Reactive Oxygen Species, Aging, and Antioxidative Nutraceuticals

J. Lee, N. Koo, and D.B. Min ABSTRACT: The important roles of reactive oxygen species in diseases related to aging and the necessity and benefits of antioxidative nutraceuticals in the prevention of diseases and promotion of healthy aging have been extensively reported in recent years. Oxygen is an essential component of living organisms. The generation of reactive oxygen species such as superoxide anion, hydrogen peroxide, hydroxyl radicals, and singlet oxygen is inevitable in aerobic metabolism of the body. Reactive oxygen species cause lipid oxidation, protein oxidation, DNA strand break and base modification, and modulation of gene expression. In the past several years, unprecedented progress has been made in the recognition and understanding of roles of reactive oxygen species in many diseases. These include atherosclerosis, vasospasms, cancers, trauma, stroke, asthma, hyperoxia, arthritis, heart attack, age pigments, dermatitis, cataractogenesis, retinal damage, hepatitis, liver injury, and periodontis, which are age-related. The body protects itself from the potential damages of reactive oxygen species. Its first line of defense is superoxide dismutases, glutathione peroxidases, and catalase. Scientists have indicated that antioxidant nutraceuticals supplied from daily diets quench the reactive oxygen species or are required as cofactors for antioxidant enzymes. Nutraceuticals play significant roles in the prevention of a number of age-related diseases and are essential for healthy aging. Epidemiological studies also reported the relevance of antioxidative nutraceuticals to health issues and the prevention of age-related diseases. Health-conscious consumers have made antioxidative nutraceuticals the leading trend in the food industry worldwide in recent years.

Aging is the accumulation process of diverse detrimental changes in the cells and tissues with advancing age, resulting in an increase in the risks of disease and death (Harman 2000). Aging is influenced by many factors, including lifestyle, environmental conditions, and genetic disposal (Spiteller 2001). With increasing age, the oxidation products from lipids, nucleic acids, proteins, sugars, and sterols are found to increase (Ashok and Ali 1999). The main causes of the aging process seem to be related to reactive oxygen species and free radicals, such as superoxide anion, hydrogen peroxide, hydroxyl radicals, and singlet oxygen. Mitochondria, which consume more than 90% of the oxygen in aerobic living organisms, are the main reactive oxygen species and free radical source. Oxygen in mitochondria is reduced to water by 4 sequential steps (Ames and others 1993). Perhydroxyl radical (HO2·) or its ionized form, superoxide anion (·O2–), is the first reduced intermediate of oxygen. Hydrogen peroxide (H2O2) and hydroxyl radical (·OH) are inevitable intermediates from oxygen to water reduction steps in body. Approximately 1% to 5% of the oxygen consumed by mitochondria is reduced and converted to these reactive oxygen species (Ames and others 1993).
Author Lee is currently with the Dept. of Food Science and Technology, Seoul National Univ. of Technology, Seoul, Korea. Author Koo is currently with the Dept. of Food and Nutrition, DaeJeon Univ., DaeJeon, Korea. Author Min is with the Dept. of Food Science and Technology, The Ohio State Univ., 2015 Fyffe Road, Columbus, OH 43210. Direct inquiries to author Min (E-mail:

Harman (2000) suggested that initially generated superoxide anion and hydrogen peroxide are the main reactive oxygen species causing the oxidation of cells and tissues. Superoxide anion itself is not a strong oxidant, but it reacts with protons in water solution to form hydrogen peroxide, which can serve as a substrate for the generation of hydroxyl radicals and singlet oxygen (Stief 2003). Hydroxyl radicals are strong oxidants and can abstract a hydrogen atom from any carbon–hydrogen bond and oxidize the compound. For example, linoleic acids are mainly located in glycerolipids and phopholipids of cell membranes; therefore, cell membranes are easily oxidized and lose their functionality during the aging process. Prooxidative enzymes such as lipoxygenase can generate free radicals (Spiteller 2001). Lipoxygenase can react with free forms of fatty acids, which can be released from glycerides by membranebound phopholipase A2. Environmental sources, such as ultraviolet (UV) irradiation, ionizing irradiation, and pollutants, also produce reactive oxygen species (Halliwell 1997). Injured cells and tissues can stimulate the generation of free radicals (Spiteller 2001). Reactive oxygen species can be formed in foods through lipid oxidation and photosensitizers exposed to light (Boff and Min 2002). Nonenzymatic lipid oxidation requires the presence of free forms of bivalent metal ions such as copper and iron, which are not common for healthy adults (Gutteridge and Halliwell 1993). It has been assumed that free forms of irons are generated by the decompositions of ion-containing natural sources such as hemoglobin and ferritin (Halliwell 1997). Enzymatic and nonenzymatic antioxidant systems in the body,

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aromatic compounds or compounds with carbon–carbon multiple bonds undergo addition reactions with hydroxyl radicals. polypeptides. Reactive oxygen species or free radicals in biological systems can be formed by prooxidative enzyme systems. cancers. regulate the balance of reactive oxygen species with antioxidants (Thomas 1995. highly reactive. asthma. and lipid oxidation products support the fact that reactive oxygen species and free radicals are involved in the aging process (Halliwell 1997. 3. Mitochondria generate energy using 4 electron chain reactions. Nitric oxide acts as a neurotransmitter and an important mediator of the immune response (Fang and others 2002). Hydrogen peroxide is a COMPREHENSIVE REVIEWS IN FOOD SCIENCE AND FOOD SAFETY—Vol. Oxygen-centered radicals are superoxide anion (·O2–). The resulting radical can undergo further reactions. cataractogenesis. catalase. Hydrogen peroxide is the least reactive molecule among reactive oxygen species and is stable under physiological pH and temperature in the absence of metal ions. A free radical exists with one or more unpaired electron in atomic or molecular orbital. retinal damage. gene expression. such as reaction with oxygen. Reactive oxygen species in biological systems are related to free radicals. initiate the oxidation of polyunsaturated fatty acids (PUFA). glutathione peroxidase. hyperoxia. Enzymes such as amino acid oxidase and xanthine oxidase also produce hydrogen peroxide from superoxide anion. heart attack. DNA. to give peroxyl radical. Superoxide anion may serve as a cell growth regulator (Halliwell 1997). As aging proceeds. alkoxyl radical (RO·). including the activation of nuclear transcription factors. including atherosclerosis. Stief 2003). Consumption of fruits and vegetables containing high amounts of antioxidative nutraceuticals has been associated with the balance of the free radicals/antioxidants status. and glycoxidation (Halliwell 1997. inflammation. and a defense mechanism to target tumor cells and microbial infections (Simon and others 2000). arthritis. so that in competition addition is often favored. nitric dioxide (NO2·). The superoxide anion can react with nitric oxide (NO·) and form peroxynitrite (ONOO –). Simon and others 2000). or so-called oxidative stress. Hydroxyl radicals also add readily to double bonds. The age-associated increases in oxidized proteins. or decompose to phenoxyltype radicals by water elimination. Nitric oxide is one of the most widespread signaling