EXNO:01

DATE:

SAMPLING OF MILK
PURPOSE:
1. To get representative sample of entire batch of milk and milk product
2. To get correct results of the ingredient to be estimated
3. To become familiar with the different procedures to be followed in
collecting milk for analysis

APPARTUS:
1.
2.
3.
4.

Dipper for fluids

Agitator/Plunger for mixing
Milk sampler
Sample bottles

DIPPERS
A dipper of the shape and size shown in figure below is suitable for collecting
samples. The capacity of the sample containers shall be such that they are
almost completely filled by the sample taken by the dipper.

Dipper for taking samples

AGITATOR/ PLUNGER:

Agitators (also called plungers) for mixing milk need to be large enough to
produce adequate mixing. In view of the different shapes and sizes of
containers, no specific design of agitator can be recommended for all purposes,
but the design should be such that damage of the inner surface of the container
is avoided during mixing.
For mixing liquids in buckets or cans, an agitator of the design and dimensions
shown in figure 1 is suitable. The length can be adjusted to the depth of the can.
An agitator of the design and dimensions shown in figure 2 is suitable for use
for larger vessels (e.g. road and farm tanks).

Figure 1: Agitator (plunger) for cans and buckets

(in mm)

Figure 2: Agitator (plunger) for road, rail and

farm tanks (in mm)

SAMPLE CONTAINERS:
Sample containers should adequately protect the sample and not affect the test
results. Appropriate materials include glass, some metals (e.g. stainless steel)
and some plastics (e.g. polypropylene). The containers should preferably not be
transparent, but if they are transparent they should be stored in a dark place.

Containers and closures should be dry, clean and either sterile or suitable for
sterilization by one of the methods described below.
The shape and capacity of the containers depend on the particular requirements
of sampling, and could be e.g.100, 150 or 250 ml. It is desirable to avoid air
space by filling the bottles to the top, leaving however sufficient space to allow
for expansion of the rubber stopper.
Single-service plastic containers as well as aluminium foil of adequate strength
(sterile and non-sterile) and suitable plastic bags, with appropriate methods of
closure, may be used.
Containers other than plastic bags should be securely closed either by a suitable
stopper or by a screw cap of metal or plastic material. If stoppers are used, they
should be made from non-absorbent, odourless material. Do not use cork
stoppers or caps with cork seals on containers for microbiological examinations.
Sample bottles, which are to be examined for flavour should be closed with
greaseproof, non-absorbent stoppers to avoid change of odour or taste.

Sampling procedures:
Sampling should be performed by an authorized, properly trained, person. That
person shall be free from any infectious disease. Sampling for microbiological
examination shall always be undertaken by an experienced person. Samples for
microbiological examinations should be taken before other examinations, and
using aseptic techniques and sterilized equipment and containers.
It is important to obtain representative samples of the product. The following
procedure can be followed for sampling of raw milk.
1) Wash and dry hands, keep hands clean during sampling operation.
2) Mix milk thoroughly, by inverting, stirring or plunging the container for at
least 5 minutes. If the volume is small, it can be poured to and from one product
container to another of the same volume. Any milk fat adhering to the neck and
under the shoulder of the can shall be well mixed with the remainder of the

milk. Milk churns (fat separates) easily at 26.5 to 29.5C and agitation near this
temperature should be avoided.
3) Check milk temperature and record it on the sample container label.
4) Take the sample as soon as possible after mixing. Make sure the size of the
sample is sufficient for all necessary tests.
5) Seal each sample container airtight immediately after filling.
6) Label the samples with all the necessary information (see below).
7) Place sample in sample case and cool the sample when necessary.
8) Rinse equipment used for sampling after use.
9) Samples should be taken in duplicate.
Samples can be taken for
1. Physico-chemical examination
2. Microbial examination
3. Sensory evaluation

1. Physico-chemical examination
1) Should be examined as fresh as possible since important changes may
take place within a short time.
2) Samples should be preserved if long time lapses are anticipated.
3) Sample should have the same composition as that of bulk.

