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ANTIOXIDANTS
IN
FOOD
Presented by
EZHIL . C

Antioxidants in Food Industry

INDUSTRY

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WHAT IS ANTIOXIDANT ?
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Antioxidant is a substance which act against oxidation

( anti +oxidant )
It is defined as any substance which prevent ,retard or inhibit
oxidation.
Oxidation combination of oxygen which other elements 0r removal
of hydrogen ion or removal of electron from an element.
Reduction it is process of removal of oxygen or addition of
hydrogen ion or addition of electron to an element

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Antioxidant is a reducing agent (donates electron, thereby causes another reactant to
be reduced ).
How does it act against oxidation?
-Hence it is a reducing agent ,it donates electron to the
oxidizing agent.
-it get oxidized itself and prevent other element from
been oxidized.
Note :All antioxidants are reducing agent but all reducing
agents are not antioxidants.

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Need for Antioxidants in food industry

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Success of food product depends on consumer acceptability

Taste,Flavor and odour are important factors for a consumer

judge the product
Causes of deterioration -microbial destruction
- Rancidity of fats and oils
Microbial destruction can be prevented by preservation and processing

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Rancidity can be prevented by freezing ,packaging under vacuum

These method are expensive and not completely satisfactory
There comes a need for alternative cost effective method
usage of antioxidants
First used during World War II for food preservation

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Definition
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In food industry,
Antioxidants are defined as any substance which
is capable
of delaying ,retarding or preventing the development of
rancidity
or other flavour deterioration due to oxidation

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Types based on Functionality

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Two types -primary

( scavenging free radicals)

-secondary ( inhibit the pro-oxidant which

produce free radicals)
Primary Antioxidant
Free radicals - they are highly reactive particle
- produced by auto oxidation of lipids
- occurs at excited state by metal catalysis or by light

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Working mechanism
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O2

Lipid

Free radical

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Antioxidant

electron

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..
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Secondary antioxidant
Pro oxidant substance which can oxidize any element
and in turn produce free radicals
Antioxidants combine with pro oxidant and form stable compounds
Thus no free radicals are produced

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Types based on production

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General classification - Natural

- Artifical
Natural
Extract from natural raw materials
Variable composition
Ex - Citric acid,Vitamin E ,tocopherol
Note :Nowadays most of natural antioxidant are also synthesized
with same molecular structure for uniform composition

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Artifical

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Produced by chemical reaction

Pure and of constant composition
Ex-butylated hydroxyanisole (BHA) , butylated
hydroxytoluene (BHT).
Note: Even though many natural and synthetic compounds have antioxidant
properties, only a few of them have been accepted as generally recognised
as safe (GRAS).

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Mechanism
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Antioxidants inhibit the oxidation by four mechanism

Hydrogen donation by the antioxidant

Electron donation by the antioxidant
Addition of the component(oxidizing element) within the aromatic ring
of the antioxidant
Formation of complex between the oxidizing compound with the
antioxidant

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Usage in food Products

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Ascorbic acid and citric acids prevention
of oxidative discoloration of
meats ,fruits.

They are also used against discoloration due to enzymatic oxidation

Water soluble Rosemary extracts are used in sauces, marinades and salad
dressings (Giese,1996)
Oil soluble Rosemary extracts used in lipid based foods.
This is stable at higher temperature and very active even at a
concentration of 0.02%

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Rice bran oil proposed as antioxidant spray for snacks. It removes

objectionable colour and flavours.(Taylor et al.,1996)
It contains several antioxidants such as tocopherol, tocotrienols, oryzanol and triterpene alcohols
It is also used to preserve edible oils containing high linolenic acids such as soyabean ,wheat grem
oil ,rapeseed oil and canola
The green tea catechins solution is used in products such as pie crusts, dressings, meats,snacks
and pastries

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Wood smoke containing 2,6-dimethyoxyphenol,

2,6-dimethylphenol and 2,6-dimethyloxy -4- ethylphenol are used as antioxidant.
Phenolic compounds such as BHA (butylated hydroxyanisole),
BHT (butylated hydroxytoluene) are used in vegetable oils.
TBHQ (Tertiary butylated hydro quinone) has an excellent ability to retard oxygen
absorption in french fries ,potato chips
Combination gives more antioxidant property than used separately synergism
.Buck (1981),coulter (1988)

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Case study Paprika(Ricardo Gomez et

al)
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Paprika is a natural colourant extracted from pepper

There was a serious outbreak of food poisoning by
microorganism (Sjoberg et al,1991)
To reduce the microbial load heat treatment was employed to
attain commerical sterility.
Super heated water vapour of 150-160 C was used,followed by
rapid cooling

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Microbial stability was attained but there was loss of colour

