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Antioxidants
By Aya Mohammed and Youssor Ridha
MSc students at Alnahrain University
What is free radicals
atoms with an unpaired number of electrons formed when oxygen
interacts with certain molecules, once formed these highly reactive
radicals can start a chain reaction
There are many different types of free
radicals, for example:
• superoxide anion (O2-)
• hydroxyl radical (OH )
• hydrogen peroxide (H2O2)
• singlet oxygen (1O2 )
Where do free radicals come from?
• normal metabolism
• Environmental pollutants ( radiation, herbicides, pesticides )
• smoking
What is Antioxidant ?
• A molecule that inhibit the oxidation of other molecules
• i.e. remove free radicals
TYPES OF ANTIOXIDANTS
Based on chemical nature
• Enzymatic : peroxidase , Catalase
• Non enzymatic : Vitamin E, Vitamin C, Uric acid, α- Lipoic acid
Based on Defense Mechanism
• Preventive Antioxidants : Peroxidase, Catalase
• Radical scavenging Antioxidants : Vitamin C and Carotenoids.
Natural
Sources of
Antioxidants
Terpenes
tetraterpenoid
carotenoids
Lycopene
Terpenes
tetraterpenoid
carotenoids
B-carotene
(Vitamin A)
polyphenols
flavonoids
Rubin
Ascorbic acid
(vitamin C)
Flavonoids
Catechin +
Epicatechin
Tomato Carrot Cocoa Beans
Orange,
Strawberry,
Pepper, Broccoli
Mode Of Action
Free Radicals Vs Anti-Oxidants
Free radicals damage cell membrane
• Initiation :lipid peroxidation process
by stealing electron from cholesterol
that attached to phosphate head of
phospholipid in cell membrane lead
to change head properties and initiate
destabilization of the Membrane
• Chain reaction of free radicals : chain
propagation
• Termination : Erosion of membrane
Anti-oxidants prevent phospholipids
peroxidation + avoid cell membrane
damage
• Scavenging initiating radicals
• Chain breaking reaction : prevent chain
propagation
• Able to repair oxidizing radicals directly
1- Cell Membrane
Free Radicals
• Steal electron from the sugar part of
nucleotide (oh) in 3 end of each
nucleotide
• Break in DNA chain causing
mutation aging and cancer
2- DNA
Anti-oxidants
• Reducing the free radicals
• Scavenge free radicals
Mode Of Action
Free Radicals Vs Anti-Oxidants
Free radical
production
𝑜2
−
, 𝐻2 𝑂2
OH
Tissue damage
Antioxidants
Scavenging initiating radicals
( enzymatic anti-oxidants)
Chelating transition metals
( non enzymatic anti-oxidants)
Repair mechanism
(prevent chain propagation)
Transition
metals
F𝑒2+
,
C𝑢2+
Evaluating Anti-oxidant Activity
Assay Principle
DPPH
Antioxidant reaction with an organic
radical
ORAC
Antioxidant reaction with peroxyl
radicals
DPPH Assay
free radical in nature
with purple color
DPPH + Anti-oxidant DPPHH
Hydrogen donor Reduced form with
yellow color
rapid, simple and inexpensive in comparison to other methods
With a UV spectrometer
DPPH shows a strong absorption maximum at 517 nm (purple)
Therefore, the antioxidant effect can be easily evaluated by the decrease of UV absorption from 517 nm ( yellow
color).
Anti-oxidant activity =
OD (control ) – OD ( sample )
OD (control )
x 100
ORAC Assay “Antioxidant Power”
Peroxyl O𝐻−2 + Fluorescein fluorescence
Peroxyl O𝐻−2 + Fluorescein + Anti-oxidant fluorescence
By preventing the
oxidation of
fluorescein
oxidation of fluorescein
Free radical Probe
At the end we use antioxidant in :
• Anti-aging cream
• Sunblock
• Supplement
Drug candidate
• Tirilazad is an antioxidant steroid derivative that inhibits the lipid
peroxidation that is believed to play a key role in neuronal death in
stroke. It demonstrated activity in animal models of stroke, but human
trials demonstrated no effect
Thank You

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Antioxidants

  • 1. Antioxidants By Aya Mohammed and Youssor Ridha MSc students at Alnahrain University
  • 2. What is free radicals atoms with an unpaired number of electrons formed when oxygen interacts with certain molecules, once formed these highly reactive radicals can start a chain reaction
  • 3. There are many different types of free radicals, for example: • superoxide anion (O2-) • hydroxyl radical (OH ) • hydrogen peroxide (H2O2) • singlet oxygen (1O2 )
  • 4. Where do free radicals come from? • normal metabolism • Environmental pollutants ( radiation, herbicides, pesticides ) • smoking
  • 5.
