Milk Analysis Methods For Dairies of Pakistan

Raw Milk Sampling Reference #

Used for: Practical implementation, for study and Issue date:
for reference.
Defination and explanation:
Sampling plays an important role in the subsequent analysis. So, taking representative sample
especially in case of liquid milk from any container mainly depends upon proper mixing of milk
in the container that in turn depends upon the appropriate agitation (technique and time).
Excessive mixing could lead to the deterioration of milk quality due to the disruption of milk fat
globules.
The International Association of Food Protection (IAFP, 1994) defines adequate agitation as the
degree of agitation which, at full tank capacity, results in a variation in fat content of the milk in
the tank of not more than ± 0.1% wt/wt as determined by an official AOAC milk-fat test.
Principal:
True representative sample can only be taken from true material, excluding any foreign material
that directly effect the composition of true material, like ice, and where there occurs phase
separation of material (like in case of milk), proper mixing is essential prior to take sample. The
sample thus taken should be handled properly in the laboratory otherwise all the efforts
precautions might go wasted.
Scope and application:
The described procedure can be used to take true representative sample from whole lot of milk in
any container (road tanker, farm cooling tank/silos and/or cans), so as to get accurate and precise
results from subsequent assay.
Procedure:
Removal of ice (where required)
Ensure the proper removal of ice from milk tanker/tank/silo/can where required..

Homogeneous mixing of milk:

For road tanker
The milk should be adequately agitated prior to sampling. Generally speaking, adequate agitation times are
five minutes for tanks under 1,000 gallons (3,800 L) and 10 min for tanks [of] 1,000 gallons or more.
Additional agitation time may be required under certain conditions." (DPC, 1991)
According to (OMAF, 1983) milk can be thoroughly agitated with plunger for 5 minutes.
For Farm cooling tanks
Agitate milk in a farm holding cooling tank for at least 5 min immediately before sampling.
Agitate contents of farm storage tanks of more than 5,700 L (1,500 gal) for 10 min, or as required
by the manufacturer.(APHA, 1992).
The earliest references we could find to what has become the commonly prescribed 5-min
standard were in Davis (1951) and Goss (1953), who both advised that milk in farm bulk tanks
must be thoroughly mixed with a sufficiently large plunger or by allowing a motor-driven agitator
or air compressor to run for several (usually 5) minutes.
For milk in cans:
For milk stored in single cans, the milk can be stirred with an up and down, circular motion of a
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Milk Analysis Methods For Dairies of Pakistan

disk-shaped stirring rod; the disc should be placed on the bottom of the can, pulled up on the near
side of the can, and then pushed down on the far side (Davis, 1951; Goss, 1953).

Sample handling in laboratory:

Uniform composition of milk collected in sampling bottle/pails/plastic beaker can be achieved by
pouring the milk from one bottle/pails/plastic beaker to another. Alternatively, the milk is mixed
with a dipper (Goss, 1953).
Calculations: Nil.
Repeatability & Reproducibility: NA
Precautions:
1. Dip your sampler to the center of the portion while taking sample.
2. Don’t take sample, if ice is present in any portion of tanker even if in empty portion.
3. Before taking sample, make sure that milk has been mixed thoroughly.
4. Clean the plunger and sampler with water after taking sample.
5. Prior to sampling make sure that beaker, sampler and plunger are neat & clean.
6. Check the tanker’s each portion carefully. If all the portions have equal quantity of milk
then take composite sample (take equal quantity of milk from each portion), otherwise
perform portion wise sampling. The sample should carry 500 – 800 ml of milk.
7. Prior to take sample, rinse the milk sampler (knoppy) and beaker with milk being
sampled.
8. After filling of knoppy with milk take care that all the milk is transferred from knoppy to
beaker.
9. If composite sample does not meet the given standards, perform portion wise sampling.
Hygiene requirements:
After performing taking the sample clean and wash all the accessories and place them at their
proper and designated places.
Reference:
1. OMAF. 1983. The Milk Act and Regulations of Ontario. Ontario Ministry of Agriculture
and Food, Toronto, ON.
2. APHA. 1992. Standard Methods for the Examination of Dairy Products. American Public
Health Association, Washington, DC.
3. Goss, E. F. 1953. Pages 2–9 in Techniques of Dairy Plant Testing. The Iowa State College
Press, Ames, IA.
4. Davis, J. G. 1951. Pages 17–23 in Milk Testing: The Laboratory Control of Milk, Dairy
Industries Ltd., London, UK.
5. IAFP. 1994. 3-A Sanitary Standards for Farm Milk Cooling and Holding Tanks, Number
13-09 International Association of Food Protection, United States Public Health Service,
The Dairy Industry Committee. Dairy Food Environ. Sanitation. 14:106–114.

Temperature Determination of liquid milk Reference #

Used for: Practical implementation, for study and for Date:
reference.
Definition and explanation:
The amount of heat energy present in the specific amount of milk describes the pretreatments of
milk as chilling of milk, storage of milk. Lesser the temperature of milk more will be its storage

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Milk Analysis Methods For Dairies of Pakistan

time depending upon the quality of milk.

Principle:
Thermometer worked on following principle:
Fluid expands on heating and contract on cooling. This expansion and contraction of fluid
depends upon the heat energy present in fluid i.e. more the energy in fluid more will it expand
and vise versa.
Temperature measurement by thermometer depends upon following principle:
Energy transferred from higher energy level to lower energy level, The heat energy from any
substance (either liquid or solid or gas) is transferred to fluid present in thermometer either or
from fluid of thermometer to substance by conviction or conduction or radiations results into
contraction and expansion of fluid in thermometer which is measured on scale corresponds to
temperature of any substance. Reading of thermometer scale will indicate the temperature of
sample in °C or °F.
Scope and application:
This method can be used to determine the temperature of fresh milk in field and at reception to
determine the chilling requirements of fresh milk. In factory temperature determination of liquid
milk is used for various reasons.
Standard: < 10oC for raw and pasteurized milk.
Reagents / Chemicals Required: Nil
Apparatus Required: 1. Thermometer -10 to110°C (Zeal) or any other
available
2. Glass, plastic or SS Beaker
Procedure:
1. Thoroughly mix the sample to get representative sample.
2. Dip thermometer bulb in it in such a way it should not touch the bottom and walls of
beaker.
3. Allow to stay thermometer in sample for at least for time that thermometer column no
more move upward or down ward.
4. Mercury thermometer, gives same reading if stays sometime outside the sample but
Alcoholic thermometer needs immediate reading of column as column rises or drops
readily outside the sample and cannot make spot or point measurement.
Calculations: NIL
Repeatability & Reproducibility: Not Applicable
Precautions:
1. Note the temperature immediately after taking representative sample.
2. Always ensure that thermometer being used is calibrated and signed.
3. Always use the alcoholic thermometer.
Hygiene Requirements:
1. Wash the thermometer after noting temperature and dry it with tissue paper for next time
usage.
2. Place all the beakers and thermometers used to their specific place after washing/cleaning.
3. Clean your work bench with clean cloth piece and alcohol where necessary.

