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Detection of Adulterants in Milk and Milk Products

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Detection of Adulterants in Milk and Milk Products

AMIT KUMAR1, UPENDER2*
1
Ph.D. Scholar, Division of Animal Reproduction,
Indian Veterinary Research Institute, Uttar Pradesh, India
2
M.V.ScScholar,DivisionofVeterinaryPublichealthandEpidemiology,
IndianVeterinaryResearchInstitute,UttarPradesh,India

Corresponding author: [email protected]

T he milk is considered as a valuable food

for all the human beings including the
children and adults. The milk is rich in easily
along with the optimum quality of the product. It
is a highly perishable commodity hence; it
should be consumed within a definite span of the
digestible nutrients and provides highly nutrient time or otherwise should be preserved with a
diet. The world population is increasing day by suitable preservative.
day which creates an alarming situation for the In today’s world, there are a lot of traders or
adequate supply of the milk to each individual vendors re there which are indulged in the

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 Date Published: 02 December, 2021 3:56 PM IST
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fraudulent practices of milk sale by addition of Nitrate test: The water always carries nitrate
various substances termed as “adulterants” with ions in its composition while the milk is deficit in
a purpose of making economic gain. The term it. The nitrate can be detected by the
adulterant refers to the substance which is diphenylalanine test. The nitrate ion is present in
intentionally added to the lot with an inherent aim large amount in polluted water samples. The
of debasing the equality of the final product. The method consists of the addition of 1ml of the
food adulteration refers to the act of intentionally diphenylalanine along with 100 ml of the
debasing the quality of the food offered for sale sulphuric acid which is poured slowly along the
either by admixture or substitution of inferior walls of the test tube in 5 ml of the milk. Mix all the
substance or by removal of some ingredient contents slowly and wait for the color
also. It takes into account only intentional not appearance. The blue color at the junction of the
incidental. The adulteration of the milk and milk sulphuric acid and milk indicates the presence of
products is directly proportional to the demand the nitrate ions in the milk which indicates the
and supply principle which is ultimately adulteration of the milk with the water.
connected with the economic principle. The Determination of freezing point: The
adulteration of the milk is punishable under freezing point of the normal milk gets lower down
section 420 of Indian Penal Code. These when water is added in it. Actually, the freezing
adulterants will increase the solid content of the point of any solution depends upon the presence
milk and thus provide more economic gain to the of the solutes in the milk. The solute
sales person. The most common adulterants concentration after the addition of the water will
along with their detection methods are described decrease which leads to the decrease in the
in this section. freezing point of the milk. The normal freezing
Reduction of the fat: The reduction of the point of the milk is 0.5470C. The freezing point of
fat can be achieved via two ways: the milk can be detected with the help of the
Hortvet cryoscope equipment. The percentage
Addition of the water: It is the most common
of the added water(W) can be calculated using
fraudulent practice being followed in Indian
the following formula,
context by the milkman. They do so to increase
the amount of the product but the quality of the W = 100(T-T’)/T, where, T is the average
water is always doubtful. The water may carry a freezing point of the normal milk(0.5300CC) and
lot of the harmful microbial and bacterial T’ is the true freezing point of the given sample.
contaminants which affects public health. The Skimming of the milk: The term skimming
presence of the water in the milk can be detected refers to the removal of the fat content from the
by following methods- milk. The fat is lighter than water, thence the
specific gravity of the milk increases. The fat
Measurement of the specific gravity: The
content of the milk can be detected using
specific gravity of the milk will decrease on the
addition of the water content in the milk which Gerber butyrometer method: This method
proves to be a better indicator of the adulteration was devised by Gerber et. al. It works on the
of the milk. The specific gravity has been principle of the action of the sulphuric acid in the
determined by the Quevenne’s lactometer. The milk along with the amyl alcohol followed by
normal specific gravity of the cow milk is 1.028 – centrifugation at 2000 rpm for 5 minutes leading
1.030 while of the buffalo milk is 1.030-1.032. to collection of the fat column at top. The
The lower specific gravity indicates addition of sulphuric acid will liberate the heat which helps
in melting of the fat. The amyl alcohol will cause
the water in the milk.
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clear demarcation between fat and other agents are cane sugar, starch, gelatin, calcium
contents of the milk. The fat deficiency can be carbonate and milk powder etc. to increase the
calculated as 100(3.0 F)/3 where F is the fat% of solid content of the milk.
the milk sample. Detection of the Starch: The starch is a
Babcock test: It is less costly as compared to homopolysaccharide which is having high
above method and more accurate. It depends specific gravity. It can be detected by addition of
upon the action of the sulphuric acid on the milk the 1.0 ml of the 5% solution of the iodine. If
proteins. The heating and centrifugation cause starch is present in the milk sample, the iodine
the fat to separate and float as a separate layer molecules get chelated with starch pockets. It
on the top of milk column. The procedure has yields blue color complex formation.
been carried out in Babcock bottle. Detection of gelatin: The gelatin is added to
Rose-Gottileb test: In this, the milk sample is
the milk asa thickener. Take 10 ml milk in a large
