MILK PHYSICO-

CHEMICAL TESTS
Dairy Technology Laboratory
SALT TEST-Liquid Milk
◦ Apparatus:- Burette, Pipette, Dropper Bottle, Beaker, 100ml & 1000ml flasks
◦ Reagents: - 0.1 N Silver Nitrate solution, 10% Potassium Chromate Solution.
◦  
◦ 0.1N Silver Nitrate solution
◦ Take N/10 Merck ampule of silver nitrate.
◦ Cut it from both sides.
◦ Pour solution from ampule to the 1000ml volumetric flask
◦ Make the volume upto 1000 ml by adding distilled water.
◦ Shake it well.
◦ 10% Potassium Chromate Solution
◦ Weigh 10gm potassium chromate.
◦ Dissolve it in 100ml of distilled water.
Procedure
Take 9 ml of well mixed sample in a 100 ml beaker with the help of a pipette.
◦ Add 3 - 4 drops of Potassium Chromate Solution, being used as an indicator.
◦ Titrate the sample against 0.1N AgNo3.
◦ Before titration make sure that level of 0.1N AgNo 3 solution in burette is at “0” mark.
◦ Add 0.1N Silver Nitrate Solution drop by drop from burette until light brown colour appears.
◦ This is the end point and no more Silver Nitrate Solution should be added.
◦ Note Silver Nitrate solution reading from burette.
◦ Calculate the percent salts as below:
◦ Salt% = volume of 0.1N Silver Nitrate solution used x 0.065
Milk Fat
◦ Introduction 
◦ Fat content of milk /cream of dairy animals is one of the important factors in determining the price of the
milk/cream. Butter/fat % of milk or amount of milk produced is considered while calculating the ration
of the lactating animal. 
◦ Testing of butterfat is important to know in order to make correct adjustments of the butterfat percentage
in standardized milk and milk products. 
FAT TEST - LIQUID MILK
◦ Apparatus: - Butyrometer, Pipette (10.94ml), Butyrometer stopper, Butyrometer (Breakdown of fat)
◦ Stand, Centrifuge Machine, Automeasures, Water bath, Stopper Key.
◦ Reagents: - Sulphuric acid, (Disrupt the emmulsion), Iso Amylalcohol, (Separate fat and water)
◦ Procedure:-
◦ Warm milk sample to 40 °C and thoroughly mix it.
◦ Cool the same sample to 20º C.
◦ Measure out 10 ml of Sulphuric acid in a Butyrometer.
◦ Add 10.94 ml of milk sample and 1 ml of Iso- Amyl Alcohol into Butyrometer.
◦ Contents should be added in the stated order.
◦ Insert stopper in the Butyrometer.
◦ Mix the contents well by inverting the Butyrometer several times.
◦ Place Butyrometer in centrifuge machine (1100 rpm) for 3 minutes in case of raw milk and 10 minutes in case of homogenized milk.
◦ Note the fat column in the neck of the Butyrometer.
◦ Express the results as percentage of fat in milk.
Precautions

◦ Sample temperature 20°C.

◦ Sample suction from centre of beaker at normal speed.
◦ Sample pouring into butyrometer at normal speed along wall of butyrometer.
◦ Volume adjustment with water for readable level.
◦ Butyrometer inverting for proper mixing of contents.
◦ Take reading at eye level in front of light & within 20sec.
◦ Do slow fat level up / down to get in readable area.
◦ Ensure use of PPEs during analysis in Lab.
MILK DENSITY 