Reference: General procedure adopted in dairy industries

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Milk Analysis Methods For Dairies of Pakistan

Clots on Boiling (COB) test of liquid milk Reference #

Used for: Practical implementation, for study and Date:
for reference.
Definition and explanation:
Acidity decreases the heat stability of milk. The clot-on- boiling test is used to determine whether
milk is suitable for processing, as it indicates whether milk is likely to coagulate during
processing (usually pasteurization). It is performed when milk is brought to the processing plant
— if the milk fails the test it is rejected.
The test measures the same characteristics as the alcohol test but is somewhat more lenient. It has
the advantage that no chemicals are needed. However, its disadvantage is that at high altitude
milk (and all liquids) boils at lower temperature and therefore the test is even more lenient.
Principle:
Acid developed in milk due to microbial activities during procuring, handling and transportation
of milk. This developed acid in milk make milk protein casein more heat sensitive so such milk
with more acidity (than natural milk) when boiled coagulate. On the other hand if milk sample
contains more whey proteins as globulin or albumins, such milk also show coagulation after
heating/ boiling as whey proteins are more heat sensitive than casein under natural acidic
condition of raw milk.
Scope and application:
This method is used to detect too advanced acidification in fresh milk that makes milk completely
unfit for purchase in field and accepting at reception (in field and at plant) and is also used in
factory to determine the heat stability of any kind of fluid milk for heat processing in factory.
Standard: COB should be - ve for all kinds of fluid milk.
Reagents Required: NIL

Apparatus Required: 1. Test tube

2. Spirit lamp or Bunsen burner
3. Wooden test tube clamp
4. Pipette 10 mL or any available.
Procedure:
1. Take 4-5 ml sample in test tube with the help of a pipette.
2. Then heat the sample on Bunsen burner or spirit lamp to boiling point.
3. After boiling tilt the test tube gently and observe.
Observations: Presence of clot/ precipitate will indicate COB +ve.
Repeatability & Reproducibility: Give same results for same kinds of samples.
Precautions:
1. Thoroughly mix the milk to get representative sample.
2. Glass ware used should be properly washed and cleaned.
Hygiene Requirements:
1. Wash and clean all glass ware, use detergent where required.
2. Place each and every chemical and glassware after use to their respective place.
3. Cover the each reagent bottle after using chemical/reagents.
4. Clean your work bench after performing analyses.
Reference:
1. LYONS, J.; O'SHEA, M.J., 1950. Page 161 in Commercial methods of testing milk and

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Milk Analysis Methods For Dairies of Pakistan

milk products, Cork Univ. Press, B.H. Blackwell Ltd., Oxford.UK.

2. Davis, J. G. 1951. Page 128 in Milk Testing: The Laboratory Control of Milk, Dairy
Industries Ltd., London, UK.

Alcohol Precipitate Test (APT) Reference #

Protein stability Test of liquid Milk
Used for: Practical implementation, for study and Date:
for reference.
Defination and explanation:
It is based on instability of the proteins when the levels of acid and/or rennet are increased and
acted upon by the alcohol. Also increased levels of albumen (colostrum milk) and salt
concentrates (mastitis) results in a positive test.
Principle: Protein of milk and alcohol compete for water available in milk. The higher the
concentration of alcohol used; greater will be it’s power to attract water. Consequently less water
will be available to keep the casein in suspension.
Scope and application:
This method is used to detect too advanced acidification in fresh milk in filed and at reception
both in field and at plant and is used to determine the heat stability of protein of fluid milk for
heat processing in factory.
Standards:
Raw milk should be - ve at least at 55 % Ethanol solution (V/V)
Reagents Required: Ethanol solution 68%, 65 % ,60% and 55 % (v/v)
Apparatus Required: 1. Pipette (10 mL)
2. Test tubes
3. Glass Petri dishes (where required)
Procedure:
1. Thoroughly mix the milk to get representative sample.
2. Take 2 ml milk sample with the help of pipette in a clean test tube.
3. Add 2 ml ethanol solution of required strength and mix, without shaking.
Observations: Presence of precipitate will be considered as APT +ve.
Repeatability & Reproducibility: Give same results for same kinds of milk.
Precautions:
1. Strength of Ethanol used should be clearly marked on each type used.
2. Pipette and Petri dishes / test tubes should be clean no accumulation of milk residue in it.
3. Light arrangement should be proper for proper visibility.
4. Make sure that none of the chemicals being used is expired.
Hygiene Requirements:
1. After performing the test clean and wash all the related apparatus and glassware.
2. Place all the chemicals in their specific place by properly labeling and covering.
3. Necessary instructions should be written about chemicals (hazardous) where required.
4. In case of any spillage or breakage, immediately clean the workbench carefully.
5. After working with solutions always wash the hands with soap.
Reference:
1. Roder, G., Grundzuge der Milchwirtschaft und des Molkereiwesens. Hamburg
u.Berlin,Verlag Paul Parey (1954) 680.
2. Sommer, Hugo H. 1938. Page 147 in Market Milk and Related Products. Madison.

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Milk Analysis Methods For Dairies of Pakistan

Wisconsin.