treated with the ammonia and ethyl alcohol. The test tube along with the 10 ml of the mercuric
ammonia causes the dissolution of the protein nitrate solution in the same test tube. Shake the
while the ethyl alcohol causes the precipitation test tube well. Add 20ml of the water and shake
of the proteins. The fat is extracted with the the mixture and wait for 5 minutes. Then after
diethyl ether and petroleum. This method is filter the solution and obtain the opalescent
mainly used for milk products and as reference filtrate which indicates the presence of the
method. gelatin. Collect this filtrate in the separate test
Adam’s method: This method consists of the tube and equal amount of the saturated picric
spreading the milk on an absorbing paper and acid solution. Wait for 2 minutes, it yields yellow
then extracting the fat from it by a fat solvent precipitate which indicates the appreciable
through a process of distillation. amount of the gelatin.
Skimming and watering: In this adulteration Detection of the Cane sugar: Take about 15
method, the fat is removed and sufficient water is ml of the milk sample in a sterile clean test tube
added to the milk to make the specific gravity at a and add 1ml of the HCl along with 0.1 g of the
normal level. The skimming of the milk has also resorcinol powder in the same test tube. Boil the
been done to extract the cream separately and solution for a few minutes. The presence of the
fetching the double profit from it. For determining cane sugar is indicated by the development of
the above adulteration, we usually employ the the red color.
calculation of the corrected lactometer reading Detection of the sucrose: The presence of
followed by application of the specific formulae the sucrose can be detected by the help of the
for TS% determination, such as resorcinol test. It involves the addition of 0.1 g of
Richmond %TS : 0.25G+1.2F+0.14 the resorcinol to the 10ml of the milk. Shake the
test tube mixture and wait for some time. The red
Babcock : 0.25G+1.2 F
color of the test tube will show the presence of
Fleischman’s : 0.25G+1.2F+0.25 the sucrose.
Where, G is corrected lactometer reading and Detection of the milk powder: the milk
F% is fat percentage. powder has been added to the milk. The milk
Addition of thickening agents: powder has been added to increase the SNF
The thickening agents leads to increase in the content of the milk. We start with the 10 ml of the
viscosity of the milk. The various thickening milk in a test tube followed by addition of 1 drop
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of the formalin in the test tube. Mix it well and The sample positive for urea indicates the
incubate the test tubes at 600C for distinct yellow color while slight yellow indicates
approximately 10 minutes. The presence of negative.
peculiar odor indicates the presence of the milk Using DMAB reagent: Take 1ml of the milk
powder in the test sample. In negative control, in a test tube. Add 1ml of 1.6%(w/v) DMAB
there will not be any such odor. reagent and mix it well. The distinct yellow color
Detection of the Calcium carbonate: indicates the urea addition to the milk sample
Calcium carbonate is a chemical which is mainly while slight yellow color is negative for urea test.
used as bleaching agent in the farms and it has a Detection of sulfate salts: The presence of
caustic action on live tissues. We proceed for its the sulfate salts(Zinc sulfate, ammonium sulfate,
detection by taking10 ml of the milk in a test tube magnesium sulfate etc.) to raise the SNF of the
followed by addition of 1 ml of the concentrated milk have been carried out using the barium
hydrochloric acid in it. The presence of the chloride. The reagents include the barium
calcium carbonate has been indicated by the chloride and TCA. Take 10 ml of the milk sample
presence of the effervesces. in a test tube of 50 ml capacity. Add 10ml of TCA
Detection of the sodium carbonate(rosalic solution in it. Filter the coagulated milk through
acid test): It is named as rosalic acid test after Whatman filter paper grade 42. Take 5 ml of the
the name of the indicator rosalic acid. First of all, clear filtrate and add few drops of the barium
take 5 ml of the milk sample in a test tube, then chloride solution. Observe for any visible ppt. in
after we have to add 10ml of the alcohol and the milk. The positive sample shows the
rosalic acid solution(1:10) and mix it well. The formation of the milky-white precipitates.
sodium carbonate acts as acidity neutralizers in Detection of hydrogen peroxide: The
the solution. The end result of the pink color for hydrogen peroxide is a preservative but as per
the mixture indicates the presence of the sodium FSSAI rules, it is not permitted to be added in the
carbonate in the milk sample. milk. Hence, if it is present in the milk, then, it is
We can also go along with the modified said to be adulterated. The reagents needed for
method in which we have to do the manual its detection include para-phenylenediamine
shaking of the test tube after addition of the reagent(2.0%). Take about 2.0 ml of the milk in a
rosalic acid in it. test tube and add 2.0ml of the raw milk in a test
Detection of the urea/synthetic milk: The tube. Add 2 drops of the 2.05 of para-
urea is present in traces in natural milk but it phenylenediamine reagent. Mix it well and
constitutes a major part in the synthetic milk observe for blue color for positive samples and
formulations which is above the prescribed white color for pure milk sample.
leveland cause toxicological hazards to the Detection of detergent in the milk: Take 5
consumers. The detection of the urea can be ml of the milk sample and add 0.1ml of
carried out of any methods- bromocresol purple solution (0.5%) in it. Mix the
Using DMAB&TCA method: Take equal solution. The violet color of the test tube solution
amount of milk and 24% TCA(tricholoroacetic indicates the presence of the detergent in the
acid) in a glass stoppered test tube. Mix it and milk while faint violet indicates the negative
filter it with Whatman no.42 filter paper. Take 3 ml result.
of the filtrate in another test tube and add 3ml of Detection of saccharin: The saccharin is a
1.6% dimethylamino-benzaldehyde reagent. sugar which has sweet taste and it has been