◦ Introduction 
◦ Milk has specific gravity or density. It can be determined lactometer. Middle men add water in milk to
increase its volume so as to get high price of the same pure milk. It is way to deceive the buyers. Every
body know that middle men add water in milk. Buyers therefore assure the addition of water in milk. 
◦ The lactometer test is designed to determine the change in density of such watered milk. If it is tested
along with butterfat test, it enables us to calculate the milk total solids (% TS ) and solids not fat (SNF). 
Lactometer Reading For SNF
Calculation in Liquid Milk
◦ Apparatus: - Lactometer (20 °C), Measuring cylinder 250/500 ml, Thermometer, Beakers.
◦ Reagents: - None
◦  
Procedure:-
◦ Warm the representative sample of milk to 40ºC by constantly stirring.
◦ Cool down the sample to 20°C.
◦ Thoroughly mix the sample by pouring gently from one beaker to another.
◦ Then pour milk into a cylinder and allow the milk to flow over the top until no foam is left at top of the cylinder.
◦ Gently lower the Lactometer into the milk sample and wait till it comes to rest.
◦ When lactometer comes to rest, read lactometer degree on the scale at the surface level of milk.
◦ While reading the degree of lactometer, the temperature should be 20ºC.
◦ Percentage of solid non fat and total solids can be calculated by the following formula.
◦ (LR/4)+ (0.22xFat) +0.72 =% SNF
Precautions:
◦ Gently insert the lactometer into sample.
◦ Use cleaned plastic cylinder.
◦ Milk sample to flow over the cylinder.
◦ Vertical level of cylinder.
◦ Reading observation within 30 seconds after movement of lactometer stopped.
◦ Ensure use of PPEs during analysis in Lab.
◦  
DETERMINATION OF SEDIMENTS
IN MILK
◦ Apparatus : 15ml solubility index tubes, Gerber Machine, 10ml pipette
◦ Reagents : NIL
◦ PROCEDURES:
◦ Take 15 ml milk sample in solubility index tube from well mixed sample with help of glass pipette.
◦ Centrifuge in Gerber Machine for 10 minutes at 40~45ºC.
◦ Gently drain the milk from tube without disturbing the sediments if any at the bottom.
◦ Fill the tube with cleaned water and stay for 2~3 minutes.
◦ After comparing the results with standard sample, mark the finding as +ve, ++ve, +++ve.
◦ A good milk gives no visible dirt/sediments at bottom of tube.
◦  
◦ +ve = Sedimentation up to 0.2ml
◦ ++ve = Sedimentation 0.2 to 0.4ml
◦ +++ve = Sedimentation 0.4 to 0.6ml
◦  
Sugar test
◦ Introduction 
◦ Generally sugar is mixed in the milk to increase its solids not fat contents i.e. to increase the lactometer
reading of milk 
SUGAR (SUCROSE) TEST FOR
MILK
◦ Apparatus : Water bath, Test Tube, Pipettes 1ml, Stop watch
◦ Reagents :
◦ Nephthol 10% : Dissolve 10g of Nephthol in Ethanol and make volume up to 100ml with
Ethanol in 100ml volumetric flask.
◦ HCl concentrated : Commercially available.

PROCEDURES: (Molisch's test)

◦ Take 0.5ml of thoroughly mixed sample in test tube.
◦ Add 3 drops of 10% Nephthol solution in it.
◦ Add 3ml of Conc. HCl in it.
◦ Place the test tube in boiling water (water bath) for 10 seconds
◦ Let the test tube contents cool down.
◦ Violet deep color after 5 minutes indicates the presence of sugar.
To detect presence of Antibiotics

◦ Apparatus : Autoclave, Incubator, Petri Dishes, Pipette, Sprit Lamp, Wire Loop, Sampling Bottle, Filter paper disc, Forceps
◦ Reagents :
◦ Nutrient Agar : Prepare as per instruction of manufacturer and store at 55ºC
PROCEDURES:
◦ Add 10 to 13ml of sensitivity agar (Nutrient Agar) in petri dish and allow solidifying.
◦ Take pure bacterial strain of Staphylococcus Aureus or Bacillus Stereothermophilis.
◦ With the help of wire loop pick a single colony from pure strain culture & make a lawn on solidified plate.
◦ Dip the pre sterilized filter paper disc in milk sample with help of sterilized forceps, stay in milk for maximum 30 sec.
◦ Put the disk on solidified agar having bacterial strain cultured.
◦ Place the Petri dishes in inverted position in incubator at 35~37ºC in case of Staphylococcus aureus and at 54~56ºC in case of bacillus
stereothermophilis for 12 to 18 hours.
◦ After incubation period examine the zone of inhibition around the paper disc.
◦ This zone of inhibition shows the presence of antibiotics in sample.
STARCHES GIVE BLUE COLOR WITH IODINE
 Starch Test of Milk
Apparatus: - Water bath, Thermometer, Beaker 500-ml, Pipettes, Petri-dish.
Reagents: - 0.1N Iodine Solution.
0.1N Iodine Solution
1.Weigh 12.69gram iodine.
2.Dissolve it in 1000ml Ethanol to prepare 0.1N solution.
Procedure:-
1. Warm up 4-5 ml milk sample in a test tube near to boil.
2. Pour it in glass Petri dish and add 3 to 4 drops of 0.1N Iodine solution.
3. Mix the contents and observe black particles.
4. Dark Blue or Black colour/particles indicate the presence of starch.
Precautions:
1.Heating temperature must be up to 75~80°C
2.Use sucker for sample.
3.Ensure use of PPEs during analysis in Lab.
SODIUM BI-CARBONATES 

◦ Introduction 
◦ Sodium bi-carbonates or other neutralizers such as sodium hydroxide, sodium carbonate etc, are used to
avoid curdling of milk during transportation. Addition of neutralizers is generally prohibited. 
CARBONATE/BICARBONATE TEST
FOR MILK
◦ Apparatus : Pipette 10ml, Test Tube, Test Tube Stand
◦ Reagents :
◦ Rosalic Acid 1%: Dissolve 1g of Rosalic acid in ethanol and make volume up to 100ml with
ethanol 95% in volumetric flask.
◦ Ethanol 95% : Commercially available.
PROCEDURES:
◦ Take 5ml of thoroughly mixed sample in test tube.
◦ Add 5ml of 95% Ethanol in it and shake well.
◦ Add 3 drops of Rosalic Acid 1% and mix
◦ A pinkish colouration indicates the presence of Carbonate / Bi-Carbonates, record the test as detergent
+Ve.