International Journal of Chemical Studies 2017; 5(4): 613-617

 
 
 
 
 
 
 
 
 
 
P-ISSN: 2349–8528 
E-ISSN: 2321–4902
IJCS 2017; 5(4): 613-617 Adulteration of milk and its detection: A review
© 2017 JEZS
Received: 05-05-2017
Accepted: 06-06-2017
D Maheswara Reddy, K Venkatesh and C Venkata Sesha Reddy
 
D Maheswara Reddy
Assistant Professor, Department Abstract
of Livestock Products Milk adulteration is a global concern and social problem. It exists both in the backward and advanced
Technology, College of countries. Developing countries are at higher risk due to lack of monitoring and policies. Consumption of
Veterinary Science, Proddatur, adulterated milk causes serious health hazards leads to fatal diseases and a great concern to the food
Sri Venkateswara Veterinary industry. Increased demand, growth in competition in dairy industry and financial gain makes the some
University, A.P. India. producers to adulterate the milk. This paper presents a detailed review of common milk adulterants as
well as different methods to detect the adulterants and their health hazards.
K Venkatesh
IIIrd BVSc & A.H., College of Keywords: Milk Adulteration, Detection Techniques, Electrical methods, Health Hazards
Veterinary Science, Proddatur,
Sri Venkateswara Veterinary
University, A.P. India. Introduction
Milk is considered to be the ‘ideal food’ because it is having abundant nutrients required for
C Venkata Sesha Reddy both infants and adults. It supplies body building proteins, bone forming minerals, health
IIIrd BVSc & A.H., College of giving vitamins, minerals and furnishes energy giving lactose, milk fat and also supplying
Veterinary Science, Proddatur,
Sri Venkateswara Veterinary
certain essential fatty acids. Milk adulteration came into global concern after breakthrough of
University, A.P. India melamine contamination in Chinese infant milk products. Unfortunately milk is being very
easily adulterated throughout the world and significantly worse in developing and
underdeveloped countries due to the absence of adequate monitoring and lack of proper law
enforcement. Apart from the ethical and economical issue, it also creates health hazards. Most
of the times, the adulteration is intentional to make greater profit, but sometimes it may be due
to the lack of proper awareness. Possible reasons behind it may include- demand and supply
gap, perishable nature of milk, low purchasing capability of customer and lack of suitable
detection tests (Kamthania et al., 2014) [9]. Chemical adulterants are used for different
purposes. The common adulterants are sugar, water, salt, starch, chlorine, hydrated lime,
sodium carbonate, formalin, ammonium sulphate, H2O2 and non-milk proteins etc. To meet the
deficit of milk, some people are preparing synthetic milk by mixing urea, caustic soda, refined
oil and common detergents which has poisonous effect. Hence, the current review highlights
the milk adulterants, their detection and their hazards on health of consumers.

Typical Adulterants and their health hazards on humans

A) Water
Water is the most common adulterant added to increase the volume of milk which in turn
decreases the nutritive value of milk. But if contaminated water is added to milk, it is a serious
health concern to the milk consuming community.

B) Melamine
Melamine is added to milk and milk powder to increase protein content falsely. It causes renal
failure and deaths in extreme cases (Cheng et al., 2010) [2].

C) Urea
Urea is added to milk to provide whiteness, increase the consistency of milk, increase non-
Correspondence protein nitrogen content and for levelling the contents of solid-not-fat (SNF) as are present in
D Maheswara Reddy natural milk. Urea is also used to prepare synthetic milk. Health hazards associated are acidity,
Assistant Professor, Department indigestion, ulcers and cancers. Urea is harmful to heart, liver especially for kidneys as the
of Livestock Products
kidneys have to do more work to remove urea from the body (Kandpal et al., 2012) [10].
Technology, College of
Veterinary Science, Proddatur, Ammonia in milk develops regression, loss of acquired speech and sensory disturbances.
Sri Venkateswara Veterinary
University, A.P. India
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International Journal of Chemical Studies
 
