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Contents
y Oxidation
y Definition

Antioxidants y Free radicals

y Lipid oxidation
y Introduction
Pham Van H
Hung,
ng PhD y Mechanism
y Effects of lipid oxidation to human health
y Antioxidants
y What are antioxidants?
y Mechanism of antioxidants
y Sources of antioxidants
y Application of antioxidants in foods
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Oxidation What is free radical?

4

… is a chemical reaction that transfers ¾ is an atom, molecule, or ion that

electrons or hydrogen from a substance to has unpaired valence electrons
an oxidizing agent.
agent or an open
p electron shell.
… Oxidation reactions can produce free
radicals.

The radical derived from α-tocopherol

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Types of Free Radicals Reactive Oxygen Species - Ground-state oxygen (3O2)

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… Reactive Oxygen Species

† Triplet oxygen (3O2) … Oxygen is the most important factor on the development of
† Singlet Oxygen (1O2) lipid oxidation
† Superoxide (O2-)
… Ground state oxygen is itself a radical, with two unpaired
† Hydroperoxyl radical (HO2)
electrons each located in a π* antibonding orbital
† Hydroxyl radical (-OH)

† Ozone (O3) … Ground-state or triplet oxygen is not very reactive

… Peroxyl radical (ROO.)
… Alkoxyl radical (RO.) … Can be activated by the addition of energy, and transformed
… Iron-oxygen complexes (ferryl and perferryl radicals) into reactive oxygen species
… Thiyl radicals (RS.)
… Nitric oxide (.NO)

Singlet oxygen (1O2) Superoxide anion (O2-)

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… Formed from triplet oxygen

… Formed in almost all aerobic cells
… Conversion of oxygen to singlet state can be accomplished
by photosensitization in the presence of suitable sensitizers, … Not reactive enough to abstract hydrogen from lipids
such as chlorophyl, or heme pigments myoglobin or … Cannot enter the hydrophobic interior of membrane
h
hemoglobin
l bi or by
b their
th i derivatives
d i ti because of its charged nature
… Has a pair of electrons with opposite spins
… Can produce hydrogen peroxide (Dismutation)
… Involved in hydroxyl radical formation
… Can also react with nitric oxide (NO.) to produce
peroxynitrate (OONO-)

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Hydroperoxyl radical (HO2.) Hydroxyl radical (OH·)

9 10

… The most reactive oxygen species known: site specific reaction

… Protonation of O2- yields the HO2.
… Can be produced by high-energy ionizing radiation

.) H2O ·OH + ·H + H2 + H2O2 + H3O+ + e-aq

… Hydroperoxyl radical (HO2 is more reactive than (ionizing radiation)
superoxide
id andd can enter membrane
b fairly
f i l easily.
il … Fe2+-dependent decomposition of H2O2 (Fenton reaction)
Fe2+ + H2O2 Fe3+ + ·OH + OH-
Fe2+ + H2O2 ferryl? Fe3+ + ·OH + OH-

Lipid (R.), peroxyl (ROO.) and alkoxyl radical (RO.) Free radical sources
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… Exogenous
… Very strong reactivity and can abstract hydrogen atom from lipids
… Can be formed from the lipid radicals by iron complexes † Foods
… ROOH + Fe3+-complex -----> ROO. + H+ + Fe2+complex † Air pollutants
… ROOH + FFe2+-complex RO + OH- + Fe
l -----> RO. F 3+-complex
l † Radiation
… Endogenous
† Metabolism (mitochondria and peroxisomes)
† Detoxification
† Immune cells

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Lipid oxidation Mechanisms to induce lipid peroxidation

… Photoxidation
… Oxidative deterioration of lipids containing any
number of carbon-carbon double bonds † Singlet oxygen is involved
† Fatty acids † Require sensitizers: chlorophyll, porphyrins, myoglobin,
† Cholesterol riboflavin, bilirubin, rose bengal, methylene blue...
… Enzymatic Oxidation
… Produces various primary and secondary by-
products that influence food quality † Cyclooxygenase and lipoxygenase catalyze the
reactions between oxygen and polyunsaturated fatty
… Oxidized foods can cause oxidative stress in acids
biological systems and cause diseases … Autoxidation
† Free Radical Chain-reaction