molecules and participates in every cellular and organ function in the body. air pollutants. resulting in the hydroxylated free radicals. Reactive Oxygen Species Reactive oxygen species can be classified into oxygen-centered radicals and oxygen-centered nonradicals. and periodontis (Figure 1) (Cohen and others 2000. The prevalent free radical states. or singlet oxygen (2 ·O2– + 2H+ → H2O2 + O2) in living systems (Stief 2003). which helps to minimize the oxidative stress in the body and to reduce the risks of cancers and cardiovascular diseases (Kaur and Kapoor 2001). age pigments. The barrier to the addition of hydroxyl radicals to double bonds is less than that of hydrogen abstraction. vasospasms. reducing oxygen to water. In general. Hydrogen peroxide is highly diffusible and crosses the plasma membrane easily. and DNA. lipid oxidation. liver injury. and the ability to remove deleterious reactive oxygen species and free radicals decreases. and energized molecules. Hydrogen peroxide (H2O2) Figure 1—Clinical conditions involving reactive oxygen species 22 Hydrogen peroxide can be generated through a dismutation reaction from superoxide anion by superoxide dismutase. Other reactive species are nitrogen species such as nitric oxide (NO·). proteins. The superoxide anion plays an important role in the formation of other reactive oxygen species such as hydrogen peroxide. Some of the electrons escaping from the chain reaction of mitochondria directly react with oxygen and form superoxide anions (Harman 2000). which can generate toxic compounds such as hydroxyl radical and nitric dioxide (ONOO– + H+ → ·OH + ·NO2) (Halliwell 1997). and water-soluble vitamin C. 2004 . When a hydroxyl radical reacts with aromatic compounds. oxidized DNA. stroke. a hydroxyl radical abstracts a hydrogen atom from the weakest C–H bond to yield a free radical (Korycka-Dahl and Richardson 1978). trauma.CRFSFS: Comprehensive Reviews in Food Science and Food Safety including superoxide dismutase. the efficiency of antioxidant defense systems lowers. Oxygen-centered nonradicals are hydrogen peroxide (H2O2) and singlet oxygen (1O2). proteins. hydroxyl radical. hepatitis. and peroxyl radical (ROO·). sterol oxidation products. Benign functions of free radicals have been reported. irradiation. Singlet oxygen can attack various pathogens and induce physiological inflammatory response (Stief 2003). Superoxide anion is an initial free radical formed from mitochondrial electron transport systems. resulting in hydroxycyclohexadienyl radical (Padmaja and Madison 1999). hydroxyl radical (·OH). carotenes. it can add on across a double bond. Wickens 2001). In saturated compounds. Free radicals are generally unstable. smoking. Clinical studies reported that reactive oxygen species are associated with many agerelated degenerative diseases. dermatitis. lipid-soluble vitamin E. The resulting radicals can react with oxygen and generate other free radicals. Rikans and Hornbrook 1997). Packer and Weber 2001). especially thiamine and guanosine (Ashok and Ali 1999). and sterols. Superoxide anion (·O2–) Superoxide anion is a reduced form of molecular oxygen created by receiving one electron (Figure 2). Hydroxyl radicals react with lipid. Hydroxyl radical (OH) Hydroxyl radical is the most reactive free radical and can be formed from superoxide anion and hydrogen peroxide in the presence of metal ions such as copper or iron (·O2– + H2O2 → ·OH + OH– + O2). Hydroxyl radicals have the highest 1-electron reduction potential (2310 mV) and can react with everything in living organisms at the 2nd-order rate constants of 109 to 1010/M/s (Korycka-Dahl and Richardson 1978). and peroxynitrite (OONO–) (Halliwell and others 1995. even though there are nonradical compounds in reactive oxygen species such as singlet oxygen and hydrogen peroxide. Reactive oxygen species also have been known to induce apoptosis of cells (Simon and others 2000).

having more than 1000 mV of standard reduction potential (Decker 1998). . Enzymatic formation Figure 2—Molecular orbitals of singlet oxygen and superoxide anion Prooxidative enzymes. Singlet oxygen Peroxyl and alkoxyl radicals Singlet oxygen is a nonradical and excited status. which can be formed from peroxynitrite-mediated reactions with amino acids. Singlet oxygen can be formed from hydrogen peroxide. 12–. can deplete the concentration of ascorbic acid and uric acid. 2003). Nitric dioxide adds to double bonds and abstract labile hydrogen atoms initiating lipid peroxidation and production of free radicals. Nitric oxide. polluted air and smoking (Noguchi and Niki 1999). It also oxidizes ascorbic acid (Papas 1999a). viruses. resulting in chain reaction. In the human organism. has been found in age-associated tissues (Knight 1999).Antioxidative nutraceuticals . Hydrogen peroxide can degrade certain heme proteins. weak oxidizing and reducing agent and is thus regarded as being poorly reactive. with therapeutic potency against various pathogens such as microbes. Irradiation of UV light or the presence of transition metal ions can cause homolysis of peroxides to produce peroxyl and alkoxyl radicals (ROOH → ROO·+ H·. Yamazaki and others 1999). Hydrogen peroxide can produce singlet oxygen through reaction with superoxide anion or with HOCl or chloroamines in living systems (Stief 2000. Nitrotyrosine. 2004—COMPREHENSIVE REVIEWS IN FOOD SCIENCE AND FOOD SAFETY . including NADPH–oxidase (Babior 1999). Peroxyl and alkoxyl radicals are good oxidizing agents. There are 3 major mammalian lipoxygenases: 5–. and initiate lipid peroxidation (Halliwell 1996). Oxidation of cholesterol by singlet oxygen results in formation of 5 –OOH (3 –hydroxy– 5 –cholest–6–ene–5–hydroperoxide). singlet oxygen has been known to be involved in cholesterol oxidation (Girotti and Korytowski 2000). Peroxynitrite (OONO–) can cause direct protein oxidation and DNA base oxidation and modification acting as a “hydroxyl radical-like” oxidant (McVean and others 1999). Compared with other reactive oxygen species. Lipoxygenase needs free PUFA. Oxidation and degradation of cholesterol by singlet oxygen was observed to be accelerated by the co-presence of fatty acid methyl ester. 3. The electrons in the antibonding orbitals of singlet oxygen are paired. lipoxygenase can convert PUFA into hydroperoxides (Spiteller 2001). Lysolecithins change the cell membrane structures. Takayama and others (2001) reported that metastable phosphatidylcholine hydroperoxides present in the living organism produced singlet oxygen during their breakdown in the presence of Cu 2+ in the dark. and 15– 23 Vol. Nitric oxide and nitric dioxide Nitric oxide (NO·) is a free radical with a single unpaired electron. Nitric oxide itself is not a very reactive free radical. or with HOCl or chloroamines in cells and tissues (Stief 2003). Once Fe2+ oxidized to Fe3+. the reaction between lipid radicals and oxygen. which are not present in healthy tissue. NO–synthase (Stuehr and others 1990). They can abstract hydrogen from other molecules with lower standard reduction potential. but the overproduction of NO is involved in ischemia reperfusion. and free PUFA are oxidized to form lipid hydroperoxides. Decomposition of alkyl peroxides (ROOH) also results in peroxyl (ROO·) and alkoxyl (RO·) radicals. Some peroxyl radicals break down to liberate superoxide anion or can react with each other to generate singlet oxygen (Halliwell and Gutteridge 1985). ROOH + Fe3+ → ROO· + Fe2+ + H+). for example. Peroxynitrite appears to be an important tissue-damaging species generated at the sites of inflammation (Papas 1999a) and has been shown to be involved in various neurodegenerative disorders and several kidney diseases (Knight 1999). Nitric oxide is formed from L-arginine by NO synthase (Fang and others 2002). singlet oxygen is rather mild and nontoxic for mammalian tissue (Stief 2003). The significance of peroxynitrite as a biological oxidant comes from its high diffusibility across cell membranes (Knight 1999). Very often the alkyl radical formed from this reaction can react with oxygen to form another peroxyl radical. Lipoxygenase with Fe2+ ion is inactivated status. and 6 –OOH (Foote 1991. Peroxyl radicals (ROO·) are formed by a direct reaction of oxygen with alkyl radicals (R·). . 