Natural antioxidant extract of rosemary was mixed with final
proportion in 1% level.
It prevents carotenoids in paprika from losing the colour
The extracts antioxidant activity is from a series of phenolic
compounds such as carnosic acid ,carnosol ,rosmarinol and
rosmarin (Escalante et al,2001)

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Antioxidants on heating
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In foods antioxidants can be lost during food processsing such as

sterilisation, pasteurization, dehydration and cooking
During thermal treatment, beside nutritent loss ,chemical reaction
such maillard reaction takes place.
The non nutrient component such as maillard reaction
components (MRPs) are formed.
MRP act as antioxidants (i.e as chain breaker & oxygen
scavengers) (Maria cristina et al,1997)

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Case study MRPs(Tuaro et al,1996)

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A 6 Brix tomato puree was tested

Aliquots of tomato + virgin olive oil mixture were introduced into
100ml screw capped bottles and heated at 95 C upto 6 hrs.
Ascorbic acid was taken as standard.
Antioxidant activity was determined .(The ability of a compound
or a mixture of compounds to quench peroxyl radical produced
by 2,2azo-bis(2 amidinopropane)dihydrochloride is measured
spectophotometrically at 443 nm)
Antioxidant activity for every 2hrs is measured for every 1 hr.
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Second level
Third level
Relationship between
Antioxidant
Fourth level
and heating
time
Fifth
level

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activity

Result from the study

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Although natural antioxidants are lost during heating , overall antioxidant
property has increased.
Due to the development of new antioxidant components MRPs.
MRPs play an important role in food stability and hence gaining importance
Note: To prevent natural antioxidants from destruction due to heat,
encapsulation with suitable binding materials are done.

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Role of antioxidants in Human health

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They are the soldiers or missile defense system of your body,
preventing damage from molecular bombs called free radicals.
Oxygen reacts with food to give energy
This reaction releases harmful byproduct called free radicals
They are highly reactive forms of oxygen that have a missing electron

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To get the missing electron, they damage the healthy cell and its DNA
Free radicals are capable of causing heart disease, cancer ,aging etc
Anti-oxidants combine with free radicals to form more stable compounds
Thus damage to the body can be prevented

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Mechanism of aging
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Oxidative stress

Free radical ,

healthly cells

Antioxidants combine with free radicals and stop this chain reaction of damaging healthy cells

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Methods to determine antioxidants

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FRAP (Ferric reducing/antioxidant power),first test used to determine antioxidants.

This assay depends on the reduction of ferric tripyridyltriazine (Fe3+ -TPTZ) complex to
ferrous tripyridyltriazine (Fe2+ -TPTZ)
Fe2+ -TPTZ has an intensive blue colour and can be measured by 593 nm
spectrometrically.
Here the ability of the compound Fe3+ to produce Fe2+ is measured.

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Fe2+ is a pro oxidant ,it can react with H2O2 to produce free radicals (OH )
Why the ability to reduce Fe3+ is defined total antioxidant power?
Since almost all antioxidants such as ascorbic acid,uric acid can reduce Fe3+
This test is misleading ,since all the reductants that are able to reduce Fe3+
are considered antioxidants

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TRAP Assay
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The Total Radical Trapping assay of Wayner
et al was the most widely
used method of the last decade.
This method uses 2,2 azobis (2-amidinopropane)dihydrochloride
(AAPH) as peroxy radical producer
Plasma is used as oxidizable material, oxidation is inhibited by the
internal antioxidant Trolox
(6-hydroxyl-2,5,7,8 tetramethylchroman -2-carboxylic acid).
This value is measured and kept as standard and other antioxidants
ability is measured and compared with the standard

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ORAC Method
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Oxygen Radical Absorbance Capacity (ORAC) is an exciting and

revolutionary new test tube analysis for antioxidant (Glazers
laboratory)
It is an improved form of TRAP assay
This analytical procedure measures the ability of a food,
vitamin, nutritional supplement, or other chemicals to protect
against the attack by free radicals, or to act as an Antioxidant

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This assay is based on generation of free radical using AAPH

(2,2-azobis 2-amido propane dihydrochloride)
b-phycoerythrin (b-PE) was used as target oxidizable material.
It is fluorescent material,it losses its fluorescence on oxidation.
Addition of antioxidant prevent oxidation of b-PE
Measurement of decrease in fluorescence in presence of free
radical scavengers was noted.( Hong et.al., 1996)

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Trolox Equivalent (TE) was determined using Trolox as standard

The ORAC value is then calculated from the Trolox Equivalent and
expressed as ORAC units or value
The higher the ORAC value, the greater the Antioxidant Power

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Regulations for antioxidants in food