% Acidity Determination of liquid milks (Th°) Reference #

Used for: Practical implementation, for study and Date:
for reference.
Definition and explanation:
The acidity of fresh milk (natural acidity) is due to phosphates, casein and whey proteins, citrates
and carbon dioxide. The natural acidity of milk is 0.12 - 0.15%. Figures higher than this signifies
developed acidity due to the action of bacteria on milk sugar.
Bacteria that normally develop in raw milk produce more or less of lactic acid which contributes
towards the developed acidity in milk. The percentage of acid present in dairy products at any
time is a rough indication of the age of the milk and the manner in which it has been handled.
Fresh milk has an initial acidity due to its buffering capacity.
Total acidity of milk is sum of natural and developed acidity in milk and is analyzed as titratable
acidity.
Percent Treatable Acidity (%TA) is the percent acid in a sample based on titration data. More
specifically, it is usually defined in terms of the phenolphthalein end point (pH 8.3), which is
really an approximation of the true equivalence point (the point where all the acid is converted to
base). However, from the stand point of TA, the volume of base required to titrate to the end point
or equivalence point is what is of importance, and generally there is virtually no difference
between these volumes.
There are two fundamentally different methods of expressing acidity: (a) treatable acidity
expressed as percent lactic acid, and (b) hydrogen ion concentration or pH. The former measures
the total acidity but does not measure the strength of the acids. The pH indicates the strength of
the acid condition.
Principle: Acid-base Titration
Phenolphthalein is colorless in acidic media. The base added in the sample neutralizes the acid
present in sample and addition of one more extra drop base tends the whole media to become
basic and phenolphthalein present turns pink as it gives this color in basic media.
Scope and application:
This is a quantitative method used to determine the total titratable acidity of fresh milk in degree
thornic (Th°) at reception at plant, reception in area and in factory and also can be used to
determine the acidity of other fresh milk products in factory.
Standard: 0.08-0.14 for raw milk
1. 1.0 % Phenolphthalein indicator-
2. 0.1 N NaOH solution.
Reagents Required: 3. Distilled water
1. Weighing Balance( accurate to 0.1 g) or any available
2. Burette or acidimeter (25ml- graduated in 0.1 mL divisions)
3. Pipette 10mL or any available
Apparatus Required: 4. Glass Beaker (100 ml or any other)
5. pH meter with electrode
6. Buffer solutions (pH 4.01 and 7.01)
Procedure:
1. Weigh 9 mL (approximately equal to 9 gm) sample in a 100ml beaker.

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Milk Analysis Methods For Dairies of Pakistan

2. Add 0.5 mL (3-4 drops) of phenolphthalein indicator and swirl to mix.

3. Fill burette with 0.1 N NaOH up to 0 mL mark on top and allow it to drain until all air
bubbles have been removed.
4. Refill burette and drain until meniscus is at 0 mL mark.
5. Allow NaOH to drip into the beaker with continuous stirring titrate to a permanent pink
color the phenolphthalein end point is at pH 8.3.
6. Read level of NaOH in burette by standing at eye level with meniscus and read number of
milliliters of NaOH delivered.
Alternative Method:
1. weigh 9 g sample in a beaker
2. Use a pH meter and electrode. Stir sample with electrode while adding NaOH.
Stop addition of NaOH at pH 8.3. At pH 8.3 end point al the hydrogen is ionized.
Acidity is expressed as % Lactic acid.
1mL of 0.1 N NaOH = 0.009 g of lactic acid
% acidity = mL of 0.1 N NaOH usedx0.0009x100
Weight of sample
Calculations: % acidity = mL of 0.1 N NaOH used x 0.1
Repeatability & Reproducibility: + 0.01
Precautions:
1. Thoroughly mix the milk to get representative sample.
2. Solutions and indicator should be standardized.
3. Glassware being used should be clean and dry.
Hygiene requirements:
1. After performing the test clean and wash all the related apparatus and glassware.
2. Place all the chemicals by proper covering to their specific place after each test
3. Clean your work bench with clean cloth piece and alcohol (where necessary) after
working.
4. In case of any spillage or breakage, immediately clean the workbench carefully.
5. After working with solutions always wash the hands with soap.
Reference:
1. Standard Methods for the Examination of Dairy Products, 14th Edition, 1978, American
Public Health Association, Washington, D.C. Pages 355-357.
2. Atherton H.V. & JA. Newlander.1987. Page 250 in Chemistry and testing of dairy
products.AVI publishing Co.,Westport,U.S.A.

Salt (chloride) determination of Fluid Milk Reference # WBI/QAD/07-a

Used for: Practical implementation, for study and Date:
for reference.
Definition and explanation:
To increase the lactometer reading of milk just like sugar common salt i.e. sodium chloride is
added to milk as an adulterant this may also come in milk through ice used to chill the milk
during storage and transportation.
Principle:
Silver nitrate added reacts with salts (chloride) in the sample to form silver chloride and nitrates
of sodium or potassium that result in light brown color in media containing potassium chromate
as an indicator.

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Milk Analysis Methods For Dairies of Pakistan

Scope and application:

This method can be used to determine the % salt and chlorides of all kinds of fluid milk at
reception at plant and in factory.
Standard: 0.22-0.25 % for Raw and pasteurized milk at 9 %SNF.
Reagents Required: 1. 0.1N AgNO3 solution
2. 10 % Potassium chromate solution
1. Beaker (100 ml or any available)
Apparatus Required: 2. Pipette10mL or any available
3. Auto Burette or pipette 10 mL or any available
4. Dropper bottle/indicator bottle
Procedure:
1. Take 9 ml of well mixed sample in a beaker with pipette.
2. Add 2-3 drops of Potassium chromate indicator.
3. Mix the sample thoroughly.
4. Titrate it against 0.1 N AgNO3 solutions till light brown end point.
5. Note the reading of burette for silver nitrate used.
% Salt = No. of mL of AgNO3 used x 0.065
Calculations: % Salt (as chloride) = Burette reading x 0.065 x St. SNF
SNF % of sample
Repeatability & Reproducibility: ± 0.02
Precautions:
1. Thoroughly mix the milk to get representative sample.
2. All glass ware used should be properly cleaned.
3. Rinse pipette and beaker with sample before using.
4. Continuously agitate the contents of beaker during titration.
Hygiene Requirements:
1. After performing the test clean and wash all the related apparatus and glassware.
2. Place all the apparatus and glassware used to their specific place after washing/cleaning.
3. Clean your work bench with clean cloth piece and alcohol (where necessary).
Reference:
Sommer, Hugo H. 1938. Pages 144 and 648 in Market Milk and Related Products. Madison.
Wisconsin.

Lactometer Reading for SNF calculation and specific

Reference #
gravity determination of liquid milk.
Used for: Practical implementation, for study and for
Issue date:
reference.
Definition and explanation:
Addition of water to milk can be a big problem where we have unfaithful farm workers, milk
transporters and greedy milk hawkers. Any buyer of milk should therefore assure himself/herself
that the milk he/she purchases is wholesome and has not been adulterated. Milk has a specific
gravity. When its adulterated with water or other materials are added or both misdeeds are
committed, the density of milk change from its normal value to abnormal. The lactometer test is
designed to detect the change in density of such adulterated milk. Carried out together with the
Gerber butterfat test, it enables the milk processor to calculate the milk total solids (% TS ) and

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Milk Analysis Methods For Dairies of Pakistan

solids non fat (SNF).