5 –OOH (3 –hydroxy– 5 –cholest–6–ene–5–hydroperoxide). However. Lipoxygenase generates free radicals. This reaction is frequently observed in the propagation stage of lipid peroxidation. and cancer cells (Stief 2003). Peroxynitrite Reaction of NO and superoxide anion can generate peroxynitrite (O2– + NO· → OONO–). Nitric dioxide (NO2·) is formed from the reaction of peroxyl radical and NO. exposed in human blood plasma. Membrane-bound phospholipase produces PUFA and lysolecithins. Peroxynitrite is a cytotoxic species and causes tissue injury and oxidizes low-density lipoprotein (LDL) (Halliwell 1997). Hydrogen peroxide can generate the hydroxyl radical in the presence of metal ions and superoxide anion (·O2– + H2O2 → ·OH + OH– + O2) (Halliwell 1997). or the cytochrome P–450 chain (Stief 2000). and neurodegenerative and chronic inflammatory diseases such as rheumatoid arthritis and inflammatory bowel disease. The molecular orbital of singlet oxygen is shown in Figure 2. to release iron ions. singlet oxygen is both a signal and a weapon. which reacts with superoxide anion. 6 –OOH. can generate reactive oxygen species. such as hemoglobin. Aromatic alkoxyl and peroxyl radicals are less reactive than respective open chain radicals because of the delocalization of electrons in the ring.

Peroxynitrite (ONOO–) oxidizes essential –SH groups of proteins. and inflammation. 2–oxohistidine. protein–pro24 Figure 3—Oxidation mechanisms of proteins (a) and protein carbonyl formation from aldehydes (b) COMPREHENSIVE REVIEWS IN FOOD SCIENCE AND FOOD SAFETY—Vol. including disease. human urine. which suggests that this enzyme may be involved in the in vivo formation of oxidized lipids (Knight 1999). and nucleic acids leading to structural modification. 2004 . which are ubiquitous in body systems. and the generation of carbonyl derivatives of amino acids are some examples of protein modifications (Berlett and Stadtman 1997). 4–Hydroxy–2–nonenal from lipid oxidation can react with protein lysine. 3. in vitro exposure of enzymes to reactive oxygen species induces changes in enzymatic activity. Nedeljkovic and others 2003). Modification of protein is mainly initiated by hydroxyl radicals. As age processing increases. which are associated with aging (Figure 1) (Packer and Weber 2001). Halliwell 1997. and its protonated form (HO2–) determines the pathways of protein oxidation processes. the oxidization of histidine to 2–oxohistidine in the metalbinding site of proteins. immune.CRFSFS: Comprehensive Reviews in Food Science and Food Safety lipoxygenases (Hari and others 2000). and molecular mobility of lipids. leading to the oxidation of amino acid side chains. Clinical studies reported that reactive oxygen species are associated with many degenerative diseases. Protein oxidation and aging Reactive oxygen species can attack proteins and produce carbonyls and other amino acid modifications. inborn processes. phospholipid. Nitrotyrosines. including methionine sulfoxide. Stadtman 2000). and oxidative modification of LDLs has been reported to be involved with the development of atherosclerosis and cardiovascular disease (Frei 1995). and NADH:NAD+ tend to shift to more prooxidant values (Halliwell and others 1995. NADPH: NADP+. and ischemia reperfusion injury (Stadtman 1992. Free radical theories are based on the chemical characteristics of free radicals. number of SH groups. liver disease. These enzymes can oxidize arachidonic acid. Oxidative stress is caused by antioxidant-deficient diets or by increased production of reactive oxygen species by environmental toxins such as those caused by smoking or by inappropriate activation of phagocytes such as with chronic inflammatory disease (Halliwell 1997). Low-density lipoproteins are complicated structures. and resistance to thermodenaturation decrease. transport systems. heat stability. platelets. For example. Induction of 3–chlorotyrosine from tyrosine by hypochlorous acid. is synthesized from L–arginine by NO synthetase in mitochondria and causes oxidation of protein. Free radicals or reactive oxygen species in the body can cause lipid oxidation. Berlett and Stadtman 1997). and free radicals. oxidized DNA such as 8–hydroxydeoxyguanosine. superoxide anion. Oxidized cholesterol or fatty acid moieties in the plasmatic LDL can develop atherosclerosis (Rikans and Hornbrook 1997. DNA strand break and base modification. and oxidative damaged proteins with carbonyl modifications and loss of protein –SH group. immune dysfunction. The availability of oxygen. enzyme activities. and proteolysis susceptibility similar to those that occur during aging (Berlett and Stadtman 1997). and cystein residues (Figure 3b). and protein nitration of tyrosine or tryptophane residues (Virag and others 2003). tein cross linkage. products of reactive nitrogen species on tyrosine. have been detected in atherosclerotic lesions. A serious imbalance between reactive oxygen species and antioxidants causes oxidative stress. and old animals are more susceptible to protein damage during oxidative stress such as hydro- Reactive Oxygen Species and Aging Theories explaining aging processes are diverse. and reactive oxygen species can accelerate lipid oxidation (Boff and Min 2002). 15–Lipoxygenase has been identified within atherosclerotic lesions. and leukocytes and airway cells. protein oxidation. both involving generation of free radicals (Halliwell 1997). the oxidation of thiol groups. and protein peroxides. atherosclerosis. histidine. The increases of lipid oxidation products are found in diabetes. Girotti 1998. Cell membranes are phospholipid bilayers with extrinsic proteins and are the direct target of lipid oxidation (Girotti 1998). As lipid oxidation of cell membranes increases. atherosclerosis. into hydroperoxyeicosatetraenoic acid. Peroxynitrite-induced protein modifications include protein oxidation on methionine. can react with the free amino group of proteins. and body fluids from patients with chronic inflammatory diseases. Spiteller 2001). increase several times and the ratio of redox couples such as glutathione:oxidized glutathione. and cardiovascular systems. Aging is associated at least partly with oxidative modification of proteins. the polarity of lipid-phase surface charge and formation of protein oligomers increase. Malonaldehyde. Malonaldehyde from lipid oxidation reacts with protein amino groups. apoplexy. respectively (Hari and others 2000). cysteine. and 15–lipoxygenases are in leukocytes and lymphocytes. Protein oxidation affects the alterations of signal transduction mechanisms. a PUFA rich in the central nervous system. Generally accepted aging theories are based on either developmentally programmed aging or damage-accumulation aging. and modulation of gene expression. A high level of lipid oxidation products can be detected in cell degradation after cell injury or disease. tryptophane or tyrosine residues. one of the lipid oxidation products. 12–. Lipid oxidation and aging Lipid oxidation is a free-radical chain reaction. NO·. environment. and protein fragmentation (Figure 3a) (Berlett and Stadtman 1997. which is an intracellular messenger in the nervous. The primary localizations of 5–. which induce dysfunction of immune systems.