Mechanism of lipid oxidation Mechanism of lipid oxidation

Initiation:
… The overall mechanism of lipid oxidation …

RH + O2 -->R· + ·OH
consists of three phases: R· + O2 --> · + ROO·
RH is any unsaturated fatty acid
† ((1)) initiation,, the formation of free radicals;; R· is a free radical
… Propagation:
ROO· + RH --> R· + ROOH
† (2) propagation, the free-radical chain ROOH is a hydroperoxide, one of the
ROOH--> RO· + HO· major initial oxidation products that
reactions; and decompose to form compounds
responsible for off-flavors and odors.
… Termination:
† (3) termination, the formation of nonradical R· + R· --> RR
products. R· + ROO·--> ROOR
ROO· + ROO· --> ROOR + O2

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Mechanism of lipid oxidation Undesirable Effects of Food Oxidation

• Flavor Quality Loss Rancid flavor

Consumer Acceptance
• Nutritional Quality Loss Essential Fatty Acids
Vitamins
• Health Risks Growth Retardation
Heart Diseases
DNA Mutation and Break
… This reaction scheme is capable of generating aldehydes, ketones, alcohols
and hydrocarbons. Many of the volatile compounds formed during lipid • Functionality Polymerization of Lipid and
oxidation originate through similar dismutations. Proteins

Effects of lipid oxidation to meat (food) products Production of toxic compounds

… A major cause of quality deterioration • Many secondary by-products of lipid oxidation are
† Develop rancidity in raw or fatty tissues potential carcinogens
† Produces WOF in cooked meats • Hydroperoxides are known to damage DNA
† Oxidized flavors in oils
• Carbonyl compounds may affect cellular signal
… Loss of functional properties transduction
… Loss of nutritional values • Aldehydes: 4-HNE and MDA
… Formation of toxic compounds
• Epoxides and hydrogen peroxide by-products are
… Forms colored products
known carcinogens

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Consequences of Lipid Peroxidation
… Structural changes in membranes
† Alter fluidity and ion channels
† Alter membrane-bound signaling proteins
† Increase membrane permeability
… Form lipid oxidation products adducts/crosslinks with
non lipids
† e.g., proteins and DNA
… Cause direct toxicity
† e.g., 4-hydroxynonenal, MDA
… DNA damage and mutagenesis
Consequences of DNA Oxidation
… DNA adducts/forms AP sites/strand break
† Mutation
† Initiation of cancer
… Stimulation of DNA repair
† Can deplete energy reserves
† Imbalanced induction of DNA repair enzymes
† Induction of error prone polymerases
† Activation of other signaling pathways
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Consequences of Protein Oxidation
… Oxidation of catalytic sites on proteins
† Loss of function/abnormal function
… Formation of mixed sulfide bonds
† Protein-protein linkages (RS-RS)
† Protein-GSH linkages (RS-GSH)
† Alterration in 2o and 3o structure
… Increased susceptibility to proteolysis
Pathological Conditions that Involve
Oxidative Stress
… Inflamation
… Atherosclerosis
… Ischemia/reperfusion injury
… Cancer
… Aging
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Effects of oxidation to human health Minimization of Lipid Oxidation

¾ If a compound inhibits the formation of free alkyl

radicals in the initiation step, or if the chemical
compound interrupts the propagation of the free
radical chain, the compound can delay the start or
slow
l th chemical
the h i l reaction
ti rate
t off lipid
li id oxidation.
id ti
¾ The initiation of free radical formation can be delayed
by the use of metal chelating agents, singlet oxygen
inhibitors, and peroxide stabilizers.
¾ The propagation of free radical chain reaction can be
minimized by the donation of hydrogen from the
antioxidants and the metal chelating agents.

Antioxidants Characteristics of Antioxidants

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™ is a molecule that inhibits the oxidation ¾ The major antioxidants currently used in foods are
monohydroxy or polyhydroxy phenol compounds with
of other molecules. various ring substitutions. These compounds have
low activation energy to donate hydrogen. The
¾ Th
The radicals
di l can start
t t chain
h i reactions
ti =>
> When
Wh resulting
lti antioxidant
ti id t free
f radical
di l does
d nott initiate
i iti t
the chain reaction occurs in a cell, it can cause another free radical due to the stabilization of
delocalization of radical electron.
damage or death to the cell.
¾ The resulting antioxidant free radical is not subject to
¾ Antioxidants terminate these chain reactions by rapid oxidation due to its stability.
removing free radical intermediates, and inhibit ¾ The antioxidant free radicals can also react with lipid
other oxidation reactions. free radicals to form stable complex compounds.