Specific gravity is the relation between the mass of a given volume of any substance and that of
an equal volume of water at the same temperature.
Since 1 ml of water at 4°C weighs 1 g, the mass of any material expressed in g/ml and its specific
gravity (both at 4°C) will have the same numerical value. The specific gravity of milk averages
1.032, i.e. at 4°C 1 ml of milk weighs 1.032 g. 
Since the mass of a given volume of water at a given temperature is known, the volume of a given
mass, or the mass of a given volume of milk can be calculated from its specific gravity. For
example, one litre of water at 4°C has a mass of 1 kg, and since the average specific gravity of
milk is 1.032, one litre of average milk will have a mass of 1.032 kg.
Principal:
The function of the Lactometer/hydrometer is based on Archimedes principle that a solid
suspended in a liquid will be buoyed up by a force equal to the weight of the liquid displaced.
Thus, the lower the density of the substance, the lower the lactometer/hydrometer will sink.
In light liquids like kerosene, gasoline, and alcohol, the hydrometer must sink deeper to displace
its weight of liquid than in heavy liquids like brine, milk, and acids. In fact, it is usual to have two
separate instruments, one for heavy liquids, on which the mark 1.000 for water is near the top,
and one for light liquids, on which the mark 1.000 is near the bottom of the stem.
Scope and Application:
This method is rapid and only approximate and is convenient to use in conjugation with Gerber
and Babcock methods for fat. This most is suitable to determine the SNF contents and specific
gravity (by using formulae) of fresh milk at reception (in field and at plant) can also be used to
determine SNF and specific gravity of fresh milk and other liquid fresh milk products in factory
but it is preferable to determine total solid contents of liquid milks by drying in factory.
Reagents Required: Nil
Glass Ware & Equipment 1. Calibrated thermometer (0 – 110° C) or any available.
Required: 2. Calibrated Lactometer (with 14 – 42 graduations)
Quenne or any available.
3. A Plastic Cylinder (250 ml) at least 4 mm greater in
diameter than the bulb of lactometer.
4. Ice Box
5. Hot Water Bath
6. SS Beaker
Procedure:
1. Take about 500 ml milk sample in SS Beaker. Heat the sample at 40-45° C for about five
minutes and then cool down to 20° C.
2. Rinse the cylinder with some milk sample and drain it.
3. Rinse the Lactometer with the milk sample.
4. Now fill the cylinder with the milk sample completely.
5. Place the Lactometer over the cylinder and leave it gently.
6. Add some more milk in cylinder to remove the foam.
7. Wait for about 20 – 30 seconds till the Lactometer become stable, and then note the
reading (LR).
Calculations: SNF=(LR/4)+0.72+(0.22 X Fat % of sample)
Specific gravity= LR/1000+1
Repeatability & Reproducibility: ±0.5
Precautions:

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Milk Analysis Methods For Dairies of Pakistan

1. Prior to test make sure that all glassware and equipments are ok and neat & clean.
2. Lactometer must not touch the walls of cylinder.
3. Foam must be removed over the sample.
4. Temperature of milk sample must be maintained at 20° C.
Hygiene requirements:
After performing the test clean and wash all the related apparatus and glassware.

Reference:
1. FAO Manual of Food Quality Control, 14/8, page 12-1986-determination of total solid
(Rapid method) milk.
2. Sommer, Hugo H. 1938. Page 179 in Market Milk and Related Products. Madison.
Wisconsin.

Fat Determination of liquid milk (Gerber Reference #

method)
Used for: Practical implementation, for study and Date:
for reference.
Definition and explanation:
11 ml pipette (which delivered about 10.90 ml milk) was used in the original Gerber method.
Volume used in some countries are as follow:
1. India, 10.75 ml
2. The Netherlands, 10.66 ml
3. Hungary, 10.8 ml
4. UK., 10.94 ml
5. USA., 11 ml
New stopper of natural rubber absorb fat to some extant. Results are in gm fat/100 gm of milk.
Principle:
Everything in milk except the fat dissolves in sulphuric acid. The fat floats to the top. The
centrifuge ensures complete separation with no bubbles in the fat, iso-amylalcohol ensures the
complete separation of fat, and the fat content can be measured using the graduations on the
butyrometer.
Scope and application:
Gerber method is quick, quantitative method to determine the fat contents of fresh milk at
reception at plant and in factory can be used for liquid milks fat determination. This method is not
suitable for formalin preserved samples. This method can also be used in field by using manual
centrifugal machine but in this case result may vary.
Standard: Raw milk: 3.5-8.5 %
1. Isoamyl alcohol (sp. Gravity 0.081 ± 0.01)
Reagents Required: 2. Sulfuric acid (d=1.816 ± 0.003 g/ml, 90 – 91% purity with clear
color).
1. Calibrated Butyrometer ( 8 % )
2. Calibrated Pipette (10.94mL)
Apparatus 3. Key Stopper
Required: 4. Auto measure 10 mL with stand or pipette 10 mL.
5. Auto measure 1 mL with stand or pipette 1 mL.
6. Water bath

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Milk Analysis Methods For Dairies of Pakistan

7. Centrifuge machine (1100 rpm, 65°C temperature)

8. Calibrated Thermometer
Procedure:
1. Take about 500 – 800 ml of milk sample in a SS Beaker. Heat the sample at 40-45° C
and then cool down to 20 o C by continuously agitating.
2. Take 10 ml sulphuric acid in butyrometer with the help of Auto-measure.
3. Mix the milk sample well and rinse the pipette with the milk sample.
4. Now fill the pipette with sample above the mark.
5. Pour the samples from the pipette so that the upper meniscus of the sample reaches to
the mark.
6. Now pour the filled pipette into the Butyrometer gently making an angle of 45° with
butyrometer neck in 30-40 seconds.
7. Add 1 ml iso-amyl alcohol with the help of Auto-measure or pipette.
8. Insert the stopper into Butyrometer with the help of Stopper Key.
9. Mix the contents well by inverting butyrometer 3- 4 times and further shaking if
required.
10. Centrifuge the Butyrometer (by placing in such a way that graduated column remains
upward) in Gerber Machine for 3 min. for raw and 10 minutes for UHT milk at 65°C
and 1100 rpm.
11. Remove the Butyrometer from the Gerber Machine and immediately read the lower
meniscus of fat column in butyrometer.
12. Fat %age of sample directly correspond to the reading in butyrometer column.
Appearance of the Test
1. The colour of the fat column should be straw yellow.
2. The ends of the fat column should be clearly and sharply defined.
3. The fat column should be free from specks and sediment.
4. The water just below the fat column should be perfectly clear.
5. The fat should be within the graduation.
Problems in test results
Curdy tests:
 Too lightly coloured or curdy fat column can be due to:
 Temperature at milk or acid or both too low.
 Acid too weak.
 Insufficient acid.
 Milk and acid not mixed thoroughly.
Charred tests:
Darkened fat column containing black speck at the base is due to:
 Temperature of milk-acid mixture too high.
 Acid too strong.
 Milk and acid mixed too slowly.
 Too much acid used.
 Acid dropped through the milk.
Calculations: Nil.
Repeatability & Reproducibility: ± 0.05
Precautions:
1. Use gloves & goggles while handling acid.
2. Prior to test make sure that all glassware and equipments are calibrated and neat & clean

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no fat accumulation in any of glassware.

