ANTIOXIDATIVE PLANT

CONSTITUENTS

Nur Permatasari

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 Effect of Reactive Oxygen Species on
Degenerative Diseases

Gastro intestinal Eye Skin Heart

• Hepatitis • Cataractogenesis • Dermatitis • Heart attack
• Liver injury • Retinal damage • Age pigment

Teeth Joints
• Periodontis Reactive Oxygen Species • Arthritis

Vessels Multiorgan failure Brain Lung

• Atherosclerosis • Cancer • Trauma • Asthma
• Vasospasms
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• Hyperoxia
 BIOMOLECULES

dangerous functions
activated/reactive (reactive oxygen species,
biomolecules free radicals)*

ANTIOXIDANTS
useful functions
(oxidative phosphorylation, membrane protein DNA
photophosphorylation, defense, damage damage damage
biosynthesis, etc.)

cell growth & mutations

maintenance premature
aging/degerative cancer
disease
 Oxidant : RO•, •O2-
Antioxidants HO•, HOO•, H2O2
ROO• , 1O2

RO•, ·O2-
HO· HOO·
H2O2,
ROO•,farm1
1
O2, 4
 Damaging reactions of free radicals

- Lipid peroxidation
Cell membranes are a rich resource of PUFA, which are
readily attacked by oxidising agents; a process that is
called lipid peroxidation disruption of hydrophobic
nature of membranes fragmentation and loss of
membrane-bound enzymatic activities
– Protein damage
Proteins & nucleic acids seem to be less susceptible
than PUFAs to free radicals  affect enzyme, receptor
- DNA damage
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Oxidising radicals readily attack DNA --> leading to
strand breaks  mutations
 Biologically significant reactive oxygen /
nitrogen species

Reactive Oxygen Species

O2·⁻ Superoxide radical
·OH Hydroxyl radical
ROO· Peroxyl radical
H2O2 Hydrogen peroxide
1
O2 Singlet oxygen
NO· Nitric oxide
ONOO⁻ Peroxynitrite
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HOCl Hypochlorous acid 6
 Major Reactive Oxygen Species

Superoxide (O2·*)
- formed when oxygen is reduced by the transfer of a
single electron to its outer shells
- the major source of superoxide in-vivo is the electron
leakage that results from the electron transfer chain of
the mitochondria
- On its own it isn't particularly damaging. However, the
superoxide radical anion appears to play a central role
as other reactive intermediates are formed from it.
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 Hydrogen peroxide
(H2O2)
-2O2·*+ 2H* H2O2 + O2
- not a free radical but falls in the category of reactive
oxygen species.
- It is an oxidising agent that is not particularly reactive
but its main significance lies in that it is the main source
of hydroxyl radicals in the presence of transition metal
ions
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 Hydroxyl radical (·OH)
- Haber-Weiss reaction
O2·- + H2O2  O2 + ·OH + OH-

- Fenton reaction
H2O2 + Fe2+  ·OH + OH- + Fe3+

- is an extremely reactive oxidising radical that will react

to most biomolecules

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 Sources of free radicals
(Endogenous sources)
- Autoxidation
Of the molecules that undergo autoxidation come
catecholamines, haemoglobin, myoglobin, reduced cytochrome
C and thiol…
- Enzymatic oxidation
xanthine oxidase (activated in ischemia-reperfusion),
prostaglandin synthase, lipoxygenase, aldehyde oxidase…
- Respiratory burst
a term used to describe the process by which phagocytic cells
consume
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between 70 and 90% of this oxygen consumption can be
- Subcellular organelles
Organelles such as mitochondria, chloroplasts, microsomes,
peroxisomes and nuclei have been shown to generate O2·*

The leaks in mitochondrial electron transport system allow O2

to accept a single electron forming O2·*
CoO*

CoQH2
CoQ
CoQ*
O-*

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- Transition metals ions
Iron and copper play a major role in the generation of free
radicals injury and the facilitation of lipid peroxidation.
- Ischemia reperfusion injury
During ischemia two factors occur, first the production of
xanthine and xanthine oxidase are greatly enhanced. Second,
there is a loss of both antioxidants superoxide dismutase and
glutathione peroxidase.

