GHEE CHEMISTRY

Karuna Meghwal
Assistant Professor
MIDFT, Mehsana
Content
 Ghee-introduction
 Composition
 Legal standards
 Method of preparation
 Cow ghee & Buffalo ghee differences
 Colour of ghee
 Flavour
 Texture of ghee
 Fat constants
 Oxidation types and prevention
 Introduction
 According to FSSR-2011, ghee means the pure
heat clarified fat derived solely from milk or curd
or from desi (cooking) butter or from cream to
which no colouring matter or preservative has
been added.
 Ghee could be in liquid, semisolid and some time
in solid state based on the storage temperature.
 Ghee made from buffalo milk is whitish with
greenish tinge and that of cow milk is golden
yellow colour.
 Usually prepared form cow milk, buffalo milk or
mixed milk.
 Composition
 Ghee is a complex lipid of glycerides (majorly triglycerides), free
fatty acids, phospholipids, sterols, sterol esters, fat soluble vitamins,
carbonyls, hydrocarbons, carotenoids, (only in ghee derived from
cow milk).
Constituents Cow ghee Buffalo ghee
Fat (%) 99-99.5 99-99.5
Moisture (%) <0.5 <0.5
Carotene (µg/g) 3.2-7.4 (2.5-12.5) -
Vitamin A (IU/g) 19-34 17-38
Cholesterol (mg/100g) 302-362 209-312
Tocopherol (µg/g) 26-48 18-31
FFA (%) 2.8 2.8
Agmark standards for ghee

Parameters Special Grade General Grade Standard Grade

Baudouin Test Negative Negative Negative
Butyro- 40.0- 43.0 40.0-43.0 40.0-43.0
refractomer readin
g at 40C
Reichert Meissl val Not less than 28.0 Not less than Not less than 28.0
ue 28.0
Polenske value 1.0-2.0 1.0-2.0 1.0-2.0
Moisture content Not more than Not more than Not more than
0.3% 0.3% 0.3%
Percentage of Free Not more than 1.4 Not more than Not more than
Fatty Acid (as oleic 2.5 3.0
acid)
The word ‘Regional’ shall be printed on each label used on a package of the ghee not
conforming to the normal physical and chemical constants 

Parameters Special Grade

Winter Summer
Baudouin Test Negative Negative
Butyro-refractomer reading at 40C 41.5- 43.0 42.5 - 45.0
Reichert Meissl value Not less than 23.0 Not less than 21.0
Polenske value 0.5- 1.2 0.5 - 1.0
Moisture content Not more than Not more than
0.3% 0.3%
Percentage of Free Fatty Acid (as oleic Not more than 1.4 Not more than 2.5
acid)
AGMARK Ghee Specifications

 Grade designation marks for ghee- The grade designation mark

shall consist of a label specifying the name of the commodity,
grade designation and bearing a design consisting of an outline
map of India with the word AGMARK and the figure of rising
sun with the words produce of India and resembling the design
as set out as follows.
Agmark replica

Grade Letter and circular

border colour

Special Red
General Green
Standard Chocolate
As per FSSR, 2011
Fatty acids in milk fat

 Milk fat contains at least 500 fatty acids and fatty acid
derivatives with 4 – 20 or more carbon atoms in their chain.
 The fatty acid may saturated or unsaturated and usually
contains an even number of carbon atoms.
 Composition of fatty acid also varies between buffalo milk fat
and cow milk fat.
Composition of buffalo milk and cow milk fatty acid

Fatty Acid (%) Buffalo milk fat Cow milk fat

Butyric (C4:0) 4.4 3.2
Caproic (C6:0) 1.5 2.1
Capric (C8:0) 1.3 2.6

Caprylic (C10:0)
Lauric (C12:0) 1.8 2.8
Myristic (C14:0) 10.8 11.9
Palmitic (C16:0) 33.1 30.6
Stearic (C18:0) 12.0 10.1
Oleic (C18:1) 27.2 27.4
Linoleic (C18:2) 1.5 1.5
Linolenic (C18:3) 0.5 0.6
Method of preparation

 Concentration of milk fat in the form of cream or butter.

