Peroxyl Radical-trapping Antioxidant Research Articles

Vitamin E protects from lipid peroxidation during winter stress in the seagrass Cymodocea nodosa

Main conclusionAdjustments in the antenna size and α-tocopherol contents provide protection from sustained damage in leaves of a seagrass, while low vitamin E contents appear to be enough to protect rhizomes (which appear to be more cold tolerant than leaves).Despite low temperatures can adversely affect the proper growth and development of marine angiosperms, by, among other processes, increasing reactive oxygen species production and causing oxidative damage to lipid membranes, the role of vitamin E in seagrasses, such as Cymodocea nodosa has not been explored thus far. Here, we aimed to better understand the possible role of this chain-breaking (peroxyl radical-trapping) antioxidant in response to low temperatures, and most particularly in relation to the occurrence of photo-inhibition and lipid peroxidation. Low temperatures caused an important desiccation of leaves, but not of rhizomes, which were much more tolerant to cold stress than leaves. Cold stress during winter was associated with chlorophyll loss and transient photo-inhibition, as indicated by reversible reductions in the Fv/Fm ratio. Adjustments in pigment antenna size and vitamin E contents per unit of chlorophyll during winter may help protect the photosynthetic apparatus from sustained photo-inhibitory damage and lipid peroxidation events in leaves. Rhizomes also accumulated significant amounts of vitamin E, although to a much lesser extent than leaves, and kept protected from lipid peroxidation during winter, as indicated by malondialdehyde contents, a product from secondary lipid peroxidation. It is concluded that vitamin E can help protect both leaves and rhizomes from lipid peroxidation, although cold stress during winter can cause transient photo-inhibition of the photosynthetic apparatus, in C. nodosa.

Analytical Methods Used in Determining Antioxidant Activity: A Review.

The study of antioxidants and their implications in various fields, from food engineering to medicine and pharmacy, is of major interest to the scientific community. The present paper is a critical presentation of the most important tests used to determine the antioxidant activity, detection mechanism, applicability, advantages and disadvantages of these methods. Out of the tests based on the transfer of a hydrogen atom, the following were presented: the Oxygen Radical Absorption Capacity (ORAC) test, the Hydroxyl Radical Antioxidant Capacity (HORAC) test, the Total Peroxyl Radical Trapping Antioxidant Parameter (TRAP) test, and the Total Oxyradical Scavenging Capacity (TOSC) test. The tests based on the transfer of one electron include the Cupric Reducing Antioxidant Power (CUPRAC) test, the Ferric Reducing Antioxidant Power (FRAP) test, the Folin–Ciocalteu test. Mixed tests, including the transfer of both a hydrogen atom and an electron, include the 2,2′-Azinobis-(3-ethylbenzothiazoline-6-sulfonic acid (ABTS) test, and the [2,2-di(4-tert-octylphenyl)-1-picrylhydrazyl] (DPPH) test. All these assays are based on chemical reactions and assessing the kinetics or reaching the equilibrium state relies on spectrophotometry, presupposing the occurrence of characteristic colours or the discolouration of the solutions to be analysed, which are processes monitored by specific wavelength adsorption. These assays were successfully applied in antioxidant analysis or the determination of the antioxidant capacity of complex samples. As a complementary method in such studies, one may use methods based on electrochemical (bio)sensors, requiring stages of calibration and validation. The use of chemical methods together with electrochemical methods may result in clarification of the operating mechanisms and kinetics of the processes involving several antioxidants.

Red/Green Currant and Sea Buckthorn Berry Press Residues as Potential Sources of Antioxidants for Food Use.

The potential for using extracts of press residues from black, green, red, and white currants and from sea buckthorn berries as sources of antioxidants for foods use was investigated. Press residues were extracted with ethanol in four consecutive extractions, and total Folin–Ciocalteu (F–C) reactive material and authentic phenolic compounds were determined. Radical quenching capability and mechanisms were determined from total peroxyl radical-trapping antioxidant capacity (TRAP) and oxygen radical absorbance capacity (ORAC) assays and from diphenylpicrylhydrazyl (DPPH) kinetics, respectively; specific activities were normalized to F–C reactive concentrations. Levels of total F–C reactive materials in press residue extracts were higher than in many fruits and showed significant radical quenching activity. Black currant had the highest authentic phenol content and ORAC, TRAP, and DPPH reactivity. Sea buckthorn grown in northern Finland showed extremely high total specific DPPH reactivity. These results suggest that berry press residues offer attractive value-added products that can provide antioxidants for use in stabilizing and fortifying foods.

