Project semester midway report

Submitted in partial fulfillment of the

requirement for the seventhsemester
of
Bachelor of Technology in Biotechnology

DEPARTMENT OF BIOTECHNOLOGY
THAPAR INSTITUTE OF ENGINEERING &
TECHNOLOGY , PATIALA

Submitted by: Prince

Roll no - (701800036)
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 INTRODUCTION TO ORGANIZATION

 The haryana state Dairy Corporation was formed in 1970 which continued
active functioning till 31.03.1977.There after its business was taken over by
Federation to set up based on Anand Pattern. Since1.04.1992 the
Federation has leased out the Plants to the Milk Unions:-

State Milk
Federation
STATE LEVEL

Milk Producer's cooperative union

District level

Milk Producer's coperative Society

village Level

 As part of stringent quality measures, milk required for processing VITA

products is procured from Dairy Cooperative Societies only. It is ensured
that the milk is transported to chilling centres and plants in clean and
sterilized milk cans as quickly as possible.
 All quality measures as per Standard of Bureau of Indian Standards/Ag
mark are being applied before the products are marketed. Well-equipped
laboratories are functioning in the chilling centres and milk plants to
maintain ideal quality standards.

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  It set-up as a three tier system with Federation as the apex body at the
State Level, Milk Unions at the District Level and Cooperative Societies at
the village level. The organization has continuously advanced towards its
coveted objectives well defined in its by-laws.

Main Objectives of organization:

a. To ensure quality milk procurement at remunerative price coupled with

improved animal productivity for reducing cost of milk production for
sustainable growth of the milk producers.

b. To provide quality extension services at the door steps of milk producers.

c. To ensure Quality Assurance in procurement, processing & marketing and

enhancing efficiency in the entire value chain.

d. To ensure acquisition, development and retention of competent

manpower.

e. To improve fiscal management by optimizing the returns on capital.

f. To ensure continual modernization in the entire value chain through

effective leveraging of technology.

g. To embrace innovation in the entire value chain for consumer delight.

h. To ensure continual expansion of distribution.

i. To enhance brand equity

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THE SIRSA DISTRICT COOPERATIVE MILK PRODUCERS’ UNION LTD.

 Milk Union Sirsa registered during 1970 and started functioning on Anand
Pattern from Sept.’1977. It purchases fresh Milk twice a day directly from
Milk Producers by organising Milk Producers’ Co-operative Societies (MPCS)
at village level in entire sirsa Distt. Presently, The Sirsa District Cooperative
Milk Producer’s Union Limited, VITA SIRSA Dairy is a front-line Milk Plant in
Co-operative Sector having installed handling capacity of 150,000 litres of
Milk per day.

 343 functional milk producer co-operative societies formed at village level

are supplying about average of 1.5 lakh litres of Milk per day. This milk
procurement goes to about 2 lakh litres in a day during peak flush season.
Fresh milk is procured directly from milk producers through village level
societies twice a day. The main function of this plant is to provide best
quality of Pasteurised Liquid Milk to the consumers in SIRSA and adjoing
cities.

 Apart from Liquid Milk supply (Full Cream Milk, Std. Milk, Toned Milk,
Double Toned Milk & Skimmed Milk), this Plant also manufacture Ghee,
Paneer, Curd, Plain Lassi, Pinni, Spiced Lassi, Sweet Lassi , milk powder.

 Being a viable unit with robust financial health, this plant pays dividend to
all member societies out of its earning every year.

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 FLOW CHART OF MILK RECEPTION

Tanker

Weigh Bridge

Sampling

Sample approved Sample not approved

Unloading

Manual Automatic

Pumped to Deaeration Vessel

Storage Tank

Duplex Filters

Raw Milk Chiller (Milk Chilled to 3°C)

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 Pumped to RMST (Raw Milk Storage Tanks)
SPECIFICATION TO GUIDE THE RMRD

Alcohol test : Negative

Acidity : 0.11 to 0.14%

Adulteration : Negative

R.M. Value : 28-32

COB : Negative

Sodium content : less than 450 ppm

B.R. reading : 40-43

P.V. Value : 1-2

TESTS PERFORMED IN LABORATORY ON RAW MILK RECEPTION:

1) FAT test (By Gerber method)

2) SNF test (by lactometer)
SNF=CLR/4+0.22F+0.29
CLR= corrected lactometer reading at 27º C
F= fat of milk by Gerber method
3) Adulteration test (negative)
4) BR reading of milk (40-43 at 40ºC)
5) Sodium test of milk (upto 450 ppm)
6) RM value of milk (28-32)
7) Quantity: To be checked
8) Acidity

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9) MBRT: for good quality raw milk is 2 hrs
Raw Milk Reception Dock

 VITA follows a strict system of quality assurance to maintain a high quality

of milk provided to the consumer.

