Free radicals and antioxidants in food and in vivo: What they do and how they work

A wide variety of oxygen free radicals and other reactive oxygen species can be formed in the human body and in food systems. Transition metal ions accelerate free‐radical damage. Antioxidant defenses, both enzymic and nonenzymic, protect the body against oxidative damage, but they are not 100% efficient, and so free‐radical damage must be constantly repaired. Nonenzymatic antioxidants are frequently added to foods to prevent lipid peroxidation. Several lipid antioxidants can exert prooxidant effects toward other molecules under certain circumstances, and so antioxidants for food and therapeutic use must be characterized carefully. Methods of measuring oxidative damage and trapping free radicals in vivo are briefly discussed. Such methods are essential in checking proposals that increased intake of food‐derived antioxidants (such as antioxidant vitamins) would be beneficial to humans.     

黄酮类化合物保护低密度脂蛋白氧化作用和减少动脉粥样硬化作用

食用含黄酮类物质食物将导致它们在血浆和组织中出现,摄入量和心血管的疾病成反比关系可能是与黄酮类减少了低密度脂蛋白氧化作用。黄酮类对动脉低密度脂蛋白细胞间接氧化作用可通过对他们在脂蛋白动脉的细胞累积来测定,如巨噬细胞。黄酮类可清除反应的氧、氮种类,螯合金属离子和节制低密度脂蛋白联合抗氧化作用,减少低密度脂蛋白氧化作用。通过抑制细胞氧合酶(如烟酰胺、腺嘌呤、二核甙酸、磷酸盐,减少氧化酶的形成)或通过启动细胞抗氧化剂(如谷胱甘肽系统)也能减少巨噬细胞氧化压力。 因此,黄酮类植物是有效的天然的抗氧化剂,既能保护反抗动脉细胞和脂蛋白类脂过氧化作用,又能有效的减少动脉粥样硬化的形成。     

Effects of Oolong Tea Supplementation on Lipid Peroxidation of Athletes at Rest and Post-exhaustive Exercise

ABSTRACT: Oolong tea contains polyphenolic catechins that can act as antioxidants and improve blood lipid status. The purpose of this study was to investigate the effect of a daily intake of Oolong tea for 30 d on the cholesterol profiles, lipid peroxidation level, and superoxide dismutase activity in athletes before and after exhaustive exercise. Twenty-two male rugby players served as the experiment subjects. They were divided into 2 groups: the tea-supplement group and control group. The results showed that there was a significant increase in total plasma cholesterol for both groups after training. There were no significant differences in the high-density lipoprotein (HDL) cholesterol level and low-density lipoprotein (LDL) cholesterol level after tea supplementation. However, ingestion of Oolong tea resulted in significantly lower resting and post-exhaustive exercise level of plasma malondialdehyde, and significantly lower resting level of superoxide dismutase activity. The results suggest that supplementation with Oolong tea for rugby players can decrease oxidative stress, and this can be explained by the decrease of lipid peroxidation level in cooperation with Oolong tea supplementation against the free radicals.     

鲜切果蔬活性氧产生和抗氧化体系代谢的研究进展

新鲜果蔬经切割后,诱导组织产生O2―?、H2O2和?OH等,均可破坏植物组织中正常的活性氧代谢平衡。但是,鲜切处理同时诱导果蔬启动抗氧化系统而主动防御。本文就鲜切果蔬O2―?、H2O2等的产生,超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)等抗氧化关键酶活性以及抗坏血酸、多酚化合物、还原型谷胱甘肽(GSH)等抗氧化物质的研究进展进行综述,以期为鲜切果蔬针对机械伤害的响应机制方面的研究提供一定的参考。     

Alleviation effect of heat-treated and in vitro gastrointestinal digested soymilks on AAPH-induced oxidative stress in human erythrocytes: Digested soymilks alleviate oxidative stress