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Mechanism of Antioxidants Reaction of antioxidants with radicals

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R• + AH RH + A•
¾ Hydrogen donation to free radicals by antioxidants.
RO• + AH ROH + A•
¾ Formation of a complex between the lipid radical
and the antioxidant radical (free radical acceptor). ROO + • AH ROOH + A•

… Prevents the transfer of electron from O2 to

organic molecules R• + A• RA

… Stabilizes free radicals RO • + A •

ROA
… Terminates free radical reactions
ROO• + A• ROOA

Antioxidant + O2 Oxidized Antioxidant

Tocopherol and Oxygen Reaction Kinds of Antioxidants

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CH3 H2 α- tocopherol
Natural antioxidants:
OH H2
CH3 CH3
1. Vitamins A, E, C
CH3
O
O2 2. Polyphenols: Flavone, isoflavone, Quercetin,
( C H 2 ) 3C H ( C H 2 ) 3C H ( C H 2 ) 3C H ( C H 3) 2
CH3
Epicatechin Sesamol
Epicatechin,

CH3
CH3 CH3 CH3 Synthetic antioxidants:
O CH2 C H 2 C ( C H 2) 3C H ( C H 2) 3 C H ( C H 3 ) 2 1.Butylated Hydroxy Anisole (BHA)
OH 2.Butylated Hydroxy Toluene (BHT)
CH3 O
3.Propyl Gallate (PG)
CH3
α- tocoquinone 4.Tertiary Butyl Hydroquinone (TBHQ)

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Natural antioxidants Vitamin A (and carotenoids)

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… Functions: … Food sources:

† Normal vision † Liver
† Protects from infections † Fish oil
Isoflavone
Flavone † Regulates
g immune † Eggs
gg
system † Fortified milk or other
† Antioxidant foods
(carotenoids) † Red, yellow, orange,
Epicatechin and dark green
veggies (carotenoids)
Quercetin
Tocopherol Sesamol

Vitamin E Vitamin C
… Functions: … Sources: … Functions: † Heals cuts and wounds
† Antioxidant, may lower † Vegetable oils † Helps produce † Protects from infection,
risk for heart disease † Foods made from oil collagen (connective boosts immunity
and stroke, some types (salad dressing, tissue in bones, muscles, † Antioxidant
of cancers margarine) etc) … Sources
† Protects fatty acids † Nuts † Keeps capillary walls,
† Citrus fruits
and vitamin A † Seeds
blood vessels firm
† Other fruits, veggies
† Helps body absorb
† Wheat germ
iron and folate
† Green, leafy veggies
† Healthy gums

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Polyphenols Natural antioxidants

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… Many common foods contain rich sources of polyphenols which have antioxidant
properties. … Should not cause off flavors or colors
… Spices, herbs, and essential oils are rich in polyphenols in the plant itself and shown
with antioxidant potential in vitro. … Must be lipid soluble
… Red wine is high in total polyphenol count which supplies antioxidant quality.
… Must be non toxic
… Deeplyl pigmented
i d fruits
f i lik cranberries,
like b i bl b i
blueberries, plums,
l bl kb i
blackberries,
raspberries, strawberries, blackcurrants, figs, cherries, guava, oranges, mango, grape … Should have carry through properties
juice and pomegranate juice also have significant polyphenol content.
… Typical cooked vegetables rich in antioxidants are artichokes, cabbage, broccoli, … Must be cost-effective
asparagus, avocados, beetroot and spinach.
… Nuts are a moderate source of polyphenol antioxidants. Typical nuts are pecans,
walnuts, hazelnuts, pistachio, almonds, cashew nuts, macadamia nuts and peanut
butter.
… Sorghum bran, cocoa powder, and cinnamon are rich sources of procyanidins, which
are large molecular weight compounds found in many fruits and some vegetables.