 The milk is received from local societies at village level, where chilling
centres are installed.

 On the basis of some main tests the milk is graded into categories.

Grading of Raw Milk

 On the basis of physical condition and some test the milk is graded into four
categories:

Types of Raw Milk Grades

Fresh raw Milk 1

Sour Raw Milk 2

Curd 3

Dirty Milk 4

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 In the Plant there are three main labs:

• Dock Lab

• Quality Control (Q.C.) Lab

• Microbiological Lab

• AGMARK lab

FUNCTIONS OF DIFFERENT LABS

DOCK LAB

1. Grading of incoming raw milk

2. Raw milk testing
3. Testing of outgoing pasteurized milk
4. Testing of processed milk stored in storage tank or silo
5. Cream testing
6. Buttermilk testing
7. Standardization of fluid milk
8. Checking of strength of detergent in crate washer

QUALITY CONTROL LAB

1. Sensory evaluation of milk and milk products

2. Checking the composition of milk products
3. Hardness of water
4. Checking the quality of reagents, packaging materials, glass wares for testing
5. Checking efficiency of waste water treatment
6. Controlling the cleaning of dairy equipments as well as sanitation of plant.

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MICROBIOLOGICAL LAB

1. Checking the bacteriological quality of various types of milk & milk products.
2. Determine the types and number of microflora in air,water& various
diryequipments pipelines.
3. Testing of keeping of milk & milk products.

AGMARK LAB

Checking quality of ghee so as to conform AGMARK Standards

TESTS PERFORMED IN LAB

Dock Lab

In Dock Lab raw milk, pasteurized milk, butter milk, Skim milk are checked w.r.t.
the chemical quality.

Various tests are performed over milk in this lab.

 Acidity Test
 MBRT Test
 Fat and SNF Test
 Adulteration Test

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 ACIDITY TEST

Bacteria that normally develop in raw milk produce more or less of lactic acid. In
the acidity test the acid is neutralised with Sodium hydroxide and the amount of
alkaline is measured. From this, the percentage of lactic acid can be calculated.

Procedure
• Take 10 ml of sample(can be milk, lassi, cream, dahi etc.) in 10 ml conical flask.

• Hold the flask against white surface.

• Add 4-5 drops of phenolphthalein to the sample and stir gently

• Titrate against N/10 NaOH solution

• Continue until the appearance of a light pink colouring which persists for 10-20
seconds
and down the volume of NaOH used.

So, %Acidity = volume of NaOH used (in ml)/10

Result :

Sample No. Acidity

1. 0.19

2. 0.15
3. 0.17

The milk with acidity 0.15 is the best milk.

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 MBRT TEST

It is indirect method of estimation of total bacterial content of milk. Instead of

counting bacteria directly, a correlation is made between the time required to
reduce dye to colorless in milk.

Procedure
• To 10 ml of milk in a sterilized test tube add 10 ml of methylene blue dye. Plug
with a

sterilized cork, invert the tube to mix the contents.

• Incubate at 37°C in a water bath.

• Note down the time in which the colour of dye gets disappear and that will
give the MBRT time.

ESTIMATION OF FAT CONTENT

Fat can be determined using Gerber’s method or using a milkoscan

Gerber Method

Principle: When a definite quantity of sulphuric acid and amyl alcohol are added
to a definite volume of milk, the proteins will be dissolved and the fat will be set
free which remains in liquid state due to heat produced by the acid. The amyl
alcohol used facilitates the separation of a clear fat column. On centrifugation, fat
being lighter will be separated on top of the solution.

Procedure for estimation of fat% of milk and lassi:

• Take about 10 ml of H2SO4(10% dilute for milk & 20% for lassi)in Butyrometer.

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• Add slowly 10.75 ml sample (can be lassi or milk) in it.

• Add 1 ml isoamyl alcohol and adjust level with distilled water.

• Cap butyrometer with lock stopper.

• Centrifuge for about 5 minutes.

• Take reading of the % fat by reading the butyrometer.