To investigate the potential protective effect of raw and heat-treated soymilks after gastrointestinal digestion against chemical oxidative stress induced by 2,2′-azobis(2-amidinopropane) dihydrochloride (AAPH) on human erythrocytes, soymilk was subjected to heat treatment and in vitro gastrointestinal digestion. The inhibition rate of hemolysis, generation of reactive oxygen species (ROS), concentration of malondialdehyde (MDA), reduced glutathione (GSH) and oxidized glutathione (GSSH) and the enzyme activity of total superoxide dismutase (SOD) and cellular glutathione peroxidase (GPx) were evaluated as the biomarkers of oxidative status. Hemolysis of erythrocytes induced by AAPH was significantly inhibited by pretreatment with the digested raw soymilk (DRS) and digested heat-treated soymilk (DHS). Moreover, heat treatment prior to gastrointestinal digestion improved the inhibition effect of soymilk on erythrocytes hemolysis. The soymilk treated at 95 °C showed the highest inhibition rate, followed by 121 °C and 143 °C, revealed that the increase of temperature caused the decrease of hemolysis inhibition rate of DHS. Preincubation with the digested soymilks reduced the accumulation of MDA in erythrocytes, indicating the inhibition effect of the digested soymilks on lipid peroxidation. Results revealed that DRS and DHS alleviated the hemolysis of erythrocytes and lipid peroxidation resulted from oxidative stress by suppressing the accumulation of ROS, reducing the increase of SOD activity and decrease of non-enzymatic antioxidant GSH and enzymatic antioxidant GPx activity. Compared with raw soymilk, heat treatment improved the protective effect of the digested soymilk on erythrocytes against oxidative stress via enhancing the free radicals scavenging activity instead of improving the inhibition effect on the generation of free radicals.     

Effect of polyphenolic compounds from Coriandrum sativum on H2O2-induced oxidative stress in human lymphocytes

Polyphenolic compounds are widely distributed in plants and known to be excellent antioxidants in vitro. They have the capacity to reduce free-radical formation by scavenging free radicals and protecting antioxidant defences. The present study evaluated the antioxidant potencies of polyphenolic compounds from a spice, Coriandrum sativum against hydrogen peroxide-induced oxidative damage in human lymphocytes. Pretreatment with polyphenolic rich fractions protected human lymphocytes against H₂O₂-induced oxidative damage. H₂O₂ treatment significantly decreased the activities of antioxidant enzymes, such as superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase, glutathione-S-transferase and caused decreased glutathione content and increased thiobarbituric acid-reacting substances (TBARS). Treatment with polyphenolic fractions (50 μg/ml) increased the activities of antioxidant enzymes and glutathione content and reduced the levels of TBARS significantly. Observed reduction in the level of lipid peroxides showed a decreased tendency of peroxidative damage. We conclude that, under these experimental conditions, polyphenolic compounds effectively suppress hydrogen peroxide-induced oxidative stress.     

The antioxidant effect of a diet rich in Maillard reaction products is attenuated after consumption by healthy male adolescents. In vitro and in vivo comparative study

BACKGROUND: Maillard reaction products (MRPs) are widely consumed as a part of the human diet. A 2 week randomised two‐period crossover trial to determine whether MRP intake affects the antioxidant defence system in male adolescents (11–14 years, n = 18) was carried out using two diets rich and poor in MRPs (brown diet, BD, and white diet, WD, respectively). Fasting blood samples were collected after the dietary intervention periods to measure oxidative status. The in vitro antioxidant activity of the diets was also assessed. RESULTS: The BD had stronger in vitro antioxidant activity to scavenge free radicals and greater ability to reduce lipid peroxidation. However, in the in vivo assay, markers of oxidative damage (serum thiobarbituric acid‐reactive substances and erythrocyte hydroperoxides) and antioxidant defence parameters (serum antioxidants and enzymatic activities of catalase, superoxide dismutase and glutathione peroxidase) were unchanged after the dietary treatments. Only treatment of biological samples with external oxidants revealed higher values of the antioxidant capacity after consumption of the MRP‐rich diet. CONCLUSION: In spite of the higher antioxidant activity of the BD shown in vitro, consumption of diets rich in MRPs does not seem to modify oxidative status in healthy male adolescents. However, a protective effect against induced oxidation was shown. Copyright © 2008 Society of Chemical Industry     

抗氧化肽延缓油脂氧化作用机制研究进展

油脂的氧化会引起食品质量劣变,影响其风味、色泽以及营养成分,寻找延缓油脂氧化的方法始终是产业发展的研究热点。抗氧化肽属于新型的抗氧化剂,然而其作用机制尚未完全了解。为推进肽在油脂体系中作用机制的认识,基于油脂氧化机理,系统梳理了抗氧化肽对油脂氧化各阶段诱因的抑制机制,主要归结于其多种功能的复杂相互作用,这些功能包括抑制活性氧生成,清除自由基,螯合促氧化金属离子,降低过氧化物的反应性。抗氧化肽清除自由基的抗氧化效果主要依赖其氨基酸残基组成,其中非极性氨基酸残基通过自由基加成作用,延缓油脂氧化起始期;极性不带电氨基酸残基通过将氢原子转移,形成抗氧化自由基来延迟油脂氧化的增长期;极性带正电荷氨基酸残基可以吸引负电荷自由基,如超氧阴离子自由基;相反地,极性带负电荷氨基酸残基可以通过排斥负电荷自由基从而减少金属离子促油脂氧化作用。     

