AntiAnti-oxidants

M Zaky

2014-11-10

CONTOH BAHAN ALAM SEBAGAI

ANTIOKSIDAN

MANGGIS (MANGOSTEEN)
SPIRULINA
KUNYIT
TEH
KEDELAI (ISOFLAVONES)
MENGKUDU
BIJI ANGGUR
BLUEBERRY
RASPBERRY
WOLFBERRY

2014-11-10

What Do We Concern Frequently in Our Life?

Can We Slow Down the Clock of Aging?

2014-11-10

What is Aging ?
Aging is mainly due to the cumulative result of free
radical oxidative damage of the cells and tissues of
the body that arises primarily as a result of aerobic
metabolism.
The structural features that develop with aging are
Lipid oxidation of membranes
Formation of age pigment
Cross linkage of proteins
DNA damage
Mitochondrial function decline
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Aging Mechanism
Aging is a complex multifactorial process in
which free radical oxidative damage plays an
very important role, but free radical oxidative
damage may not be the exclusive mechanism
of physiological decline.

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Free Radicals
Free radicals are produced during various
metabolic processes
Environment can increase free radicals Free
radicals react with lipids (membranes),
proteins and DNA
Can cause extensive cell damage
Damage repair of DNA can lead to mutation
Mutation can lead to cancer
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Reactive Oxygen Species

Oxygen centered radicals : superoxide anion (O2-),
hydroxyl radical (HO), hydroperoxyl radical
(HOO), peroxyl radical (ROO)
Oxygen centered non-radicals : hydrogen peroxide
(H2O2), singlet oxygen (1O2)
2014-11-10

Antioxidants are classified into five types :

Primary antioxidant s: those compounds, mainly phenolic
substances, that terminate the free radical chains in lipid
oxidation.
For example :
1. Natural antioxidant : tocopherols,
Synthetic antioxidant butylated hydroxyanisole (BHA), butylated
hydroxytoluene (BHT), etc, belong to this group, and function
as a electron donors.
2. Oxygen scavengers, which react with oxygen, and can thus
remove it in a closed system. The regeneration of phenolic
antioxidants, an entirely different mechanism. For example,
ascorbic acid (vitamin C), ascorbyl palmitate etc.
2014-11-10

3. Secondary antioxidants which function by decomposing

the lipid hydroperoxides into stable end products. For
example, dilauryl thiopropionate and thiodipropionic
acid.
4. Enzyme antioxidants which function either by removing
dissolved oxygen. For example, superoxide dismutase,
catalase, glutathione peroxidase, etc.
5. Chelating agents which chelate metallic ions such as
copper and iron that promote lipid peroxidation through
a catalytic action. The chelates are sometimes referred
to as synergist since they greatly enhance the action of
phenolic antioxidants. For example, citric acid, amino
acids, EDTA, etc.
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Initiating
Event
R

R-OO-H
Lipid Hydroperoxide

Initiation

Carbon-centered
Free Radical

O2

Chain
Propagation

Reaction
R-H

ROO
Peroxyl Radical

H H

TO

=
Polyunsaturated fat
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TOH

R-OO-H
Lipid Hydroperoxide

Termination
via
Antioxidant
11

O2

H2O +
R

R
.
Hydrogen Abstraction

1. Initiation
R

OO

OH
R

+
R

2. Propagation

+
R

OO

OOH
R

3. Termination
+
R

R
R

Lipid crosslinkage
R

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Prooxidants
R3C. Carbon-centered

Free Radicals:
Radicals
 Any species capable of independent
existence that contains one or more
unpaired electrons
 A molecule with an unpaired electron
in an outer valence shell

Non-Radicals:
 Species that have strong oxidizing
potential
 Species that favor the formation of
strong oxidants (e.g., transition
metals)
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R3N. Nitrogen-centered
R-O. Oxygen-centered
R-S. Sulfur-centered
H2O2 Hydrogen peroxide
HOCl- Hypochlorous acid
O3

Ozone

1O
2

Singlet oxygen

ONOO- Peroxynitrite
Men+

Transition metals

13

Exogenous sources of free radicals

Radiation
UV light, x-rays, gamma rays

Chemicals that react to form peroxides

Ozone and singlet oxygen

Chemicals that promote superoxide formation

Quinones, nitroaromatics, bipyrimidiulium herbicides

Chemicals that are metabolized to radicals

e.g., polyhalogenated alkanes, phenols, aminophenols

Chemicals that release iron

ferritin
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Reactive Oxygen Species

The 1- 5 % consumed oxygen is converted to reactive
oxygen during normal aerobic metabolism.