The effect of cryotherapy on total antioxidative capacity in patients with active seropositive rheumatoid arthritis.

Patients with rheumatoid arthritis (RA) have increased oxidative stress, decreased antioxidant levels, and impaired antioxidant capacity. Cold treatments are used to relieve joint inflammation and pain. Therefore, we measured the effect of cold treatments on the antioxidative capacity of RA patients with active disease. Sixty patients were randomized to (1) whole body cryotherapy at -110°C, (2) whole body cryotherapy at -60°C, or (3) local cryotherapy. Each treatment was given three times daily for 7 consecutive days in addition to the conventional rehabilitation. Blinded rheumatologist evaluated disease activity before the first and after the last cryotherapy. We collected plasma samples daily immediately before the first and after the second cryotherapy and measured total peroxyl radical trapping antioxidant capacity of plasma (TRAP), which reflects global combined antioxidant capacity of all individual antioxidants in plasma. Baseline morning TRAP levels (mean, 95% CI), adjusted for age, body mass index, disease activity, and dose of prednisolone, were 1244 (1098-1391)µM/l in the local cryotherapy, 1133 (1022-1245)µM/l in the cryotherapy at -60°C, and 989 (895-1082)µM/l in the cryotherapy at -110°C groups (p=0.006). After the first treatment, there was a rise in 1-h TRAP of 14.2 (-4.2 to 32.6)µM/l, 16.1 (-7.4 to 39.6)µM/l, and 23.6 (4.1-43.2)µM/l, respectively. The increase was significant in the whole-body cryotherapy -110°C group (p<0.001) but not significant between the groups (p=0.78). When analyzed for the whole week, the daily morning TRAP values differed significantly between the treatment groups (p=0.021), but there was no significant change within each treatment group. Whole-body cryotherapy at -110°C induced a short-term increase in TRAP during the first treatment session with but not during other treatment modalities. The effect was short and the cold treatments did not cause a significant oxidative stress or adaptation during 1week.

Effect of simulated gastrointestinal digestion on the phenolic compound content and in vitro antioxidant capacity of processed Cowpea (V. unguiculata) cultivars

ABSTRACTThe aim of this study was to analyze the phenolic content and antioxidant activity of five cowpea cultivars after processing and in vitro digestion. Raw cowpea samples showed a significant decrease in total phenolic content (TPC) when compared with the processed samples, however an increase was subsequently observed in digested samples. The antioxidant activity determined using, DPPH, ABTS, ferric reducing antioxidant power (FRAP) assay, and total peroxyl radical-trapping antioxidant parameter showed a similar trend to the phenolic content with a significant decrease in activity upon processing and an increase after digestion. In conclusion, all cowpea cultivars showed a high TPC content as well as an increased antioxidant activity after digestion indicating the potential health benefits which cowpea could provide to consumers. Therefore, this study shows that in vitro digestion improves the digestion and absorption of beneficial components of processed cowpea at the intestinal level.

Effects of extra virgin olive oil and fish oil on lipid profile and oxidative stress in patients with metabolic syndrome

ObjectiveThe aim of this study was to verify if extra virgin olive oil and fish oil have a synergistic effect on lipid and oxidative stress parameters in patients with metabolic syndrome (MetS). MethodsThis intervention study included 102 patients (81 women and 21 men) with MetS (mean age 51.45 ± 8.27 y) from the ambulatory center of the University Hospital of Londrina, Paraná, Brazil. Patients were randomly assigned to one of four groups: Patients in the control group (CG) were instructed to maintain their usual diet; the second group (fish oil group [FO]) received 3 g/d of fish oil ω-3 fatty acids (10 capsules); the third group (extra virgin olive oil group [OO]) received 10 mL/d of extra virgin olive oil at lunch and dinner; and the fourth group (fish oil and extra virgin olive oil group [FOO]) received 3 g/d of fish oil ω-3 fatty acids and 10 mL/d of extra virgin olive oil. MetS related markers and oxidative stress were measured at baseline and after 90 d. ResultsDifferences across treatment groups showed a statistically significant decrease (P < 0.05) in total cholesterol (TC) and low-density lipoprotein cholesterol (LDL-C) when FOO was compared with CG and OO, respectively. Hydroperoxides showed a significant decrease (P < 0.05) when FOO was compared with CG, whereas there was an increase in total peroxyl radical-trapping antioxidant potential/advanced oxidation protein products (TRAP/AOPP; P < 0.05) in FOO when compared with FO. In relation to baseline values, there was a significant decrease (P < 0.05) in LDL-C values, and TC/high-density lipoprotein cholesterol (HDL-C) and LDL-C/HDL-C indexes in FOO. There was also a decrease (P < 0.05) in hydroperoxides, in AOPP and in AOPP/TRAP index in FOO, and an increase (P < 0.05) in TRAP/AOPP index in FOO and in TRAP/uric acid ratio in OO. ConclusionThe present study provides evidence that increased dietary ω-3 polyunsaturated fatty acids and extra virgin olive oil have beneficial synergistic effects on lipid metabolism and oxidative stress in patients with MetS.