Initiation of lipid peroxidation in biological systems

The direct oxidation of PUFA by triplet oxygen is spin forbidden. The data reviewed indicate that lipid peroxidation is initiated by nonenzymatic and enzymatic reactions. One of the first steps in the initiation of lipid peroxidation in animal tissues is by the generation of a superoxide radical (see Figure 16), or its protonated molecule, the perhydroxyl radical. The latter could directly initiate PUFA peroxidation. Hydrogen peroxide which is produced by superoxide dismutation or by direct enzymatic production (amine oxidase, glucose oxidase, etc.) has a very crucial role in the initiation of lipid peroxidation. Hydrogen peroxide reduction by reduced transition metal generates hydroxyl radicals which oxidize every biological molecule. Hydrogen peroxide also activates myoglobin, hemoglobin, and other heme proteins to a compound containing iron at a higher oxidation state, Fe(IV) or Fe(V), which initiates lipid peroxidation even on membranes. Complexed iron could also be activated by O2- or by H2O2 to ferryl iron compound, which is supposed to initiate PUFA peroxidation. The presence of hydrogen peroxide, especially hydroperoxides, activates enzymes such as cyclooxygenase and lipoxygenase. These enzymes produce hydroperoxides and other physiological active compounds known as eicosanoids. Lipid peroxidation could also be initiated by other free radicals. The control of superoxide and perhydroxyl radical is done by SOD (a) (see Figure 16). Hydrogen peroxide is controlled in tissues by glutathione-peroxidase, which also affects the level of hydroperoxides (b). Hydrogen peroxide is decomposed also by catalase (b). Caeruloplasmin in extracellular fluids prevents the formation of free reduced iron ions which could decompose hydrogen peroxide to hydroxyl radical (c). Hydroxyl radical attacks on target lipid molecules could be prevented by hydroxyl radical scavengers, such as mannitol, glucose, and formate (d). Reduced compounds and antioxidants (ascorbic acid, alpha-tocopherol, polyphenols, etc.) (e) prevent initiation of lipid peroxidation by activated heme proteins, ferryl ion, and cyclo- and lipoxygenase. In addition, cyclooxygenase is inhibited by aspirin and nonsteroid drugs, such as indomethacin (f). The classical soybean lipoxygenase inhibitors are antioxidants, such as nordihydroguaiaretic acid (NDGA) and others, and the substrate analog 5,8,11,14 eicosatetraynoic acid (ETYA), which also inhibit cyclooxygenase (g). In food, lipoxygenase is inhibited by blanching. Initiation of lipid peroxidation was derived also by free radicals, such as NO2. or CCl3OO. This process could be controlled by antioxidants (e).(ABSTRACT TRUNCATED AT 400 WORDS)     

Neuroprotective effect of Decalepis hamiltonii roots against ethanol-induced oxidative stress

The neuroprotective potential of the aqueous extract of the roots of Decalepis hamiltonii (D. hamiltonii root aqueous extract-DHRAE) was studied against ethanol-induced oxidative stress in the rat brain. Ethanol, single dose (5 g/kg body weight), induced oxidative stress in the rat brain which was evident from the increased lipid peroxidation and protein carbonylation, reduced glutathione, and suppressed activities of antioxidant enzymes such as superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase, and glutathione-S-transferase. Pretreatment of rats with multiple doses of DHRAE, 50 and 100 mg/kg b.w., for 7 consecutive days significantly prevented the ethanol-induced oxidative stress. DHRAE, as such, boosted the antioxidant status of the rat brain. The neuroprotective potential of DHRAE can be attributed to the known antioxidant constituents or its interaction with antioxidant response elements (AREs) which needs to be ascertained.     