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Effect of Reactive Oxygen Species on

Degenerative Diseases
Gastro intestinal

Eye

Skin

Heart
Heart attack

Hepatitis

Cataractogenesis

Dermatitis

Liver injury

Retinal damage

Age pigment

Teeth

Joints

Periodontis

Vessels
Atherosclerosis
Vasospasms
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Reactive Oxygen Species

Multiorgan failure
Cancer

Arthritis

Brain

Lung

Trauma

Asthma

Stroke

Hyperoxia
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How to Slow Down Aging?

Only 9% of Americans eat the recommended five
servings of fruits and vegetables per day, therefore
there is a great opportunity to improve health and
slow down aging process by increasing
consumption of fruits and vegetables.
The antioxidants in apple keep doctors away.

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Pro-oxidants
Irradiation -- -ray, x-ray, and ultra violet rays
Inflammation and macrophages
Argininie nitro oxide synthetase, peroxide nitrites
Autoxidation of electron transport carriers
Air pollutants
Metal catalyzed oxidation systems
Lipid peroxidation
Glycation and glycoxidation products
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Antioxidants
Proteins and Enzymes:
Superoxide dismutase, catalase, glutathione peroxidase,
glutathione transferase, glutathion reductase, thiol specific
peroxidase, methione sulfoxide reductase

Metabolites and Vitamins:

Vitamin C and E, Bilirubin, Uric Acid, GSH/GSSG.
NADH/NAD, Mg, Mn, Zn, Lipoic Acid and Carotenoids

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Antioxidant Mechanisms
Hydrogen donating compounds

Singlet oxygen quenchers

Enzymes
Metal chelators

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20

Singlet Oxygen Formation

Enzyme
3O +
2

Endoperoxide

Ozone

H2O2+ HO2 2014-11-10

1O
2

O2 .- + O2 -.

Sensitizer

H2O2 + O2 .-

.OH + O2 .21

Oxidative Stress
The increase of the reactive oxygen species which leads
to oxidative damage in

Antioxidant

foods

or body.

Reactive
Oxygens

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RO, O2-

Antioxidants

HO, HOO, H2O2

ROO , 1O2

RO, O2HO HOO

H2O2,
ROO, 1O2,
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Various Cosmetics Rich in

Antioxidant

Strating Material
Process
Antioxidant Enrichment
In vitro Antioxidant Evaluation
Antioxidant Stability

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Biological Activities of Phytochemicals

Antioxidants

Carotenoids tocopherol
catechins
quercetin
kaempferol genistein
daidzein

Anticancer Agent s

Isoflavones lignans

DNA repair Agents

Vanillin cinnamaldehyde
coumarin anisaldehyde

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Naturally Occurring Tocopherols

CH3

-tocopherol

phytyl tail

HO

CH3
O

H3C

CH3

CH3

2R

CH3

-tocopherol

CH3

4R

H CH3

CH3

8R

HO

phytyl tail
O
CH3

CH3
HO

-tocopherol

phytyl tail
O

H3C

CH3

CH3
HO

phytyl tail

-tocopherol
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O
CH3

CH3

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Tocopherol as an antioxidant
R + TOH RH + TO

R1
HO

Phythyl Tail
R2

.O

CH 3
CH 3

CH 3

(CH 2 )3 CH (CH 2 )3 CH (CH 2 )3 CH (CH 3 )2

R3
CH 3

Chromane Head

CH 3

CH 3

R1

R2

R3

CH 3

CH 3

CH 3

CH 3

CH 3

CH 3

CH 3

CH 3

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Tocopherol Functions
Specific role in a required metabolic
function has not been found
Major function appears to be as a fatsoluble, chain-breaking antioxidant
Peroxyl radical scavenger
Protects polyunsaturated fatty acids
(PUFAs) within membranes and
lipoproteins
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Resonance of Antioxidant Radicals

OH
C (CH 3) 3

OCH 3
ROH , ROOH

RO , ROO
O.