D) Detergents number of antibiotic resistant, interference in intestinal flora
Detergents are added to emulsify and dissolve the oil in water and some of them (such as sulfamethazine residues) may have
giving a frothy solution, the characteristic white colour of carcinogenic properties. It may also cause tissue damage. It
milk (Singuluri & Sukumaran, 2014) [18]. They enhance the interferes in the bacterial fermentation process which
cosmetic nature of milk. Detergents cause gastro – intestinal produces important losses in fermented products. Very low
complications. amount of penicillin residue is a potential cause of urticaria
(Das et al., 2016) [3].
E) Hydrogen peroxide (H2O2)
Hydrogen Peroxide is added to milk to prolong its freshness, J) Food colours
but peroxides damages the gastro intestinal cells which can Many food colorants are also added to improve the
lead to gastritis and inflammation of the intestine. H2O2 appearance and have hazardous effects on health.
disturbs the antioxidants in the body disturbing the natural
immunity hence increasing aging. K) Milk powder
Sometimes milk powder as an adulterant is added in fresh
F) Starch milk. This is done for economic advantage when a country
Starch is used to increase solid-not-fat (SNF) and if high has milk powder in excess or subsidy is provided for dried
amounts of starch are added to milk, this can cause diarrhoea powder milk (Guan et al., 2005) [4].
due to the effects of undigested starch in colon. Its
accumulation in the body may prove very fatal for diabetic L) Non-Milk Proteins & fats
patients (Singuluri & Sukumaran, 2014) [18]. Apart from the Milk, milk powder and other dairy products are often
starch, wheat flour, arrowroot, rice flours are also added. adulterated by low priced non-milk proteins such as soy, pea
and soluble wheat proteins. Bovine rennet whey powder is
G) Sugar sometimes mixed in milk powder (Haasnoot et al., 2006) [5].
Generally sugar is mixed in the milk to increase the solids not Sometimes milk fat is replaced by fat from other sources
fat content of milk i.e. to increase the lactometer reading of which may also pose a risk to human health. Since milk fat is
milk, which was already diluted with water. very expensive, some manufacturers of milk and dairy
products remove milk fat for additional financial gain and
G) Neutralizers compensate it by adding non-milk fat such as vegetable oil
NaOH is often used in synthetic milk to neutralize the acidic (e.g., sunflower oil) (Jha and Matsuoka 2004) [8].
effect. Synthetic milk is a common problem in India which is
prepared by adding urea, caustic soda, refined oil and M) Low valued Milk
common detergents. Caustic soda contains sodium, acts as Milk is adulterated by mixing lower valued milk to the higher
slow poison for those suffering from hypertension and heart valued milk. For example often goat milk is adulterated with
ailments. Caustic soda deprives the body from utilizing lysine, cow milk for greater profit. It has been found that health
an essential amino acid in milk, which is required by growing hazards related to this practice are not well defined (some
babies. Such artificial milk is danger for all, but is more may have allergy in cow milk) but from commercial and
harmful for pregnant women. As a substitute of milk fat, ethical point of view this is a great concern in food industry
refined oil is mixed; and to dissolve the oil in water and to (Das et al., 2016) [3]. Low priced cow milk is often added in
give a frothy solution, detergents are used. Carbonates and the milk of ewes, goats, buffalos (Haasnoot et al., 2004) [6]
bicarbonates are added to milk which can cause disruption in and in sheep milk (Pappas et al., 2008) [13].
hormone signalling that regulate development and
reproduction (Manual of Methods of Analysis of Foods: Milk N) Preservatives
and Milk Products 2005) [12]. Carbonates in milk produce Development of micro organism spoils the milk and spoiled
gastrointestinal problems including gastric ulcer, diarrhoea, milk is not good for health. Boric acid, Formalin, Sodium
colon ulcer and electrolytes disturbance. carbonate (Na2CO3), Sodium bicarbonate (NaHCO3),
Salicylic acid, Benzoic acid, Sodium azides can preserve the
H) Chlorine milk for long time and has poisonous effect which can lead to
Chlorine is added to compensate the density of the diluted death (Poisonous milk for pupils;1905) [14]. It develops
milk after addition of water. Chlorinated milk can cause abdominal pain, diarrhoea, vomitions and other poison related
clogging in arteries and develop heart problem (Hattersley, symptoms.
2000) [7]. Chloride in the milk disturbs the acid base balance in
the body and also blood pH. O) Whey
Addition of liquid-whey is a very common practice to
I) Antibiotics increase the volume of milk. Low priced rennet whey is
Antibiotics are used mainly to treat a variety of diseases and sometimes mixed with liquid milk and milk powder. Whey,
80% of veterinarians use antibiotics for treatment of mastitis rennet whey solid are often found in milk (Recio et al.; 2000)
[19]
disease. These antibiotics in the form of antimicrobial . Addition of rennet whey solid in UHT milk causes
residues are found in milk. Presence of tetracycline, aromatic lowering of blood pressure. Some businessmen, for greater
amines, gentamicin residue after mastitis treatment, neomycin profit, use cheap muriatic acid to prepare whey which causes
residues, sulfamethazine residues, chloramphenicol residues, serious health problem.
aflatoxin M1 contamination etc. are also a deep concern as
milk adulterants (Das et al., 2016) [3]. Intramammary infusion P) Pesticides
of antibiotics for mastitis therapy is a major reason for milk Pesticides are also used to kill the microorganisms present in
contamination. Residues of these drugs in milk poses serious milk and to resist its further growth or in other words it is
health hazards such as allergic reactions, increase in the
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International Journal of Chemical Studies
 
used to preserve milk. Their presence in milk poses serious Detection of milk adulteratants: Various milk adultrants
health hazards due to its toxicity or carcinogenicity. and the method used to detect those adulterants are presented
in table 1.