The etiology of oxidative stress in the various species of animals,a review

Oxidative stress is the consequence of an imbalance of pro‐oxidants and antioxidants leading to cell damage and tissue injury. The exhaustion of antioxidant systems is one of the reasons for the occurrence of oxidative stress, which results in avalanche production of reactive oxygen species (ROS) or free radicals. High oxidative stress is common in organs and tissues with high metabolic and energy demands, including skeletal and heart muscle, liver and blood cells. Stress arises in animals in response to unavoidable or adverse environmental conditions. In the external environment, which affects the body of the cow, there are four main groups of stressors: physical, chemical, biological and psychological. Physical stressors include fluctuations in ambient temperature as well as mechanical injuries. High ambient temperature is one of the factors affecting the productivity of cows. Biological stressors are conditioned by errors and irregularities in habits. Both of these phenomena have an adverse impact on both the resistance of animals and fertility and are the etiological agent of oxidative stress. Various mechanisms may be responsible for metal‐induced oxidative stress: direct or indirect generation of ROS, depletion of glutathione and inhibition of antioxidant enzymes are well known for all redox‐active and redox‐inactive metals. © 2014 Society of Chemical Industry     

Studies on the antioxidative activity of red pigments in Italian-type dry-cured ham

Aqueous phosphate buffer extracts and acetone/water extracts of pigments from Parma ham were assessed as antioxidants by (1) electron spin resonance spectroscopy using a spin probing technique to evaluate their efficiencies as scavengers of free radicals, and (2) by electrochemical measurement of oxygen depletion rate in an aqueous methyl linoleate emulsion to evaluate their efficiencies as chain-breaking antioxidant, and using both methods, compared with the effect of apomyoglobin and nitrosylmyoglobin. Aqueous phosphate extracts and acetone/water extracts of Parma ham pigment both scavenged a semi-stable nitroxide radical (Fremy's salt), and both extracts reduced the rate of oxygen consumption for lipid peroxidation (initiated by metmyoglobin) very efficiently. For apomyoglobin no antioxidative capacity was observed, and the heme moiety of the pigment(s) of Parma ham were concluded to have antioxidative properties. The more lipophilic pigment, as extracted by acetone/water, had the most significant effect, and its ability to inhibit lipid oxidation was further tested in a model food system based on cooked pork. The lipid oxidation was increasingly inhibited by increasing additions from 0.12 ppm to 0.24 ppm Parma ham pigment, and the pigment protected -tocopherol against degradation in a concentration dependent manner.     

Antioxidant activity of natural vitamin E extracted from soybean sludge as assessed by chemiluminescence

The antioxidant activity of natural vitamin E (VE) towards superoxide anion, hydroxyl radical and lipid free radicals was investigated using a chemiluminescence technique. VE was extracted from soybean sludge, where it was present at a concentration of 600 g kg^?1 as determined by HPLC. Superoxide anions, hydroxyl radicals and lipid free radicals were generated by the pyrogallol autoxidation system, the Fenton reaction system and the AAPH‐induced γ‐linolenic acid peroxidation system respectively. VE rapidly scavenged hydroxyl radicals and lipid free radicals. The efficient concentration (EC₅₀) of VE against both was 0.1 mg ml^?1. A reaction time of 6 s was adequate to scavenge hydroxyl radicals and a reaction time of 24 s was enough to scavenge lipid free radicals. However, VE scavenged superoxide anions at a relatively low rate, and the extent of scavenging was less than 20% even after 3 min at a VE concentration of 4.3 mg ml^?1. © 2001 Society of Chemical Industry     

欧李多酚清除自由基活性研究

采用大豆卵磷脂脂质模型系统、低密度脂蛋白体系和牛血清白蛋白介质三个不同的评价体系考察欧李多酚的清除自由基和抗氧化活性。研究表明,欧李多酚的脂质过氧化和蛋白质氧化抑制呈剂量和时间依赖性方式,总酚与抗氧化能力之间的相关性较好,能完全抑制由过氧自由基和铜离子诱导早期的共轭二烯(CD)的生成,延长脂质氧化诱导期,有效地抑制由过氧自由基诱导的牛血清白蛋白氧化。欧李多酚能通过淬灭自由基的方式抑制因自由基所致的生物膜、脂蛋白和血清白蛋白氧化损伤。     

Antioxidants and prevention of chronic disease

The generation of reactive oxygen species (ROS) and other free radicals (R) during metabolism is a necessary and normal process that ideally is compensated for by an elaborate endogenous antioxidant system. However, due to many environmental, lifestyle, and pathological situations, excess radicals can accumulate, resulting in oxidative stress. Oxidative stress has been related to cardiovascular disease, cancer, and other chronic diseases that account for a major portion of deaths today. Antioxidants are compounds that hinder the oxidative processes and thereby delay or prevent oxidative stress. This article examines the process of oxidative stress and the pathways by which it relates to many chronic diseases. We also discuss the role that endogenous and exogenous antioxidants may play in controlling oxidation and review the evidence of their roles in preventing disease.     