 Heat clarification of fat rich milk portion and thus
reducing the amount of water to less than 0.5%.
 Removal of the curd content in the form of ghee residue.
 There are five methods of ghee making:
 Desi or Indigenous Method
 Direct Cream Method
 Creamery Butter Method
 Prestratification Method
 Continuous Method
 Prestratification Method

 Butter is produced from aged cream of 38 to 40% fat using

continuous butter making machine or batch churn.
 Butter is then transferred to butter melter, and melt at 80°C.
 Molten butter is kept undisturbed in a ghee kettle or boiler at a
temperature of 80-85°C for 30 min.
 In ghee kettle, stratification of mass takes place, product
stratifies into 3 distinct layers.
 Denatured protein particles (curd particles) and impurities are
collected on top layer and floats on surface.
 Middle layer consists of clear fat and bottom layer consists of
buttermilk serum carrying 80% of moisture and 70% of solids-
not-fat contained in butter.
 The bottom layer is then carefully removed without disturbing
the both top and middle layers.
 Middle layer, largely consists of fat is heated to 114±2°C along
with top layer of floating curd particles and denatured protein.
 This step is necessary to develop characteristic ghee aroma.
 Milder flavour ghee can be produced, since most of the curd
content is removed before final clarification temperature of
ghee.
Ghee- fat
 Ghee majorly consists of milk lipids and richest source of
milk fat of all Indian dairy products.
 The constituents of ghee tend to vary with the method of its
manufacture.
 Chemically ghee is a complete lipid of glycerides, 97-98%
triglycerides.
 Small amount of di-and mono-glycerides are also present in
traces.
 Cow milk ghee is different from buffalo milk ghee in
terms of its composition

 Fatty acid composition of buffalo milk ghee also varies from cow
milk ghee.
 The amount of butyric acid is significantly higher in buffalo than
in cow ghee.
 The levels of short chain fatty acids caproic to myristic are
significantly higher in cow than buffalo ghee where as levels
of palmitic and stearic are higher in buffalo than in cow ghee. 
 Ghee- colour
 Ghee made from buffalo milk is white (lack of carotenoids) with
greenish tinge and that made from
 The characteristic colour of buffalo fat has been attributed to
tetrapyrozole pigments- biliverdin and bilivubin.
 This pigment is conjugated to a protein in milk, but is released during
the manufacturing process of ghee making.
 Imparting yellowish-green colour to buffalo ghee.
 Ghee made from cow milk is yellow‐greenish or mostly golden
yellow
 Because of beta‐carotene (3.2–7.4 µg/g) and chlorophyll.
Granulation

 Granulation is important criterion of quality; higher temperature

of clarification gives better grain size due to high phospholipids
content.
 Completely melted ghee on cooling to prevailing Indian
temperatures, can assume the form of large, coarse grains
suspended discretely or in clusters in a liquid phase.
 The process of crystallization is initiated with the formation
suitable nuclei.
 Rate of cooling strongly influences the rate nucleus formation.
 Cont..
 Stirring or agitation and seeding (at the rate of 1 – 3%)
encourage the nucleus formation.
 For better granulation, ghee should be slowly cooled to
28°C in 2-3 hours time and agitation is required during
granulation to form smaller granules.
 The partly granular form assumed by ghee is primarily
due to certain content of glycerides of higher melting
saturated fatty acids, especially palmitic & stearic.
 Thus buffalo milk ghee show predominant granulation
than cow milk ghee.
Changes during preparation of Ghee

 Water gets evaporated and fat present in the cream or butter

getting concentrated (curd particles (MSNF present in cream
or butter) starts settling at the bottom during clarification
process.
 Flavour formation in ghee happens during fermentation of
cream and during clarification process. 
 Colour development and granulation also happens during
clarification for the subsequent packaging of ghee.
 Ghee has a long keeping quality; it can be stored for 6 to
12 months under ambient temperature provided proper
packaging and filling
 Factors Influencing Keeping Quality of Ghee

 Temperature of storage: Higher the temperature of storage.

Lower will be the keeping quality and vice versa.
 Initial moisture content: Higher the initial moisture content.

The lower the keeping quality and vice versa.

 Initial acidity: Higher the initial acidity, lower the keeping

quality and vice versa.

 Exposure to metals: When ghee comes in contact with metals

especially iron and copper, its keeping quality gets reduced.

Since these is an act as catalytic agents for oxidation.
 Exposure to light: Greater exposure to sunlight causes

oxidation of ghee and thus reduces the shelf life.

 Method of packaging: Higher the air-content in the head-space

the lower will be the keeping quality.

