Grape epicatechin conjugates prevent erythrocyte membrane protein oxidation

Epicatechin conjugates obtained from grape have shown antioxidant activity in various systems. However, how these conjugates exert their antioxidant benefits has not been widely studied. We assessed the activity of epicatechin and epicatechin conjugates on the erythrocyte membrane in the presence and absence of a peroxyl radical initiator, to increase our understanding of their mechanisms. Thus, we studied cell membrane fluidity by fluorescence anisotropy measurements, morphology of erythrocytes by scanning electron microscopy, and finally, red cell membrane proteins by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Our data showed that incubation of red cells in the presence of epicatechin derivatives altered membrane fluidity and erythrocyte morphology but not the membrane protein pattern. The presence in the medium of the peroxyl radical initiator 2,2'-azobis(amidinopropane) dihydrochloride (AAPH) resulted in membrane disruptions at all levels analyzed, causing changes in membrane fluidity, cell morphology, and protein degradation. The presence of antioxidants avoided protein oxidation, indicating that the interaction of epicatechin conjugates with the lipid bilayer might reduce the accessibility of AAPH to membranes, which could explain in part the inhibitory ability of these compounds against hemolysis induced by peroxidative insult.     

Ability of surface-active antioxidants to inhibit lipid oxidation in oil-in-water emulsion

Lipid oxidation in dispersed lipids is prevalent at the oil-water interface where lipid hydroperoxides are decomposed into free radicals by transition metals. Free radical scavenging antioxidants are believed to be most effective in lipid dispersions when they accumulate at the oil-water interface. The surface activity of antioxidants could be increased by their conjugation to hydrocarbon chains. In this study, p-hydroxyphenylacetic acid (HPA) was conjugated with either a butyl or dodecyl group. The HPA conjugates were more effective at decreasing interfacial tension than unconjugated HPA, indicating that they were able to adsorb at lipid-water interfaces. However, free HPA was a more effective antioxidant than butyl and dodecyl conjugates in Menhaden oil-in-water emulsions as determined by both lipid hydroperoxides and thiobarbituric acid reactive substances. The increased antioxidant activity of free HPA could be due to its more effective free radical scavenging activity and its higher concentration in the lipid phase of oil-in-water emulsions in the presence of surfactant micelles where it can act as a chain-breaking antioxidant.     

Redox-related cytotoxic responses to different casein glycation products in Caco-2 and int-407 cells

Sugar-casein glycation products (GPs) were generated by Maillard reaction (MR) with different monosaccharide sources [e.g., glucose (Glc), fructose (Fru), and ribose (Rib)] and prolonged heating (e.g., 27 days at 55 degrees C) to produce Maillard reaction products (MRPs) that varied in opponent (L, a, b) color measurement and changes in pH, available lysine, and amino-sugar ratio. Theses results signified different rates of three sugar and casein glycation. Sugar-casein GPs from aldohexose, ketohexose, and aldopentose sugar sources were recovered on day 18 of heating and compared for bioactive properties using human embryonic intestinal cell (Int-407) and adenocarcinoma cell (Caco-2) lines. Glu- and Fru-casein GPs produced significant (p < 0.05) decreases in antioxidant superoxide dismutase (SOD), glutathione peroxidase, and glutathione reductase enzyme activities in the Int-407 cell line, whereas no effect on antioxidant enzymes was obtained from Rib-casein GP. Moreover, the Caco-2 cell antioxidant enzyme status was not affected by the presence of sugar-casein GPs, regardless of sugar source. The reduction in antioxidant enzyme activity of Int-407 cells by Glu and Fru- casein GPs corresponded to a significant (p < 0.05) reduction in Int-407 cell viability. In contrast, no change in Caco-2 cell viability was observed with sugar-casein GP. This finding demonstrates that the noted variable cytotoxic, sugar specific effects of casein GP were related to reductions in critical antioxidant enzyme activities. Moreover, the source of intestinal cell line was an important factor to show the effect of sugar-casein GPs on redox-related cytotoxicity.     