Antioxidant Modulation of F2-Isoprostanes in Humans: A Systematic Review

F2-isoprostanes are a biomarker of lipid peroxidation, and their measurement has emerged as a reliable approach to assess oxidative stress. However, dietary intervention studies in humans have provided contrasting results following supplementation with antioxidant-rich foods or supplements. In this paper, we have systematically reviewed the evidence about the effect of supplementation with antioxidant-rich foods and galenic antioxidants on isoprostanes levels in humans. Moreover, the association with nonenzymatic antioxidant capacity (NEAC), a biomarker of endogenous antioxidant status, has also been investigated.

MEDLINE database was searched using the terms “(isoprostane* OR isoP OR iso-PGF OR epi-PGF) AND (intervention* OR consumption* OR administration* OR supplementation*),” with limits activated “humans” and “English.” Abstracts and full texts were screened, from which were selected human intervention studies reporting isoprostanes measurement in biological fluids. The total of the studies carried out with antioxidant-rich foods and antioxidant galenic supplements was 113, reporting 154 interventions. Results suggest that dietary antioxidants modulate successfully the levels of isoprostanes in less than 45% of the interventions. A correspondence between the effect on isoprostane and NEAC has been evidenced, and this correspondence suggests the importance of measuring different biomarkers to obtain a better outline of the redox events following supplementation.     

Vitamin E supplementation during the dry period in dairy cattle. Part II: oxidative stress following vitamin E supplementation may increase clinical mastitis incidence postpartum

The aim of this study was to evaluate, retrospectively, which physiological states influenced the effect of vitamin E supplements during the dry period on the level of oxidative stress at 2 wk antepartum. Furthermore the effect of oxidative stress at 2 wk antepartum on the risk of clinical mastitis in early lactation was investigated. Cows experience oxidative stress around calving. Vitamin E is able to decrease oxidative stress by scavenging free radicals. Normally, vitamin E radicals formed when vitamin E reacts with free radicals are regenerated by a network of other antioxidants, termed the “vitamin E regeneration system” (VERS). In case of vitamin E supplementation, VERS should be sufficient to regenerate formed vitamin E radicals; if not, oxidative stress might increase instead of decrease. Additionally, the level of oxidative stress and vitamin E might be important physiological states to evaluate before supplementation. In a clinical trial, 296 cows on 5 farms were randomly divided into 2 groups, supplemented with a mineral mix between dry off and calving that supplied 3,000 or 135 IU/d, respectively. Blood samples collected at dry off and 2 wk antepartum were analyzed for vitamin E, reactive oxygen metabolites, ferric-reducing ability of plasma, glutathione peroxidase, and malondialdehyde. Cows were allocated retrospectively into 8 subgroups based on the level of oxidative stress, vitamin E, and VERS status at dry off. To evaluate whether differences in physiological states at dry off influenced the effect of vitamin E supplementation on the level of oxidative stress, group effects (supplemented vs. control) were studied with Student's t-test for all 8 subgroup at 2 wk antepartum. Differences in physiological states at dry off influenced the effect of vitamin E supplements. In 2 insufficient VERS subgroups, the supplemented group had higher levels of free radicals at 2 wk antepartum compared with the control group. Relative risk calculation was used to study the effect of oxidative stress at 2 wk antepartum on the incidence of mastitis in the first 100 d of lactation. Higher levels of oxidative stress at 2 wk antepartum were related to higher risk of clinical mastitis. In conclusion, not every dry cow responded well to high vitamin E supplementation. This subgroup analysis provides a possible explanation for the unexpected adverse effects observed in the clinical trial.     