2. Microbial examination
1) Careful sampling is required to prevent contamination
2) Milk sample should be drawn aseptically in sterilized bottle with a
sterilized cap using sterilized pipette.
3) Sample should be tested immediately or it must be kept in ice.
4) No preservative should be added to the sample.
A) Sampling of milk from a single container : The operations
involved are mixing and immediately taking of the sample. Mixing can
be done by using a plunger which should be moved up and down for
about 10 times. If it is small container it should be poured from one
vessel for the other for mixing.
B) Sampling milk from different containers: Pour all the milk into
the vat and collect a single sample. Collect samples from different
containers in such quantities proportional to the quantities of milk and
mix and finally draw the required quantities of sample.
For Ex: 1ml - 1 litre- container
2mi - 2 litre- container

C) Sampling from rail and road milk tankers: Mix the milk
thoroughly using a large plunger. Insert the plunger through the man hole
and move it forward, downward and backwards and each time bring it to
surface. Mix for 10- 15 min and take sample using a dipper.
D) Sampling from storage tanks: Mix the milk thoroughly by
mechanical agitator or bubbling

Sample labels:
Samples should have a label attached with the following information:
Identification of the product
Nature of the product
Identification number
Name / signature of the person responsible for taking the samples.

Example of a label for a milk sample

Milk Producer .
Reference No......................................
Sample of (1)................................................................................................
Taken by (2)..................................................................................................
................................................................................................................
Time and temp. of sampling...................................................................
Date of sampling.....................................................................................

PRECAUTIONS:
1. Make milk homogenous by thorough mixing before sampling.
2. Agitators or plungers for mixing should be sufficient size and weight to
bring about thorough mixing and easy to move rapidly in milk by
operator
3. Milk churns easily at around 27C, avoid taking sample at this
temperature to avoid foaming, whipping and churning.
4. Collect 250ml of milk approximately from different samples to be mixed
together later.
5. Milk samples from bottled milk or tetra packs/poly packs be withdrawn at
random.
6. Milk bottle kept at low temperature for some time, be heated in water
bath up to 40C to melt the fat prior to mixing and sampling.
7. Avoid violent shaking to prevent foaming, whipping and churning.

PLATFORM TESTS
Objectives:

For examination of milk by adopting rapid test for acceptance / rejection of

incoming milk.
Platform tests include the tests for judging the quality of the raw milk.
These are:
(a) Organoleptic evaluation (OE),
(b) Clot on boiling test (COP),
(c) Alcohol test (AT),
(d) Sediment test (ST)
(e) Alizarin alcohol test
(f) Bromothymol blue test
The milk is collected from various sources and transported to milk scheme
for processing, marketing and distribution. Large quantity of milk is
supplied to the plant through different agencies, so that is subjected to check
for its suitability. Hence it is essential to examine the milk by using different
platform tests.

Material Required:
Milk cans or containers.

Apparatus:
i. Thermometer.
ii. Dipper
iii. Plunger.

Procedure:
Organoleptic Evaluation:
Examine the given sample of milk and record your observations on
following aspects.

Odor / Smell:
1. It can be judged within few seconds.
2 Remove the lid of can hold it inverted and raised to the nose and inhale the
smell.
3. Record the odor / smell as normal or abnormal.
4. Milk should be free from any off flavor like feed, fishy, barny etc.

General Appearance:
Note whether the milk is clear or any visible dirt or foreign matter has gained
entrance in milk. If so describe its nature for detecting the amount of dirt, in
the milk, conduct the sediment test if necessary.

Colour:
Observe the colour of milk as white, light yellow; record whether color of
milk is normal or abnormal. (Abnormal colours are reddish, bloody, bluish
etc.)

Consistency:
Record the consistency of milk as normal, watery, thick, ropy, and slimy.

Temperature:
Note down the temperature of milk at the time of receiving. It should be
below 50 F.

Precaution:
1. Raw milk should be tested.
2. Curdled / spoiled milk should be separated.
3. Temperature of milk while receiving at milk plant should be 5C.

Clot on boiling test:

Principle: The heating of milk in an advanced state of souring (acidity of
more than 0.20%) or abnormal milk (colostrums or mastitis milk) will
result in clotting.

Equipment and materials:

a) Test tubes (15.0 x 1.0 cm, preferably with a mark at 5 ml).
b) Source of heating, e.g. a boiling water bath or a flame.

Procedure:
1.Put test tubes with about 5 ml of milk in heating source for up to 5
minutes.
2.Rotate the tubes in an almost horizontal position and examine the film of
milk or side of the test-tube for any precipitated particles.
Interpretation:The acidity of milk that gives a positive test is generally
above
0.22 % (as % lactic acid) or has an abnormally high percentage of
protein like colostrum milk. Such milk cannot stand the heat treatment in
processing and is therefore not suitable for distributing as liquid milk or for
processing. Such milk must therefore be rejected. Please take note that at
high altitude milk boils at a lower temperature. This test is not very sensitive
to slightly sour milk.

