidation, not from the effect on probe decay.     

Antioxidant content and free radical scavenging ability of fresh red pummelo [Citrus grandis (L.) Osbeck] juice and freeze-dried products

The antioxidative phytochemicals in various fruits and vegetables are widely recognized for their role in scavenging free radicals, which are involved in the etiology of many chronic diseases. Colored fruits are especially considered a quality trait that correlates with their nutritional values and health benefits. The specific aim of this study was to investigate the antioxidants in the juice and freeze-dried flesh and peel of red pummelo and their ability to scavenge free radicals and compare them with those in white pummelo juice. The total phenolic content of red pummelo juice extracted by methanol (8.3 mg/mL) was found to be significantly higher than that of white pummelo juice (5.6 mg/mL). The carotenoid content of red pummelo juice was also significantly higher than that in white pummelo juice. The contents of vitamin C and delta-tocopherol in red pummelo juice were 472 and 0.35 mug/mL, respectively. The ability of the antioxidants found in red pummelo juice to scavenge radicals were found by methanol extraction to approximate that of BHA and vitamin C with a rapid rate in a kinetic model. The ability of methanol extracts of freeze-dried peel and flesh from red pummelo to scavenge these radicals was 20-40% that of BHA and vitamin C effects. Fresh red pummelo juice is an excellent source of antioxidant compounds and exhibited great efficiency in scavenging different forms of free radicals including DPPH, superoxide anion, and hydrogen peroxide radicals.     

The chemistry behind antioxidant capacity assays

This review summarizes the multifaceted aspects of antioxidants and the basic kinetic models of inhibited autoxidation and analyzes the chemical principles of antioxidant capacity assays. Depending upon the reactions involved, these assays can roughly be classified into two types: assays based on hydrogen atom transfer (HAT) reactions and assays based on electron transfer (ET). The majority of HAT-based assays apply a competitive reaction scheme, in which antioxidant and substrate compete for thermally generated peroxyl radicals through the decomposition of azo compounds. These assays include inhibition of induced low-density lipoprotein autoxidation, oxygen radical absorbance capacity (ORAC), total radical trapping antioxidant parameter (TRAP), and crocin bleaching assays. ET-based assays measure the capacity of an antioxidant in the reduction of an oxidant, which changes color when reduced. The degree of color change is correlated with the sample's antioxidant concentrations. ET-based assays include the total phenols assay by Folin-Ciocalteu reagent (FCR), Trolox equivalence antioxidant capacity (TEAC), ferric ion reducing antioxidant power (FRAP), "total antioxidant potential" assay using a Cu(II) complex as an oxidant, and DPPH. In addition, other assays intended to measure a sample's scavenging capacity of biologically relevant oxidants such as singlet oxygen, superoxide anion, peroxynitrite, and hydroxyl radical are also summarized. On the basis of this analysis, it is suggested that the total phenols assay by FCR be used to quantify an antioxidant's reducing capacity and the ORAC assay to quantify peroxyl radical scavenging capacity. To comprehensively study different aspects of antioxidants, validated and specific assays are needed in addition to these two commonly accepted assays.     

Antioxidant activity of knotwood extractives and phenolic compounds of selected tree species

The antioxidant potency and the radical scavenging capacity of superoxide and peroxyl radicals were assessed for 13 hydrophilic knotwood extracts of commercially important wood species, or fractions thereof, as well as for five pure wood-derived lignans and the flavonoid taxifolin. The chemical composition of the knotwood extracts was determined by gas chromatography combined with mass spectrometry. Most of the investigated wood species were rich in hydrophilic extractives (10-20% of the dry wood) with one or a few compounds dominating in each extract. All extracts had a high antioxidative potency and/or radical scavenging capacity as compared to the well-known antioxidants Trolox and butylated hydroxyanisole. The pure wood-derived lignans and taxifolin also had a high antioxidative potency and/or radical scavenging capacity. However, the antioxidant potency and/or radical scavenging capacity of several of the hydrophilic knotwood extracts were higher than that of the dominating compounds in pure form.     