ESTIMATION OF S.N.F. CONTENT BY LACTOMETER

Principle: The method is based on the Archimedes Principle, which states that
when a solid is immersed in a liquid it is subjected to upward thrust equal to the
weight of liquid displaced by it and acting vertically upwards. Lactometers are
variable immersion type hydrometers and calibrated beforehand with liquid of
known specific gravity.

Procedure:
• Warm milk in lactometer jar at about 40˚C then cool at 15.5˚C.

• Immerse lactometer in jar and take C.L.R.

% SNF = CLR/4+0.2×fat+0.29

Fat% & S.N.F.% can also be estimated by using milkoscan before which the
sample is heated to a temperature of 40ºC

Observations:

Sample Fat C.L.R. SNF(%)

No.
1. 4.25 29.4 8.49

2. 3.5 26.4 7.59

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 3. 6.7 31 9.38

Sample 1 and 2 are of Packaged milk and Sample 3 is Raw milk

DETECTION OF MAJOR ADULTERANTS IN MILK

• To overcome the increasing need of milk and milk products and to make
maximum profit there is increase in adulteration cases.
• In adulteration test we used raw milk in 2 test tubes for each test, one of
them containing only raw milk and the other one was added with the
tested material.

TESTS FOR DETECTION OF ADULTERANTS

ALCOHOL TEST

The alcohol test is used for rapid assessment of stability of milk to processing.This
test gives an indication of the mineral balance of milk. The test aids in detecting
abnormal milk such as colostrum, milk from the animal in the late lactation, milk
from animal suffering from mastitis and milk in which mineral balance has been
disturbed.

Procedure:

• Take 5.0 ml of milk in a test tube using graduated pipette.

• Add equal amount of 70% ethyl alcohol
• Close the mouth of the test tube with thumb and mix the contents well by
inverting the test tube several times.

Result :
Formation of any flakes on the wall of the test tube indicates positive test.

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 FORMALINE TEST
Procedure:

• Take 2.0 ml of milk in a test tube using graduated pipette.

• Add equal amount of 90% H2SO4 containing traces of Feric Chloride from the
side of test tube slowly.
Result:
 Voilet ring formed (+ve)
 Brown colour (-ve)

DETECTION OF HYDROGEN PER OXIDE

Procedure:

 Take 10 ml of milk in a test tube using graduated pipette.

 Add 10 to 20 drops of vanadium pentoxide reagent
 Mix them properly
 Note the colour of sample

Result: Appearance of pink or red colour (+ve)

DETECTION OF HEAT STABILITY

Procedure:
 Take 10 ml of milk in a test tube using graduated pipette.
 Add 0.6 ml HCl (N/10)
 Mix well
 Boil for 5 minutes in boiling water

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Result:
 Formation of clots (+ve)
 No formation of clots (-ve)

DETECTION OF DETERGENT
Procedure:
 Pipette 1ml milk in 15ml test tube.
 Add 1ml dye solution followed by addition of 2ml chloroform vortex for 15
sec and centrifuge at 1100RPM for 3 minutes.
 Note the intensity of blue colour in lower and upper layer.

Result:
 More intense blue colour in lower layer (+ve)
 More intense blue colour in upper layer (-ve)

DETECTION OF MALTOSE

Procedure:
 Take 5ml milk and boil it.
 Add 0.5ml drops of 5% lactic acid.
 Cool and filter.
 Add 3 drops of 1% iodine solution in filterate.

Result:
 If colour changes to brown ( +ve)
 If colour remain golden yellow (-ve)

DETECTION OF NEUTRILIZER

In milk, sodium hydroxide, sodium carbonate and sodium bicarbonate are added
by adulterators to neutralize the developed acidity in milk.

Procedure:

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  Take 2 ml Rosalic acid solution.
 Add 2 ml milk sample.
 If pink color appears, it indicates added neutralizer.

DETECTION OF SUGAR

Procedure:
 Take 2 ml of 0.5% Resorcinol solution.
 Add 2ml milk sample.
 Place it in boiling water bath for 5 min.

Result:
 Red colour (+ve)
 No change (-ve)

DETECTION OF GLUCOSE

Take strip for glucose detection and dip it in milk. If blue color persists, it indicates
positive glucose.

DETECTION OF SALT
Procedure:
 Take 2ml of Silver Nitrate solution.
 Add few drops of potassium chromate and 2ml milk sample.
 Yellow color indicates positive salt test.

