Antioxidants

• An antioxidant is a molecule that inhibits

the oxidation of other molecules. Oxidation is
a chemical reaction involving the loss of
electrons or an increase in oxidation state.
Oxidation reactions can produce free radicals.
In turn, these radicals can start chain
reactions. When the chain reaction occurs in
a cell, it can cause damage or death to the
cell. Antioxidants terminate these chain
reactions by removing free radical
intermediates, and inhibit other oxidation
reactions.
 Food
Nutrients Non-nutrients

Energy, building Factors regulating

materials metabolism Phyto-
chemicals
•pigments
•Antioxidants
Lipid Protein Carbo- Water Vitamins Minerals
hydrate Fibers

Zn, Se Other food

EFA & EAA & C, E, b-Car
components
non-EFA non-EAA Glucose

Functional food or
Neutraceuticals
Cys GSH
 ROS (reactive oxygen species)
Free radicals Particals, which are not
superoxide, O2 · - free radicals
hydroxyl radical, OH hydrogen peroxide,
· H2O2 (Fenton´s
peroxyl, ROO · reaction)
alkoxyl, RO · hypochlorous acid,
HClO
hydroperoxyl, HO2 ·
ozone, O3
singlet oxygen, 1O2
 Ten Leading Causes of Death in
the United States (%)
1. Heart diseases (32)* 6. Pneumonia & Influenza (4)
2. Cancers (23)* 7. Diabetes Mellitus (3)*
3. Strokes (7)* 8. AIDS (2)
4. Lung Diseases (5) 9. Suicide (1)
5. Accidents (4) 10. Liver Disease (1)
• indicates diet-related diseases (65%)
• >90% of disease incidence involves free radical damage
 Antioxidants
• Prevents the transfer of electron from O2 to
organic molecules
• Stabilizes free radicals
• Terminates free radical reactions
 Antioxidant
The substance present in low concentrations relative to the oxidizable
substrate that significantly delays or reduces oxidation of the
substrate.
They reduce the effect of dangerous oxidants by
binding together with these harmful molecules,
decreasing their destructive power.
They can also help repair damage already
sustained by cells.
They may be considered as the scavengers of free
radicals.
• During this reaction the antioxidant sacrifices itself by
becoming oxidized.
• However, antioxidant supply is not unlimited as one
antioxidant molecule can only react with a single free
radical. Therefore, there is a constant need to replenish
antioxidant resources.
Free Radical Defense System

• Antioxidant Enzymes
• Antioxidant Quenchers
• Antioxidant from Foods –
nutrients/non-nutrients
 Classification of antioxidant
Ⅰ. According to their location
a) Plasma antioxidants:
– ascorbic acid (Vitamin C), bilirubin, uric acid, transferrin,
ceruloplasmin, β-carotene;
b) Cell membrane antioxidants:
– α-tocopherol (Vitamin E)
c) Intracellular antioxidants:
– superoxide dismutase (SOD),
– catalase,
– glutathione peroxidase (GPx)
 Classification of antioxidant
Ⅱ. According to their nature and action
a) Enzymatic antioxidants:
– SOD, catalase, GPx, glutathione reductase
b) Non-enzymatic antioxidants:
– Nutrient antioxidants:
β-carotene, α-tocopherol, ascorbic acid,
– Metabolic antioxidants:
bilirubin, uric acid, ceruloplasmin, ferritin,
transferrin, albumin, glutathione
Reactive Oxygen Species (ROS)

Oxygen-derived free radicals :

e.g., Superoxide and hydroxyl radicals

Non-free radical: Hydrogen peroxide

Antioxidant Mechanisms
 Antioxidants
• Enzymes:
– Superoxide dismutase
– Catalase
– Glutathione system (glutathione, NADPH, reductase,
peroxidase & selenium)

• Vitamins:
– Vitamin C (ascorbic acid)
– Vitamin A and β-carotenes
– Vitamin E

• Trace elements:
– Selenium
 Enzymatic antioxidant
1. superoxide dismutase (SOD)
2O2·⁻+ 2H+ SOD H O + O
2 2 2

SOD is present in essentially every cell in the body which actually

represented by a group of metalloenzymes with various prosthetic
groups. SOD appears in three forms:
a) Cu-Zn SOD: in the cytoplasm with two subunits
b) Mn-SOD: in the mitochondrion
c) Cu-SOD: extracellular SOD
This is the first line of defence to protect cells from the
injurious effects of superoxide.
 Enzymatic antioxidant
2. catalase, CAT
2H2O2 catalase 2H2O + O2

Catalase, iron dependent

enzyme, is present in all body
organs being especially
concentrated in the liver and
erythrocytes. The brain, heart
and skeletal muscle contains
only low amounts.
 Enzymatic antioxidant
3. glutathione peroxidase, GPx

GPx is a selenium-dependent enzyme.

