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Lecture - 8 (Milk Adulteration & Its Detection)
Lecture - 8 (Milk Adulteration & Its Detection)
IMPORTANT
TERMINOLOGIES
ADULTERATION:
Making the article impure by the addition of some of
the legally prohibited substances into a more
valuable product or the subtraction into a more
valuable component.
ADULTERATION OF MILK:
• Addition of water.
• Addition of separated milk.
• Addition of thickening agents when milk is watered, it becomes
deficient in density and viscosity which may be restored by
addition of substances like gelatin, cane sugar, starch, glucose.
• Partial skimming.
• Unusual adulterants, a few new types of adulterants like urea,
melamine, ammonium sulphate etc. have come up.
Prevalence of Milk Fraud in Chinese market and its
relationship with Fraud vulnerabilities in the Chain
• 1. Study aimed to survey the prevalence of ultra high temperature (UHT) processed milk samples suspected of being
adulterated in Chinese market.
• 2. For this 12 UHT milk samples from five different brands were purchased from local supermarket in Beijing in Nov 2018
as reference sample.
• ✓ Four label as premium quality (protein content>3.4%) from north of china.
• ✓Four of were normal quality also producing from north.
• ✓ Remaining four were of normal quality and produced in south.
• 3. So, accordingly 3 milk pools were prepared first one is mixture of premium quality (Pool A), second one was the mixture
of normal quality(Pool B) from north and the third one was mixture of normal quality(Pool C) from south of China. The
ratio of four milk samples in each pool was 1:1:1:1
• 4. Three milk pools of 100g, were prepared to which adulterant was added at several concentration.
• 5. Twelve reference sample and three milk pools were from major producers considered as control sample(ie. Non
adulterated) .These 15 samples comprised the training set.
• 6. Now 52 commercial UHT milk samples were purchased from local market in Beijing and e-commerce(across china)
during winter of 2018/2019 (test set). Same adulterants and same adulteration level applied.
Prevalence of Milk Fraud in Chinese market and its
relationship with Fraud vulnerabilities in the Chain
• 7. A total of 24 adulterant were used categorized into five group named protein rich adulterant, Nitrogen based
adulterant, Carbohydrates based adulterant, preservative and water. Both single and combined adulteration test were
conducted.
• 8. In Combined adulteration 40g water added in 100g of milk pool sample then one of adulterants from either protein,
CHO, nitrogen based added to diluted milk pool to increase apparent protein content with 40% w/w, same with other two.
• 9. Ultimately a total of 288 single adulterated sample and 51 combined adulterated samples prepared These 339
adulterated sample were considered as adulterant test set.
• 10. Milk scan FT120 equipment used to measure milk composition. It is based on FTIR technology and reporta series of
milk compositional parameter namely protein, fat, lactose, total solid(TS), solid non fat(SNF) content, density and freezing
point depression (FPD).
• 11. All measured compositional features of premium milk pool (pool A) were higher than the normal milk pool from both
North and South (pool B & C).
• 12. The protein content of control sample (3.4%- 3.7%) w/w was higher than that of raw milk collected from Chinese
Holstein cattle which is 2.9-3.3% w/w.
• 13. Fat and lactose content of Chinese sample is 4.0% and 5.1% respectively. While in Dutch sample it is low about 3.8%
and 4.7% respectively.
Milk
Adulteration and
Its Toxicological
Effects
■ Cane sugar
■ Starch
■ Urea
■ Sodium chloride
ADULTERANTS ■ Neutralizers
IN MILK ■ Quaternary ammonium compounds
■ Anionic detergents
■ Skimmed milk powder
■ Formalin
■ Hydrogen peroxide
■ Boric acid and borates
• Formalin is added in milk to preserve it for longer
period.
• Cough, dyspnea, asthma (one case); redness, in eyes and skin, headache; nausea, vomiting, lung
damage, inflammation.
UREA
The presence of ammonium sulphate increases the
lactometer reading.
• For the purpose to activate Lacto peroxidase system, which destroy the bacterial
population by its antibacterial effect.
• Lacto peroxidase (LP) system is naturally present in raw milk that prevents the
bacterial multiplication due to its bacteriostatic effect. By activating the LP-system,
shelf life of milk can be increased for 8 to 12 hours.
Side Effects of Hydrogen Peroxide
• Eye exposure to dilute hydrogen peroxide may cause extreme irritation and pain but is unlikely to cause
permanent damage.
• Dilute hydrogen peroxide has been reported to cause serious gastrointestinal side effects.
• An overuse of H2O2 can cause damage to DNA cells, and prevent them from replicating, hence leading to
premature aging.
• The use of hydrogen peroxide can cause your skin to dry out or rather drains away the moisture content in skin,
which is required to keep your skin looking young and supple.
• Vitiligo is a disease that is marked by the loss of skin pigment, which affects a lot of people. Studies show that it
is due to the high levels of hydrogen peroxide in the blood and epidermis.
• It can cause irritation in gums and teeth, since H2O2 is a highly reactive compound.
Sugar
• Sugar can increase the levels of glucose in the blood much higher than complex carbohydrates in
a glucose tolerance test can.