Antioxidant activity, cytotoxicity, and DNA information of Glossogyne tenuifolia

This study investigates the antioxidant activity and cytotoxicity of Glossogyne tenuifolia extract on various cancer cell lines. The 5.8s DNA of G. tenuifolia was isolated, and the species of this plant was confirmed by NCBI's DNA database. G. tenuifolia was then extracted with ethanol and separated into several fractions using the partition procedure with water, n-butanol, and ethyl acetate (EA). Among these, the EA fraction most significantly affected the activity of DPPH(*) and superoxide anion scavenging. Additionally, only the EA fraction exhibited cytotoxicity on breast cancer cells (MCF-7 and MDA-MB-231) and liver cancer cells (Hep G2 and Hep 3B). Next, the EA fraction was further separated by column chromatography, and 15 fractions were obtained. Three effective components were isolated and identified separately from the active fractions: oleanolic acid (OA) from fraction 6, luteolin from fractions 8-10, and luteolin-7-glucoside from fraction 12. The test of these three compounds on scavenging activity of DPPH(*) and superoxide anion indicates that luteolin had the highest antioxidant activity, whereas the effect of OA was negligible. Additionally, a synergistic effect between luteolin and luteolin-7-glucoside was observed. Kick-out experiments showed that the activities were vanished or decreased. Especially on MDA-MB-231 and MCF-7 cells, the cytotoxicity completely disappeared when luteolin was eliminated from fractions 8-10. These findings demonstrate that luteolin plays a crucial role in the inhibition of the growth of hepatoma cancer cell lines. Fraction 3, which did not contain luteolin, luteolin-7-glucoside, and oleanolic acid, had cytotoxicity on MDA-MB-231, MCF-7, Hep G2, Hep 3B, and A549, which implies that this fraction contained some other effective ingredients and requires further study. The investigation is currently underway in our laboratory.     

Immunomodulatory activity of a new family of antioxidants obtained from grape polyphenols

We examined the potential antioxidant activity and the immunopharmacological activity of new epicatechin conjugates obtained by depolymerization of grape polymeric flavanols in the presence of cysteamine or cysteine and with or without gallate. The compounds studied were (-)-epicatechin (1), cysteinyl-epicatechin (2), cysteamine-epicatechin (3), (-)-epicatechin gallate (4), cysteinyl-epicatechin gallate (5), and cysteamine-epicatechin gallate (6) When incubated with an erythrocyte suspension, flavanols protected the erythrocyte membrane from hemolysis induced by 2,2'-azobis(2-amidinopropane) dihydrochloride, an azo free-radical initiator. All the epicatechin derivatives tested were more efficient as antioxidant than epicatechin. The most potent antioxidant was compound 6. The compounds were tested for their capacity to modulate IL-1beta and IL-6, which are the main cytokine factors influencing the acute phase of the inflammatory response. (-)-Epicatechin and its related compounds inhibited the production of IL-1beta and IL-6 in whole blood incubated in the presence of Escherichia coli lipopolysaccharide. The most efficient inhibitor of cytokine formation was compound 3.     

Activity and concentration of polyphenolic antioxidants in apple: effect of cultivar, harvest year, and storage conditions

Consumers' increasing interest in the relationship between diet and health is a sign for food producers to pay more attention to potential health-protecting compounds in new product development and food processing. From a production chain perspective the choice of the raw material that is used is important for the health-protecting potential of the end product. Four apple cultivars (Jonagold, Golden Delicious, Cox's Orange, and Elstar), which can be used as fresh apples or in processed apple products, were compared with regard to flavonol, catechins, phloridzin, and chlorogenic acid concentrations and antioxidant activity. Jonagold apples possessed the highest flavonoid concentration and the highest antioxidant activity. To study seasonal differences, apples from three different harvest years were analyzed, but in three cultivars no effect on flavonoid concentration and antioxidant activity was observed. Long-term storage, both at refrigerator temperature and under controlled atmosphere conditions, was found not to influence flavonoid concentration or antioxidant activity.     