 Preservation of Ghee
• Ghee is very much susceptible to oxidation.
• Reaction of oxygen with unsaturated fat, aggravated by metallic
contamination or sunlight, is a major cause for spoilage.
• To extend the shelf life of the product antioxidants can be added.
• Butylated Hydroxy Anisole (BHA) at concentration of not
exceeding 0.02% can be added.
• Use of ghee residue to extend the shelf life ghee has been a
practice.
• Phospholipids present in ghee acts as antioxidants and preserve the
ghee.
• It is reported that some curry leaves and betal leaves were also
tried to extend the shelf life of ghee
 Oxidation (Rancidity)
• Physico-chemical change in the natural properties of the fat
leading to the development of unpleasant odour or taste or
abnormal colour particularly on ageing/storage after exposure to
atmospheric oxygen, light, moisture, bacterial or fungal
contamination and/or heat
• Saturated fats resist rancidity more than unsaturated fats that
have unsaturated double bonds
Types of rancidity:
 Hydrolytic rancidity

 Oxidative rancidity

 Ketonic rancidity
 Hydrolytic rancidity
• It results from slight hydrolysis of the fat by lipase from
bacterial contamination leading to the liberation of free fatty
acids and glycerol at high temperature and moisture
• Volatile short-chain fatty acids have unpleasant odor

O O
CH2 O C R1 H2C OH R1 C OH
O
Lipase O
R2 C O C H HO C H + R C OH
2
O
O
CH2 O C R3 3 H2O H2C OH
R3 C OH
Triacylglycerol Glycerol Free fatty acids
(volatile, bad odor)
 Oxidative rancidity
• It is oxidation of fat or oil catalyzed by exposure to oxygen,
light and/or heat producing peroxide derivatives which on
decomposition give substances, e.g., peroxides, aldehydes,
ketones and dicarboxylic acids that are toxic and have bad odor
• This occurs due to oxidative addition of oxygen at the
unsaturated double bond of unsaturated fatty acid of oils
• Decomposition products of hydroperoxides are responsible for
off flavour in fat rich products
• Hydroperoxides are odourless
Mechanism of oxidation of fat
Primary oxidation

• Primary oxidation products are formed by the reaction of

an alkyl radical, which is formed by reaction to light or
heat (initiation), with oxygen to form a peroxy free
radical.
• The peroxy free radical reacts with an unattacked
unsaturated fatty acid to form a fat hydroperoxide and an
alkyl free radical (propagation).
• This product is tasteless and odourless.
• The reaction continues until there is a depletion of oxygen
or when a fatty radical reacts with a stable antioxidant
radical or when two unstable radicals react (termination).
Primary oxidation involves three steps, namely initiation,
propagation and termination, called autoxidation

Initiation:
 RH  R◦ + H (Abstraction of Hydrogen from fatty acid)
 R◦ + O2 ROO◦ (Addition of O2)
 RH (Unsaturated lipid), R◦ (Free radical), ROO◦ (peroxy radical)
 
Propogation:
 ROO◦ + RH  ROOH + R◦ (Abraction of hydrogen from another fatty acid and
formation of free radicals)
 ROOH (Hydroperoxides)
Termination When two free radicals react
 R◦ + R◦  RR
 R◦ + RO◦  ROR
 R◦ + ROO◦  ROOR
 2RO◦ + 2ROR◦  2ROOR + O2
 ROO◦ + ROO◦  ROOR + O2
Hydroperoxide formation

30
Secondary oxidation

• Secondary oxidation products are formed when the

hydroperoxides decompose to secondary oxidation products
as a result of heating, radiation, or the presence of heavy
metals (Cu, Fe) and other radical initiating agents.
• The secondary oxidation products are formed by either
peroxide scission alone or simultaneous peroxide and chain
scission.
• Chain scission leads to short-chain volatiles such as
aldehydes, ketones, alcohols and acids, which cause the
characteristic off-flavours and odours of rancid fats and oils.
Ketonic rancidity

• Free fatty acids are present in milk fat either inherently or

due to hydolysis of glycerides
• These free fatty acids are more prone to degradation by
molds etc. through β-oxidation
• Deterioration brought about by fungi/molds eg. Aspergillus
Niger
• Ketones, fatty aldehydes, short chain fatty acids and fatty
alcohols are formed
• Moisture accelerates ketonic rancidity
 Hazards of rancidity

1. Products of rancidity are toxic, i.e., causes food poisoning

and cancer
2. Rancidity destroys the fat-soluble vitamins (vitamins A, D,
K and E)
3. Rancidity destroys the polyunsaturated essential fatty acids
4. Rancidity causes economical loss because rancid fat is
inedible
Prevention of rancidity

1. Avoidance of the causes (exposure to light, oxygen,

moisture, high temperature and bacteria or fungal
contamination)
2. By keeping fats or oils in well-closed containers in cold,
dark and dry place (i.e., good storage conditions)
3. Removal of catalysts such as lead and copper that catalyze
rancidity
4. Addition of anti-oxidants to prevent rancidity eg. BHA,
BHT, tannins and hydroquinones.
1. Most common natural antioxidant is vitamin E
 Antioxidants