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 Sources of free radicals
(Exogenous sources)

- Drugs (aminoglycoside, paracetamol etc)

- Radiation
- Tobacco smoking
- Metal (aluminium, lead,arsenic etc)
- Gases
- Others ( alcohol, halogenated hydrocarbon)
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 Mode of action of antioxidants

1. Catalycally remove ROS (enzymes)

2. Minimize the avaibility of pro oxidant
( transferin, metal chelation)
3. Protect biomolecules against ROS
(heat shock protein)
4. Low-molecular-mass agents that
scavenge ROS (gluthatione, uric
acid,bilirubin ,vitamin)
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 Antioxidants

Enzymes Antioxidant Role Remarks

Superoxide dismutase Dismutates O2·⁻ Contains Manganese
(SOD) to H2O2 (Mn.SOD)
Mitochondrial Contains Copper &
Cytoplasmic Zinc (CuZnSOD)
Extracellular Contains Copper
(CuSOD)
Catalase Dismutates H2O2
to H2O

Glutathione Removes H2O2 Selenoproteins

peroxidase (GSH.Px) and lipid (contains Se2+)
peroxides Primarily in the cytosol
also mitochondria
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Uses GSH
Vitamins Alpha Breaks lipid peroxidation Fat soluble
tocopherol Lipid peroxide and O2·⁻ and ·OH vitamin
scavenger

Beta carotene Scavenges ·OH, O2·⁻ and peroxy Fat soluble

radicals vitamin
Prevents oxidation of vitamin A
Binds to transition metals

Ascorbic acid Directly scavenges O2·⁻, ·OH, and Water soluble

H2O2 vitamin
Neutralizes oxidants from
stimulated neutrophils
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vitamin E
 NAC

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Vitamin E
 Family of related compounds
 tocopherols & tocotrienols
 tocotrienols less widely distributed than
tocopherols - considered of less nutritional
importance
 4 major forms based on # & position of
methyl groups on ring - , , , 
 all-rac tocopherol

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 CH 3
HO H 3C CH 3
H 3C H H
2 -Tocopherol
H 3C O 4' 8' CH 3 Activity
CH 3
CH 3 roughly
corresponds to
number of
methyl groups

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Absorption &Transport
 Absorbed in the lumen intestine
 Tocopherol in chylomicrons equilibrates with
other lipoproteins
 Transport in lipoproteins
 Uptake in chylomicron remnants by liver
 Secreted in VLDL from liver (via tocopherol
transfer protein)
– site of enrichment:  vs , , 
– distributes to tissues
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Uptake and Storage
 Uptake into tissues - pathways
– in LDL via LDL receptors (apo B)
– lipoprotein lipase: hydrolysis of
chylomicrons and VLDL
– other mechanisims?
 No specific storage site, but most vitamin E in
adipose tissue, liver, and muscle

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Excretion
 major - feces
 not absorbed, secreted from enterocytes, lost
with enterocytes, biliary secretion

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Biochemical Function
 Deficiency in animals many symptoms
– species specific
– other compounds (e.g. Se, methionine)
sometimes effective in relieving deficiency
signs
 maintenance of membrane integrity
 lipid solubility -> direct protection of cell
membranes

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Mysterious Interactions With Selenium
Explained!
 Selenium is part of glutathione peroxidase
– metabolizes lipid hydroperoxides
– explanation for amelioration of vitamin E
deficiency by selenium
 Glutathione is:
– a tripeptide (-glutamyl-cysteinyl-glycine)
– explanation for amelioration of vitamin E
deficiency by sulfur amino acids

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 -oxidation

Peroxide
catalysis of
ROH GSSG
peroxidation NADPH

GSH Peroxidase GSH Reductase

(Se) (riboflavin)
Vitamin E
PUFA ROOH GSH
(RH) NADP

Peroxidation with
free radical
damage sulfur amino acids

Cellular
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free radical 26
damage
Deficiency
 Rare in humans
 Fat malabsorption disorders can lead to
deficiency
– adults: celiac disease, pancreatitis, biliary
cirrhosis
– genetic diseases: cystic fibrosis, others

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Deficiency - 2
 Deficiency signs
– neuropathological changes, esp. spinal cord
degeneration
– swollen, distrophic axons
– accumulation of organelles, esp. mitochondria,
neurofilaments
– reason unknown
 Incr risk for athersclerosis, cancer & cataract
formation?
 Oral administration of vit E perday,the adult
RDA :10mg/day (men), 8mg/day (women)
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Deficiency - 3
 Premature infants susceptible to deficiency
– poor status
– hemolytic anemia: decreased RBC survival
without increased production
– hypothesis: low vit. E increased
hemolysis due to oxidative damage
– results of supplementation controversial