Antioxidative activity of heterocyclic compounds found in coffee volatiles produced by Maillard reaction

Typical heterocyclic compounds substituted with various functional groups found in Maillard reaction products were examined for antioxidant activity. Pyrroles exhibited the greatest antioxidant activity among all heterocyclic compounds tested. All pyrroles inhibited hexanal oxidation by almost 100% at a concentration of 50 microg/mL over 40 days. Addition of formyl and acetyl groups to a pyrrole ring enhanced antioxidative activity remarkably. Pyrrole-2-carboxaldehyde, 2-acetylpyrrole, 1-methyl-2-pyrrolecarboxaldehyde, and 2-acetyl-1-methylpyrrole inhibited hexanal oxidation by >80% at 10 microg/mL. Unsubstituted furan exhibited the greatest antioxidant activity among furans tested. Addition of all functional groups used in this study to furan decreased antioxidative activity. The antioxidant activity of thiophene increased with the addition of methyl and ethyl groups, but the addition of formyl or acetyl groups to thiophene decreased antioxidant activity. Thiazoles and pyrazines were ineffective antioxidants at all concentrations tested. Reaction of all heterocyclic compounds with hydrogen peroxide resulted in the formation of various oxidized products.     

Activity and concentration of polyphenolic antioxidants in apple juice. 1. Effect of existing production methods

Apples are an important source of flavonoids in the human diet. The effect of processing apples into juice on polyphenolic antioxidant content and activity is described. Raw juice obtained from Jonagold apples by pulping and straight pressing or after pulp enzyming had an antioxidant activity that was only 10 and 3%, respectively, of the activity of the fresh apples. The levels of flavonoids and chlorogenic acid in the juice were reduced to between 50% (chlorogenic acid) and 3% (catechins). Most of the antioxidants were retained in the pomace rather than being transferred into the juice. Apparently, most of the antioxidant compounds are absorbed to the solid matter of the pomace. In apple juice, 45% of the total measured antioxidant activity could be ascribed to the analyzed antioxidants. For three apple cultivars tested (Elstar, Golden Delicious, and Jonagold), the processing methods had similar effects. The results indicate that processing can have a major impact on the bioactivity of products.     

Effect of processing on the phytochemical profiles and antioxidant activity of corn for production of masa, tortillas, and tortilla chips

The phytochemical profiles (total phenolics, anthocyanins, ferulic acid, carotenoids) and antioxidant activities of five types of corn (white, yellow, high carotenoid, blue, and red) processed into masa, tortillas, and tortilla chips were studied. The nixtamalization process significantly (p < 0.05) reduced total phenolics and antioxidant activities when compared to raw grains. Nixtamalized grains exhibited higher concentration of free phenolics and soluble conjugated ferulic acid and had lower concentrations of bound phenolics and ferulic acid than unprocessed grains. Among processed products, there was little difference in the phytochemical contents and antioxidant activities. Among types of corn, the highest concentrations of total phenolics, ferulic acid, and antioxidant activity were observed in the high-carotenoid genotype followed by the regular yellow counterpart. The white corn contained the lowest amount of total phenolics and antioxidant activity. The pigmented blue corn had the highest anthocyanin concentration followed by the red counterpart. These findings suggest that lime-cooking significantly reduced the phytochemical content of nixtamalized products but released phenolics and ferulic acid.     

Structural identification of new curcumin dimers and their contribution to the antioxidant mechanism of curcumin

As a part of the research project on the elucidation of the chain-breaking antioxidant mechanism of natural phenolics against the oxidation of food components, curcumin, a main turmeric pigment, was investigated. A relatively high concentration of curcumin gave three dimers as radical termination products in addition to the coupling products with curcumin and the lipid hydroperoxide. The structural analysis of these dimers and quantitative analysis of their production rates revealed that radical-radical termination mainly occurred at the 2-position of curcumin. The contribution of the pathway for production of these dimers to the antioxidant mechanism of curcumin was estimated from the concentration-dependent data of the antioxidant activity and formation rates of these termination products. The A-A termination (dimer formation) was estimated to contribute at least about 40% of the entire antioxidant process against ethyl linoleate oxidation.     