• Antioxidants are compounds or systems that delay autoxidation

by inhibiting formation of free radicals or by interrupting
propagation of the free radical by one (or more) of several
mechanisms:
• Scavenging species that initiate peroxidation,
• Chelating metal ions such that they are unable to generate reactive
species or decompose lipid peroxides,
• Quenching •O2− preventing formation of peroxides,
• Breaking the autoxidative chain reaction, and/or
• Reducing localized O2 concentrations
 The addition of antioxidants can retard autoxidation, as they are
free radical scavengers and by interrupting the chain reaction
prevent or slow down the propagation of oxidation.
 Synthetic antioxidants used are phenols such as BHA, BHT, TBHQ
and propyl gallate.
 Tocopherols (Vitamin E) naturally present in oil act as natural
antioxidants.
 The most effective antioxidants are those that interrupt the free
radical chain reaction.
 Usually containing aromatic or phenolic rings,
• Antioxidants donate H• to the free radicals formed during
oxidation becoming a radical themselves.
• These radical intermediates are stabilized by the resonance
delocalization of the electron within the aromatic ring and
formation of quinone structures
• Antioxidant is represented by AH.
• AH  Aº + H
• RH  Rº + H or Rº + O2  ROOº ( Initiation of oxidation process)
• Rº + AH  RH + Aº
• ROOº + AH  ROOH + Aº
Secondary antioxidants
 Reduce the rate of chain initiation by various
mechanisms such as scavenging oxygen, decompose
hydroperoxides to non-radical species, binding to metal
ions, absorb ultraviolet (UV) radiation or deactivate
singlet oxygen.
The requirements for an ideal antioxidant

• Safe in use
• Does not impart odour, flavour or colour to the product
• Must be readily incorporated in the product
• Be effective at low concentrations
• Should survive processing procedures, cooking and frying
• Economic to use.
 Antioxidants types

• Natural antioxidants:
• α-tocopherol, β-carotene
• Betel leaf (hydroxyl charcoal) and drumstick leave or other leaves or
fruit etc (gallic acid) addition during ghee manufacture extend the shelf
life of product
• Synthetic antioxidants:
• BHA (Butylated hydroxyl anisole)
• BHT (Butylated hydroxyl toluene)
• NDGA (Nor dihydroguaretic acid)
• TBHQ (tert butyl hydroxyquinone)
• Hydroquinone
• Catechol
• Pyrogallol etc
Synergists

• There are some compounds which by themselves possess

very little antioxidant activity but are capable of significantly
increasing the activity of primary antioxidants.
• As described in the mechanism of auto-oxidation, metal ions
play an important role in promoting oxidation.
• The synergists act by capturing or binding up the metal ions
and thus prevent them from taking part in any deteriorating
activities.
• The synergists could also function by regerating the spent
antioxidant or by preventing the antioxidant from stabilizing
the hydroperxide free radical.
• Common naturally occurring synergists are
phospholipids, ascorbic acid, citric acid and phosphoric
acid.
• Ethylene diamine tetra acetic acid (EDTA) is a very
strong sequestering agent and is capable of binding up
metal ions.
• Most synergists are poly basic acids.
• The synergists are capable of retarding autoxidation at
low levels but if present in higher levels may even
promote or accelerate auto-oxidation.
Pro-oxidants
• Pro-oxidants are compounds that initiate, facilitate, or
accelerate lipid oxidation. 
• Transition metals, such as iron and copper ions, are common
catalysts of lipid oxidation. 
• Pro-oxidants in oil have a detrimental affect on oil stability.
• Metals act as pro-oxidants by electron transfer whereby they
liberate radicals from fatty acids or hydroperoxides as in the
following reactions.
 Analysis and identification of fats/oils
(Fat constant)
Fat constants or numbers are tests used for:

1. Checking the purity of fat for detection of adulteration

2. To quantitatively estimate certain properties of fat
3. To identify the biological value and natural characteristics of fat
4. Detection of fat rancidity and presence of toxic hydroxy fatty
acids
Fat constants
• Iodine Number- It is the number of grams of iodine absorbed by 100
gm of fat under specified conditions. This constant is a measure of the
unsaturated linkages present in a fat
• Saponification Number- It is equivalent to number of milligrams of
potassium hydroxide require to saponify 1 gm of fat.
• Reichert-Meissl Number- Number of millilitres of decinormal sodium
hydroxide or aqueous alkali solution required to neutralize the steam
volatile water soluble fatty acids distilled from 5 gm of fat under the
precise conditions.
• Polenske value- Number of millilitres of decinormal sodium
hydroxide or aqueous alkali solution required to neutralize the steam
volatile water insoluble fatty acids distilled from 5 gm of fat under the
precise conditions.