3. During filling of butyrometer take care that no acid falls on the outside of the butyrometer.
4. Always mix the contents in butyrometer very carefully better to cover the butyrometer
with piece of cloth.
5. Make sure that stopper is properly fixed in butyrometer.
6. Read the fat% as early as possible.
7. While starting centrifuge machine make sure that it is properly balanced.
8. Stir the sample gently and continuously while heating and cooling.
9. Always read the meniscus accurately.
10. Use calibrated Gerber Machine, Thermometer & pipette.
11. In some Butyrometers water needs to be added or Stopper needs to be adjusted to maintain
the level of Fat Column.
12. Note the reading immediately because esterification of iso-amyl alcohol starts if remains
at 65°C for more time and it results in high fat reading.
13. After reading, always remove the stopper & contents from the butyrometer under tap
water to avoid any suffocation from acid alcohol vapors, while the butyrometer is in
straight position to avoid any kind of intact with its contents.
14. In case of any breakage, make its entry in breakage record.
15. When any butyrometer is broken inside the machine, during its cleaning always use
gloves or cloth so that to prevent any injury caused by glass pieces.
16. Water bath should be handled carefully during working, always take care that hands don’t
touch the electric connections.
17. Always turn off the electric supply of the Gerber machine, water bath etc. before cleaning.
Hygiene requirements:
1. After performing the test clean and wash all the related apparatus and glassware.
2. Place all the beakers and thermometers used to their specific place after washing/cleaning.
3. Clean your work bench with clean cloth piece and alcohol (where necessary) after
working.
4. In case of any spillage or breakage, immediately clean the workbench carefully.
5. After working with solutions always wash the hands with soap.
Reference:
1. Davis, J. G. 1951. Page 59 in Milk Testing: The Laboratory Control of Milk, Dairy
Industries Ltd., London, UK.
2. FAO Manual of Food Quality Control, 14/8, page 8-1986-determination of milk fat by
Gerber method.

Detection of Glucose adulteration in liquid Reference #

milk. (by Barfoed reagent)
Used for: Practical implementation, for study and Date:
for reference.
Defination and explanation:
Glucose is mostly added in fresh milk by the vendors of milk to increase its lactometer reading so
to increase the SNF contents of milk either to adjust the fat/SNF ratio of milk or to prepare
fabricated milk.
Principle: Glucose reduces the soluble blue copper ion (Cu+2) to insoluble reddish brown
copper oxide (Cu+1), that precipitates and settles down giving following colors with increased

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Milk Analysis Methods For Dairies of Pakistan

concentration of sugar/glucose. Green – yellow – orange – Reddish brown – Brick Red.

Scope and application:
This is a qualitative method used to determine the adulteration of all kinds of milk by glucose
(corn syrup) either liquid or solid at any fresh milk reception set up either in factory or in field.
Standard: Milk should be glucose – ve except where it is being used as an ingredient.
Reagents Required: Barfoed reagent
1. Test tube (16 X 160 mm)
Apparatus Required: 2. Boiling water bath
3. Pipette
4. Stop watch
Procedure:
1. Take 2.5 mL sample to be tested in a test tube and add 5 ml of Barfoed reagent.
2. Place the test tube in water bath at boiling temperature for 3.5 minutes.

Observations: If brick red color precipitate is found in test tube bottom

after. 10 minutes the test is +ve.
Repeatability & Reproducibility: Give same results for same kind of milk.
Precautions:
1. Thoroughly mix the milk to get representative sample.
2. Stay time in water bath should not more than 3.5 min. with more time slight +ve will
change into severe + ve glucose.
3. Water bath should be handled carefully during working, always take care that hands don’t
touch the electric connections.
Hygiene Requirements:
5. Wash and clean all the glass ware after use.
6. Cover each reagent bottle after using chemical/reagents in test.
7. Clean your work bench after performing analyses.
8. In case of any spillage or breakage, immediately clean the workbench carefully.
9. After working with solutions always wash the hands with soap.
Reference:
1. Ervin J., 1985. Spot Test Analysis: Clinical, Environmental, Forensic, and Geochemical
Applications, vol 75.John Wiley & Sons Inc
2. Mittel, S.B. and Roy, M.K. 1974. XIX 1st Dairy Congress, IE 432-3.

Detection of Glucose in Fresh milk by Glucose Reference #

strips
Used for: Practical implementation, for study and Issue date:
for reference.
Definition and explanation:
Glucose is mostly added in fresh milk by the vendors of milk to increase its lactometer reading so
to increase the SNF contents of milk either to adjust the fat/SNF ratio of milk or to prepare
fabricated milk. Using glucose strips for determination of glucose adulteration in milk is quicker
and easy method.
Principle:
Glucose concentrations are indicated by a color change form yellow to bluish green of test zone/
portion of glucose strip.

Page 13 of 23
Milk Analysis Methods For Dairies of Pakistan

Scope and Application:

This method can be used to determine the adulteration of all kinds of milk by glucose (corn
syrup) either liquid or solid at any fresh milk reception set up either in factory or in field.
Procedure:
1. Take Fresh, Well mixed milk sample in a clean dry beaker.
2. Remove one test strip from the container.
3. Immerse test strip in sample for approximately one sec.
4. After removing the test strip from the sample, put lateral edge of strip on the paper.
5. Wait for 30 seconds.
6. Find a match or select the color value found on the container that comes as close as
possible to the test field and note the results.
7. Finish the evaluation within 60 sec. after immersion the strip into the sample.
8. Submit the results observed within the above time limits, since the test filed may change
its color intensity after this period.
Calculations: Nil.
Repeatability & Reproducibility: NA
Precautions:
1. Do not touch reaction zone /test field of strip.
2. Close container tightly after removing test strip.
3. Test field must be submerged in sample.
4. Do not put the strip down on the paper.

Hygiene requirements:
After performing taking the sample clean and wash all the accessories and place them at their
proper and designated places.
Reference:
Manufacturer Literature.

Sodium contents of milk by Ion Selective Electrode Reference #

Used For: Study, practical implementation and for Date:

reference.
Definition and explanation:
To increase the lactometer reading of milk just like sugar common salt i.e. sodium chloride is
added to milk as an adulterant this may also come in milk through ice used to chill the milk
during storage and transportation.
Principle:
The sodium ion selective electrode (ISE) consists of sodium containing glass membrane, which is
highly selective to Na+ ions. Based on Na+ ion activity in a given solution, the ISE generate an
electric potential across the membrane vs. a reference electrode. The membrane potential, E, is
thus related to the activity of Na+ by the nernst Equation.
E = Eo + S log (A)
E = measured electrode potential
Eo= reference potential
A = Na+ in solution
S= electrode slope
Scope and application:

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Milk Analysis Methods For Dairies of Pakistan

Description of a rapid direct potentiometer method that measures sodium in fresh milk and
reconstituted milk powder using an ion-selective electrode and a potentiometer.