Antioxidant activity of peptides obtained from porcine myofibrillar proteins by protease treatment

Hydrolysates obtained from porcine myofibrillar proteins by protease treatment (papain or actinase E) exhibited high antioxidant activity in a linolenic acid peroxidation system induced by Fe(2+). Hydrolysates produced by both papain and actinase E showed higher activities at pH 7.1 than at pH 5.4. The antioxidant activity of the papain hydrolysate was almost the same as that of vitamin E at pH 7.0. These hydrolysates possessed 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity and chelating activity toward metal ions. Antioxidant peptides were separated from the papain hydrolysate by ion exchange chromatography. The acidic fraction obtained by this method exhibited higher activity than the neutral or basic fractions. Antioxidant peptides in the acidic fraction were isolated by high-performance liquid chromatography on an ODS column and shown to possess the structures DSGVT, IEAEGE, DAQEKLE, EELDNALN, and VPSIDDQEELM. The DAQEKLE peptide showed the highest activity among these peptides.     

祁白术多酚酶法提取工艺优化及其抗氧化、抑制黑色素合成活性

为考察祁白术多酚(polyphenols from Atractylodes macrocephala Koidz grown in Qimen, AMP)的可利用性,采用响应面分析法优化酶法提取工艺,并研究AMP的抗氧化、抑制黑色素合成活性。结果表明,在料液比1:30 g·mL^-1、酶解时间20 min的条件下,当纤维素酶添加量为1.35%、酶解温度为44℃、pH值为4.7、搅拌转速为670 r·min^-1时,AMP提取量最高,为26.58±0.23 mg·g^-1。统计学分析显示,所选响应面模型拟合较好,优化后的提取工艺条件合理可行。AMP具有较好的抗氧化活性,其总还原力、对1,1-二苯基-2-三硝基苯肼(DPPH)和2,2-联氮-二(3-乙基-苯并噻唑-6-磺酸)二铵盐(ABTS)自由基清除作用随其浓度的增加而增强。当AMP浓度为0~0.02 mg·mL^-1时,对B16细胞无毒性作用。与α-MSH模型组相比,AMP显著下调了细胞内酪氨酸酶活性(P<0.05),并呈浓度依赖性地抑制细胞内黑色素的合成(P<0.05);当AMP浓度为0.02 mg·mL^-1时,对细胞内酪氨酸酶活性及黑色素合成抑制率分别为30.11%、43.35%,阳性对照熊果苷(0.1 mg·mL^-1)对细胞内酪氨酸酶活性及黑色素合成的抑制率分别为22.03%、39.77%,表明AMP对黑色素生成的抑制效果强于熊果苷。本研究结果为祁白术多酚的综合开发利用提供了依据。     

复合抗氧化剂浸渍处理提升冻藏罗非鱼片品质

为了提高罗非鱼片的冻藏稳定性,以硫代巴比妥酸值(thiobarbituric acid,TBA)、Ca2+-ATPase活性、盐溶性蛋白含量、挥发性盐基氮(total volatile basic nitrogen,TVB-N)含量及质构等为试验指标,进行了维生素E、异抗坏血酸钠及茶多酚等抗氧化剂溶液浸渍处理对冻藏罗非鱼片品质影响的单因素试验,并在单因素试验的基础上进行抗氧化剂复配的正交优化试验。获得了不同的抗氧化剂浸渍浓度对冻藏罗非鱼片品质的影响规律,以主观赋权法对TBA值、Ca2+-ATPase活性、盐溶性蛋白含量、TVB-N和质构(硬度和弹性)5个指标分别赋权后计算各指标的综合评分,确定了各抗氧化剂的较佳浸渍浓度及最佳复合抗氧化剂的组合配方。结果表明,维生素E溶液和异抗坏血酸钠溶液的较佳质量浓度均为6 g/L,茶多酚溶液的较佳质量浓度为4 g/L,在较佳质量浓度条件下各品质指标综合评分与蒸馏水浸泡60 min的对照组相比分别提高了29.11%,18.46%和46.28%。参照成本分析与较优品质结合优化组合的复合抗氧化剂的配方为:异抗坏血酸钠质量浓度为4 g/L、维生素E为质量浓度2 g/L和茶多酚质量浓度为3 g/L,经此配方处理后的冻藏罗非鱼片对应的品质指标分别为TBA值0.1248 mg/kg,Ca2+-ATPase活性3.12μmol/(mg·h),盐溶性蛋白质量分数为77.72 mg/g,均优于单一抗氧化剂的作用,与蒸馏水浸泡60 min的对照组相比,各指标提升幅度为:Ca2+-ATPase活性提高52.20%,盐溶性蛋白质量分数提高16.78%,TBA值下降46.07%。研究结果为复合抗氧化剂的开发及其在冻藏水产品中的应用提供参考。     

