Title of Invention	A METHOD FOR THE DETECTION OF CHEMICAL ADULTERANTS IN SYNTHETIC MILK AND READY TO USE KIT FOR THE DETECTION OF SYNTHETIC MILK.
Abstract	The present invention relates to a ready to use kit for the detection of chemical adulterants in synthetic milk and also a method for the detection of urea, detergent and sodium bicarbonate.

Full Text

Field of the Invention The present invention relates to a method for the detection of chemical adulterants in synthetic milk. The present invention also provides for a ready- to use kit for the detection of chemical adulterants in synthetic milk. This invention can be used for both for household and dairies. Background of the Invention Synthetic milk is prepared by emulsifying vegetable oils with appropriate amount of detergents, urea, caustic soda, sugar, salt etc. which are dissolved in water and blended with the detergent emulsified oil. This mixture is then mixed with natural milk in varying proportions. The dairy industry employs various checks, but adulterers manage to get away because synthetic milk is an excellent imitation of natural milk. Milk fat is mimicked by vegetable oil, the nitrogen component in milk protein is mimicked by urea and the white colour is got by shampoo. Detergents are added to make it frothy. Synthetic milk consumption leads to kidney failure and liver damage. Now a days synthetic milk has become a public health hazard specially in our country. Therefore, it is a major concern not only for a common man but for dairy industry as well. Synthetic milk is used for chemically synthesized milky liquid adulteration of natural milk. A number of tests are currently employed by the dairy industry to check this adulteration. A common test used for detection of adulteration in synthetic milk is MAT which, is a biophysical method in which the sample is subjected to centrifugation followed by microscopic examination for detection of micro coagulation of milk proteins and detection of non milk fat (emulsified oil). Highly sensitive and specific for detection of synthetic milk adulteration in milk supply. This test can replace a number of other chemical tests routinely pet formed for quality assurance of milk. A lot of work has been done in this area for the detection of synthetic milk in dairy milk by Manish M Paradkar, Rekba S Singhal and Pushpa R Kulkarni. For instance, "an approach to the detection of synthetic milk in dairy milk: Detection of detergents" reports on the methods to detect and estimate anionic detergents, which are important ingredients of synthetic milk formulations, based on the extraction of a detergent-methylene Blue complex in chloroform, which can be subsequently quantified by measuring the optical density at 653 nm. A base value of 0.52 for optical density was found to represent pure cows' milk. Any result above this value suggested the presence of an anionic detergent in milk. The work by Paradkar (Manish M.) et al in "an approach to the detection of synthetic milk in dairy milk: detection of urea" reports a qualitative test for the detection of externally added urea (as synthetic milk) using soybean urease. The quantitative detection of synthetic milk in dairy milk could be achieved by this method based on the a' value of L, a, b1 scale of the HunterLab colorimeter and also by the diacetyl complexation method. Both methods are sensitive enough to detect amounts as low as 2% synthetic milk added to dairy milk, corresponding to 20 mg of added urea per 100 ml. Another method for the detection of detergent has been reported in International Journal of Dairy Technology, Volume 54 Issue 1 Page 36 - February 2001 in "An approach to the detection of synthetic milk in dairy milk: effect of the addition of synthetic milk on the flow behaviour of pure cow milk" by Manish M Paradkar, Rekha S Singhal and Pushpa R Kulkarni. According to this report, the flow behaviour of cow milk, synthetic milk and their blends was evaluated with a Haake rheometer. Changes in flow behaviour, were believed to be due to detergent, an important constituent of synthetic milk, which emulsifies the vegetable fat in die aqueous phase. The results are of value in detecting very low levels (1-10%) of synthetic milk in cow milk Yet in another report of Manish M Paradkar, Rekha S Singhal and Pushpa R Kulkarni, in the international journal of dairy technology (February 2001) in "An approach to the detection of synthetic milk in dairy milk: detection of vegetable oil and sodium bicarbonate" discloses a high-performance thin layer chromatography (HPTLC) method for the detection of groundnut oil and palm oil, the main constituents of synthetic milk.. The minimum detectable level of the oils was 80 mg/100 ml. The rosolic acid qualitative test was found to be sensitive enough to detect levels as low as 20 mg/100 ml sodium bicarbonate in milk National Dairy Development Board also developed a kit for the detection of the chemical components of the synthetic milk. The kit costs around US $130 and is out of the reach of common people. However, there is no method, which is sufficient to check most of the major chemical components. Most of the tests are out of the reach of common people as they are not cost effective and require sophisticated instruments. No such other kit is available in the market. An overview of the literature related to synthetic milk shows that the detection of synthetic adulteration in milk requires sophisticated instruments and technical expertise. Also these methods are not approachable to common people. There is therefore a need to develop methods in order to obtain rapid, easy and cost effective detection of major harmful synthetic components present in milk. 