DETECTION OF UREA
Procedure:
 Take 2ml urea detecting agent (DMAB-Dimethyl Amino Benzaldehyde
reagent) and add 2ml milk.
 Deep yellow color indicates positive urea test.

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 PHOSPHATASE TEST

Alkaline Phosphatase is an enzyme which is naturally present in milk, but is

destroyed at a temperature just near to the pasteurization temperature. Alkaline
Phosphatase test is used to indicate whether milk has been adequately
pasteurized or whether it has been contaminated with raw milk after
pasteurization.

Principle:Based on the principle that the alkaline phosphatase enzyme in raw milk
liberates phenol from a disodium para-nitro phenyl phosphate and forms a yellow
coloured complex at alkaline pH. The intensity of yellow colour produced is
proportional to the activity of the enzyme.

Procedure:
 Take 1 ml of milk sample.
 Put 5 ml of Phosphatase solution.
 Incubate it in water bath at temperature of about 37˚C.
 Check for colour change after 15 minutes.
 If colour changes then the milk is not pasteurized.

In the image Test tube on left side with pale yellow colour is showing
pasteurised milk which is having no alkaline phosphate activity and test tube on
right side is showing colour change in case raw milk having alkaline phosphate
activity.

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 ESTIMATION OF BR VALUE OF MILK SAMPLE

Vegetable oils, hydrogenated fats, animal body fats and their blends have been
reported as adulterants in milk and milk fat products. The elevation of butyro-
refractometer (BR) reading of milk fat by addition of different vegetable oils is
used to detect adulteration of milk. . It should be between 40-43 for milk fat with
no adulteration. The average BR reading of groundnut, cottonseed, corn, sesame,
sunflower, mustard, soya and castor oils are 59.81, 62.43, 64.50, 64.88,
65.25,65.64, 66.00 and 73.00, respectively.

Procedure:

 Take Al tubes and fill it with sample.

 Then keep these in centrifuge so that cream layer is formed on top.
 Then take the cream with the help of spatula and heat cream so as to make
ghee.
 Then put drop of ghee in butyrometer.
 Note down the value of BR reading.

ESTIMATION OF SODIUM CONTENT OF MILK SAMPLE

Milk is being adulterated with sodium to increase S.N.F. So Sodium content is

checked by ISE sodium analyser. Sodium analyser is calibrated at 100 ppm & 1000
ppm of sodium. Sodium content of milk should be between 350-450ppm.

Procedure for estimation of sodium content of milk:

Take 45 ml of milk and add 5 ml of ISA & then dip the probe of sodium analyzer in
beaker. ON the stirrer & wait till reading is stable , when ready is displayed . Note
down the reading.

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 RM (REICHERT-MEISSAL) VALUE

It is the number of millilitres of 0.1 N sodium hydroxide or alkali solution required

to neutralize the steam volatile, water soluble fatty acids distilled from 5g fat
under precise conditions specified in the method.

This value is quite significant for milk fat, since it primarily measures the butyric
acid and caproic acid content in given fat. When compared to other fats this value
for milk fat is high. So this helps in identification of milk fat from other fats.

Value should be 28 to 32.

• Centrifuge the sample for 15 min.

• Separate the cream and heat it to make ghee.
• Take 5g of ghee in flask.
• Add 20 g glycerol, 2ml of 50% NaOH and 6 to 8 glass beads.
• Heat clarify the sample.
• Add 93 ml boiling distilled water followed by 50 ml 1 N H2SO4.
• Fix it with still head to distillation apparatus.
• Mildly heat the flask so as to form the golden ring on top.
• After that heat it properly so that distillation starts and 110 ml distillate is
collected in round bottom flask.
• Distillate is filtered and chilled.
• Titrate it against N/10 NaOH using phenolphthalein indicator.
• RM Value = (ml of NaOH used – 0.4) x 1.1

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 QUALITY CONTROL LAB

 In quality control lab sensory evaluation of milk and milk products is done,
in which there is testing of shelf life.

 A check is put over cleanliness of dairy equipments and overall sanitization

of plant.

 The products produced in the plant are checked for their quality and shelf
life before they are sent to the market.

 Some of the main tests done in the labs are:

1. Hardness of water.
2. Curd acidity test.
3. Moisture test for paneer.
4. Fat test for paneer.
5. Total solid test for kheer.
6. Ghee test/ Free fat acidity.