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Toxicity
 one of least toxic vitamins known
 at extremely high levels, vitamin E may
potentiate anticoagulation effect of drugs
(Coumadin) & may decr platelet adhesion
 pro-oxidant

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Carotenoids
• About 600 identified so far
• Most familiar is yellow-orange pigments of
carrots, -carotene
• Two major groups: carotenes and
xanthophylls

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Carotene Groups

Caroteniods

carotenes xanthophylls

ά-carotene Β-carotene crypto- lutein zeaxanthin

xanthin

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Absorption &Transport
 Absorbed in the lumen intestine depend on
the presence of bile acid and absorbale fat
 Transported via lymphatics to the liver and
circulate in association with lipoprotein

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Biochemical Function
 To be metabolized to retinoids (ά-carotene Β-
carotene , cryptoxanthine)
 To acts as antioxidant ( protective against
light-induced skin damage in patient with
porphyria)
 Anti cancer effects? (in vitro yes, in vivo
uncertain)

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Ascorbic acid (vitamin C)
 Biochemical function: cofactor for at least
eight enzymes
 At the tissue level, a major function 
collagen synthesis ( vit C deficiency can lead
scurvy)
 The antioxidant properties protect NO,
protect against age-related cataract

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Absorption & Excretion
 Absorbed in the lumen intestine via an
energy dependent process (saturable)
 The daily intake > 100mg  excreted, in
adult RDA: 60 mg/daily
 The renal threshold for ascorbic acid :
1.5mg/dl. Urinary axcretion of oxalate and
urate  1000mg

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Toxicity
 Megadosage treatment  formation of
kydney stone and rebound scurvy
 Pro oxidant / acts as reductant to the iron
(be aggravated in disease and traumatic
injury)?

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 Antioxidative Nutraceuticals

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• Nutraceuticals are naturally derived,
bioactive compounds that have health
promoting, disease preventing or medicinal
properties.

• Nutraceuticals can be delivered in the

form of food (functional foods) or as a
dietary supplement or in both forms.

• Nutraceutical industry is a fast growing

industry.
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Antioxidative Nutraceuticals ( preventive antiox, chain
breaking)

• Vitamin A, C, and E
• Plant phenols (OH / two OH group attached to benzene ring) 
flavanol (epicatechin, catechin), flavanones (naringin, taxifolin),
flavonols (kaempferol, quercetin), flavones/isoflavons (genistein,
daidzein), anthocyanidins , phenylpropanoids (caffeic acis, p-
coumaric acid)
• Thiol molecules
• Terpenoids (prevent lipid perox in mit, mit function)
• Lipoic acid

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Effects of plant phenol

• Scavenging alkoxyl and peroxyl radicals

• Chelation of metals
•. Oestrogenic effect / anti estrogen (isoflavones)

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 Flavonoids terpenoids
quercetin

NAC

ROOH

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phenolic terpenoids
 Nutraceuticals in Orange

OCH3
Hesperidin OCH3 3'
4'
CH3O O 1'B
8 1
7
Tangeretin 6
A C
5 4
CH3O
OCH3 O

OCH3
Nobiletin OCH3 3'
OCH3
B 4'
CH3O O 1'
8 1
7
6
A C
5 4
CH3O

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OCH3 O
43
Contain hesperidin, tangeretin, nobiletin, and limonene
 Anthocyanin
Quercetin

Beta-Carotene

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Lycopene 44
 Quercetin
Resveratrol

Lycopene

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Hesperidin
Broccoli
 Contains beta-carotene, lutein,
quercetins, sulphoraphane, and indoles

Beta-Carotene

farm1 Lycopene 46
Soybean

R2 O

R1 O
OH

Isoflavone R1 R2
Genistein OH OH
Genistin OH O-glucose
Daidzein H OH
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Daidzin H O-glucose
Herb and Spice

OH CH3 OH CH3
HO HO
CH3 O CH3
HOOC C

H3C CH3 H3C CH3

Carnosoic Acid Carnosol

OH
HOOC

O OH

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OH Rosemarinic Acid
Tea

OH

OH
OH

O HO O
HO OH
OH

OH
OH
OH
OH

Epicatechin Epigallocatechin
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Wine

Red wine decreases coronary heart disease mortality

in some part of France. Please drink up to two
glasses !!!

The coumaric, cinnamic, caffeic, and ferulic acids

catechin, epicatechin and quercetin in are good
antioxidants.
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