Comparative evaluation of the antioxidant potential of infant cereals produced from purple wheat and red rice grains and LC-MS analysis of their anthocyanins

Cellular oxidative damage by endogenous and exogenous sources of free radicals and reactive oxygen species is a particular threat in infants. Antioxidant protection is normally achieved through a balance between pro-oxidants and endogenous and/or dietary antioxidants. Comprehensive research is required on optimization to achieve good antioxidant protection through infant foods, in particular, the commercially available infant cereals. This study therefore investigated the properties of whole purple wheat, unpolished red rice, and partially polished red rice before and after processing to produce infant cereals. Total phenolic content (TPC), total anthocyanin content (TAC), oxygen radical absorbance capacity (ORAC), individual anthocyanin components, and cellular antioxidant activity were measured. Home-made and laboratory-made pigmented infant cereals differed in that the latter required longer exposure to higher temperature and enzymatic hydrolysis. Home-made and laboratory-made unpolished red rice infant cereals showed higher total phenolic contents and peroxyl radical scavenging activity than home-made and laboratory-made purple wheat infant cereals; however, the latter had higher TAC. Pigmented infant cereals generally had higher TPC, TAC, and ORAC than the commercial ones (p < 0.05). Anthocyanins were identified in whole purple wheat, but they were not detected in unpolished red rice. C-Glycosyl apigenin was found in both whole purple wheat and unpolished red rice. Processing significantly decreased anthocyanin and C-glycosyl apigenin contents (p < 0.05). Purple wheat infant cereals had higher cellular antioxidant activity than unpolished red rice ones (p < 0.05). Whole purple wheat infant cereals showed higher antioxidant activity than the commercial infant cereal, suggesting a possibility of improving infant antioxidant status by incorporating this grain in their diet.     

Contribution of proanthocyanidins to the peroxy radical scavenging capacity of some Italian red wines

Highly reactive radicals, ROO(*), were generated from 2, 2'-azobis[2-(2-imidazolin-2-yl)propane] and linoleic acid. The ROO(*) scavenging capacity of some Italian red wines was evaluated following the changes in oxygen consumption. Under the experimental conditions the time course of oxygen consumption shows two typical behaviors: trolox-like (class I) and gallic acid-like (class II). Usually the time course of wine was similar to that of gallic acid. The rate of oxygen consumption was found to decrease exponentially with the amount of wine or gallic acid added to the test solution. On this basis the capacity of red wines to scavenge peroxy radicals was expressed as content of gallic acid (S(GA)). The S(GA) values were found to be correlated to the amount of total proanthocyanidins and total polyphenols of some Italian red wines (p < 0.01). The proanthocyanidins extracted from seeds were shown to make a major contribution to the peroxy radical scavenging capacity of red wines, whereas, interestingly, the chemical class of the low molecular weight tannins reactive to vanillin did not correlate with the S(GA) values.     

Antioxidant properties of pearled barley fractions

Two barley varieties (Falcon and AC Metcalfe) were separated by pearling into seven fractions and subsequently extracted with 80% methanol. The extracts, after solvent removal, were evaluated for their radical scavenging efficacy using Trolox equivalent antioxidant capacity (TEAC). The radical scavenging capacity of the extracts was determined using 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical, oxygen radical absorbance capacity (ORAC(FL)), and superoxide radical assays and a photoinduced chemiluminescence technique. In both barley varieties the outermost fraction (F1) yielded the highest phenolic content. In general, Falcon had a significantly higher total phenolic content than AC Metcalfe. A similar trend was observed for TEAC, DPPH, and superoxide radical scavenging capacities of the extracts. The contents of water-soluble antioxidants of Falcon and AC Metcalfe were 1.15-12.98 and 2.20-12.25 micromol of Trolox equiv/(g of defatted material), while the corresponding lipid-soluble counterparts varied from 1.44 to 4.70 micromol of alpha-tocopherol equiv/(g of defatted material). Phenolic acids, namely, vanillic, caffeic, p-coumaric, ferulic, and sinapic acids, were identified by HPLC in barley fractions.     

Analysis of Genotype,Environment, and Their Interaction Effects on the Phytochemicals and Antioxidant Capacities of Red Rice (Oryza sativa L.)

Fourteen red rice varieties were planted in two locations during summer (Hangzhou) and winter (Hainan) to study the effect of genotype and environment on the phytochemicals and antioxidant capacities of rice grain. B‐type proanthocyanidins in red rice were detected by LC‐MS/MS and quantified by using the vanillin assay. Analysis of variance showed that total phenolic content (TPC), total flavonoid content (TFC) and 2,2′‐azino‐bis‐(3‐ethylbenzothiazoline‐6‐sulfonic acid) (ABTS) radical scavenging capacity were mainly affected by environmental factors, which accounted for more than 60% of the total variance. However, total proanthocyanidin content (TPAC) and 1,1‐diphenyl‐2‐picrylhydrazyl (DPPH) radical scavenging capacity were equally affected by both genotype and environment. The genotype × environment effects were significant for all traits. The pairwise correlations among TPC, TFC, TPAC, ABTS, and DPPH were also significant (r > 0.900, P < 0.001). Principal component analysis identified the genotypes that had higher contents of antioxidants and more stability across environments. This study showed that indirect selection of a simple trait (i.e., TPC) is an effective way to select rice high in antioxidant capacity in breeding programs. This study also suggests that rice should be produced specifically in a certain environment for the end user to minimize the variation in the functional properties and maximize their contents.     

