Title of Invention

"PROCESS FOR THE DETECTION OF SYNTHETIC MILK"

Abstract This invention relates to a process for the detection of synthetic milk comprising the step of: a) subjecting the micro centrifuge tube containing sample of milk to the step of centrifugation at 8000 to 10,000 rpm for 10 to 12 minutes at room temperature to obtain fat layer on the top of the cream layer; b) taking small amount of fat from said fat layer on a glass slide and adding solution A as herein described c) the fat changes to red colour oil fat indicating the presence of non milk fat, d) again subjecting centrifuge tube containing milk sample to the step of centrifugation of 3800 to 4200 rpm for 4 to 6 minutes at room temperature, e) decanting the centrifuged milk of step (d) to recover pellet, f) suspending the pellets of step (e) in solution B as herein described, g) adding solution C to suspension of pellets of step (f) tp find the adulteration in milk.
Full Text This invention relates to a diagnostic kit for detection of synthetic milk.
Adulteration of milk supply with chemically synthesized milky liquid termed as "Synthetic milk" has become a public health hazard and major concern for the dairy industry. Synthetic milk is prepared by emulsifying vegetable oils with appropriate amount of detergents. Urea, caustic soda, sugar, salt etc. are dissolved in water and blended with the detergent emulsified oil. This mixture is mixed with the natural milk supply in varying proportions. A number of tests are currently employed by the dairy industry to check this adulteration. However, it has been a common experience that inspite of all the testing, the problem of milk adulteration continues to grow unchecked.
The process as such known in the art for the detection of synthetic milk comprises a number of chemical testing methods wherein different chemicals are used for detection of the synthetic milk. There are number of disadvantages associated with the conventional process used for the detection of synthetic milk.
One of the main disadvantage is that the sensivity of the chemical tests is poor to determine as to whether the particular adulterant has been added or not, since the number of chemicals which could be added to increase solids not fat (SNF) value of the milk is numberous.
Another disadvantage is that the conventional
process of checking the presence of vegetable oils by
Refractometry becomes ineffective by using a mixture of
different oils to give the B R value close to B R valve
of milk fat and thus it becomes difficult to know as to
whether the vegetable oils are added in the milk.
Therefore the main object of this invention is to propose the diagonstic kit useful for detection of synthetic milk present in the natural milk.
Another object of this invention is to propose a diagnostic kit having a plurality of invented reagents used for the detection of synthetic milk.
Yet another object of this invention is to propose a process for the detection of synthetic milk which is a quick sensitive and specific test for the detection of adulteration of natural milk by synthetic milk.
According to this invention there is provided a process for the detection of synthetic milk comprising the step of :
a) subjecting the microcentrifuge tube containing sample of milk to the step of centrifugation at 8000 to 10,000 rpm for 10 to 12 minutes at room temperature to obtain fat layer on the top of the cream layer;
b) taking small amount of fat from said fat layer on a glass slide and adding solution A as herein described
c) the fat changes to red colour oil fat indicating the presence of non milk fat,
d) again subjecting centrifuge tube containing milk sample to the step of centrifugation of 3800 to 4200 rpm for 4 to 6 minutes at room temperature,
e) decanting the centrifuged milk of step (d) to recover pellet,
f) suspending the pellets of step (e) in solution B as herein described,
g) adding solution C to suspension of pellets of step (f) to find the adulteration in milk.
In accordance with this invention the diagonstic kit has microcentrifuge tubes of the capacity of 2 ml placed in a kit box. Glass slides and cover slips are provided in said box for use during the testing of the adultered milk. These features do not constitute the inventive features of the present invention. However, according to this invention a plurality of reagents are provided in the kit box which are being used for the detection of synthetic milk. The reagents comprises for example solution A, solution B and solution C. Solution A is named as Sudan III dye solution and comprises 0.8 to 1.2% Sudan III dye dissolved in 100 ml of 95% ethanol and 100 ml of
glacial acetic acid in the form of a saturated solution. The solution so obtained is filtered to get said solution A. Solution B comprises 0.9 of 0.15M Sodium Chloride (Nacl) dissolved in the distilled water and solution C comprises 0.03 to 0.05X Trypan blue dissolved in phosphate buffered saline of the pH 7.2. Besides the above items other instruments like microscope, tooth picks, Microcentrifuge (Remi, R12C or equivalent) are required to perform the process for the detection of synthetic milk.
In accordance with the process of the present
invention 1.5 ml of the milk sample is taken in the 2 «1
microcentrifuge tubes and the microcentrifuge is
subjected to the step of centrifugation at 8000 to
10,000 rpm for a period of 10 to 12 minutes at the room
temperature. At the end of centrifugation the tubes Are
observed in light for a distinct fat layer on
the top of the cream layer. A small amount of the
fat layer is taken out without touching the cream
layer and is put on a glass slide. A
drop of the solution A is put on the elide. After a few
minutes the slide is observed for presence of red colour oil droplets. The presence of such droplets demonstrate
the presence of non milk fat in the sample and thus the adulteration of non milk fat in the milk sample is
suspected.
Similarly another milk sample of i.S ml is
taken in to a 2 ml microcentrifuge Tube is subjected to the step of centrifugation at 3800 to 4200 rpm for 4 to
6 minutes at the room temperature. After centrifugation the milk sample is subjected to the step of decanting to
recover the pellet. The pellet so recovered after decanting is suspended in 0.05 ml of solution B. If the
pellet is not suspended easily in the solution B and also is of large size relative to the normal milk
pellet, the quality of the milk is suspected. In the normal milk sample the pellet suspends easily and nicely
in solution B. A drop of suspension of the pellet is aken on the glass slide and mixed with one drop of
solution C.After putting a cover slip, the suspension is
mined under the microscope for the presence of coagulated mass of protein which takes blue stain. No such stained mass is observed in case of the normal milk even after storage of milk for 48 hr at 15C.
The observed protein coagulation is due to addition of different adulterants to the milk supply. Presence of clot like structures confirms that milk has been adulterated.
The kit and the process for the detection of synthetic milk is herein described further in the following example. EXAMPLE
1.5 ml of the milk sample is taken in 2 ml microcentrifuge tube and subjected to centrifugation at 9000 RPM, for 10 minutes at room temperature. (Twenty four such samples can be processed in one run). At the end of centrifugation, the tubes are observed in light for a distinct fat layer on top of the cream layer. Using the free end of a tooth pick, a small amount from the top layer (without touching the cream layer) is taken and
put^ on a glass slide. A drop of solution A is put on the slide. After a few minutes, the slide is observed
for presence of red coloured oil droplets. Presence of any droplets will demonstrate the presence of non milk
fat in the sample.
Normal buffalo or cow milk when tested, even
after storing for 24 hrs at room temperature or at 37 C, is found to be negative for above described non milk fat
droplets.
One and half ml of milk is again subjected to
centrifugation at 4000 RPM for 5 minutes. After centrifugation, the pellet is recovered by decanting the
milk sample. The pellet is then examined after suspending in 0.05 ml of solution B. In a normal milk
sample, the pellet suspends nicely. On the contrary, in the samples where the pellet is not suspended easily and
is of large size relative to the normal milk, the quality of the milk is suspected.
A drop of suspension of the pellet is taken on the glass slide and mined with one drop of solution
C. After putting a cover slip, the suspension is examined under the microscope for the presence of coagulated mass of protein which takes blue stain. No such stained mass is observed in case of the normal milk even after storage of milk for 48 hr at 15C.
The observed protein coagulation is due to addition of different adulterants to the milk supply. Presence of clot like structures confirms that milk has been adu1terated.