竹盐酿造酱油对H2O2诱发LLC-PK1细胞氧化损伤的保护作用

目的：研究竹盐酿造酱油乙醇提取物对H2O2诱发猪肾近曲上皮小管细胞（pig proximal tubular cell）LLC-PK1氧化损伤的保护作用。方法：以不同质量浓度（10～200 μg/mL）的竹盐酿造酱油乙醇提取物预培养LLC-PK1细胞24 h后,换用含500 μmol/L H2O2的DMEM细胞培养液继续培养4 h建立细胞氧化损伤模型。四甲基偶氮唑蓝（methyl thiazolyl tetrazolium,MTT）法检测细胞生存率,硫代巴比妥酸比色法测定细胞内丙二醛（malondialdehyde,MDA）含量,比色法测定细胞内过氧化氢酶（catalase,CAT）、超氧化物歧化酶（superoxidedismutase,SOD）、谷胱甘肽过氧化物酶（glutathione peroxidase,GSH-Px）活性和谷胱甘肽（glutathione,GSH）含量。结果：经不同质量浓度竹盐酿造酱油乙醇提取物预处理24 h后,受损细胞的生存率上升,细胞内MDA生成量减少,ROS水平显著降低。同时,细胞内抗氧化酶（CAT、SOD和GSH-Px）的活性及其mRNA转录水平,GSH含量也较未经竹盐酿造酱油乙醇提取物处理的受损细胞增加。结论：竹盐酿造酱油乙醇提取物可以通过提高细胞内抗氧化酶系的活性和抗氧化物质GSH的含量而有效地对抗H2O2诱发的LLC-PK1细胞氧化损伤。     

Use of the synthetic superoxide dismutase/catalase mimetic EUK-134 to compensate for seasonal antioxidant deficiency by reducing pre-existing lipid peroxides at the human skin surface

The generation of reactive oxygen species (ROS) in UV-exposed skin is believed to contribute to the photoaging process. The stratum corneum (SC) contains a variety of enzymatic and non-enzymatic antioxidants to protect against various environmental sources of free radicals. We have previously shown a seasonal variation in SC catalase activity with strong deactivation in sun-exposed skin in the summer, whereas SC superoxide dismutase (SOD) activity remained intact in those conditions. This potentially leads to the local overproduction of H(2)O(2). The oxidized lipid squalene hydroperoxide accumulates at the surface of sun-exposed skin in the summer and upon exposure to ultravoilet A (UVA) doses as low as 0.1 J cm(-2) and adequate protection against excessive lipid peroxidation at times of UV exposure should be aimed for. We have been using the induction of lipid hydroperoxides at the skin surface by a single dose of UVA (1 J cm(-2)) as a model system to evaluate the protective effect of antioxidants in vivo. Topical treatment with the synthetic SOD/catalase mimetic molecule (EUK-134) 1 h before UVA exposure reduced the level of lipid peroxides at the surface of UVA-exposed skin but also baseline peroxide levels on non-irradiated skin were reduced in a dose-dependent fashion. In contrast to alpha-tocopherol, EUK-134 even reduced the level of lipid peroxides at the surface of UVA-exposed skin when it was applied after irradiation. We confirmed that this salen-manganese complex was able to reduce squalene hydroperoxide levels in vitro, suggesting peroxidase-like activity towards organic peroxides. These data support the concept that the synthetic SOD/catalase mimetic EUK-134 might be able to compensate for seasonal deficiencies in antioxidant defense capacity at the skin surface, thereby contributing to an optimal protection of the skin against the accumulation of oxidative damage.     

Assay of the antioxidant capacity of foods using an iron(II)-catalysed lipid peroxidation model for greater nutritional relevance

The formation of free radicals by the iron-catalysed Fenton reaction is a major cause of oxidative damage in the body. Here a common assay of antioxidant capacity, inhibition of the β-carotene-linoleic acid model of lipid peroxidation, has been modified by the addition of ferrous iron (final concentration 36 μmol/l), which makes the rate of oxidation of the lipids occur 25 times faster. Such an assay can simulate the oxidative damage to membrane lipids and low density lipoproteins occurring in the body in the presence of free iron. It thus may be nutritionally more relevant than traditional chemical assays of antioxidant capacity, as it measures pre-emptive antioxidant activity, i.e. activity which prevents free radicals being formed in the first place. Pre-empting their formation is likely to be more protective than scavenging of free radicals. The relative antioxidant activity of some food products found using this new assay was very different from that found using a radical-scavenging assay. Vitamin C, at 280 mg/l, was found to be 60 times better than blackcurrant puree in scavenging free radicals, but only one eighth as good as the blackcurrant puree in preventing iron-catalysed lipid peroxidation.