The effect of lipid peroxidation products on reactive oxygen species formation and nitric oxide production in lipopolysaccharide-stimulated RAW 264.7 macrophages

Lipid peroxidation induced by oxidants leads to the formation of highly reactive metabolites. These can affect various immune functions, including reactive oxygen species (ROS) and nitric oxide (NO) production. The aim of the present study was to investigate the effects of lipid peroxidation products (LPPs) – acrolein, 4-hydroxynonenal, and malondialdehyde – on ROS and NO production in RAW 264.7 macrophages and to compare these effects with the cytotoxic properties of LPPs. Macrophages were stimulated with lipopolysaccharide (0.1 μg/ml) and treated with selected LPPs (concentration range: 0.1–100 μM). ATP test, luminol-enhanced chemiluminescence, Griess reaction, Western blotting analysis, amperometric and total peroxyl radical-trapping antioxidant parameter assay were used for determining the LPPs cytotoxicity, ROS and NO production, inducible nitric oxide synthase expression, NO scavenging, and antioxidant properties of LPPs, respectively. Our study shows that the cytotoxic action of acrolein and 4-hydroxynonenal works in a dose- and time-dependent manner. Further, our results imply that acrolein, 4-hydroxynonenal, and malondialdehyde can inhibit, to a different degree, ROS and NO production in stimulated macrophages, partially independently of their toxic effect. Also, changes in enzymatic pathways (especially NADPH-oxidase and nitric oxide synthase inhibition) and NO scavenging properties are included in the downregulation of reactive species formation.

Solid-phase extraction of berries’ anthocyanins and evaluation of their antioxidative properties

Solid-phase extraction (SPE) was used to obtain anthocyanin-rich extracts from five berry species: chokeberry, elderberry, black currant, blackberry and blueberry. During SPE more than 94.4% of the sugars and more than 88.5% of the acids present in the crude extracts were separated. The SPE resulted in 90–95.6% anthocyanins recovery. The antioxidative properties of the anthocyanin-rich extracts were tested by measuring their oxygen radical absorption capacity (ORAC), hydroxyl radical averting capacity (HORAC), total peroxyl radical trapping antioxidant parameter (TRAP), scavenging of nitric oxide and inhibition of lipid peroxidation. Elderberry extract revealed the highest ORAC value of 5783 μmol TE/g. Chokeberry extract was the most potent inhibitor of lipid peroxidation and had the highest TRAP value of 4051 μmol TE/g. Blueberry extract had the highest HORAC result – 1293 μmol GAE/g and was the most powerful scavenger of NO. The high antioxidant activity according to all antioxidant assays revealed opportunities to apply these preparations as antioxidants.

Effect of polyunsaturated fatty acids on the reactive oxygen and nitrogen species production by raw 264.7 macrophages