Table 1: Different milk adulterants and the method used to detect those adulterants.
S. No Type of Adulterant Method used to detect adulterant
Sequential Injection Analysis (SIA)
Flow Injection Analysis (FIA)
1 Chlorine
Potentiometric detection
Conductometric sequential injection analysis
Electrical conductivity
BRT Test (Test kit)
Spot Test
Penzyme milk test, SNAP test and LACTEK test (kits)
Chromatography (HPLC)
2 Antibiotics
Liquid chromatography mass spectrometry
Somatic cell count (SCC)
Screening test
Biosensor assay based on surface plasmon resonance (SPR)
E-Nose
Fluorescence spectroscopy
Analysis of triacylglycerols using gas liquid
Chromatography NIR spectroscopy
3 Non-milk proteins Electrical conductivity and capacitive reactance
Reversed Phase HPLC method in combination with fluorescence detector
Sulfate capillary electrophoresis and chromarography
E-nose
Optical biosensor (BIACORE 3000) tool
Duplex polymerase chain reaction
Electrophoretic, chromatographic and PCR techniques
Gas chromatography
ELISA and PCR techniques
4 Low-valued milk Reverse-phase high performance liquid chromatography and
urea-polyacrylamide gel electrophoresis
TaqMan real time PCR
E-tongue
HPLC method
Sandwich IgG ELISA
5 Milk powder FAST (Fluorescence of advanced maillard products and soluble tryptophan)
6 Color Color Capillary electrophoresis
Conductivity
Impedance
Capacitance
Piezoelectric transducer
7 Preservatives
Impedimetric
E-Tongue
Thermoacoustic analysis
Rosalic acid test
8 Neutralizers Conductivity or pH measurement
Potentiometric biosensor
pH measurement
Durable NH+
4 sensitive CHEMFET based sensor
9 Urea pH sensitive field effect transistor
Manometric biosensor
Ion selective electrode
Calorimetric method
Biosensors
Reverse phase HPLC method
Capillary Electrophoresis
ELISA
Fourth derivative spectroscopy
10 Whey/Liquid whey
Blot immunoassay method
Phosphor partition
NIR spectroscopy
Immunochromatographic assay
Frequency admittance measurements
E-nose
Electrical conductivity
11 Water
Ultrasonic transmitter receiver system
NIR measurement
Freezing point osmometry and freezing point cryoscopic method
Source: Das S, Goswami B and Biswass K 2016. Milk Adulteration and Detection” A Review. Sensor Letters, 14:4-18 [3].

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International Journal of Chemical Studies
 
Table 2: Different electrical methods to detect milk adulteration.
S. No. Electrical Method
1 Potentiometric Sensors
2 Conductance measurement
3 Electrical Conductivity method
4 Ultrasonic Detectors
5 E-nose
6 E-tongue
7 Capacitance growth curve
8 Piezoelectric sensor
9 Impedance probe
Source: Das S, Goswami B and Biswass K 2016. Milk Adulteration and Detection” A
Review. Sensor Letters, 14:4-18 [3].

Table 3: Rapid qualitative detection of different edible adulterants in milk

Limit of
Adulterant Procedure Observation Reference
detection
(Sharma et al.
2012) [15]
Take 10 ml milk sample in a test tube. Add 5 ml conc.
Appearance of red color indicates the 0.2% (Kamthania et
Sugar HCl and 0.1 g resorcinol. Place the test tube in water
presence of added sugar. (w/v) al. 2014) [9]
bath for 5 min.
(Singh et al.
2012)
(Sharma et al.
2012) [15]
Take 3 ml milk sample in a test tube. After boiling it
Appearance of blue color indicates the 0.02% (Singh et al.
Starch thoroughly, cool it to room temperature. Add 2 to 3
presence of starch. (w/v) 2012) [17]
drops of 1% iodine solution.
(Kumar et al.
1998) [11]
Take 1 ml of milk sample in a test tube. Add 1 ml of
modified Barford's reagent. Heat the mixture for exact 3 Immediate appearance of deep blue color 0.1% (Sharma et al.
Glucose
min in a boiling water bath. Rapidly cool under tap indicates the presence of glucose. (w/v) 2011) [16]
water. Add 1ml of phospomolybdic acid and shake well.
Appearance of yellow color indicates the
Take 5 ml of milk sample into a test tube. Add 1 ml of
Common presence of added salts, whereas, brick 0.02% (Sharma et al.
0.1 N silver nitrate solution. Mix the content thoroughly
salt red color indicates the milk free from (w/v) 2012) [15]
and add 0.5 ml of 10% potassium chromate solution.
added salt.
Dilute the milk 1/10. Put a drop of diluted milk on the (Kamthania et
centre of a glass slide. Now place a drop of Hansa test Curdy particles develop within half a al. 2014) [9]
Buffalo milk
serum (duly preserved) on the drop of milk and mix minute in milk containing buffalo milk. (Singh et al.
together with a glass rod or clean tooth pick. 2012) [17]
The development of orange colour, it
Skim milk indicates the milk is adulterated with Singh et al.,
Add nitric acid drop by drop in to the test milk sample.
powder skim milk powder. Samples without 2012 [17]
skim milk powder shows yellow colour.