O
C (CH 3) 3

OCH3

C (CH 3) 3

O C H3

O
C (CH 3) 3

C (CH 3) 3

.
2014-11-10 O C H

O C H3

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Antioxidant Mechanism of Tocopherol

Transfer of phenolic hydrogen to reactive

oxygen species

Scavenging of singlet oxygen

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Terpenes
Common examples and plant parts:
Cinnamon in bark
Peppermint in hairs on leaves
Lemon - in the rind
Rose in the petals
Ginger in the rhizome
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CAROTENOIDS

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Quenching Mechanism of - Carotene

on Singlet Oxygen

O2 + 1-Carotene Energy transfer

-Carotene

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O2 + 3-Carotene

Radiationless Energy transfer

-Carotene

33

Shikimic Acid Natural Products

CH3

CH2

Eugenol

Anethole

CH3O
OMe

OH
CHO
CO2CH3
OH

Cinnamaldehyde
2014-11-10

Methyl Salicylate
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Shikimic Acid Flavones

OH
HO

HO

OH O

OH O

An Isoflavone

A Flavone

OH

OH

HO

O
An Anthocyanidin
OH

OH
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ANTHOCYANINS
Anthocyanidin
R1
R1
R2
+
O

HO

R3
OH

OH

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R2

R3

Pelargonidin H

OH

Cyanidin

OH

OH

Delphinidin

OH

OH

OH

Peonidin

O-Me OH

Petunidin

O-Me OH

OH

Malvidin

O-Me OH

O-Me

36

Soybean
CO

O
R2

O
1

OH

Isoflavone

R1

R2

Genistein

OH

OH

Genistin

OH

O-glucose

Daidzein

OH

Daidzin

O-glucose

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Tea
OH
OH

OH

HO

HO

OH

OH

OH
OH

Epicatechin
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OH

OH

Epigallocatechin
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1. Ferric Thiocyanate method

or

Ferric Reducing Antioxidant Power (FRAP)

or Cupric Reducing Antioxidant
H H
ROOH

2 Fe

++

2 H

C C

2 Fe

+++

OH OH

2 Fe

+++

6 CNS

Fe [ Fe (CNS)6 ]

Chromophore (500 nm)

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2. By DPPH (1,1-Diphenyl-2-Picrylhydrazyl)
(Free Radical Scavenging )

NO2

NO2

NO2

N N
NO2

NO2

H
N N
NO2

DPPH
Chromophore (532 nm)

Assesment of radical scavenging activity : the active

constituents will bleach the purple of DPPH giving a
white-yellowish color
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Oxidation Mechanism

Two types of oxidation mechanism in

cosmetics:
Auto-oxidation proceeds in the presen
ce of oxygen via a radical-chain mech
anism
Non-radical oxidation proceeds in the
presence of ozone, single oxygen, etc
.
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AUTO OXIDATION

Auto-oxidation is a radical chain reactio

n occurring due to the presence of 20%
oxygen in the air and it is the most impo
rtant reaction to be noted to prevent the
oxidation of cosmetics. Auto-oxidation
response is accelerated or inhibited by
many factors including heat, light (uv),
metal ions, water, proteins, etc.
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Prevention of oxidation

Suppress the initiation and chain transf

er reaction phases.
Initiation: Storage in a cool location, u
sing UV absorbents, Using Chelating A
gent
Addition of compounds to prevent gen
eration of radicals: Tocopherol, BHT,
BHA, Gallic Acid esters, NDGA (nordih
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ydroguaiaretic
acid)

43

Sesquestering Agents

Metalic ions can cause changes in pe

rfume and color and can also promot
e oxidation of oily raw materials. Also
block the action of pharmaceutical ag
ent and can cause transparency to b
e lost through precipitaion, for lotions
etc. Compounds used to deativate th
ese metallic ions are called sequester
ing agents.

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Sesquestering Agents
Sodium Edetat (EDTA)

Phosphoric Acid

Citric Acid

Ascorbic acid

Succinic acid

Gluconic acid

Sodium polyphosphat

Sodium metaphosphate
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