As oxidative stress increases. Glutathione peroxidase is the most important hydrogen peroxide–removing enzyme existing in the membrane. the level of the prooxidants against antioxidants increases and the aging process accelerates. and others 1996. dietary supplements. Catalase is involved in cellular detoxification and can convert hydrogen peroxide into water and oxygen (Figure 5). are much higher in long-living species than in short-living ones. Hydrogen-donating nutraceuticals Antioxidative nutraceuticals. convert reactive oxygen species into nonreactive oxygen molecules. Antioxidative nutraceuticals can be antioxidative enzymes. herbal products. depending on the food sources. The functional food market has increased because of the fast growth of the older generation in the United States and their concerns about health-beneficial foods (Dillard and German 2000). and close locations to the reactive oxygen species-producing systems.Antioxidative nutraceuticals . Antioxidative enzymes Antioxidative Nutraceuticals Nutraceuticals or functional foods are any food or food ingredients that may provide beneficial health effects beyond the traditional nutrients they contain (Wildman 2001a). and mutagenesis and/or carcinogenesis can be induced. hydrogen donating compounds. . catalase. However. which can donate hydrogen atoms Figure 4—Formation of 8-hydroxydeoxyguaosine from the reaction of guanosine and hydroxyl radical Figure 5—Antioxidant enzymes and their reaction mechanisms 25 Vol. Consumption of optimal amounts of vitamin A and E increased the average life expectancy of animals (Duthie Antioxidant enzymes. It has been reported that levels and activities of antioxidant enzymes. Proteins showing antioxidant properties are listed in Table 1. slow the aging process. genetically engineered “designer” foods. histones. metal chelators. 3. and processed products such as cereals and soups. the DNA repair system using glycosylase is not enough. catalase. Mitochondrial DNA is susceptible to oxidative damages because of the lack of protective protein. and chemical structures (Wildman 2001b). Nutraceuticals range from isolated nutrients. respectively. nutritional supplements. and singlet oxygen quenchers. If reactive oxygen species and free radicals are the major causes of aging processes. gen peroxide than are younger animals (Agarawal and Sohal 1993). 2004—COMPREHENSIVE REVIEWS IN FOOD SCIENCE AND FOOD SAFETY . antioxidative nutraceuticals can reduce the level of reactive oxygen species and free radicals. Nutraceuticals can be grouped in different ways. If oxidative stress is too great. including superoxide dismutase. The concentration of vitamin E in elderly people (older than 65 years) is lower than that in younger adults. . Hydroxyl radical oxidizes guanosine or thymine to 8–hydroxy–2–deoxyguanosine and thymine glycol. and glutathione peroxidase/reductase. which changes DNA and leads to mutagenesis and carcinogenesis (Figure 4) (Ames and others 1993). concentration of oxidized DNA base increases in humans with chronic inflammatory diseases such as rheumatoid arthritis or under oxidative stresses such as smoking (Halliwell 1997). Altered DNA can be repaired by DNA glycosylase. Antioxidative nutraceuticals can inhibit or slow the formation of free alkyl radicals in the initiation step and interrupt the free-radical chain reactions in the propagation step during lipid oxidation. mode of action. There are 2 types of SOD: a magnesium-containing SOD and a copper-zinc–dependent SOD. Teoh and Davies 2002). A low level of oxidative base damage in DNA is found in the cells of a healthy person. DNA strand breaks and modification and aging Table 1—Antioxidative enzymes Proteins Superoxide dismutase Catalase Glutathione peroxidase Glutathione disulfide reductase Glutathione-S-transferase Methionine sulfoxide reductase Peroxidase Functions Superoxide removal Hydroperoxide removal Hydroperoxide removal Oxidized glutathione reduction Lipid hydroperoxide removal Repair oxidized methionine residues Decomposition of hydrogen peroxide and lipid hydroperoxide Mitochondria and nuclei have their own DNA. Glutathione disulfide reductase is a flavoprotein that permits the conversion of oxidized glutathione (GSSG) to reduced glutathione (GSH) by the oxidation of NADH to NAD+ (Figure 5) (Papas 1999c). Superoxide dismutase (SOD) converts superoxide anion into hydrogen peroxide and oxygen. and glutathione peroxidase. 8–Hydroxy–2–deoxyguanosine has been used as a biological marker for oxidative stress (Kasai 1997). Nutraceuticals are also known as medical food. including superoxide dismutase. and increase life span. and dietary supplements.