Standard:
Reagents/Chemicals Required: 1. Nitric acid (65%), sodium chloride (extra pure)
2. Triethanolamine,
3. ISA solution
4. Electrode rinsing solution
5. Electrode storage solution
6. Sodium standard solution
Apparatus Required: 1. Magnetic stirring bar.
2. CSCE with BNC connector.
3. pH meter with BNC connector.
4. Plastic lab. Ware.
5. Volumetric flask.
Sample Preparation:
Fresh Milk:
Take 50 mL of fresh milk into a 100 mL beaker. Adjust pH of milk sample to above 9 by addition
of ISA solution (approx. 1.7 mL).
Milk Powder:
Weigh 5 g of powder in a 100 mL volumetric flask and add 60 mL warm wate (60°C) and mix
well.
Cool it to room temperature, adjust pH to above 9.0 by adding approx. 1.7 mL of ISA solution
and make up to the mark with water.

Instrument Check:
1. Make sure that the pH meter is operating the mV mode. Perform an electrode slope check
before commencing analysis of sample solution the change in potential observed when the
sodium concentration changes by a factor of10 should be 56±2 mV.
Calibration Procedure:
As per brochure of Equipment
Procedure 1:
1. Transfer 50ml sample solution into a 100 ml beaker.
2. Add 1.7 ml ISA solution check that the pH>9.0.
3. Immerse the electrode into the beaker and measure under slow magnetic stirring. It takes
10-20 seconds for the reading to stabilize; in the case of the Hach meter, an acoustic signal
is emitted and “ready “ will be display on the right side of the LED display.
4. Note the mV reading and remove the electrode.
5. Rinse the electrode thoroughly with rinsing solution and keep the electrode between
measurements in the electrode storage solution (5.3).
Procedure 2:
Following is the procedure for Sodium determination of the raw milk
1. Take sample and heat it till temperature 20-25°C.
2. Take 50-60 ml of this sample in 100 ml plastic beaker.
3. Place that beaker on the magnetic stirrer and start stirring it (100-rpm aprox.
Not more than it).

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Milk Analysis Methods For Dairies of Pakistan

4. Add 1 ml ISA solution in the sample.

5. Remove the electrode of sodium meter form the storage bottle, wash it with tap
water and dip the electrode in the sample (keep 1-1.5 cm distance of electrode
bulb from the bottom of the beaker).
6. Meter start reading the value in gm.
7. Wait until the reading is stable.
8. Note the reading as ppm (parts per million) of sodium e.g. if reading is 0.425 gm then it is
425 ppm and if the reading is 1.125 gm then it is 1125 ppm.
9. Always record the result at 9 % SNF contents e.g. if the vehicle having SNF
6.645 % and its sodium contents are 425 ppm then calculate the sodium
contents at 9 % SNF in the following way
425/6.645 X 9 = 575 ppm

Observations/Calculations Calculat the sodium content of the product as follow if procedure

: 1is used,
Fresh MILK, in mg/l:
F. {(C.D)-B}

MILK Powder, in mg/kg

V. {(C.D)-B}/m
Where
F= 1.034 Factor to compensate dilution of milk with ISA solution)
V= Volume of analytical solution in mL (normally 100 mL)
C=Conc. Sodium in analytical solution, in mg/L
D= Dilution factor if required
B= conc. Of sodium in blank solution, in mg/L
m=mass of milk powder analyzed, in gram
Repeatability & The difference between two independent single test results, within
Reproducibility: a short interval of time should not exceed 5% of their average
value
Precautions:
1. Always use plastic beaker for this test.
2. Sodium meter require calibration after at least 48 hrs or when needed.
3. Always store the electrode after proper washing and cleaning in its storage
bottle when not in use.
4. Reading stabilizes in 3-4 minutes but some times take more or less time
depending upon the sample condition.
5. If reading is not stabilizing then contact the under signed.
Reference:
1. Instruction Manual, hach Model 50310 Combination Sodium calomel Electrode (pp 1-30).
2. AOAC Official method 976.25. Sodium in Foods for Special Dietry Use. Ion Selective
Electrode Method. AOAC 1995.
3. Ion Selective Electrodes, Honold, F. & camman, K., Chapter 4, Pg 75. In: Rapid methods
for Analyses of Food and Food Raw Material. (Ed.) baltes, W. technomic Publishing Co.,
Lancaster, basel, 1990.

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Milk Analysis Methods For Dairies of Pakistan

Detection of Urea adulteration in liquid Milk Reference #

Used for: Practical implementation, for study and Date:

for reference.
Definition and explanation:
Urea is added in milk as preservative, Urea has strong buffering capacity so it resists the change
in pH due to developed acidity in milk by microbial activities. Urea also increase the lactometer
reading of milk and hence SNF contents. The milk adulterated with urea is some what bitter in
taste.
Principle: Urea is hydrolyzed by urease enzyme. During this, ammonia liberated will form
ammonium hydroxide (base) with water.  Phenol red indicator changes its color in basic medium
(pH reaches 8.4 by liberation of ammonium hydroxide. the tube will turn from an orange color to
a pink color.
Scope and application:
This is a qualitative, colorimetric, visual comparison method for the detection of presence of urea
and urea like compounds at any fresh milk reception set up either in factory or in field.
Standard: Urea should be –ve for all kinds of fluid milk.
Reagents Required: 1. Urease solution (0.2 %, freshly prepared)
2. Phenol red indicator.
1. 10 ml Pipette or any available.
Apparatus Required: 2. Incubator
3. Test tube (16 X160 mm)
Procedure :
1. Take 5 ml of sample in two test tubes namely A and B each.
2. Add 5 mL urease solution in each of the test tubes by using pipette.
3. Add 1 - 2 drops of phenol red indicator in both tubes.
4. Place test tube A in incubator at 40ºC for 15 min. while other will remain in shelf
5. Observe the color of solution in test tube A and compare its color with that of B to
evaluate status of milk.
Observations: o Color changes from B urea +ve
o Color remains same as that of B urea - ve.
Repeatability & Reproducibility: Give same results for same kind of milk.
Precautions:
1. Thoroughly mix the milk to get representative sample.
2. Make sure that pipette used should be free from any milk residue before taking sample in
it.
3. Sample should be incubated at 40 o C.
4. Urease solution should be freshly prepared (not use more than 3-4 days).
5. Only observe the change in color after incubation period as at initial stage color change
may be due to original high pH of milk.
6. In case of any breakage, make its entry in breakage record.
Hygiene requirements:
1. After performing the test clean and wash all the related apparatus and glassware.
2. Properly cover chemicals.
3. Clean your work bench after each test performed.