Polyunsaturated fatty acids (PUFAs) can affect various functions of the immune system including inflammatory responses. An oxidative burst of phagocytes accompanied by reactive oxygen species (ROS) and reactive nitrogen species (RNS) formation is one of the phagocyte functions that could be modulated by PUFAs. To investigate the effects of omega-3 (alpha-linolenic, docosahexaenoic, eicosapentaenoic) and omega-6 (arachidonic, linoleic) PUFAs on lipopolysaccharide (LPS)-stimulated ROS and RNS production by the murine macrophage cell line RAW 264.7. Murine peritoneal macrophages RAW 264.7 were stimulated with LPS (0.1 microg/ml) and treated with 0.1-100 microM omega-3 or omega-6 PUFAs for either 8 (ROS production) or 20 h (RNS production). The cytotoxicity of PUFAs was evaluated by an ATP (adenosine triphosphate) test after both 8 and 20 h of treatment with PUFAs. Changes in ROS production by LPS-treated macrophages subsequently activated with phorbol myristate acetate (PMA) or opsonized zymosan particles (OZP) were determined by luminol-enhanced chemiluminescence, whilst the production of RNS was determined as the concentration of nitrites in cell supernatants (Griess reaction). Changes in inducible nitric oxide synthase (iNOS) expression were evaluated by Western blot analysis. The antioxidant properties of PUFAs were tested by TRAP (total peroxyl radical-trapping antioxidant parameter) assay. All PUFAs in 100 microM concentration except eicosapentaenoic acid decreased ROS production. The effect was most significant when docosahexaenoic acid was used. Arachidonic acid decreased PMA-activated ROS production even in 1 and 10 microM concentrations. On the other hand, 10 and 100 microM eicosapentaenoic acid potentiated ROS production. As concerns RNS production, all the fatty acids that were tested in a concentration of 100 microM decreased iNOS expression and nitrite accumulation. Fatty acids had no significant effect on the viability and proliferation of RAW 264.7 cells. The TRAP assay confirmed that none of the tested PUFAs exerted any significant antioxidant properties. High concentrations of PUFAs of both omega-3 and omega-6 groups can inhibit ROS and RNS formation by stimulated macrophages. The expression of iNOS can also be inhibited. This effect, together with the absence of antioxidant activity and cytotoxic properties, indicates that PUFAs can participate in the regulation of enzymes responsible for reactive species production.

Alterations in plasma antioxidants during reperfusion of the ischemic small intestine in rats

The aim of this study was to evaluate the contribution of three plasma antioxidants (albumin, uric acid, SH groups) to the plasma total peroxyl radical-trapping antioxidant capacity (TRAP) in 2 and 4 h of intestinal reperfusion in rats. TRAP increased significantly both after 2 and 4 h of reperfusion. Neither albumin nor SH groups contributed significantly to this increase. TRAP was strongly influenced by the increase in uric acid concentration and also probably by the cell destruction caused by oxidative stress. Since the TRAP increase was accompanied by an increase in the level of thiobarbituric acid reactive substances (TBARS, a marker of lipid peroxidation), we can conclude that even such a large increase in TRAP is not sufficient to prevent the progression of lipid peroxidation and oxidative stress.

Acute and long‐term effects of winter swimming and whole‐body cryotherapy on plasma antioxidative capacity in healthy women

The effects of severe cold stress on total peroxyl radical trapping antioxidant capacity of plasma (TRAP) were studied in two groups of healthy women: a whole‐body cryotherapy group (WBC, n = 10) and a winter swimming group (WS, n = 10). The biovariability of TRAP values was also analysed. The WBC group was exposed to −110°C for 2 min, whereas the exposure for the WS group lasted 20 s in ice‐cold water. Sessions were organized three times per week for 12 weeks. Blood specimens were collected at 2, 4, 8 and 12 weeks at rest, 2 and 35 min after the cold exposures and at the corresponding times without cold exposure on a separate day. Conventional methods were used to determine TRAP values. The between‐subject variation was 13.6% and the within‐subject variation 6.4%. The index of individuality was 0.46, and the index of heterogeneity was 0.079. These results indicate a marked heterogeneity among subjects. During the first 4 weeks, the mean TRAP value significantly increased at 2 min after cold exposure in the WBC group, returning to baseline 35 min after the exposure. Similar changes were observed in the WS group. However, all changes due to cold were relatively mild (<5%). After 4 weeks no changes in TRAP values after the cold exposures were noticed and no long‐term changes in basal TRAP values were observed. In the main, regular WBC and WS do not seem to be harmful as far as plasma antioxidative capacity is concerned.

Effects of quercetin on liver damage in rats with carbon tetrachloride-induced cirrhosis.

Flavonoids are reported to exhibit a wide variety of biological effects, including antioxidant and free radical-scavenging activities. Evidence of oxidative reactions is often associated with various chronic disease processes characterized by accumulation of connective tissue. This study was aimed to investigate the protective effects of chronic administration of the flavonoid quercetin (150 micromol/kg body wt/day intraperitoneally) in rats with carbon tetrachloride-induced fibrosis. In animals rendered cirrhotic by administration of carbon tetrachloride for 16 weeks, cell necrosis, fibrosis, and inflammatory infiltration were found. Histological abnormalities were accompanied by a higher hepatic content of collagen and thiobarbituric acid-reactive substances. Expression of inducible nitric oxide synthase (iNOS) was significantly increased in the liver. Treatment with quercetin during 3 weeks improved liver histology and reduced collagen content, iNOS expression, and lipid peroxidation. Those effects were associated with an increased total peroxyl radical-trapping antioxidant capacity of liver. We conclude that quercetin is effective in this model of liver damage.