which can prevent the interactions between metals and lipid intermediates. form complex ions or coordination compounds with metals by occupying all metal coordination sites and preventing metal redox cycling. H+ / Ascorbate O2/O 2–· RSSR/ RSSR–· (GSH) H2O / e–aq 2310 1600 1000 920 600 530 480 320 282 –330 –1500 –2870 a GSF = reduced glutathione. Decker and others 1999). including ferritin. Metal chelating nutraceuticals Table 2—Reaction of hydrogen-donating antioxidants with radicals a R· RO· ROO· R· RO· ROO· Antioxidant + + + + + + + AH AH AH A· A· A· O2 → → → → → → RH + ROH + ROOH + RA ROA ROOA Oxidized antioxidant A· A· A· a AH = antioxidant. To work as an antioxidant and prevent lipid oxidation. Radicals from phenolic compounds can be stabilized through resonance formation (Figure 6). ascorbic acid. Met- als can decompose the hydroperoxide to form peroxyl radical and alkoxyl radical. H+ / ROH ROO·. Metal chelators are phosphoric acid. and the concentration of peroxyl radicals is found to be the greatest of all fatty acid radicals (Frankel 1985). Fe2+ + 2 O2 → Fe3+ + ·O2– 1O2 Hydrogen peroxide can react with transition metal ions to form hydroxyl radical. For example. which react with peroxyl radicals before the PUFA react with peroxyl radicals. H+ / H2O. Antioxidant radicals are stabilized through resonance structures. respectively (Table 3) (Buettner 1993). carnosine. The availability of metal ions is determined by the concentrations of metal-binding proteins. Chain-breaking antioxidants donate hydrogen atoms to peroxyl radicals and convert them to more stable and nonradical products (Table 2) (Decker 1998. Halliwell and others 1995. The propagation step is a slow step in lipid oxidation. alkoxyl. Standard 1-electron reduction potentials of alkyl. Mn+ + H2O2 → M(n+1)+ + ·OH + OH– This reaction is dependent on the availability of transition metal ions such as copper and iron. can prevent lipid oxidation. H+ / Catechol -Tocopheroxyl·. HO· Ascorbate–·. ascorbic acid and tocopherol. the reduction potential of a free-radical scavenger should be lower than 600 mV. Table 3—Standard 1-electron reduction potential (mV) at pH 7. can donate a hydrogen atom to peroxyl radicals of PUFA before PUFA do (Buettner 1993). and ceruloplasmin (Decker 1998). which have lower standard 1-electron reduction potential (282 and 480 mV. 2004 . and accelerate the lipid oxidation at the exponential rate (Min 1998. lactoferrin. RO· = alkoxyl free radical. Ramon and Gonzalo 2002). Metal chelators can convert metal ions into insoluble metal complexes or generate steric hindrance. 3. PUFA = polyunsaturated fatty acids. Lipid oxidation in foods and in biological systems is a typical free-radical chain reaction of unsaturated fatty acids with initiation. The initiation step of oxygen oxidation requires removal of a hydrogen atom. peptides.CRFSFS: Comprehensive Reviews in Food Science and Food Safety to free radicals. and peroxyl radicals of PUFA and generate nonradical stable compounds (Table 2). and alkoxyl radicals of PUFA are 600. 26 Singlet oxygen-quenching nutraceuticals Singlet oxygen is highly reactive toward any molecules with COMPREHENSIVE REVIEWS IN FOOD SCIENCE AND FOOD SAFETY—Vol. Fe3+ (Cu2+) + → ROOH Fe2+ (Cu+) + → ROOH Fe2+ (Cu+) →+ ROO· + H+ Fe3+(Cu2+) → + RO·+ OH– Metals are also involved in the formation of singlet oxygen. R· = alkyl free radical. can scavenge free radicals and prevent lipid oxidation. ROO· = peroxyl free radical. propagation. and 1600 mV. Some reported metal-chelating proteins are shown in Table 4. citric acid. Metal chelators. 1000. H+ / PUFA Catechol·. respectively) than PUFA (600mV). H+ / -Tocopherol H2O2.0 for selected radical couplesa HO·. polyphenols such as quercetin. H+ / ROOH GS·/GS– (glutathione) PUFA·. The presence of metal can accelerate the initiation step of lipid oxidation by the mechanism of RH + Mn+ → R· + H+ + M(n–1)+. Papas 1999b). The newly generated free radicals from antioxidative nutraceuticals should be stable enough not to participate in other lipid oxidation chain reactions. Whether a compound acts as an antioxidant or a prooxidant can be determined by the standard 1-electron reduction potential (Table 3). some amino acids. Transition metals such as iron and copper play important roles in initiation and propagation steps of lipid oxidation. Figure 6—Resonance stabilization of phenolic antioxidant radicals. one type of antioxidative nutraceuticals. peroxyl. which is a reduction potential of PUFA. H+ / H2O RO·. Free-radical scavengers. and proteins such as transferrin and ovotransferrin (Decker 1995. Antioxidant radicals formed from hydrogen-donating antioxidants can react with alkyl. and termination steps.

Ascorbic acid L–Ascorbic acid is a 6-carbon lactone ring structure with 2. cereal grains. is the most abundant among tocopherols. Detailed information about the singlet oxygen-quenching mechanisms can be found in an excellent review by Boff and Min (2002). and kale (Watkins and others 1999). There are 2 types of singlet oxygen-quenching mechanisms: physical and chemical quenchings. Tocopherol has been associated with the reduction of heart disease. Tocotrienols have been shown to have anticancer activity and cholesterol-lowering ability. Tocotrienols are found mainly in palm oil. Singlet oxygen reactions with compound (A) to form oxidized products (AO2) are shown in Figure 7. 12 –trimethyltridecyl) chroman–6–ols. –Tocopherol has higher vitamin E activity and singlet oxygen-quenching ability than –. and tocotrienols have 3 double bonds at position 3 . Tocopherols are natural constituents of biological membranes. respectively (Niki 1996). . 108. 7 . –. whereas –tocopherol has better nitrogen dioxde and peroxynitrite radical-scavenging ability than –tocopherols (Gregory 1996). The –. Tocols are 2–methyl–2(4 . which have many double bonds. 7. Chemical quenching between singlet oxygen (1O2) and quencher (Q) involves the generation of an oxidized product QO2 . Tocopherols can protect PUFA within the membrane and LDL. which acts independently of the redox potential (Kumuda and Chandan 2000). 3. alkoxyl. and the antioxidant mechanisms of tocotrienols are the same as those of tocopherols. –. 2004—COMPREHENSIVE REVIEWS IN FOOD SCIENCE AND FOOD SAFETY 27 . Physical quenching converts singlet oxygen (1O2) to triplet oxygen (3O2) without production of oxidized product QO2 . Singlet oxygen quenchers should have electron-rich structures such as double bonds in the molecules to react with singlet oxygen. arthritis. –tocopherol.3– Figure 7—Singlet oxygenquenching mechanisms Figure 8—Structures of tocopherols and tocotrienols Vol. Tocopherols are typical and important antioxidants in humans. electrons or lone pairs of low ionization energy.Antioxidative nutraceuticals . and neurologic diseases. which is present at the ratio of 1 to 1000 lipid molecules. Antioxidant mechanisms of tocopherols include the transfer of a hydrogen atom at 6–hydroxyl group on the chroman ring. Tocotrienols are more mobile within the biological membrane than tocopherols and have more recycling ability and more inhibition of liver oxidation (Watkins and others 1999). and – tocopherols. such as oxidized products. saturated phytyl chain. and –tocopherols and tocotrienols differ in the number and position of methyl groups attached to the 5. and prevention of cancer (Meydani 2000). . Carotenoids. and 11 of the side chain in tocols (Figure 8). Chemical quenching is involved with the generation of intermediates. –Tocopherols can reduce the concentration of nitrogen dioxide (NO2) better than other tocopherols. and inhibit smooth muscle cell proliferation and protein kinase C activity. Physical quenching converts singlet oxygen into triplet oxygen by either energy transfer or charge transfer without generating any other intermediates. Phytyl chain in tocopherols can be fit in the membrane bilayer while active chroman ring is closely positioned to the surface. Uric acid is also a powerful quencher of singlet oxygen (Halliwell 1996). Some in vitro studies showed that tocot- rienols inhibited LDL oxidation better than tocopherols (Watkins and others 1999). and 106 (/M/s). 8 . are well-known singlet oxygen quenchers (Boff and Min 2002). and 8 of the ring structure (Gregory 1996). Efficiency of scavenging hydroxyl. Nitrogen dioxide is involved in carcinogenesis. Thioredoxin has been reported as a singlet oxygen quencher and a hydroxyl radical scavenger. and scavenging of singlet oxygen and other reactive species. Tocotrienols are potential nutraceuticals. Tocopherols and tocotrienols are very nonpolar and exist in lipid phase. Table 4—Metal-chelating proteins Proteins Ferritin Transferrin Lactoferrin Haptoglobin Ceruloplasmin Albumin Transferrin ferro-oxidase Hemopexin Functions Iron storage Iron storage Iron storage Hemoglobin sequestration Copper storage Copper storage Iron transport Stabilization of heme Antioxidative Nutraceuticals Tocopherols and tocotrienols Tocopherols consist of a chroman ring and a long. This unique structure enables tocopherols to act as effective antioxidants and to be regenerated through reaction with other antioxidants such as ascorbic acid (Papas 1999c). and peroxyl radicals by –tocopherol is approximately 1010. delay of Alzheimer’s disease. –. Tocopherols are regenerated in the presence of ascorbic acids.