Comparison of Swiss red wheat grain and fractions for their antioxidant properties

Swiss red wheat grain, bran, aleurone, and micronized aleurone were examined and compared for their free radical scavenging properties against 2,2-diphenyl-1-picrylhydrazyl radical (DPPH*), radical cation ABTS*+ and peroxide radical anion O(2)*-, oxygen radical absorbance capacity (ORAC), chelating capacity, total phenolic content (TPC), and phenolic acid composition. The results showed that micronized aleurone, aleurone, bran, and grain may significantly differ in their antioxidant properties, TPC, and phenolic acid composition. Micronized aleurone had the greatest antioxidant activities, TPC, and concentrations of all identified phenolic acids, suggesting the potential of postharvesting treatment on antioxidant activities and availability of TPC and phenolic acids. Ferulic acid was the predominant phenolic acid in Swiss red wheat and accounted for approximately 57-77% of total phenolic acids on a weight basis. Ferulic acid concentration was well correlated with scavenging activities against radical cation and superoxide anion, TPC, and other phenolic acid concentrations, suggesting the potential use of ferulic acid as a marker of wheat antioxidants. In addition, 50% acetone and ethanol were compared for their effects on wheat ORAC values. The ORAC value of 50% acetone extracts was 3-20-fold greater than that of the ethanol extracts, indicating that 50% acetone may be a better solvent system for monitoring antioxidant properties of wheat. These data suggest the possibility to improve the antioxidant release from wheat-based food ingredients through postharvesting treatment or processing.     

In vitro activity of almond skin polyphenols for scavenging free radicals and inducing quinone reductase

Observational studies and clinical trials suggest nut intake, including almonds, is associated with an enhancement in antioxidant defense and a reduction in the risk of cancer and cardiovascular disease. Almond skins are rich in polyphenols (ASP) that may contribute to these putative benefits. To assess their potential mechanisms of action, we tested the in vitro effect of ASP extracted with methanol (M) or a gastrointestinal juice mimic (GI) alone or in combination with vitamins C (VC) or E (VE) (1-10 micromol/L) on scavenging free radicals and inducing quinone reductase (QR). Flavonoid profiles from ASP-M and -GI extracts were different from one another. ASP-GI was more potent in scavenging HOCl and ONOO (-) radicals than ASP-M. In contrast, ASP-M increased and ASP-GI decreased QR activity in Hepa1c1c7 cells. Adding VC or VE to ASP produced a combination- and dose-dependent action on radical scavenging and QR induction. In comparison to their independent actions, ASP-M plus VC were less potent in scavenging DPPH, HOCl, ONOO (-), and O 2 (-) (*). However, the interaction between ASP-GI plus VC promoted their radical scavenging activity. Combining ASP-M plus VC resulted in a synergistic interaction, inducing QR activity, but ASP-GI plus VC had an antagonistic effect. On the basis of their total phenolic content, the measures of total antioxidant activity of ASP-M and -GI were comparable. Thus, in vitro, ASP act as antioxidants and induce QR activity, but these actions are dependent upon their dose, method of extraction, and interaction with antioxidant vitamins.     

Novel fluorometric assay for hydroxyl radical scavenging capacity (HOSC) estimation

A novel fluorometric method was developed and validated for hydroxyl radical scavenging capacity (HOSC) estimation using fluorescein as the probe. A constant flux of pure hydroxyl radical is generated under physiological pH using a Fenton-like Fe3+/H2O2 reaction. The generation of pure hydroxyl radicals under the experimental conditions was evaluated and confirmed using electron spin resonance with DMPO spin-trapping measurements. The hydroxyl radical scavenging capacity of a selected antioxidant sample is quantified by measuring the area under the fluorescence decay curve with or without the presence of the antioxidant and expressed as Trolox equivalents per unit of the antioxidant. The assay may be performed using a plate reader with a fluorescence detector for high-throughput measurements. The assay was validated for linearity, precision, accuracy, reproducibility, and its correlation with a popular peroxyl radical scavenging capacity assay using selected pure antioxidant compounds and botanical extracts. This method may provide researchers in the food, nutrition, and medical fields an easy to use protocol to evaluate free radical scavenging capacity of pure antioxidants and natural extracts in vitro against the very reactive hydroxyl radical, which may be linked to numerous degenerative diseases and conditions.     