Synthetic antioxidants Synthetic antioxidants

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• BHA • BHT
• Butylated hydroxyanisole is a mixture • Butylated hydroxytoluene is
of two isomers. Referred to as a also a 'sterically hindered'
'hindered phenol' because of the
proximity
p y of the tertiary
y butyl
y ggroup
p to
phenol. Susceptible to loss
the hydroxyl group. This may hinder th
through
h volatilization
l tili ti i high
in hi h
the effectiveness in vegetable oils, temperature applications.
but increase the 'carry through'
potency for which BHA is known. • Uses: Lard, shortening,
Butylated Hydroxytoluene (BHT)
vegetable oils, cereals,
• Uses: Lard, shortenings, vegetable
oils, cereals, package liners, potato Butylated Hydroxyanisole (BHA)
animal feeds, etc. Usually
products, dry soups, chewing gum, used in combination with
etc. Usually in combination with other BHA or BHT and citric acid.
primary antioxidants.

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Synthetic antioxidants Synthetic antioxidants

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… Propyl Gallate
• TBHQ †Three hydroxyl groups make it
• Tertiary-butylatedhydroquinone very reactive and lower solubility.
is an extremely potent It can chelate trace minerals
antioxidant.
a o da It had
ad bee
been used such as iron and form colored
extensively in non-food complexes. It is heat labile,
applications prior to gaining especially under alkaline
approval in food. conditions.
• Uses: Lard, cottonseed oil, Tertiary Butylatedhydroquinone † Uses: Lard, shortening, Propyl Gallate
(TBHQ)
potato chips, corn flakes vegetable oils, cereals, package
liners, animal feeds, etc. Used
alone and in combination with
BHA or PG and citric acid.

Food Antioxidants Food Antioxidants

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… Food antioxidants are all of the substances that i) Primary antioxidants

have some effect on preventing or retarding
† Terminate free radical chains and function as electron
oxidative deterioration in foods
donors
… They can be classified into a number of groups:
groups
† They include the phenolic antioxidants, butylated
† Primary antioxidants
hydroxyanisole (BHA), butylated hydroxytoluene
† Oxygen scavengers (BHT) tertiary butyl hydroquinone (TBHQ),
† Chelating agents or sequestrants propylgallate (PG) and natural synthetic tocopherols
† Enzymatic antioxidants

† Natural antioxidants

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Food Antioxidants Food Antioxidants

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ii) Oxygen scavengers iii) Chelating agents or sequestrants

† Can remove oxygen in a closed system † They remove metallic ions, especially copper and
† Most widely used compounds are Vitamin C,C and iron, that are powerfull pro
pro-oxidants
oxidants
related substances, ascorbyl palmitate, and † Citric acid is widely used for this purpose
erythorbic acid (the D-isomer of ascorbic acid)
† Amino acids and ethylene diamine tetraacetic
acid (EDTA) are examples of chelating agents

Food Antioxidants Food Antioxidants

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iv) Enzymatic antioxidants v) Natural antioxidants

† Can remove dissolved head space oxygen, such † Present in many spices and herbs
as glucose
l oxidase
id
† Superoxide dismutase can be used to remove
highly oxidative compounds from food systems

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Choices of Antioxidants Application of Antioxidants to Foods

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Different antioxidants show substantially different antioxidant effectiveness in

different fats and oils and food systems due to different molecular structures.
¾ Direct addition of antioxidants to oil or melted
We should consider the following: fat.
Safety
A i id
Antioxidant effectiveness
ff i ¾ Additi
Addition off antioxidants
ti id t to t the
th food
f d after
ft they
th
Off-odor
Off-color
are diluted in oil.
Convenience of antioxidant incorporation to foods
Carry-through effect
¾ Spraying antioxidant solution in oil on the food
Stability to pH and food processing or dipping food into antioxidant solution.
Availability
Cost
Non-adsorbable, if possible

Application of Antioxidants to Foods Antioxidants

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Pastry Cracker
Treatment
… Combinations Control 2 3
† Antioxidants are usually combined to take advantage of their differing .005 TBHQ 2 7
properties. Stability of
Bakery Products .001 TBHQ 3 10
† For example BHA may be combined with PG and citric acid. The citrate
chelates metals, the propyl gallate provides a high level of initial (AOM – .020 TBHQ 4 5
protection
i while
hil the
h BHA hhas good d carry through
h h properties.
i Days of stability) .005 BHA 8 12
.010 BHA 21 22
… Reasons for Combinations .020 BHA 27 33
† Take advantage of different properties
.005 BHT 5 10
† Allow for better control and accuracy
.010 BHT 10 14
† May provide synergistic effects
† Combinations may provide more complete distribution in some foods .020 BHT 19 21
† More convenient to handle .005 PG 2 3
.010 PG 5 6
.020 PG 3 11

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You are what you eat!

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