The entire process is driven by energy production at the cellular
level, which involves proper thyroid hormone levels, healthy
mitochondrial function, and an active pentose-phosphate metabolic
pathway.
Oxygen Radical Defense Enzyme

CuZnSOD GSH
Peroxidase

O2•¯ H2O2 H2O + O2

Fe2+
Mn SOD Catalase

OH•
 Nutrient antioxidant
1. α-tocopherol (vitamin E)

The most important lipid-soluble antioxidant

Present in all cellular membranes.
Protect against lipid peroxidation.
1. Vitamin E was shown to be stored in adipose tissue.
2. Vitamin E prevents the peroxidation of membrane
phospholipids and avoids cell membrane damage
through its antioxidant action.
 Nutrient antioxidant

2. ascorbic acid (vitamin C)

+ 2O2·⁻ + 2H+ H2O2 +

Dehydroascorbate, DHA

It is a water-soluble, antioxidant present in citrus fruits, potatoes,

tomatoes and green leafy vegetables.
It is a chain breaking antioxidant as a reducing agent or electron
donor. It scavenges free radicals and inhibits lipid peroxidation. It also
promotes the regeneration of α-tocopherol.
Vitamin C Protects against oxidation stress
by reacting with ROS
.
Ascorbate + O
-
2

H2O2 + Monodehydroascorbate

Catalase O2 + H2O
.
Ascorbate + OH
-

H2O + Monodehydroascorbate
 Glutathione-
H2O2
Ascorbate ASC Ascorbate cycle
peroxidase MDA DHA
H 2O Dehydro
Monodehydro GSH
Ascorbate
Ascorbate GSSG reductase
reductase
ASC NADPH
Glutathion
Metabolic mechanism reductase
that detoxifies NADP
Hydrogen peroxide and
Dehydroascorbate GSH
Ascorbate can also be a source of
Superoxide & Hydroxyl radicals
However pro-oxidant actions
require relatively high concentrations
of ascorbate that are unlikely to
be reached in tissues,
because of rapid renal elimination
 Ascorbate + O2

.O – + Monodehydroascorbate
2
Ascorbate + Cu2+

Cu+ + Monodehydroasacorbate

H2O2  Cu2+ + OH + .OH

–
 Nutrient antioxidant
3. carotenoids

Carotenoids consist of C40 chains with conjugated double bonds,

they show strong light absorption and often are brightly colored (red,
orange). They occur as pigments in bacteria, algae and higher plants.
 β-carotene is the most important.
• It is composed of two molecules of
vitamin A (retinol) joined together.
• Dietary β-carotene is converted to
retinol at the level of the intestinal
mucosa. It can quench singlet oxygen.
– Quenching of singlet oxygen is
the basis for it's well known
therapeutic efficacy in
erythropoietic protoporphyria (a
photosensitivity disorder).
Lycopene is responsible for color of
certain fruits and vegetables like
tomato. It also possesses antioxidant
property.
 Are carotenes protective
against Cancers and
Cardiovascular diseases

• A considerable evidence suggested an

increase in death from lung (and other)
cancer among people given
supplements of -carotene.
 Nutrient antioxidant
4. α-lipoic acids

It is vitamin-like compound, produced in the body,

besides the supply from plant and animal sources.
It plays a key role in recycling other important
antioxidants such as ascorbic acid, α-tocopherol and
glutathione.
 Other important nutrient antioxidants

Antioxidant Source
1.Coenzyme Q10 organ meats (best heart), beef, chicken
2. Selenium sea foods, meats, whole grains
3. Proanthocyanidins grape seeds
4. Catechins green tea
5. Quercetin onions, red wine, green tea
6. Ellagic acid berries, walnuts, pomegranates
 Selenium and zinc
have no antioxidant
action themselves
• Selenium and zinc are
commonly referred to as
antioxidant nutrients.
• But these elements have no
antioxidant action themselves
and are instead required for the
activity of some antioxidant
enzymes.
 Bilirubin and Uric acid
can act as antioxidants
• There is good evidence bilirubin and uric acid
can act as antioxidants to help neutralize
certain free radicals.
• Too much bilirubin, though, can lead to
jaundice, which could eventually damage the
central nervous system, while too much uric
acid causes gout.
 Metabolic antioxidant
1. glutathione, GSH

In addition to its role as a substrate in GSH redox cycle, GSH is also a

scavenger of hydroxyl radicals and singlet oxygen. GSH also has an
important role in xenobiotic metabolism.
 Metabolic antioxidant
1. Uric acid scavenge singlet oxygen and hydroxy radical
2. Ceruloplasmin inhibit iron and copper dependent lipid
peroxidation
3. Transferrin prevents iron-catalyzed radical formation
4. Albumin scavenge radicals on its surface
5. Bilirubin protects albumin bound FFA from
peroxidation
6. Haptoglobin bind to free Hb and prevent the
accelaration of lipid peroxidation
 PHYTOCHEMICALS
• Non-nutrient compounds found in plant-
derived foods that have biological activity in
the body.
• Contribute to food taste, aromas, colors and
other characteristics.
• Act as antioxidants, mimicking hormones,
and suppressing the development of
diseases.
 GREEN TEA – A WONDERFUL
ANTIOXIDANT
Primarily consumed in China, Japan,
Middle East, North Africa & North America -
Rich in polyphenol – an antioxidant

200 times more powerful than Vitamin E

Scavenges free radicals, high rate
Reduce the risk of heart diseases
Lowers LDL oxidation
Prevents Red blood Cell breakdown
Protects against digestive & respiratory
infections
Prevents cancers of colon, pancreas &
stomach
 Ideally, the balance between the
production of free radicals and
antioxidant defenses is maintained

Oxidative stress is caused by an imbalance between

the production of ROS and a ability to detoxify the
free radicals or easily repair the resulting damage.