  • 6. What is Antioxidant ? • A molecule that inhibit the oxidation of other molecules • i.e. remove free radicals
  • 7. TYPES OF ANTIOXIDANTS Based on chemical nature • Enzymatic : peroxidase , Catalase • Non enzymatic : Vitamin E, Vitamin C, Uric acid, α- Lipoic acid Based on Defense Mechanism • Preventive Antioxidants : Peroxidase, Catalase • Radical scavenging Antioxidants : Vitamin C and Carotenoids.
  • 8. Natural Sources of Antioxidants Terpenes tetraterpenoid carotenoids Lycopene Terpenes tetraterpenoid carotenoids B-carotene (Vitamin A) polyphenols flavonoids Rubin Ascorbic acid (vitamin C) Flavonoids Catechin + Epicatechin Tomato Carrot Cocoa Beans Orange, Strawberry, Pepper, Broccoli
  • 9. Mode Of Action Free Radicals Vs Anti-Oxidants Free radicals damage cell membrane • Initiation :lipid peroxidation process by stealing electron from cholesterol that attached to phosphate head of phospholipid in cell membrane lead to change head properties and initiate destabilization of the Membrane • Chain reaction of free radicals : chain propagation • Termination : Erosion of membrane Anti-oxidants prevent phospholipids peroxidation + avoid cell membrane damage • Scavenging initiating radicals • Chain breaking reaction : prevent chain propagation • Able to repair oxidizing radicals directly 1- Cell Membrane
  • 10.
  • 11. Free Radicals • Steal electron from the sugar part of nucleotide (oh) in 3 end of each nucleotide • Break in DNA chain causing mutation aging and cancer 2- DNA Anti-oxidants • Reducing the free radicals • Scavenge free radicals Mode Of Action Free Radicals Vs Anti-Oxidants
  • 12.
  • 13.
  • 14. Free radical production 𝑜2 − , 𝐻2 𝑂2 OH Tissue damage Antioxidants Scavenging initiating radicals ( enzymatic anti-oxidants) Chelating transition metals ( non enzymatic anti-oxidants) Repair mechanism (prevent chain propagation) Transition metals F𝑒2+ , C𝑢2+
  • 15. Evaluating Anti-oxidant Activity Assay Principle DPPH Antioxidant reaction with an organic radical ORAC Antioxidant reaction with peroxyl radicals
  • 16. DPPH Assay free radical in nature with purple color DPPH + Anti-oxidant DPPHH Hydrogen donor Reduced form with yellow color rapid, simple and inexpensive in comparison to other methods With a UV spectrometer DPPH shows a strong absorption maximum at 517 nm (purple) Therefore, the antioxidant effect can be easily evaluated by the decrease of UV absorption from 517 nm ( yellow color). Anti-oxidant activity = OD (control ) – OD ( sample ) OD (control ) x 100
  • 17. ORAC Assay “Antioxidant Power” Peroxyl O𝐻−2 + Fluorescein fluorescence Peroxyl O𝐻−2 + Fluorescein + Anti-oxidant fluorescence By preventing the oxidation of fluorescein oxidation of fluorescein Free radical Probe
  • 18. At the end we use antioxidant in : • Anti-aging cream • Sunblock • Supplement Drug candidate • Tirilazad is an antioxidant steroid derivative that inhibits the lipid peroxidation that is believed to play a key role in neuronal death in stroke. It demonstrated activity in animal models of stroke, but human trials demonstrated no effect