Alcohol test:
Principle: Proteins in milk that has become sour (i.e. because of lactic acid

formation) will coagulate when mixed with alcohol.

Equipment and materials:

a) Test tube.
b) Pipette.
c) 68% ethanol solution (by weight: e.g. mix 68 ml 96% alcohol with 28
ml distilled water) or 75 % ethyl alcohol by volume (density0.8675 g/ml at
27 C).

Procedure:
1) Mix equal amounts (e.g. 2 ml) of milk and ethanol solution in test tube
with the pipette.
2) Agitate by gentle movement and look for coagulation.

Interpretation:
Milk containing more than 0.21% acid and milk that is abnormal
(e.g.colostrum or mastitis milk) will not pass the test. This milk is not fit for
further processing.

Sediment Test:
Objective:
To know the extent of visible dirt present in the milk as a mark of clean milk
production.
Principle: The sediment test presents a simple, rapid and quantitative
means of indicating the cleanliness of milk with respect to visible dirt. It
need not necessarily indicate heavy bacterial contamination
Material Required: Milk

Apparatus:
i. Sediment tester: This apparatus consists of a bottle open at both ends. To
the neck of which is fastened a clip and wire guage. This inverted neck is
kept downwards in stand small cotton discs are used to retain the dirt.
ii. Lintine disc 32 mm in diameter.
iii. Sampling dipper of 500 ml. capacity.
iv. Sediment disc ratings: 0.0, 0.2, 0.5, 1.0, 2.0 mg sediment per 500 ml. of
milk.
v. Sieves

Procedure:
i. Take a milk sample from well stirred can of milk with the help of
sampling dipper.
ii. Filter the milk through properly adjusted firm link disc held in the
sediment tester, so that a filtration area of 28 mm in diameter is exposed.
iii. Remove the cotton disc from sediment tester after filternation.
iv. Compare the lintine cotton disc with the standard disc.

Observation:
Observe whether the disc are free from residue, has slight residue or heavy

residue.

Conclusion:
Classification of the disc may be as Satisfactory ,Fair,Unsatisfactory and
Very unsatisfactory or Rejection grade as compared with standard disc
prepared

Resazurin Test:
Objective:
To determine extent of bacteriological quality of milk.

Material Required:
i Milk sample
ii. Resazurin colour/solution 0.05%.

Apparatus:
i. All purpose lovibond comparator.
ii. One Resazurin colour disc from blue to white.
iii. Water path, thermostically controlled to maintain temperature of
37 C.
iv. Test tube 10 ml.
v. Pipette 10 ml. and 1 ml.

Procedure:
i. Milk the sample thoroughly by inverting from one to another container.
ii. Pour 10 ml. of milk sample in to previously sterilized test tube.
iii. Add quickly 1 ml. of Resazurin solution in the test tube.
iv. Mix the milk and dye thoroughly by inverting 2-3 times.
v. Place the tubes in the water bath at the temperature of 37.5C
only for two minutes.
vi. The tubes are then removed from the water bath.
vii. Compare the colour to test tube with standard disc until the colours are
matched under comparator.

viii. Record the number of disc/colour of disc. If colour falls

between two disc numbers record half value.
Alizarin alcohol test :
Objective:
To detect the unbalanced and acidic milk by alizarin alcohol test

Principle:
The alizarin indicator changes its color at different PH.

Apparatus:
Test tubes, 5ml pipettes.

Reagents:
0.2%Alizarin solution in alcohol

Procedure:
1. Take 5ml of milk in a test tube.

2. Add an equal volume of indicator solution and shake well.

3. Observe the color of curd flakes formed.

Observation:
Range of color

Acidity%

Brown red
Reddish brown
Yellowish brown
Brown yellow
Yellow

0.16
0.20
0.24
0.28
0.36

Bromo- Thymol blue test:

Objective:
To detect mastitis milk by Bromo-Thymol blue test.

Principle:
Test is based on that milk from infected udders is high in chloride
content and has higher PH. With normal milk BTB gives yellow color but
with abnormal milk it gives green to slightly bluish green color depending
upon degree of alkalinity.

Procedure:
1. Take 5ml of milk in a test tube.
2. Add 5 drops of 0.2%alocoholic solution of Bromo-Thymol
Blue indicator.
3. Mix well and observe the color.
\

Observation:
Green or greenish blue color indicate positive test.