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ANTIOXIDANTS
IN
FOOD
Presented by
EZHIL . C
INDUSTRY

Antioxidants in Food Industry 4/27/2010

 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.

Antioxidants in Food Industry 4/27/2010

 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

Antioxidants in Food Industry 4/27/2010

 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”

Antioxidants in Food Industry 4/27/2010

 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

Antioxidants in Food Industry 4/27/2010

 Working mechanism
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O2 Lipid Free radical Antioxidant electron

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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

Antioxidants in Food Industry 4/27/2010

 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
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)’.

Antioxidants in Food Industry 4/27/2010

 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

Antioxidants in Food Industry 4/27/2010

 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)

Antioxidants in Food Industry 4/27/2010

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 extract’s antioxidant activity is from a series of phenolic

compounds such as carnosic acid ,carnosol ,rosmarinol and
rosmarin (Escalante et al,2001)

Antioxidants in Food Industry 4/27/2010

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)

Antioxidants in Food Industry 4/27/2010

 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 2,2’azo-bis(2 amidinopropane)dihydrochloride is measured
spectophotometrically at 443 nm)

Antioxidant activity for every 2hrs is measured for every 1 hr.

Antioxidants in Food Industry 4/27/2010

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Second level
Relationship between Antioxidant
● Third level

● Fourth level
activity
and heating time
● Fifth level

Antioxidants in Food Industry 4/27/2010

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.

Antioxidants in Food Industry 4/27/2010

 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

Antioxidants in Food Industry 4/27/2010

 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

Antioxidants in Food Industry 4/27/2010

 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

Antioxidants in Food Industry 4/27/2010

 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

Antioxidants in Food Industry 4/27/2010

 ORAC Method
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Oxygen Radical Absorbance Capacity (ORAC) is an exciting and
revolutionary new test tube analysis for antioxidant (Glazer’s
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”

Antioxidants in Food Industry 4/27/2010

 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).

Antioxidants in Food Industry 4/27/2010

 Acceptable daily intake (ADI) of various
antioxidants
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ANTIOXIDANT 32
ADI (mg/kgbw)
BHA 0–0.5
TBHQ 0–0.2
gum guaiac 0–2.5 bw –body
Tocopherols 0.15–2.0
wt
Ethoxyquin 0–0.06
EDTA 2.5
citric acid not limited
ascorbic acid not limited
ascorbyl palmitate 0–1.25

propyl gallate 0–2.5

BHT 0–0.125
Phosphates 0–70.0
tartaric acid 0–30.0
Lecithin not limited
Sulphites 0–0.7
Antioxidants in Food Industry 4/27/2010
 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

Antioxidants in Food Industry 4/27/2010

 Antioxidants used in various foods
E Name Foodstuff Maximum level
number 34
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E 330 citric acid weaning foods quantum satis
E 300 l-ascorbic acid fruit- and vegetable-0.3 g/kg
E 301 Sodium l-ascorbate baseddrinks, juices 0.3 g/kg
E 302 calcium l-ascorbate Fat containing cereal-0.2 g/kg
based foods including
biscuits
E 304 l-ascorbyl palmitate fat-containing cereal 100mg/ kg individually
E 306 Tocopherol biscuits, rusks or with combination
E 307 α-tocopherol
E 308 Y-tocopherol
E 309 β-tocopherol
E 322 Lecithins 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
Antioxidants in Food Industry vegetable origin 4/27/2010
 Rules to be followed
35 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.

Antioxidants in Food Industry 4/27/2010

 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)

Antioxidants in Food Industry 4/27/2010

 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

Antioxidants in Food Industry 4/27/2010

Antioxidants rich Fresh fruits and vegetables readily
available in our local market and their antioxidant
content (ORAC38 METHOD)
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ORAC CONTENT
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Second level 750207
● Third level
210 100
● Fourth level

● Fifth level
5570
1450

Antioxidants in Food Industry 4/27/2010

 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

Antioxidants in Food Industry 4/27/2010

Consume antioxidant rich foods…..
Stay young…stay fit…..

THANK U

Antioxidants in Food Industry 42 4/27/2010

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