Oxidative status and malondialdehyde in beta-thalassaemia patients.

In beta-thalassaemia syndromes, decreased or impaired biosynthesis of beta-globin leads to accumulation of unpaired alpha-globin chains. Moreover, the iron overload in beta-thalassaemia patients generates oxygen-free radicals and peroxidative tissue injury. The aim of this study was to detect and correlate iron overload parameters with the oxidative stress and the antioxidant capability in beta-thalassaemia patients. Serum iron, transferrin saturation, serum ferritin, nontransferrin-bound iron (NTBI), levels of serum free and total (free + bound) malondialdehyde (MDA) and total peroxyl radical-trapping antioxidant parameter (TRAP) were evaluated in 21 regularly transfused beta-thalassaemia major (TM) patients, 13 untransfused beta-thalassaemia intermedia (TI) patients and 17 healthy controls. Blood from the TM patients was drawn 48 h after the last desferoxamine (20-40 mg kg(-1)) infusion and just before transfusion. Free and total MDA and NTBI levels were higher in the TM patients than in the TI. In the TM patients the free MDA levels correlated positively with serum iron (r = +0.3, P = 0.0006), whereas the total MDA correlated positively with NTBI (r = +0.45, P = 0.037). However, a negative correlation was observed between TRAP and NTBI (r = -0.4, P = 0.0006). In the TI patients there was no significant correlation between free or total MDA and TRAP or NTBI. Our results confirm the peroxidative status generated by iron overload in thalassaemia patients and highlight the rapid formation of marked amounts of free MDA despite the chelation therapy in TM patients.

End-of-life care-related publications in cardiology journals

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Does a relationship exist between the urate pool in the body and lipid peroxidation during exercise?

In this study, we investigated whether a relationship exists between the levels of urate in vivo and lipid peroxidation during exercise. Seven healthy male subjects performed exhaustive cycling exercise under the following three conditions. The levels of urate, thiobarbituric acid reactive substances (TBARS) and allantoin in plasma and urine were examined before exercise and during a 3 h recovery period. (1) Benzbromarone administration experiment: benzbromarone (an uricosuric agent) was administered orally the day before exercise. (2) IMP administration experiment: inosine 5′-monophosphate disodium salt (a precursor of urate) was administered orally the day before exercise. (3) Control experiment: no test substance was administered. The main results obtained were as follows. Plasma urate levels and total peroxyl radical-trapping antioxidant parameter (TRAP) for deproteinized plasma in the resting period significantly decreased depending on the treatment, in the order of IMP > control > benzbromarone. A significant positive correlation was evident between plasma urate levels and TRAP values for deproteinized plasma. The increase in plasma levels of allantoin was observed only in the case of IMP treatment. A significant negative correlation between plasma levels of urate in the resting period and the amounts of urinary TBARS excreted during the recovery period was recognized. These results suggest that the urate level in vivo before exercise is a factor influencing lipid peroxidation during exhaustive exercise. Furthermore, these findings support the view that urate may serve as an important free-radical scavenger in vivo.

A marathon run increases the susceptibility of LDL to oxidation in vitro and modifies plasma antioxidants.

Physical activity increases the production of oxygen free radicals, which may consume antioxidants and oxidize low-density lipoprotein (LDL). To determine whether this occurs during strenuous aerobic exercise, we studied 11 well-trained runners who participated in the Helsinki City Marathon. Blood samples were collected before, immediately after, and 4 days after the race to determine its effect on circulating antioxidants and LDL oxidizability in vitro. LDL oxidizability was increased as determined from a reduction in the lag time for formation of conjugated dienes both immediately after (180 +/- 7 vs. 152 +/- 4 min, P < 0.001) and 4 days after (155 +/- 7 min, P < 0.001) the race. No significant changes in lipid-soluble antioxidants in LDL or in the peak LDL particle size were observed after the race. Total peroxyl radical trapping antioxidant capacity of plasma (TRAP) and uric acid concentrations were increased after the race, but, except for TRAP, these changes disappeared within 4 days. Plasma thiol concentrations were reduced after the race. No significant changes were observed in plasma ascorbic acid, alpha-tocopherol, beta-carotene, and retinol concentrations after the marathon race. We conclude that strenuous aerobic exercise increases the susceptibility of LDL to oxidation in vitro for up to 4 days. Although the increase in the concentration of plasma TRAP reflects an increase of plasma antioxidant capacity, it seems insufficient to prevent the increased susceptibility of LDL to oxidation in vitro, which was still observed 4 days after the race.