Red chicories as potent scavengers of highly reactive radicals: a study on their phenolic composition and peroxyl radical trapping capacity and efficiency

Eight varieties of Cichorium genus vegetables (five heavily red colored, one red spotted, and two fully green) were investigated for their phenolic content (by HPLC and UV-vis spectrophotometry) and for their antioxidant activity. In particular, the capacity (that is, the amount of trapped peroxyl radicals) and the efficiency (that is, the amount of antioxidant necessary to halve the steady-state concentration of peroxyl radicals) were measured. All of the studied chicories are characterized by the presence of a large amount of hydroxybenzoic and hydroxycinnamic acids, whereas the red color is due to cyanidin glycosides. The presence of these phenolics in red chicories confers to them an exceptionally high peroxyl radical scavenging activity in terms of both capacity and efficiency, particularly in their early stage of growth, and makes this popular and low-cost foods comparable or superior to many foods having well-known antioxidant properties such as red wine, blueberry, and tomato.     

Comparison of the concentrations of phenolic constituents in cane sugar manufacturing products with their antioxidant activities

Polyphenol content and free radical scavenging capacity of seven kinds of sugar manufacturing products (A sugars, clear juices, syrups, massecuite, and A, B, and C molasses) were studied. Seventy-two samples were collected at different stages of the process during two sugar harvests from a local sugar factory (Bois-Rouge, La Réunion). The total phenolic content of sugar products was determined according to the Folin-Ciocalteu assay. Polyphenols of sugar products were extracted with ethyl acetate and quantified by LC-UV-ESI-MS during all of the process. ABTS and DPPH assays were applied to aqueous solution of sugar products, which exhibited interesting free radical scavenging activity. Comparatively, ethyl acetate extracts exhibited higher antioxidant activity. Multivariate analyses (principal component analysis and canonical discriminant analysis) demonstrated a significant correlation between polyphenols and antioxidant activity. Moreover, it was observed that the sugar process results in an increase of the phenolic content and the free radical scavenging capacity of the different products. These products and especially molasses proved to be a rich source of natural antioxidants and may represent an interesting alternative to synthetic food antioxidants.     

Evaluation of the antioxidant capacity of limonin, nomilin, and limonin glucoside

The antioxidant capacity (AOC) of three representative citrus limonoids, limonin, nomilin, and limonin glucoside, was examined by the oxygen radical absorbance capacity (ORAC), Trolox equivalent antioxidant capacity (TEAC), beta-carotene-linoleic acid bleaching, and 1,1-diphenyl-2-picryl hydrazyl (DPPH) radical scavenging assays. Pure compounds and proper negative (cinnamic acid) and positive (2,6-di-tert-butyl-4-methylphenol (BHT) and ascorbic acid) controls were used to remove any ambiguity in interpreting results. In all cases, limonin and nomilin gave results equivalent to those of cinnamic acid, indicating that they do not possess any inherent AOC and should not be considered antioxidants. Similar results were observed for limonin glucoside, with the exception of an anomalous result obtained from the beta-carotene-linoleic acid bleaching assay. Limonin glucoside was deemed not to be an antioxidant on the basis of the three unequivocal assays.     

Comparative study of polyphenol scavenging activities assessed by different methods

The effect of procyanidin solutions on superoxide anion radicals was studied with an enzymatic method and their EC(50) values were determined. A comparative study of the results suggested that the free radical scavenger potential of these phenolic compounds closely depends on their chemical and stereochemical structures. Oligomeric procyanidins were isolated in different fractions from grapes and wines by low- and high-pressure liquid chromatography. These compounds were found to be efficient free radical scavengers even for the weak concentrations in wines. Their activity in grapes or wines was much stronger than that of other commercially available natural antioxidants (such as ascorbic acid and gallic acid). The effect of tannins isolated from grapes on different radicals was analyzed according to three distinct methods: an enzymatic method for superoxide anion radicals (O(2)(*)(-)), a chemical method for the stable 1,1-diphenyl-2-picrylhydrazyl radical (DPPH(*)), and an immunochemical method to study the scavenging activity of seed procyanidins on DNA lesions induced by the radical HO(*).     