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added in the flavored milk and milk products Groundnut oil – 2%

mainly. First of all, we have to Curdle the aliquot Sunflower oil – 1%
of the diluted milk sample with acetic acid. Shake
Detection of the animal fat(mutton/beef):
it well and filter it. We have to add 2.0ml of the
concentrated HCl to the filtrate to acidify the Take the sample of ghee and add a mixture of
filtrate and extract it with 25ml of the ether and alcohol and acetate as reagent and heat the
wash it with 3 consecutive layers of the 5ml mixture. The slight deposition of the crystals at
water. The ether extract is a volatile compound the bottom of the test tube indicates the
which can be evaporated by putting in the water presence of the mutton or beef fat in the ghee.
bath and add 1-2 drops of water, mix well with Conclusion:
glass rod and taste a little. The characteristic The adulterants are mostly extraneous
sweet taste indicates the presence of the materials which have been added to the milk or
saccharin. milk products with an aim to increase the
Adulteration of Ghee: economic gain mainly. These are very harmful
Detection of the vegetable oils: for certain age groups mainly the young ones
and elder group population. They can be
Take a sample of ghee and add trichloro-
detected with various methods as described
methyl in it. Mix it well followed by addition of the
above which usually have to be carried out by
acetic anhydride and then-after heat the mixture.
the FSSAI at regular intervals. The adulterants
The melting point of the unadulterated ghee is
debase the intrinsic quality of the milk and milk
114-1150F while that of adulterated ghee is
products and hence strict legislations needed to
1170F.
be implement over them.
Detection of the hydrogenated
References:
oils(sesame oil):
Azad T. et al., Common milk adulteration and
For this Baudouin test has been carried out.
their detection techniques. International Journal
Take a ghee sample and add 2ml of furfural
of Food Contamination.
solution in alcohol and HCl(5ml) in it. The
crimson red color (after 5 minutes) indicates the Sharma Ranjan et al., Detection of
presence of the hydrogenated oil(sesame oil) in adulterants in milk - A laboratory manual.
the ghee. It is helpful in differentiating between National Dairy Research Institute, Karnal.
desi ghee and vanaspati ghee. The vanaspati Biswas R. et al., Adulteration in Milk – a
ghee contains 5% sesame oil. Pure ghee not growing concern. J. Dairy and Veterinary
contains sesame oil. Sciences.
The detection of the sesame oil, refined Sherikar A.T. et al., Textbook of Elements of
soyabean oil, groundnut oil and sunflower oil Veterinary Public Health. ICAR.
have been done by HPLC as recommended by Kamthania M. et al., Milk adulteration:
FSSAI with the limit of detection as follows- Methods of detection and remedial measures.
Cocconut oil – 5% International Journal of Engineering and
Refined soyabeen oil- 1% Technical Research.

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