Table 4: Rapid qualitative detection of different hazardous chemicals in milk

Limit of
Adulterant Procedure Observation Reference
detection
(Singh et al.
2012) [17]
Take 10 ml of sample and add 2 drops of paraphenylene diamine Intense blue color indicates (Kamthania et al.
H2O2 0.025%
hydrochloride solution. presence of H2O2 in milk 2014) [9]
Sharma et al.
2012) [15]
Appearance of violet or blue (Singh et al.
Take 10 m milk sample. Add 5 m conc. sulphuric acid with a little color at the junction of two 2012) [17]
Formalin 0.1%
amount of ferric chloride without shaking. liquid layers indicates the (Kamthania et al.
presence of formalin. 2014) [9]
If the turmeric paper turns red,
Borax and Take 5 ml milk sample. Add 1 ml conc. HCl. A turmeric paper is (Singh et al.
it indicates the presence of
boric acid dipped and it is dried in a watch glass at 100°C. 2012) [17]
borax or boric acid.
If the colour of milk changes to
Take 5 ml of milk in a test tube and add 5 ml alcohol followed by 4- pinkish red, then it is positive (Singh et al.
Neutralizers
5 drops of rosalic acid for sodium carbonate 2012) [17]
/bicarbonates
Appearance of distinct yellow
Take 5 ml milk sample in a test tube. Add 5 ml p-Dimethyl Amino 0.2% (Singh et al.
Urea color indicates presence of
Benzaldehyde reagent. (w/v) 2012) [17]
added urea
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International Journal of Chemical Studies
 
5 ml of hot milk is taken in a test tube and added with a suitable acid Appearance of precipitate
Ammonium 0.05% (Singh et al.
for e.g. citric acid and the whey thus separated is filtered. Collect the indicates the presence of
Sulphate (w/v) 2012) [17]
whey in another test tube and add 0.5 ml of 5% barium chloride. ammonium sulphate in milk.
Take 10 ml sample milk in a beaker. Add 10 ml mercuric chloride Appearance of blue colour
solution to it. After mixing, filter through what man indicates the presence of (Sharma et al.
Nitrate 0.2%
No 42 filter paper. Take 1 ml filtrate in a test tube and add 4 ml of nitrates. Pure milk sample will 2011) [16]
diphenyl amine sulphate or diphenylbenzidine reagent. not develop any color.
Take 5 mL milk sample in a test tube. Upon acidification with
Appearance of buff color
sulfuric acid, 0.5% ferric chloride solution is added to it drop by
Benzoic and indicates the
drop. Mix it. Five ml of milk is taken in a test tube and acidified with (Singh et al.
salicylic presence of benzoic acid
concentrated sulphuric acid. 0.5% ferric chloride solution is added 2012) [17]
acid whereas that of violet color
drop by drop and mixed well. Development of buff colour indicates
indicates salicylic acid.
presence of benzoic acid and violet colour indicates salicylic acid.
(Singh et al.
Pulverized Take 10 ml milk sample in a test tube. Add equal quantity of hot Appearance of pink color 2012) [17]
soap water to it, then add 1 – 2 drops of phenolphthalein indicator. indicates presence of soap. (Kamthania et al.
2014) [9]
Appearance of violet colour
Take 5 ml of milk in a test tube and add 0.1 ml of bromocresol (Singh et al.
Detergents indicates the presence of
purple solution. 2012) [17]
detergent.
Appearance of yellow color in
Coloring Take 10 mL milk sample in attest tube. Add 10 ml diethyl ether. (Batis et al.
ethereal layer indicates the
matter After shaking, allow it to stand. 1981) [1]
presence of added color.

Conclusion 9. Kamthania M, Saxena J, Saxena K, Sharma DK. Methods

On the basis of above review, we can conclude that, the milk of Detection & Remedial Measures. Int J Engg Tech Res.
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