Nonenzymatic antioxidative and antiglycative effects of oleanolic acid and ursolic acid

Oleanolic acid and ursolic acid are two triterpenes presented in several herbs. This study analyzed the content of oleanolic acid and ursolic acid in eight locally available fresh herbs, also examined several nonenzymatic antioxidant activities of these two triterpenes, and used a liposome system to evaluate the influence of temperature and pH upon the antioxidant property of these two triterpenes. The impact of these two triterpenes on the production of nonenzymatic glycative products, pentosidine and carboxymethyllysine (CML), was also evaluated. alpha-Tocopherol was used for comparison. Results showed that the content of oleanolic acid and ursolic acid in glossy privet fruit and hawthorn fruit varied from season to season and was in the range of 200-650 microg/g of fresh weight. Both oleanolic acid and ursolic acid possessed greater antioxidant activity against 2,2'-azobis-(2-amidinopropane) dihydrochloride and less antioxidant activity against 2,2'-azobis(2,4-dimethylvaleronitrile) when compared with alpha-tocopherol at equal concentration (P <0.05). At 75 and 100 degrees C, oleanolic acid exhibited greater antioxidant activity than alpha-tocopherol and ursolic acid (P <0.05). At pH 2 and pH 4, oleanolic acid and ursolic acid showed greater antioxidant activity than alpha-tocopherol (P <0.05). These two triterpenes also exhibited a dose-dependent effect in superoxide anion scavenging activity, chelating effect, xanthine oxidase inhibition activity, and reducing power (P <0.05). Oleanolic acid significantly and dose-dependently inhibited pentosidine and CML formation (P <0.05). Ursolic acid also significantly suppressed CML formation (P <0.05). These data support that these two triterpenes possessed nonenzymatic antioxidative and antiglycative properties.     

Efficient preparation of catechin thio conjugates by one step extraction/depolymerization of pine (Pinus pinaster) bark procyanidins

The skin penetrating antioxidant cysteamine derivative of (-)-epicatechin as well as other thio conjugates were efficiently obtained with high yields from pine (Pinus pinaster) bark by simultaneous one pot extraction and depolymerization using water and cysteamine hydrochloride. The influence of the concentration of bark, acid, and cysteamine, as well as the reaction time on the total conversion, was studied. The total conversion into the epicatechin and catechin conjugates was as high as 47 g/kg pine bark with 1666 g cysteamine/kg bark and 28 g/kg with 166 g cysteamine/kg bark. A fast cleanup step by absorption/desorption on XAD-16 greatly facilitated further purification of the active major component. At a pilot scale, 4beta-(2-aminoethylthio)epicatechin (1) (conversion 263 g, purity 35% by reversed phase high-performance liquid chromatography/weight) was obtained from 17 kg of pine bark after simultaneous extraction/depolymerization followed by cleanup with the polymeric resin in approximately 10 h. The results show that pine (P. pinaster) bark is a suitable source of flavanols for the preparation of active thio derivatives. Conditions are given for the fast and efficient preparation of the conjugates.     