CLAIMS
1. A process for the detection of synthetic milk comprising the step of :
a) subjecting the microcentrifuge tube containing sample of milk to the step of centrifugation at 8000 to 10,000 rpm for 10 to 12 minutes at room temperature to obtain fat layer on the top of the cream layer;
b) taking small amount of fat from said fat layer on a glass slide and adding solution A as herein described
c) the fat changes to red colour oil fat indicating the presence of non milk fat,
d) again subjecting centrifuge tube containing milk sample to the step of centrifugation of 3800 to 4200 rpm for 4 to 6 minutes at room temperature,
e) decanting the centrifuged milk of step (d) to recover pellet,
f) suspending the pellets of step (e) in solution B as herein described,
g) adding solution C to suspension of pellets of step (f) to find the adulteration in milk.

2. The process as claimed in claim 1 wherein Solution A comprises 0.8 to 1.2% Sudan III dye dissolved in 100 ml of 95% ethanol and 100 ml of glacial acetic acid.
3. The process as claimed in claim 1 wherein Solution B comprises 0.15 M sodium chloride dissolved in distilled water.
4. The process as claimed in claim 1 wherein said solution C comprises 0.03 to 0.05% Trypan blue dissolved in phosphate buffered saline of the pH 7.2.
5. A process for the detection of synthetic milk as substantially as herein described.

Documents:

1433-del-1999-abstract.pdf

1433-del-1999-claims.pdf

1433-del-1999-correspondence-others.pdf

1433-del-1999-correspondence-po.pdf

1433-del-1999-description (complete).pdf

1433-del-1999-form-1.pdf

1433-del-1999-form-19.pdf

1433-del-1999-form-2.pdf

1433-del-1999-gpa.pdf


Patent Number 243797
Indian Patent Application Number 1433/DEL/1999
PG Journal Number 46/2010
Publication Date 12-Nov-2010
Grant Date 08-Nov-2010
Date of Filing 02-Nov-1999
Name of Patentee ASHOK KUMAR
Applicant Address 15, PRAYAG SAROVAR, RAMGHAT ROAD, ALIGARH-202 001, INDIA
Inventors:
# Inventor's Name Inventor's Address
1 ASHOK KUMAR 15, PRAYAG SAROVAR, RAMGHAT ROAD, ALIGARH-202 001, INDIA
PCT International Classification Number A23C
PCT International Application Number N/A
PCT International Filing date
PCT Conventions:
# PCT Application Number Date of Convention Priority Country
1 NA