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Milk Analysis Methods For Dairies of Pakistan

4. In case of any spillage or breakage, immediately clean the workbench carefully.

After working with solutions always wash the hands with soap.
Reference:
1. Kurunegala BT-QM&R RKR/27.01.94.
2. Atherton, H.V. and J.A. Newlander. 1977. Chemistry and Testing of Dairy Products, 4th
Edn., AVI Publ., Westport, CT.

Detection of Sugar adulteration in liquid Milk Reference #

Used for: Practical implementation, for study and Date:
for reference.
Definition and explanation:
Cane sugar is most common adulterant in milk. It increases the specific gravity of milk so
increases lactometer reading. If water is adulterated to milk its lactometer reading lowers down
and to compensate this sugar is added to milk as an adulterant so that its lactometer reading
should increase. And so seller can deceive to the consumer or purchaser.
Principle:
Hydrochloric acid breaks down the cane sugar into its monosaccharide units (Glucose and
Fructose). Fructose such formed reacts with nephthol and form violet color compound.
Scope and application:
This is a qualitative, colorimetric method for the detection of presence of cane sugar, cane juice in
fresh milk at any milk reception set up. It is also applicable to heated treated milk.
Standard: Sugar should be –ve for milk except that in which it has been used as an ingredient.
Reagents Required: 1. 35 % HCl.
2. 10% naphthol
1. Test tubes (16 X 160mm)
2. Pipette 10 mL or any available
Apparatus Required 3. Test tube stand
4. Electric heater or any other heating source.
5. Glass beaker
6. Stop watch
Procedure:
1. Take 0.5 ml of sample in a test tube and add 3 drops of 10% naphthol.
2. Then add 3 ml of HCl.
3. Place the test tube in boiling water for 10 seconds.
4. Allow to stand the contents of tube for 8-10 min.
Observations: Appearance of light violet color sugar +ve.
Repeatability & Reproducibility: Give same results for same kind of milk.
Precautions:
1. Thoroughly mix the milk to get representative sample.
2. Glass ware used should be properly washed and cleaned.
3. While keeping in boiling water, keep close contact on stop watch. Time should not exceed
more than 10 sec.
Hygiene Requirements:
1. Wash and clean all glass ware, use detergent where required.
2. Place each and every chemical and glassware after use to their respective place.

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Milk Analysis Methods For Dairies of Pakistan

3. Cover the each reagent bottle after using chemical/reagents.

4. Clean your work bench after performing analyses.
Reference:
Modified method from
Valentinis, G.: “Contributo alla ricerca del saccarosio nel latte e nel latte in polvere”, in: Boll.
Lab. Chim. Prov. 5 (4) 122-126 (1954). A summary of this article can be found in: Dairy Science
Abstracts 17 703 (1955).

Detection of adulteration of liquid milk with Reference #

carbonates and bicarbonates. (Soda test)
Used for: Practical implementation, for study and Date:
for reference.
Definition and Explanation:
During storage of milk its acidity goes on increasing due bacterial development. After the acidity
of milk more than 0.17 %, milk curdles on heating it. Soda means sodium bicarbonate, which acts
as a preservative in milk. It neutralizes the developed acidity of milk.
Principle:
Ethanol added in milk sample coagulates the casein protein which is separated from milk serum.
The milk serum containing the carbonates and/or bicarbonates turns the colour of rosalic acid
added from red to pink.(color change of rosalic acid depends upon the pH of serum at neutral pH
colour of rosalic acid is red while at basic pH, due to presence of any base, colour of rosailc acid
is pink).
Scope and application:
This is a qualitative, colorimetric method for the detection of presence of carbonates, bicarbonates
and other neutralizers in fresh milk at any milk reception set up and in factory where required.
Standard: Detergent should be – ve in all kinds of fluid milk.
Reagents Required: 1. 95 % Ethanol
2. 1.0 % Rosalic acid
Apparatus Required: 1. Test tube
2. Pipette 10 mL or any available.
Procedure:
1. Take 5 ml sample in a test tube. Add 5 ml 95% ethyl alcohol in it.
2. Add 3 drops of rosalic acid and mix thoroughly.
3. Appearance of pinkish color indicates the presence of carbonates and bi carbonates in
milk.
Calculations: Not applicable
Repeatability & Reproducibility Same results of same samples
Precautions:
1. Thoroughly mix the milk to get representative sample.
2. Glassware being used should be clean and dry.
3. In case of any breakage, make its entry in breakage record.
Hygiene requirements:
1. After performing the test clean and wash all the related apparatus and glassware.
2. Place all the chemicals by proper covering to their specific place after each test.
3. Clean your work bench with clean cloth piece and alcohol (where necessary) after
working.

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Milk Analysis Methods For Dairies of Pakistan

4. In case of any spillage or breakage, immediately clean the workbench carefully.

5. After working with solutions always wash the hands with soap.
Reference:
1. Government of India Manual of methods of analysis of Foods, page 12. 2005. Rosalic acid
test for presence of carbonates.
2. Atherton, H.V. and J.A. Newlander. 1977. Chemistry and Testing of Dairy Products, 4th
Edn., AVI Publ., Westport, CT.

Detection of adulteration of starch in liquid

Reference #
Milk
Used for: Practical implementation, for study
Date:
and for reference.
Defination and explanation:
Starch is mostly added in fresh milk by the vendors of milk to increase its lactometer reading so to
increase the SNF contents of milk either to adjust the fat/SNF ratio of milk or to prepare fabricated
milk.
Principle:
Starch form a complex with iodine that ranges from blue to blackish colour depending upon the
type of starch present.
Scope and application:
This method can be used to determine the adulteration of milk with starch.
Standard: Starch test should be – ve for all kinds of milk except where used
Reagents Required: 0.1 N Iodine solution
Apparatus Required: 1. Petri dish
2. Pipette (2, 5 or 10 mL)
3. Dropper bottle
Procedure 1:
1. Take 2 mL of sample in a Petri dish using pipette.
2. Add 1-2 drops of iodine solution in it and mix/whirl thoroughly.
3. Allow it to stand. And after one minute observe/examine the bottom of the Petri dish.
Procedure 2:
1. Take 5 mL of sample in a test tube using pipette.
2. Bring to boiling conditions and allow the test tube to cool to room temperature.
3. Add 1-2 drops of iodine solution in it and mix/whirl thoroughly.
4. Allow it to stand and after one minute observe/examine the test tube.
Observations: Presence of dark blue or black grains will indicate the
addition of flour or starch in milk.
Repeatability & Reproducibility: Give same results for same kind of milk.
Precautions:
1. Thoroughly mix the sample to get representative sample.
2. Iodine solution should not be very old.
3. If there is same results (+ve) for each sample then check the sample and ensure there is not
grain formation in it. If present then replace solution with fresh.
Hygiene Requirements:
1. After performing the test clean and wash all the related apparatus and glassware.
2. Place all the chemicals in their specific place by properly labeling and covering.
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Milk Analysis Methods For Dairies of Pakistan