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The Codex Alimentarius is a collection of internationally

adopted food standards .
It permits only those antioxidants which have been
evaluated by the Joint FAO/WHO Expert Committee on
Food Additives (JECFA)
The primary objective of establishing permitted levels of
use of antioxidants in various food groups is to ensure
that the intake does not exceed the acceptable daily
intake (ADI).
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Acceptable daily intake (ADI) of various

antioxidants
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ANTIOXIDANT
BHA
TBHQ
gum guaiac
Tocopherols
Ethoxyquin
EDTA
citric acid
ascorbic acid
ascorbyl palmitate
propyl gallate
BHT
Phosphates
tartaric acid
Lecithin
Sulphites
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ADI (mg/kgbw)
00.5
00.2
02.5
0.152.0
00.06
2.5
not limited
not limited
01.25
02.5
00.125
070.0
030.0
not limited
00.7

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bw body
wt

Regulation of European Union

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E U regulation of antioxidants is stipulated by European

Parliament under Directive No. 95/2/EC of 20 February
1995 on food additives other than colours and sweeteners
There is a special position for sulphur dioxide and sulphites
They are classified as preservatives but may be used as
antioxidants when the risk of oxidation is greater than risk
of microbial spoilage

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Antioxidants used in various foods

E
Name
number

Foodstuff
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Maximum level

E
E
E
E

330
300
301
302

citric acid
l-ascorbic acid
Sodium l-ascorbate
calcium l-ascorbate

E
E
E
E
E
E

304
306
307
308
309
322

l-ascorbyl palmitate
Tocopherol
-tocopherol
Y-tocopherol
-tocopherol
Lecithins

weaning foods
quantum satis
fruit- and vegetable-0.3 g/kg
baseddrinks, juices
0.3 g/kg
Fat containing cereal-0.2 g/kg
based foods including
biscuits
fat-containing cereal 100mg/ kg individually
biscuits, rusks
or with combination

biscuits and rusks,10 g/kg

cereal based foods
E 330
citric acid
Cocoa and chocolate0.5%
E 334
tartaric acid
products
E 322
lecithins
Non-emulsified oils
30 g /kg
and fats of animal or
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Antioxidants in Food Industry vegetable origin

Rules to be followed
by Producer
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Only food grade antioxidants according to FAO/WHO limits can

be used
The food producer has full responsibility for the choice of
suitable antioxidant
They should obtain all information about the antioxidant and
declares its safety.
In most countries the antioxidants used in the product must
be labelled on the package.
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Benefits for Consumer

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Antioxidants can play a significant role towards

reducing the health hazards and decrease the
deterioration of oils,fats and foodstuffs made from
them
Extra expenditure which is about 0.04% is
significantly less when compared to the value of the
products and benefits derived out of them( Sumit
Arora et al,2004)

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Conclusion
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Recent investigations have been targeted towards identification of

novel antioxidants from natural sources because of adverse
toxicological reports on many synthetic antioxidants
There is an urgent need for standardization of evaluation methods
in order to obtain meaningful information
These are the big task for the food technologists to find ways in
discovering new natural antioxidants and determination its activity
It is also necessary to make the consumers to know why and how
the antioxidant are useful for the welfare of the society
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Antioxidants rich Fresh fruits and vegetables readily

available in our local market and their antioxidant
content (ORAC
38 METHOD)
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Second level
Third level
Fourth level
Fifth level

Antioxidants in Food Industry

ORAC CONTENT

750207
100
210
5570

1450

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References
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Maria Christina Nicoli, Monica Anese,1997.Loss and /or formation of antioxidants during
food processing and storage . Elsevier Science Ireland Ltd ,pp 71-74

Shama Rao P,J.Jaganathan,2004.Role of Antioxidants in food industries . Beverage & Food

World , June 2004,71-72

Ronald L.Prior ,Guohua Cao,1999.In vivo total antioxidant capacity and comparation of
different analytical methods. Free radical Biology & Medicine, Vol 27,pp 1173-1181

Ahmed Liyaqati S,2004.Role of antioxidants in improving the quality and shelf life of food
and food products. Beverage & Food World , July 2004,52-54

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Charles R. Caldwell,2000.A device for the semiautomatic determination of Oxygen Radical

Absorbance Capacity . Analytical Biochemistry 287,226-233

Singh.V.P,Neelam,2006.Natural antioxidants and their Health Aspects. Processed Food

Industry ,Vol-3,21-23

Joseph A. Price, Charles G. Sanny , Dennis Shevlin,2006, Application of manual assessment of

oxygen radical absorbent capacity (ORAC) for use in high throughput assay of total antioxidant
activity of drugs and natural products. Journal of Pharmacological and Toxicological Methods 54
(2006) 56 61

Codex Alimentarius General Requirements, 1995 Vol. 1A.

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Official Journal of the European Communities, 1995 38 L 61.

Jan Pokorny , Nedyalka Yanishlieva , Michael Gordon, 2001, Antioxidants

in food .Woodhead Publishing Ltd

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Consume antioxidant rich foods..

Stay youngstay fit..
THANK U

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