which are associated with coronary heart disease (Weisburger cals and regeneration of –tocopherol from the tocopheroxyl rad. monocyclic. The effects are greater in nonsmokers than in smokers. and water activity (Gregory 1996). light exposure. lycopene is acyclic. Ascorbic acid is an excellent electron donor be. and pizza is significantly related to a low incidence of prostate Ascorbic acid may be related to the prevention of some can. or epoxy groups. The basic carotenoid structural backbone consists of isoprenoid units formed either by head-to-tail or by tail-to-tail biosynthesis. The antioxidant potentials of carotenoids have been reported for the prevention of free radical initiated diseases. or bicyclic. Ascorbic acid is highly susceptible to oxidation in the presence of metal ions such as Cu2+ and Fe3+. For example. cataracts. Lycopene and –carotenes are typical carotenes whereas lutein in green leaves and zeaxanthin in corn are typical xanthophylls. Lycopene. and removal of molecular oxygen. and then L–dihydroascorbic acid by donating a 2nd hydrogen atom and electron (Figure 9). Carotenes are hydrocarbon carotenoids and xanthophylls contain oxygen in the form of hydroxyl. and the easy conversion of dehydroascorbic acid to ascor. pH. Tomato juice compounds in the gas phases and the ascorbic acid radical could with 40 mg of lycopene can reduce the endogenous levels of be prooxidant in smokers (Kaur and Kapoor 2001).bel and others 1997).otene inhibit the formation of oxidized products of LDL cholestergen. The structures of carotenoids are acyclic. ascorbic acid can act as a prooxidant under certain conditions. Ascorbic acid can donate a hydrogen atom to a tocopheroxyl radical at the rate of 2 × 105/ M/s because of the difference of 1-electron reduction potential between ascorbic acid (282 mV) and (480 mV). heart disease.ol.epidermis by inducing lipid oxidation.3– enediol. Double bonds in carotenoids are conjugated forms and usually the all trans forms of carotenoids are found in plant tissues (Figure 11). 2004 .cancer (Giovannucci and others 1995).1999). quenching of singlet oxy. keto. prostate. which results in premature cause of the low standard 1-electron reduction potential (282 aging of the skin. damage. oxygen concentra. and ascorbic acid can access easily to the antioxidant active site of tocopherols and regenerate tocopherols from tocopherol radicals (Figure 10) (Buettner and Jurkiewicz 1996). Carotenoids including lycopene and –cardrogen atom donation to lipid radicals.main carotenoid of tomatoes and tomato products. mV).multiple sclerosis. –Carotene is involved in the protection of the skin against ical species are also well known antioxidant mechanisms of deleterious effects of sunlight. Both L–ascorbic acid and L–dihyFigure 9—Sequential 1-electron oxidations of L-ascorbic acid droascorbic acid retain the vitamin C activity.3 × 1010/M/s ascorbic acid with other radicals are shown in Table 5. One mole of –carotene can quench 250 to bic acid. Giovannucci 1999). The reaction rate constants of 1000 molecules of singlet oxygen at a rate of 1. the generation of relatively stable semi-dehydroascorbic Carotenoids are the most efficient singlet oxygen quencher in acid. The phenol group of tocopherol is located near the interface of a biological membrane-water phase. Regeneration of tocopherol radicals to tocopherols by ascorbic acid has been known since the 1940s. lymphocyte DNA breakage in a group of male smokers (Pool–ZoThe antioxidant mechanisms of ascorbic acid are based on hy. tomato sauce. which is the cers. including reducing ferric iron to more active ferrous iron. –carotene is monocyclic. colon. rectal. There are primarily 2 classes of carotenoids: carotenes and xanthophylls. Carotenoids Carotenoids are a group of tetraterpenoids. Consumption of fresh tomatoes. and the common cold. has several copherol supplementation can substantially reduce oxidative health benefits including decreasing the development of cervical. (Rumsey and others 1999). including atherosclerosis. and other types of cancers (GioSmoking induces oxidative stresses from numerous free-radical vannucci and Clinton 1998. Scavenging aqueous radi. However. L–Ascorbic acid first changes to semi-dehydroascorbic acid through donating 1 hydrogen atom and electron. UV ray initiates free radical in the ascorbic acid. The kinetics of electron or hydrogen atom transfer reactions are rapid. methoxyl. and – and –carotenes are bicyclic carotenoids (deMan 1999). Ascorbic acid and to. stomach. The antioxidant activity of ascorbic acid comes from 2. 3. resulting in ascorbic acid being an excellent antioxidant. carboxyl.CRFSFS: Comprehensive Reviews in Food Science and Food Safety enediol moiety. age-related muscular degeneration.biological systems. and 28 Figure 10—Regeneration of tocopherols by ascorbic acid COMPREHENSIVE REVIEWS IN FOOD SCIENCE AND FOOD SAFETY—Vol. tion. Oxidation of ascorbic acid is also influenced by heat.