'Objects of the invention The object of the present invention is therefore to develop the rapid method for the detection of chemical adulterants present in synthetic milk. Another object is to provide a ready to use kit that provides for a rapid, easy and cost effective method for detection of major harmful synthetic components present in milk. Yet another object of the invention is to propose a method, which does not involve any instrument and any technical, know how. Still another of this invention is to develop ready to use kit, which is portable and accessible to person even at the grassroots level that uses eco-friendly natural products for the detection of the chemical components of the synthetic milk. Summary of the Invention To achieve the aforementioned objects, the present invention provides for a ready to use kit for the detection of chemical adulterants in synthetic milk comprising predetermined amount of an extract from a pulse or cereal having urease activity predetermined amount of hydrophobic solvent and - predetermined amount of dilute HC1 (0.01 - 0.1 N) This invention further provides for a rapid method for the detection of chemical adulterants in synthetic milk using the ready to use kit of the present invention. Reference to the accompanying drawings For the purpose of illustrating the present invention without any limitation thereof to the scope of the invention, reference is made to the accompanying drawings Figure 1 illustrates the presence of urea in milk using the kit of the present invention Figure 2 illustrates the presence of detergent in milk using the kit of the present invention Detailed description of the invention Synthetic milk contains emulsified vegetable oils with appropriate amount of detergents, urea, sodium bi carbonate, sugars, salts, etc. and varying amount of natural milk. The ready to use kit of the present invention involves the use of natural products and simple laboratory reagents mainly for the detection of urea, detergents and sodium bi carbonate. The kit consists of three reagents namely extracts of a pulse or cereal having urease activity, a hydrophobic solvent and dilute HC1 (0.01 - 0.1 N) for the detection of urea, detergents and sodium bicarbonate respectively. The plant sources having urease activity without any limitation thereof includes Jackbean (Canvavlia ennsiformis), Chickpea (Cicer arietinum), (Cajanas cajan), Potato (Solanum tuberosum), Barley (Hordeum vulgare), Soyabean (Glycine max), Mulbery (Morus alba) leaves and other cereals legumes. The hydrophobic solvents used for detection of detergent without any limitation thereof includes hexane or diethyl ether or any other hydrophobic solvent. Procedure of detection of chemical components in synthetic milk The method for the detection of the urea, detergent or caustic soda in synthetic milk comprises of the following steps: 1. Reconstituting dry powder of the extract of cereal or pulse in two-tablespoon water, which can be used for one week under refrigeration and adding equal amounts of milk with the reconstituted solution. An instant pink to red colour appears which shows the presence of urea in the milk. As shown in Figure 1, the amount of detergent present is as shown in table 1. 2. When one tablespoon of milk is mixed with equal amount of hydrophobic solvent, heated for Vz minute to one minute at a temperature ranging between 40-80°C and left at room temperature for some time shows that if there is no formation of a clear layer above the milk, detergent is absent whereas formation of a turbid layer shows presence of detergent The detectable limit for detergent is as low as 0.002% As shown in Figure 2, the amount of detergent present is as shown in table 2. 3. When a drop of a solution of HC1 was mixed with equal amount of milk, formation of effervescence shows the presence of sodium bi carbonate. The present invention is not to be limited in scope by the specific embodiments disclosed herein which are intended as illustrations of a number of aspects of the invention and any embodiments which are functionally equivalent are within the scope of this invention. Those skilled in the art will know, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. Table 1 Tube Amount of urea/100ml 1st Control 2ad 2mg 3rd 3mg 4th 4mg 5th 6mg 6th 8mg lOmg Table 2 Tube Amount of detergent 1st Control 2nJ 0.002% 0.004% 4th 0.006% 5th 0.008% 6th 0.01%, , , , , , , , , , , , , , , , , ,

Documents:

711-del-2004-abstract.pdf

711-DEL-2004-Assignment (29-10-2009).pdf

711-del-2004-claims.pdf

711-DEL-2004-Correspondence-Others (29-10-2009).pdf

711-DEL-2004-Correspondence-Others-(30-11-2009).pdf

711-del-2004-correspondence-others.pdf

711-del-2004-correspondence-po.pdf

711-del-2004-description(complete).pdf

711-del-2004-drawings.pdf

711-del-2004-form-1.pdf

711-DEL-2004-Form-16 (29-10-2009).pdf

711-del-2004-form-19.pdf

711-del-2004-form-2.pdf

711-del-2004-form-3.pdf

711-DEL-2004-GPA-(30-11-2009).pdf

711-del-2004-gpa.pdf

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