3. Necessary instructions should be written about chemicals (hazardous) where required.

4. Clean your work bench with clean cloth piece and alcohol where necessary.
Reference:
1. Gupta, A.K. & M.L. Varshaney. 1989. Practical Manual for Agricultural Chemistry,
Kalyani publishers. New Delhi.India. (Procedure 1)
2. I.S. 1479 (Part I)- 1960. Methods of test for dairy industry, rapid examination of milk.
(Procedure 2)

Determination of BR value or refractive index by

Reference #
using BR Machine or refrectometer.
Used for: Practical implementation, for study and for
Issue date:
reference.
Defination and explanation:
Fresh milk is adulterated with foreign fat mostly vegetable fats, in order to increase the fat
contents of fresh milk to fetch more price of specific quantity of milk or it is added in milk as a
recipe ingredient to prepare fabricated milk.
Principal:
The natural milk fat has BR value from 39-43.5. If the extracted fat sample has BR value or
refractive index value more than the prescribed limit of variability, it means the milk sample or
fat sample is adulterated with foreign fat (vegetable fat).
Scope and application:
This is a qualitative method used to detected vegetable oil adulteration in fresh milk at any kind
of reception set up and is also used in factory to determine purity of any kind of fat or oil either in
Butyro-refrective value or in refractive index.
Standard:
Raw milk must have BRV: 39-43.5
Reagents Required: 1. Ethanol absolute or any available.
Glass Ware & Equipment 1. BR machine/Refractometer
Required: 2. Water bath/ Water Circulator
3. Tissue Paper
4. Spatula/Spoon
5. Heating source
Setting up the Instrument (Indian BR machine):
Place the BR machine on the workbench with prism assembly facing towards a convenient source
of illumination such a window or a filament lamp of about 25 watts, 12 inch in front of the prism
box and on the same level. Screw the thermometer on to the nipple adaptor mounted on to the left
hand side of the lower prism box. Connect the two halves of the prism box by a loop of tube.
Connect a tube from Water Circulator exit point to the nipple on the top of the upper prism box
(as the inlet) and another tube to the thermometer nipple (as the outlet).
Sample preparation for fat:
Milk:
1. Fill the test tube with a milk sample.
2. Rotate the filled test tube in centrifugal machine for about 3-5 minutes at 1100 rpm or
more by balancing this test tube with another test tube.
3. After centrifugation remove the upper thick layer of cream from tube with the help of any
suitable spatula.

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Milk Analysis Methods For Dairies of Pakistan

4. Heats that cream in spoon on either direct flame of fire or any other heating source so that
fat is chard out from cream (with least possible water in it).
Cream:
Take some cream sample in spoon and heat it either on direct flam of fire or any other heating
source so that fat is chard out from cream with least possible water in it.
Ghee and oils:
No need to prepare sample for refractive index/BR value determination.

Procedure for BR or refrectometer reading:

1. Maintain the temperature of the water circulator to 40° C and allow the water to circulate
in the Refractometer prism assembly for about 10-15 minutes.
2. Open the prism box by means of the knob on the right hand side.
3. Clean the polished surface of the fixed prism box (lower half of the prism box) with the
help of ethanol and place 2-3 drops of fat/oil on it.
4. Close the prism box by lowering down the upper half.
5. Make the light in front of prism box on.
6. Look through the eyepiece of the instrument. There will be a borderline between the light
and dark portions crossing the oil scale. If borderline is colored, then make it clear by
vertical movement of the small level protruding from the right hand side of the body.
7. Note the reading either BR value or refractive index depending upon type of equipment.
8. Use table to interchange each value into other value.
Calculations: Nil.
Repeatability & Reproducibility: ±0.5
Precautions:
1. Prior to test make sure that all equipments are ok and neat & clean.
2. Maintain the temperature of the circulating water at 40° C throughout the whole activity.
3. Clean the polished surface of the upper and lower prism box with the help of ethanol and
tissue paper, before and after every reading.
4. Always use distilled water in water circulator and care about its level.
5. Make sure that water is properly circulating in the tubes.
6. Read all the instruction/safety measures/precautions form the manual of equipment use.
Hygiene requirements:
After performing the test clean and wash all the related apparatus and glassware and place them at
their proper place.
Reference:
1. Instruction manual of instrument.
2. Government of India Manual of methods of analysis of Foods, pages 11-12. 2005. Test for
the presence of foreign fat in milk.

Detection of fabricated fresh milk

Reference #:
Used for: Practical implementation, for study and
Issue date:
for reference.
Scope and application:
This method can be used to detect fabricated fresh milk.
Principal:

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Milk Analysis Methods For Dairies of Pakistan

Acid and heat coagulate the casein of fresh milk; the coagulated casein is separated from serum
by filtration. The brix value of filtrate indicates the total solids present in it except casein protein.
So by subtracting the value of brix from SNF of milk sample we obtained the amount of casein
that is coagulated. The lesser the amount means more the adulterants present in milk.
Reagents Required: 1. Sulphuric Acid (d=1.816 ± 0.003 g/ml, 90 – 91% purity
with clear color).
Glass Ware & Equipment 1. Glass funnel
Required: 2. Stand.
3. Filter Paper (Whatman # 42).
4. Glass Beaker 100 ml.
5. Hand Refrectometer (0-32 % brix).
Procedure:
1. Rinse the milk sample well and take 10.94 ml in a 100 ml beaker with the help of a milk
pipette.
2. Add 0.05 ml Conc. H2SO4 and mix thoroughly.
3. Heat the sample in hot water bath at 60-65°C for 5-10 seconds.
4. Filter the contents with filter paper.
5. Cool down the filtrate to 20°C.
6. Find the Brix of filtrate with the help of Hand Refrectometer and note it.
Calculations: (SNF of milk sample – Brix value) X 9
________________________________
SNF of milk sample

Results: For pure milk the resultant value must always be upto
1.25 or more, the value below 1 indicates any type of
adulteration in milk.
Precautions:
Hygiene requirements:

Reference:

Page 23 of 23