Alkyl hydroxytyrosyl ethers show protective effects against oxidative stress in HepG2 cells

Alkyl hydroxytyrosyl ethers (methyl, ethyl, propyl, and butyl ethers) have been synthesized from hydroxytyrosol (HTy) in response to the increasing food industry demand of new lipophilic antioxidants. Having confirmed that these compounds reach portal blood partially unconjugated and thus are effectively absorbed, their potential antioxidant activity was evaluated in the human hepatocarcinoma cell line (HepG2). The effects of 0.5-10 μM alkyl hydroxytyrosyl ethers on HepG2 cell integrity and redox status were assessed as well as the protective effect against oxidative stress induced by tert-butylhydroperoxide (t-BOOH). Cell viability (Crystal violet) and cell proliferation (BrdU assay) were measured as markers of cell integrity, concentration of reduced glutathione (GSH), generation of reactive oxygen species (ROS), and activity of antioxidant enzymes glutathione peroxidase (GPx) and glutathione reductase (GR) as markers of redox status and determination of malondialdehyde (MDA) as a marker of lipid peroxidation. Direct treatment of HepG2 with alkyl hydroxytyrosyl ethers induced slight changes in cellular intrinsic antioxidants status, reducing ROS generation and inducing changes in GPx and GR activities. Pretreatment of HepG2 cells with alkyl hydroxytyrosyl ethers counteracted cell damage induced by t-BOOH, partially after 2 h and completely after 20 h, by increasing GSH and decreasing ROS generation, MDA levels, and antioxidant enzyme (GPx and GR) activity. According to these results the alkyl hydroxytyrosyl ethers show clear protective effects against oxidative stress, related to their lipophilic nature, that are similar to or even higher than those of their precursor, HTy.     

Antioxidant properties of malt model systems

The aim of this study was to investigate the effect of kilning and roasting temperatures on antioxidant activity of malt model systems prepared from combinations of glucose, proline, and ferulic acid. Model systems (initial a(w) = 0.09, 6% moisture) were heated at 60 degrees C for up to 24 h, at 90 degrees C for up to 120 min, and at 220 degrees C for up to 15 min. The antioxidant activity of the glucose-proline-ferulic acid model system increased significantly on heating at 60 degrees C for 24 h or at 90 degrees C for 120 min. In contrast, the glucose-proline, ferulic acid-glucose, and ferulic acid-proline systems presented either nonsignificantly increased or unchanged antioxidant activity. The antioxidant activity of both the glucose-proline-ferulic acid and glucose-proline model systems increased significantly after heating at 220 degrees C for 10 min, followed by a significant decrease at 15 min. The data suggest that (1) at 60 degrees C, ferulic acid reacts with Maillard reaction products, resulting in a significant increase in antioxidant activity; (2) at 90 degrees C, the antioxidant activity of the glucose-proline-ferulic system comes from both ferulic acid and Maillard reaction products; and (3) at 220 degrees C, the major contributors to antioxidant activity in glucose-proline-ferulic acid and glucose-proline systems are glucose-proline reaction products.     

Effect of tocopherols in the antioxidative activity of oxidized lipid-amine reaction products

Phosphatidylethanolamine (PE), phosphatidylcholine (PC), lysine (Lys), and mixtures of them were tested for antioxidative activity in a tocopherol-stripped olive oil (TSO) and the same oil after addition of 250 microg of alpha-tocopherol g of oil/(tocopherol-added olive oil, TAO) to evaluate the role of tocopherol in the antioxidant activity of oxidized lipid-amine products. Neither PE nor PC nor Lys protected TSO when tested alone, but both PE and Lys increased the induction period (IP) of TAO. On the contrary, PE/Lys and PC/Lys mixtures, but not PC/PE mixtures, protected both TSO and TAO. These results were a consequence of both the formation of oxidized lipid-amine products, which were determined by gas chromatography-mass spectrometry after their conversion into volatile derivatives, and a synergism between alpha-tocopherol and the produced compounds. These results were confirmed by analyzing the antioxidative activity of two of the produced carbonyl-amine products: 6-amino-2-(1H-pyrrol-1-yl)hexanoic acid (1) and 2,3-dipalmitoylpropyl 2-(1H-pyrrol-1-yl)ethyl phosphate (2). The hydrophilic compound 1 was more antioxidant than the analogous lipophilic compound 2, and this antioxidative activity was observed in TAO and not in TSO. All these results suggested that antioxidative activity of carbonyl-amine products may be greatly increased with the addition of tocopherols, and those products derived from Lys are more antioxidant in bulk oils than those derived from PE.     