Antioxidants and antioxidant activity of several pigmented rice brans

This study investigated the antioxidant content and activity of phenolic acids, anthocyanins, α-tocopherol and γ-oryzanol in pigmented rice (black and red rice) brans. After methanolic extraction, the DPPH free radical scavenging activity and antioxidant activity were measured. The pigmented rice bran extract had a greater reducing power than a normal rice bran extract from a long grain white rice. All bran extracts were highly effective in inhibiting linoleic acid peroxidation (60-85%). High-performance liquid chromatography (HPLC) analysis of antioxidants in rice bran found that γ-oryzanol (39-63%) and phenolic acids (33-43%) were the major antioxidants in all bran samples, and black rice bran also contained anthocyanins 18-26%. HPLC analysis of anthocyanins showed that pigmented bran was rich in cyanidin-3-glucoside (58-95%). Ferulic acid was the dominant phenolic acid in the rice bran samples. Black rice bran contained gallic, hydroxybenzoic, and protocatechuic acids in higher contents than red rice bran and normal rice bran. Furthermore, the addition of 5% black rice bran to wheat flour used for making bread produced a marked increase in the free radical scavenging and antioxidant activity compared to a control bread.     

Chemical compositions, antioxidant capacities, and antiproliferative activities of selected fruit seed flours

Seed flours from black raspberry, red raspberry, blueberry, cranberry, pinot noir grape, and chardonnay grape were examined for their total fat content, fatty acid composition, total phenolic content (TPC), total anthocyanin content (TAC), radical scavenging capacities against the peroxyl (ORAC) and stable DPPH radicals, chelating capacity against Fe(2+), and antiproliferative activities using the HT-29 colon cancer cell line. Significant levels of fat were detected in the fruit seed flours and their fatty acid profiles may differ from those of the respective seed oils. Cranberry seed flour had the highest level of alpha-linolenic acid (30.9 g/100 g fat) and the lowest ratio of n-6/n-3 fatty acids (1.2/1). The ORAC value of the chardonnay seed flour was 1076.4 Trolox equivalents mumol/g flour, and its TPC was 186.3 mg gallic acid equivalents/g flour. These values were 3-12 times higher than the other tested fruit seed flours. Furthermore, the ORAC value was significantly correlated to the TPC under the experimental conditions (P < 0.05). These fruit seed flours also differed in their TAC values and Fe(2+)-chelating capacities. In addition, black raspberry, cranberry, and chardonnay grape seed flour extracts were evaluated for their antiproliferative effects using HT-29 colon cancer cells. All three tested seed flour extracts significant inhibited HT-29 cell proliferation. The data from this study suggest the potential of developing the value-added use of these fruit seed flours as dietary sources of natural antioxidants and antiproliferative agents for optimal human health.     

Electron spin resonance estimation of hydroxyl radical scavenging capacity for lipophilic antioxidants

Hydroxyl radical scavenging capacity estimation for lipophilic antioxidants is a challenge due to their poor solubility in aqueous radical generating and measuring systems. In this study, an electron spin resonance (ESR) method was developed and validated for its application in estimating the relative hydroxyl radical (HO*) scavenging capacity for lipophilic antioxidants under physiological pH using a Fenton Fe2+/H2O2 system for radical generation and acetonitrile as a solvent. The Fenton Fe2+/H2O2 system generates a constant flux of pure HO* under the assay conditions. The method was validated by linearity, precision, and reproducibility using selected known lipophilic antioxidants including alpha-tocopherol, lutein, beta-carotene, and BHT. The potential effects of commonly used water-miscible and water-immiscible organic solvents on the Fenton Fe2+/H2O2 HO* generating system as well as their possible interactions with the fluorescent and spectroscopic probes were also reported. In addition, the limitation of the ESR assay was described.     

Antioxidant activity of hydroxytyrosol acetate compared with that of other olive oil polyphenols

Hydroxytyrosol acetate was synthesized, and the antioxidant activity of this olive oil component was assessed in comparison with that of other olive oil components, namely hydroxytyrosol, oleuropein, 3,4-DHPEA-EA, and alpha-tocopherol in bulk oil and oil-in-water emulsions. The activity of the compounds was also assessed by scavenging of 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals. Hydroxytyrosol acetate had a weaker DPPH radical scavenging activity than hydroxytyrosol, oleuropein, or 3,4-DHPEA-EA but it had a radical scavenging activity similar to that of alpha-tocopherol. In oil, the antioxidant activity of hydroxytyrosol acetate was much higher than that of alpha-tocopherol or oleuropein, but in an emulsion 3,4-DHPEA-EA and alpha-tocopherol were more effective as antioxidants than hydroxytyrosol acetate. The antioxidant activity of hydroxytyrosol acetate was rather similar to that of hydroxytyrosol in oil and emulsions despite the difference in DPPH radical scavenging activity.