and -carotenes are carotenes. and aldehyde groups. especially the presence of oxygencontaining functional groups. Lee and others 2003). –Carotene may not donate a hydrogen atom to peroxyl radicals effectively. 2 Contrary to the singlet oxygen quenching ability of carotenoids. However. and therefore cannot act as an antioxidant. Haila and others 1997). near-infrared absorption species were not observed from xanthophylls containing hydroxyl. at least 7 conjugated bonds are required. and –carotene molecules become resonance-stabilized. . keto. hydrogen donating antioxidant activities of carotenoids are controversial. –Carotene in high oxygen concentration can act as a prooxidant rather than an antioxidant. (Foote 1976). Mortensen and others 2001. Lycopene and .Antioxidative nutraceuticals . To act as an effective singlet oxygen quencher. Figure 11—Structures of carotenoids. which has a similar standard 1-electron reduction potential of peroxyl radicals (1000 mV). – Carotene may donate electrons instead of hydrogen atom to free radicals. and lutein and zeaxanthin are xanthophylls. carbon-centered. quenching efficiency increases (Boff and Min 2002). The free radical scavenging mechanism of b–carotene has been proposed to be different from the hydrogen donating phenolic compounds (Liebler 1993. Skibsted and co-workers reported the presence of near infrared absorption species from –carotene using laser flash photolysis. –Carotene can scavenge superoxide anions with the following equation (Edge and others 1997). –Carotene radical cation can absorb near infrared energy. Not only oxygen concentration but also carotenoid concentration plays an important role in determining antioxidant or prooxidant properties. Antioxidant activity of –carotene increases at low oxygen concentrations.6 × 109 1-2 × 106 6 × 106 2 × 105 2 × 105 1 × 105 (pH 5. –Carotene can become radical cation by donating electron not by hydrogen. Depending on the redox potentials of free radicals and chemical structures of carotenoids. and become –carotene radical cation (R + –carotene → R–+ –carotene+) (Liebler 1993. . 3. Figure 12—Structures of flavonoids 29 Vol. Relatively high standard 1-electron reduction potential of –carotene radTable 5—Rate constants for reaction of equilibrium mixture of ascorbic acid/semidehydroascorbic acid/dehydroascorbic acid at pH 7. 2004—COMPREHENSIVE REVIEWS IN FOOD SCIENCE AND FOOD SAFETY . either hydrogen atoms or electrons may transfer from carotenoids to free radicals (Edge and others 1997). Burton and Ingold (1984) proposed that –carotene may react with free radicals by an additional mechanism.4 Radical HO· RO· (tert-butyl alkoxyl radical) ROO· (alkyl peroxyl radical) GS· (glutathiyl radical) Tocopheroxyl radical Ascorbate–· (dismutation) O2–· /H2O· k (/M/s) 1. and conjugated radicals.1 × 1010 1. The rate of singlet oxygen quenching by carotenoids is dependent on the number of conjugated double bonds and on the type and number of functional groups on the ring structure of the molecules (Beutner and others 2000).6) ical cation (1060 mV) (Edge and others 2000) could explain the prooxidant property of –carotene. which may donate hydrogen atoms instead of electrons to free radicals (Edge and others 1997). and as the number of conjugated bond increases. Singlet oxygen quenching mechanisms by carotenoids are physical quenching without generating oxidizing products (1O2 + 1Carotenoid → 3O + 3Carotenoid).

Isoflavones found in soybeans are aglycone forms. The most ubiquitous flavonoid is quercetin. lipoic acid. and other substituents on the 2 benzene rings. daidzein. 5. Lipoic acid plays an important role in reducing blood glucose con- Figure 13—Isoflavone structures. 7. 4 –pentahydroxy flavone. Antioxidant activity of lipoic acid can help to reduce diabetic late complication. The flavonoids including flavones. Chemical structures also affect the antioxidant activities. antiatherosclerotic. malonyl glucoside. were reported in vivo and in vitro (Naim and others 1976). Each flavonoid group is different. Isoflavones. Hwang and others (2000) suggested that isoflavones may prevent lipid oxidation through stabilizing LDL structures instead involving in lipid oxidation chain reaction. and hydroxyl groups in positions 3 and 5. whereas the flavanones have a saturated C2–C3. in simple lipid system such as liposomes. and heart (Bast and Haenen 2001). and dihydrolipoic acid. The bioactive components are isoflavones (Hendrich and others 1999). Genistein. such as high concentrations of phenolic compounds or metal ions. are glycosides with a benzopyrone nucleus. daidzein.CRFSFS: Comprehensive Reviews in Food Science and Food Safety Polyphenols Phenolic compounds or polyphenols are ubiquitous in plants with more than 8000 structures reported (Bravo 1998). are chemically related with flavonoids. which contains high content of polyphenols. 3. Soybean is the unique source of isoflavones with 1 to 3 mg/g and with 0. Antioxidant mechanisms of polyphenolic compounds are based on hydrogen donation abilities and chelating metal ions (Bravo 1998). Lipoic acids Some sulfur-containing compounds. 30 Figure 14—Structures of lipoic acids COMPREHENSIVE REVIEWS IN FOOD SCIENCE AND FOOD SAFETY—Vol. which do not have the common flavonoid structures. Flavonoids. prostate and mammary) by modulating the activity of estrogen (Hendrich and others 1999). which do not easily participate in other radical reactions. Antioxidant activities of flavonoids are influence by hydroxylation and the presence of sugar moiety (Bravo 1998). have shown antioxidant activities. depending on the number of hydroxyl. and -glucoside. a 2–3 double bond conjugated with the 4–oxo function. The chemical structure of lipoic acid is 1. After donating a hydrogen atom. Antioxidant activities of isoflavone on lipoxygenase-catalyzed lipid oxidation were dependent on the concentrations and structures of isoflavones (Naim and others 1976). Flavonoids have the most potent antioxidant activities because of the chemical structures with o–diphenolic group. German and Walzem 2000). and in more complex system such as lipoproteins (Patel and others 2001). followed by daidzein and its derivatives and glycitein and its derivatives (Hendrich and others 1999). However. flavanones. liver. Patel and others (2001) suggested antioxidant mechanisms of isoflavone as analogous of tocopherol-mediated peroxidation. flavanonols. and acetyl glucoside derivatives (Figure 13). Lipoic acid is present in meat. Consumption of soy and soy products are related with biological effects. It has been reported that phenolic compounds have antioxidant. Isoflavones are not consumed during lipid oxidation and show synergic antioxidant effects with ascorbic acid (Patel and others 2001). the most important single polyphenol group. and malonyl glucosides of each aglycone are found in soybean. flavonols. Antioxidant activities of isoflavones. is associated with a low risk of coronary heart disease (Bravo 1998. Flavononols have an additional hydroxyl group at the C3 position. antimutagenic. which can be developed through oxidative stress. Moderate consumption of red wine. The classes of phenolic compounds are shown in Table 6. The flavones have a double bond between C2 and C3. and flavanonols are saturated between C2 and C3 with a hydroxyl group at the C3 position. and glycitein. including anticarcinogenic. 3. Addition of purified forms of isoflavones inhibited copper-dependent LDL oxidation (Hwang and others 2000). acetyl glucoside. phenolic compounds act as prooxidants under certain conditions.025 to 3 mg/g soy products (Wang and Murphy 1994). including glutathione. and free-radical scavenging activities. Flavonoids are effective hydroxyl radical and peroxyl radical scavengers. Flavonoids can make complexes with metals and inhibit metal initiating lipid oxidation (Hendrich and others 1999). Soybeans contain significantly high isoflavone levels and are the major dietary source of isoflavones in humans. Epidemiologic studies showed that increased consumption of phenolic compounds reduces the risk of cardiovascular disease and certain type of cancer. Oral intake of the isoflavone genistein is associated with an increased resistance of LDL oxidation and inhibition of plasma lipid oxidation products (Wiseman and others 2000). and anthocyanins are based on the common structures of carbon skeletons (Figure 12).2–dithilane–3–pentanoic acid (Figure 14). including genistein. Lipoic acids can prevent oxidative damages of proteins. especially genistein. methoxyl. 2004 . and glycitein are aglycones. The antioxidant mechanisms of isoflavones are not clearly understood and have been suggested to be different from conventional antioxidants. Genistein and its derivatives are found in the highest content in soybeans. 3 . and high pH. Glucose linkage to aglycone reduced the antioxidant activities of isoflavone. The structural similarities of genistein and daidzein to naturally occurring estrogens suggest that these compounds may protect against hormone-dependent cancers (that is. and antihemolytic effects. phenolic compounds become resonance-stabilized radicals. and their glycoside.