Antioxidant activity of minimally processed red chicory (Cichorium intybus L.) evaluated in xanthine oxidase-, myeloperoxidase-, and diaphorase-catalyzed reactions

Minimally processed red chicory products (Cichorium intybus L. var. silvestre) were studied for their polyphenol content and antioxidant activity evaluated by using the synthetic 2,2-diphenyl-1-(2,4,6-trinitrophenyl)hydrazyl radical and three model reactions catalyzed by relevant enzymatic sources of reactive oxygen species, namely, xanthine oxidase, myeloperoxidase, and diaphorase. Products were analyzed at the time of production and after storage at 4 degrees C within either a gas permeable film or a gas barrier film. The antioxidant activity and contents of hydroxycinnamic acids and flavonoids decreased by less than 20% during storage of the minimally processed red chicory products. Total phenolics were significantly correlated with the antioxidant activity evaluated with both the synthetic radical and the enzyme-catalyzed reactions. On a molar basis, red chicory phenolics were as efficient as the reference compound Trolox in scavenging the synthetic radical. However, red chicory phenolics had a much higher inhibitory activity than Trolox in the model enzymatic systems.     

Antioxidant activity of tomato products as studied by model reactions using xanthine oxidase, myeloperoxidase, and copper-induced lipid peroxidation

The antioxidant content and activity of commercial tomato products differing in variety and processing were studied. Two procedures for extracting hydrophilic and lipophilic antioxidants, namely, two-step 0.1 M phosphate buffer (pH 3.0 and 7.4) extraction and tetrahydrofuran extraction followed by petroleum ether fractionation, were developed. Carotenoids (lycopene, beta-carotene, and lutein) and ascorbic acid were analyzed by HPLC with spectrophotometric and electrochemical detectors, respectively. Total phenolics were determined by using the Folin-Ciocalteu reagent. The antioxidant activity was studied by the following three model systems: (a) the xanthine oxidase (XOD)/xanthine system, which generates superoxide radical and hydrogen peroxide; (b) the myeloperoxidase (MPO)/NaCl/H(2)O(2) system, which produces hypochloric acid; and (c) the linoleic acid/CuSO(4) system, which promotes lipid peroxidation. Results showed that the hydrophilic and lipophilic fractions of all tomato products were able to affect model reactions, whatever reactive oxygen species and catalysts were used to drive oxidation. In the XOD/xanthine system both the hydrophilic and lipophilic fractions displayed an inhibitory activity. The hydrophilic fractions were more effective (I(50) ranging from 680 to 3200 microg, dry weight) than the lipophilic fractions (I(50) ranging from 4000 to 7750 microg, dry weight). In the MPO/NaCl/H(2)O(2) system the hydrophilic fractions inhibited oxidation (I(50) ranging from 2300 to 2900 microg, dry weight), whereas the lipophilic fractions had a lower inhibitory effect at the same concentration. Conversely, in the copper-catalyzed lipid peroxidation only the lipophilic fractions were effective (I(50) ranging from 1030 to 2100 microg, dry weight), whereas the hydrophilic fractions had a pro-oxidant effect in the same concentration range. The extent of inhibition varied according to the tomato sample in the superoxide and hydrogen peroxide generating system and in lipid peroxidation, but was substantially the same in the HClO generating system. Fresh tomato varieties differed considerably in the antioxidant activities of their hydrophilic and lipophilic fractions. Processed tomatoes showed a significantly lower antioxidant activity than fresh tomatoes in their hydrophilic fractions but had a high antioxidant activity in their lipophilic fractions. Because the oxidative reactions produced by the above-mentioned model systems are also involved in the pathogenesis of several chronic diseases, the antioxidant activity of tomato fractions might be related to their in vivo activity. Hence, these measurements may be used for optimizing tomato technologies.