HARDNESS OF WATER

This test is performed to check the hardness of water i.e. whether the water used
in various production units is suitable to use or not.

Procedure:
 Take 100ml water sample in a flask.
 Add 1-2ml NH3 buffer.
 Add 2-3 drops of indicator (Phenolphthalein) to it.
 Titrate it against 0.02N EDTA solution.
 Blue colour marks the end.

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Calculations
 Hardness =T.V. x 10
 When T.V. = 21.5 Hardness = 21.5 x 10 = 215 ppm

CURD ACIDITY TEST

Curd acidity test is a test to check the acidity in packed form of curd i.e. with
increasing time the acidity of curd increases and it becomes unfit to consume. So
to check it the test is performed. Curd Acidity test is same as in case of milk.

Procedure:
 Take 10 ml of curd sample in a china dish.
 Put about 5 -10 ml of water in the dish.
 Mix it properly.
 Add few drops of Phenolphthalein indicator in it.
 Titrate it with NaOH.
 Light Pink colour marks the end.

Observations:

Sample No. Acidity

1. 6.0
2. 7.3
3. 6.3

Result:

Sample no.2 is acidic in nature and sample no. 3 is with optimum acidity
6.

MOISTURE TEST FOR PANEER

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This test is used to check the presence of moisture in paneer in packed form, as
more moisture may lead to microbial activity.

Procedure:
 Take an empty dish and take its weight as W1.
 Put about 3g of crushed paneer in it, weigh it and mark it as W 2.
 Put the dish in oven for about 4 hrs.
 Measure the weight and mark it as W3.

Calculation:
 W2-W1=A
 W3-W2=B
 B/A x10= Moisture

Observations:

Sample W1 W2 W3 A B Moisture(%)
No.
1. 27.739 30.338 29.534 2.599 0.804 30.95

2. 33.966 38.864 36.189 4.898 2.676 54.61

3. 10.028 13.648 11.921 3.620 1.726 47.69

FAT TEST FOR PANEER

Fat test is carried out to determine the amount of fat in paneer.

Procedure:
 Take a butyrometer and tear it.
 Take paneer sample into butyrometer.
 Put 3g sample into butyrometer.

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  Put 10 ml sulphuric acid and 1ml amyl alcohol and to maintain its level we
have to add hot water.
 Then centrifuge it for 5 mins for 12000pm.
 Note the fat reading.

Calculations:
Fat% =Fat x11.26/W

Observations:

Sample No. Weight of Fat Fat (%)

Sample(g)

1 1.6 3.9 27.45

2 1.5 4.3 32.27

3 1.6 3.5 26.27

GHEE TEST/ FREE FAT ACIDITY

This is the test to check the quality of ghee/fat in terms of acidity.

Procedure:
 Take the sample butter to be tested.
 Heat the sample until it gives brown golden shades.
 Prepare a solution by adding 50 ml ethanol and 2-3 drops of
phenolphthalein.
 Now neutralize it with N/9 NaOH.
 Take about 10g of ghee.
 Mix the solution prepared in 10g of ghee.
 Heat to boil.

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  Then add NaOH to that for titration.

Observations:

Sample No. Acidity(%)

1 0.27

2 0.44

3 0.30

Result:

The normal value of acidity is 0.3 %, which indicates the good quality of ghee,
whereas the increased acidity means the rancid ghee.

EQUIPMENTS USED IN MILK PROCESSING

PASTEURIZER

It is device used to pasteurize the milk i.e. heating at 63 ○C for 30 min or 70○C for
15 sec in approved and properly operated equipment. At one end of the device
the temperature is 75-80○C and at the other end it is 4-5○C, i.e. instant cooling of
milk is done after exposure to high temperature which kills almost all germs in
milk.

HOMOGENIZER

Homogenization is a process where in fat globules are broken down to 2 microns

or less under high pressure.

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  It consist of high- pressure piston pump that forces the milk and thereby
subdivided into smaller particles of more uniform size.

CREAM SEPERATOR

It is a device which separates cream from the milk by means of a set of plates
which rotate around a central axis using centrifugal force.
 There are 135 plates in each cream separator.
 Cream and skim milk are obtained.
 After extraction cream is chilled to 4-6○C through plate chiller.

STORAGE TANK

To store the milk in different states like raw, chilled, pasteurized and
standardized.
They have capacity of 10000-20000 liters.

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