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Determination of Fat in Milk,

EXPERIMENT 4 DETERMINATION Cream, Butter, Khoa, Paneer,


Ice-cream, Milk Powder
OF FAT IN MILK
CREAM, BUTTER,
KHOA, PANEER,
ICE CREAM AND
MILK POWDER
Structure

4.1 Introduction
4.2 Objective
4.3 Experiment
Principle
Requirements
Procedure
Observation

4.1 INTRODUCTION
Fat is the most important component of milk. It provides more energy than the
energy provided by carbohydrate (lactose) and proteins taken togather. It imparts
soft texture and creamy taste to milk products. It is the source of essential fatty
acids and carrier for fat-soluble vitamins. Due to these reasons the producers and/
or sellers of milk and milk products are paid for their product on the fat basis.
Therefore, determination of fat in milk and its products is an important exercise.

4.2 OBJECTIVE
l To determine the fat content in a given sample of milk or milk product.

4.3 EXPERIMENT
i. Principle

Fat content determination in milk and its products is done by methods,


which are based on two principles: -

A. Centrifugal Separation of Fat (Volumetric Gerber Method)

The method used for rapid determination of fat in milk and its product is known
as ‘Gerber Method’. It is based on the principle of measuring the volume of fat
released from a known volume of milk or known weight of product in a specially
devised and accurately calibrated modified cylinder called butyrometer. After
taking 10 ml of the Gerber acid (Sulphuric acid with specific gravity 1.84 at
200C) in the butyrometer 10.75 ml of milk is added to it in such a way that it
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forms a layer above the acid without mixing with it. Amyl alcohol (1 ml) is added Determination of Fat in Milk,
to break the emulsion. The sulphuric acid dissolve the milk fat in it after breaking Cream, Butter, Khoa, Paneer,
Ice-cream, Milk Powder
the fat globule membrane through its coagulating effect on proteins. The fat dissolved
in the acid is separated through centrifugal force and read in the stem of
butyrometer. In case of milk powder 10 ml powder is diluted 10 times with
distilled water and 10.75 ml of diluted milk is taken in butyrometer. Incase of
khoa, paneer 5g of the sumple is taken in cheese butyrometer instead of 10.75
ml milk. Rest of the method remain the same.

B. Extraction of Fat with Organic Solvents

For extraction of fat from the sample (oil in water emulsion) Fat globules are to
be broken. The milk proteins, which form the membrane around the globules of
fat, can be coagulated with alcohol. The free fat is then dissolved in acid (conc.
HCl) or alkali (conc. Ammonia). The separated, fat is then extracted with the help
of organic solvent.

ii. Requirements

A. Gerber Method

l Butyrometer; with 6%, 8% and 10% scale for whole milk. 1% and 4% for
skimmed milk and special butyrometer for ice cream, butter, paneer or Khoya.

l Automatic measures or 10 ml pipette for Gerber acid.

l Automatic measure or 1 ml pipette for amyl alcohol.

l Milk pipette (volumetric) having 10.75 ml mark at 270C.

l Gerber centrifuge capable of being rotated at 1100 rpm.

l Gerber acid (H2SO2) specific gravity 1.84 at 270C (90- 91% by weight)

l Amyl alcohol

B. Rose Gottlieb Method:

l Fat extraction tube (Rose Gottlieb or Mojonnier tube)

l Evaporating dish/ flask 150 ml capacity.

l conical flask 150 ml capacity.

l Ammonium hydroxide solution with specific gravity 0.88 at 200C

l Ethanol 95%

l Diethyl ether (peroxide free)

l Light Petroleum ether (40-800C)

l Mixed solvent 1:1 mixture of diethyl ether and petroleum ether

iii. Procedure

A. Gerber Method: Transfer 10 ml Gerber acid to milk butyrometer with


automatic measuring device. Add 10.75ml of well-mixed sample of whole milk 21
or skimmed milk. In case of milk powder either 1 gram powder is added then it Determination of Fat in Milk,
is diluted with 10 ml of water or 10 time diluted reconstituted sample (10.75ml) Cream, Butter, Khoa, Paneer,
Ice-cream, Milk Powder
is added. In case of other products about 5-gram sample is taken in cream or
cheese butyrometer in place of 10.75 ml milk. Milk is added to butyrometer
slowly and carefully in such a way that the tip of the pipette touch the base of the
neck of the butyrometer and form a layer above the acid without dissolving in it.
Add 1 ml of amyl alcohol with auto measure and close the neck of the butyrometer
firmly by inserting the stopper so as not to disturb the content. Carefully shake
the content by applying gentle swirling motion. After thoroughly mixing the content
place the butyrometer in water bath maintained at 650C for 5 minutes. After
removing from the bath and drying the butyrometer along with its contents is
centrifuged it for 5 min at 11000 rpm. Again keep the butyrometer along with its
in the water bath for 5 minutes and read the fat percentage by inverting the
butyrometer and keeping gradually and at eye level. The reading of the
butyrometer from the two mark as the scale which corresponds to lowest point
of the clear fat meniscus and to the fat acid interface.

In case of milk products the following amount of the sample is taken and miix with
boiling or hot distilled water and rest of the method is same as in case of milk.

1. Cream: 5 grams of is taken in cream butyrometer.

2. Butter: 2.5 grams butter is taken.

3. Paneer and Khoa: 5 grams sample is taken in cheese or cream butyrometer.

4. Ice Cream: 10 grams is taken in milk butyrometer.

5. Milk Powder: 1 gram of sample is taken and 10 ml distilled water is added.

B. Rose Gottlieb Method: Weigh accurately about 10 gram well mixed sample
of milk and 5-10 grams of milk product into the Rose Gottlieb or Mojonnier
tube. Add 1.25 ml of concentrated ammonia and mix well by gentle rotating.
Add 10 ml ethyl alcohol and mix thoroughly again. Add 25 ml diethyl ether and
close the tube with pre wetted bark cork and shake well for 1 minute then add
25 ml of light petroleum ether. Shake the contents thoroughly for 30 seconds.
Then allow the tube to stand for at least 30 minutes. When there is a separation
of ether layer siphon or decant the ether layer into a clean, dry and weighed
conical flask. Repeat the extraction process using 15 ml each of diethyl ether
and petroleum ether 2 times more. The cork of the tube is washed with solvent
mixture often before each removal. Evaporate the solvent of the conical flask by
keeping it on water bath being heated. Residual fat is dried at 102 ± 20C in a hot
air oven till its weight is constant.

In case of milk products the following amount of the sample is taken and miix with
boiling or hot distilled water and rest of the method is same as in case of milk.

1. Cream: 1- 2 grams of sample is taken.

2. Butter: 1- 1.5 grams of sample is taken.

3. Paneer and Khoa: For both the products generally 2- 2.5 grams of sample is
taken. 22
4. Milk Powder: 1 gram of sample is taken and 10 ml distilled water is added. Determination of Fat in Milk,
Cream, Butter, Khoa, Paneer,
iv. Observation/ Results Ice-cream, Milk Powder

W1 − W 2
Fat % = × 100
W

Where:
W = Weight of the sample
W1 = Weight of flask and fat
W2 = Weight of flask

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