On the antioxidant activity of melatonin

Melatonin has been widely reported to be an effective antioxidant. Studies of its ability to inhibit the autoxidation of lipids in homogeneous solution and in model heterogeneous systems show that melatonin is not a peroxyl radical trapping antioxidant. In contrast, melatonin can inhibit metal ion-catalyzed oxidation processes.

Role of Bilirubin in the Regulation of the Total Peroxyl Radical Trapping Antioxidant Activity of Plasma in Sickle Cell Disease

Alpha-tocopherol and bilirubin antioxidant properties were evaluated in sickle cell disease. The total peroxyl radical trapping antioxidant activity of plasma (TRAP) was measured with a polarographic method using hemin as oxidative stress initiator. The TRAP was not correlated with plasma alpha-tocopherol concentration. The TRAP was correlated with plasma total bilirubin concentration [TRAP = 8.1 (total bilirubin) +363.7 (r = 0.67)] and the correlation was even better with the sum (alpha-tocopherol + total bilirubin) plasma concentration [TRAP = 9.1(alpha-tocopherol + total bilirubin)+ 170.5(r = 0.77)]. The alpha-tocopherol contribution in the antioxidant capacity of plasma was significant but bilirubin level acted as the limiting factor of plasma antioxidant capacity in sickle cell anemia. This result can be explained by the antioxidant property of bilirubin but also by coantioxidant activity towards oxidized alpha-tocopherol.

Effects of anthracyclin-based chemotherapy on total plasma antioxidant capacity in small cell lung cancer patients

Plasma total peroxyl radical trapping antioxidant parameters (TRAP) and their main antioxidant components (vitamin E, uric acid, protein sulfhydryl groups, and unidentified antioxidant proportions) were measured in 12 small cell lung cancer (SCLC) patients receiving combined chemotherapy consisting of vincristine, adriamycin and cyclophosphamide for SCLC. Plasma samples were collected ten times during the first two cycles of chemotherapy. There is previous evidence that many anticancer drugs exert their cytotoxity via free oxygen radicals. We hypothesized that adriamycin-induced, increased oxygen free radical production should decrease plasma TRAP as a consequence of oxidative stress. A statistically significant reduction of plasma TRAP was noted 8 hours after the first adriamycin infusion. A reduction of calculated TRAP (TRAP calc)—the sum of concentrations of individual antioxidants, corrected by their experimentally-determined stochiometric factors—appeared 3 hours after the first adriamycin infusion and continued for up to 1 week afterwards. This decrease was due to the reduction of ascorbate and urate concentrations. Total TRAP, however, recovered to initial levels after 20 hours, due to an increase in unidentified antioxidants. The second course of chemotherapy yielded a decrease in unidentified antioxidant proportions only 20 hours following the second infusion of adriamycin. These results are in accordance with previous studies showing the formation of oxidants with the use of anthracyclines. Evidence suggests that the as yet unidentified component of TRAP (UNID) increases during the oxidative stress caused by anthracycline based chemotherapy.

Serum antioxidant status in streptozotocin-induced diabetic rat.

To determine whether decrease in serum antioxidants contributes to the increased oxidative stress, we measured the antioxidant activity (AOA), total peroxyl radical-trapping antioxidant parameter (TRAP), and their component individual antioxidants in the serum of diabetic rats. AOA was assayed as the ability of serum to inhibit lipid autoxidation in brain homogenates. TRAP was assayed as the ability to delay lipid peroxidation induced by an azo initiator. Antioxidants measured were ceruloplasmin, unsaturated iron binding capacity (UIBC) and albumin as components of AOA; and uric acid, protein sulfhydryl and alpha-tocopherol as components of TRAP. AOA was decreased markedly in the diabetics due to the decrease in ceruloplasmin, UIBC and albumin. On the other hand, the directly measured TRAP in the diabetics was unaltered. Uric acid and alpha-tocopherol were increased in the diabetics. However, decrease in unidentified scavengers offset the increase brought about by those antioxidants These results suggest that the decreased metal binding capacity contributes to the increased oxidative stress in the diabetic rats.