and used for fortifying foods or vitamin supplements. including types of nutraceuticals. Esterification of tocopherols. Even when cis and trans forms of –carotene are ingested. Naturally occurring geometrical isomers of lycopene are primarily in the all-trans configuration. blackberries. Table 6—Classes of phenolic compounds Class Simple phenols Basic skeleton C6 C6 C6-C1 C6-C2 C6-C2 C6-C3 C6-C3 C6-C3 Basic structure Benzoquinoes Phenoic acids Acetophenones Phenylacetic acids Hydroxycinnamic acids Phenylpropens Coumarins Bioavailability of Antioxidative Nutraceuticals Definition of bioavailability is the amount or the percentage of an ingested nutrient that is absorbed and thus available to the body for metabolic use. Esters of tocopherols are hydrolyzed by lipases and tocopherols are absorbed in their free. and metals. 3. which is significantly lower than that of GSSG/GSH and dehydroascorbic acid/ascorbic acid. In dietary studies. Lipoic acid regenerates GSH in liver. and crowberries. Berries. Even though tocotrienols have a higher radical scavenging activity than tocopherols. Boileau and others 2002). Food processing can affect bioavailability. However. herbs. a shorter length to fit into micelles. which blocks the 6–hydroxyl group of tocopherols. contain large amounts of phenolic compounds such 31 Vol. It has been known that –tocotrienol is preferentially absorbed compared with – and –tocotrienols (Ikeda and others 1996). Lipoic acids are excellent antioxidants. Hydrolysis of flavonoid glycosides can be done by microorganism glucosidases in the colon. 2004—COMPREHENSIVE REVIEWS IN FOOD SCIENCE AND FOOD SAFETY . tartaric acid ester of caffeic acid. and anthocyanins. centration. Boileau and others 2002). glucosides are not detected in human blood and urine whereas aglycones of isoflavones are found. Sugar moiety of glycosides is an important factor of absorption and bioavailability of isoflavones (Hendrich and others 1999). they are less bioavailable after oral ingestion (Packer and others 2001). light. strawberries. metabolic regeneration. Reduced (dihydrolipoic acid) and oxidized forms of lipoic acid both act as antioxidants and scavenge the reactive oxygen species. unesterified form. showing abilities for radical scavenging. the concentration of trans –carotene in blood and tissue is higher than cis –carotene (Deming and others 2002). geometric isomers. –Tocopherol is preferentially secreted by the liver into the blood lipoprotein. not by the host mammalian. kidney. Dihydrolipoic acid is a reductant and regenerates GSH from GSSG and ascorbic acid from dehydroascorbic acid at the rate constant of 32 and 875 /M/min. Bioavailability of antioxidative nutraceuticals is influenced by many factors. even though – and –tocopherols are equally well absorbed. Clinton and others (1996) have shown that cis-isomers of lycopene represent approximately 50% of total lycopene in blood Chromones C6-C3 Anthraquinones C6-C2-C 6 Flavonoids C6-C3-C 6 and up to 80% in prostate tissues. 250 and 282 mV. A dietary study of lipoic acid showed a decrease in age-related decline in oxygen consumption and radical formation. Lipoic acids may improve age-related decline in memory and cognitive function and brainrelated ailments. and a lower tendency to aggregate (Boileau and others 1999. interaction with other antioxidants. vegetables. Foods Containing Antioxidative Nutraceuticals Fruits. including Alzheimer’s disease and Parkinson’s disease (Kramer and Packer 2001). flavon–3–ol catechin. grapes contain polyphenolic compounds such as caftaric acid. and gene regulation (Bast and Haenen 2001). The mechanisms explaining the isomerization of alltrans to cis-lycopene isomers in vivo after food consumption and the physiological importance of cis-lycopene are not fully understood (Nguyen and Schwartz 1999. It has been suggested that cisisomers of lycopene are more bioavailable than all trans-isomers. Ascorbic acid is suggested to be absorbed better than dehydroascorbic acid in humans (Gregory 1996). including blueberries. Excessive heating can promote oxidation or formation of complexes of antioxidants with carotenoids and proteins. . However. . and beverages such as tea and wine are typical foods containing various antioxidative nutraceuticals. and lung tissue and also regenerates vitamins C and E. Aglycones of isoflavone can be absorbed in the gut better than their glucoside derivatives. most likely because of the greater solubility of cis-isomers in the bile acid micelles. Heat processing breaks the carotenoid protein complexes and converts cis to trans –carotene. spices. respectively. In fruits. improvement of mitochondrial membrane potential. respectively. A tocopherolbinding protein plays an important role in this preferential incorporation with –tocopherol (Papas 1999b). processing methods. and increases of ascorbic acid and GSH levels (Hagen and others 1999). makes tocopherols more stable to oxidizing agents such as air. ingested lycopene is predominately (about 95%) in the all-trans form.Antioxidative nutraceuticals . which can affect bioavailability. Tocopherols and tocotrienols in human blood and tissues are in their free and unesterified form. The standard 1-reduction potential of lipoic acid/dihydroxy lipoic acid is –320 mV. Absorption of lycopene from fresh tomatoes and –carotene from fresh carrots is significantly lower than from tomato juice or cooked carrots. Ascorbic acid in foods is mainly (80% to 90%) in the reduced form and is absorbed in human intestine by a sodium-dependent active transport system. Major forms of isoflavones in foods are –glycosides. metal chelating. and matrices surrounding the compounds (Papas 1999b).

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