Salt Excess Salt
Consumption Has
▪
Been Linked To:
• Soap is added to milk to increase the foaming of milk and thus to have thick milk.
• These prohibited substances may be added to fresh or badly preserved milk with the
tricky purpose to correct its pH and acidity values to optimal ones, and so pretend the
milk is freshly milked or perfectly preserved.
Quaternary Ammonium Compounds
Purpose:
■ Triggering of asthma symptoms in people who already have asthma or new onset
of asthma in people with no prior asthma
Boric Acid
Purpose
■ Like formalin, boric acid and sodium borates are also common preservatives.
■ They are available in the form of either boric acid (H3BO3) or its salt as sodium
borate(Na3BO3) as commercial chemicals.
Toxicity
• Causes renal and urinary problems and even infant death when it reacts with cyanuric
acid inside the body
https://www.researchgate.net/profile/Masood-
Akhtar/publication/267566928_Adulteration_and_Microbiological_Quality_of_Milk_A_Review/links/5582487608ae12bde6e4b
7b3/Adulteration-and-Microbiological-Quality-of-Milk-A-Review.pdf
Quantitative & Qualitative test for
determination of adulteration in milk
Methods of Detecting Adulterants in Milk
In general, these classified into detection methods
Qualitative detection methods.
Quantitative detection methods.
In this, SB-ATR FTIR (Single Bounce Attenuated Total Reflectance - Fourier transform infrared
spectroscopy) has been used to quantify melamine in both liquid and powder milk.
This portable screening system based on Laser Raman spectroscopy has been developed to
quantify melamine
Using Raman band, melamine in dried milk powder has been immediately detected without
extracting melamine from milk (Okazaki et al., 2009).
5. The researchers used Illumina's Solexa
sequencing technology
This method is used to screen for microRNAs in healthy cow milk.
The researchers believe that these microRNAs can serve as biomarkers for reflecting
the quality of milk and dairy products, including milk powder.
Manipulated milk would not be able to replicate the expression levels of these
microRNAs in a standard test
However, mostly such types of detection methods are not accessible at all level of
farmer, milk collection points, or at the spot of dairy cooperatives.
Analytical methods to carry out the
control of adulteration
It is based on chromatographic techniques such as;
1. Reversed phase high performance liquid chromatography (HPLC)
2. Gas chromatography coupled to mass spectrometry (GC-MS)
3. Mid-infrared microspectroscopy
4. Nuclear magnetic resonance (NMR)
5. Matrix assisted laser desorption/ionization time of flight (MALDI-TOF)
6. Raman spectroscopy
QUALITATIVE (BIOCHEMICAL)DETECTION METHODS
For urea detection
Urea forms a yellow complex with DMAB in a low acidic solution at room temperature.
Presence of hydrogen peroxide
Take 1 ml of milk sample in a test tube. add 1 ml of the potassium iodide-starch reagent and mix well. observe the
color of the solution in the tube. blue colour will be developed in the presence of h2o2, whereas pure milk sample
remain white in color.
FOR PRESENCE OF FORMALIN
Formalin reacts with Sulphuric acid and ferric chloride to give a purple colored ring at the junction of the milk layers.
FOR DETECTION OF HYPOCHLORITE
The chloride test allows to identify any added salt water in the milk, a sophistication that the normal cryoscopic
examination does not detect. This analysis allows to quantify the concentration of salt in products such as cheese,
vegetable purees and sauces, and liquid of processing and storage of dairy products.
FOR DETECTION OF MELAMINE
Formerly in 2008, a national standard method issued by the Chinese authorities (AQSIQ and SAC 2008a) identified
HPLC as the technique for rapid determination of melamine in raw milk. The standard showed a detection limit of
0.05 mg/kg and applied to raw milk and milk-based products without additives.
FOR DETECTION OF STARTCH
Starch in milk uses iodine solution, addition of which turns the milk solution to
blue black color due to the formation of starch –Iodo complex, in the presence of starch
As per the law, use of skimmed milk powder (SMP) is not allowed for adjustment of SNF in case of sale of
cow/buffalo or mixed milk. The method is based on the fact that the coagulum obtained from reconstituted
skim milk powder by addition of acetic acid, gives intense blue colour on boiling with phosphomolybdic
acid due to certain reducing groups present in the proteins of milk powder which are able to cause
reduction of molybdenum blue resulting in formation of blue colour.
Fat is extracted from milk and its refractive index is determined and compared with that of the pure milk
fat.
Big order of Supreme Court on
packaged milk of all brands
Chief Justice of Pakistan Justice Mian Saqib Nisar, on January 13, 2018, ordered the authorities to recall
packaged milk of all brands from the markets of Karachi for testing purpose. Hearing a suo motu case
at the Karachi Registry regarding sale of substandard packaged milk, he ordered the relevant
authorities to conduct laboratory tests of packaged milk.
Effectual Gold Nanoprobe Sensor for screening Cow Milk Adulteration in goat
milk-Comparision with PCRArticle in journal agriculture science April
2021Published by Canadian center of science and education