DESC:Method for Determining Content of Foaming in Protein Supplements
Field of the Invention
The present invention relates to a method of determining the content of foaming in whey protein supplements wherein the method comprises measuring the percentage of foaming upon shaking a suspension of a protein supplement with a shaking apparatus.

Background of the Invention
Protein supplements are one of the most widely consumed supplements by athletes and physically active individuals. Amongst the ingredients used to manufacture this type of product (i.e. caseinates, whey, egg, soy, plant, hemp, and wheat proteins), whey protein is the most commercialized in the nutritional supplement market due to its high nutritional value when compared to other proteins sources. Differences in the physical-chemical composition of whey protein supplements potentially influence its nutritional effect on the human body. The nutritional quality of whey protein supplement depends on amino acid composition, bioavailability of essential amino acids, protein digestibility, and physiological utilization of specific amino acids after digestion and absorption.
Whey Protein has its own unique properties of mixing and dissolving in water and milk. These unique properties make it an accurate drink for mixing and drinking with water or milk. Proteins contain long chains of amino acids that bind the particles inside them. When mixed in a shaker firmly, it causes a breakage of all the bonds between acids. Hence, these bonds then exposed to air cause foam bubbles to arise and cause foaming.
Dispersion of gases in the liquid is referred to as foams. Protein forms pliable, cohesive films throughout the gas bubble surface which is responsible for foam stabilization. As the hydrophobic area of protein is adsorbed at the interface followed by partial unfolding during whipping or shaking. Protein adsorption causes a decrease in surface tension which in turn further facilitates the formation of the gas bubble as well as the emergence of a new interface. Film formation is associated with partially unfolded proteins.
Foaming is a parameter which directly influences consumer’s acceptance for a protein supplement. If there is too much foaming in a product then it leads to negative perception about the quality of protein among the users. Hence there is a high need for a method which accurately measure the foaming of any protein supplement. Foaming is directly related to the quality of the protein present in a nutritional supplement such as whey protein supplement. If a protein supplement product gives lesser foaming compared to other products then it is safe to conclude that the source of protein raw material is of good quality.
The inventors of the present invention have developed a user friendly and cost effective method for determining the quality of protein in protein supplements in a laboratory setting that is easy to perform with good reproducibility.

Summary of the Invention
It is an object of the present invention to provide a method for the determining the foaming percentage in a protein supplement wherein the method comprises shaking a predetermined quantity of the protein supplement suspended in a water-based liquid with a mechanical shaker at a fixed vibration for a predetermined time and transferring the suspension in a measuring apparatus and measuring the percentage of foam at the top of the suspension.
In an embodiment of the above aspect, water-based liquid is selected from the group consisting of water and milk.
In another embodiment of the above aspect, the suspension of protein supplement in water-based liquid is shaken in a shaker bottle of about 825mL capacity.
In a further embodiment, the suspension is in a quantity as per the recommended dosage/serving.
In an embodiment of the above aspect, the predetermined quantity of the protein supplement is a unit dose amount for the protein supplement.
In another embodiment of the above aspect, the suspension is transferred in a 500 ml measuring cylinder.
In a further embodiment of the above aspect, the foaming percentage is calculated using the formula:
Foam % = [A/(A+B)]/100
A = Foam Volume (ml)
B = Liquid Volume (ml) without foam layer
In another embodiment, the protein supplement is whey protein supplement.
In another embodiment, the mechanical shaker is a horizontal mechanical shaker.
Detailed Description of the Invention
Scientifically, foam is a mass of small bubbles in the liquid, usually formed by agitation or fermentation. The gas bubbles are segregated from each other by an elastic liquid wall. Foam basically forms by trapping gas particles in a liquid. As a result, trapped gas comes out in the form of bubbles known as foam. The foam bubble’s diameter ranges from 1 micrometer to several centimeter. The foam density relies on the size of the bubble and thickness of foam wall. Few bigger size bubbles form a light foam while dense foam is formed by numerous small size bubbles. If the number of the bubbles is less and bigger in size it forms a light foam. If the size of the bubble is less and more in number it will form a dense foam. Some common examples are whipped cream, ice cream, cake, bread, aerated drinks, etc.
The lesser the amount of foam, the greater the amount of protein in the protein supplement. Foams are unacceptable or undesirable in certain food for example during processing of certain fruit juices, vegetable oils, syrups, heating of liquids, evaporation processes, drying of food, etc. Anti-foaming agents are used as excessive foaming results in a reduced rate of processing, packaging and losses in the product volume.
The test method of the invention can indicate the quality of protein in any protein sample and several marketed protein supplement products were tested by this novel method. Table 1 provides the results of testing the foaming capacity of some commercially available protein supplements.
This method is fast, reliable and consumer friendly which can be done quickly in a laboratory setting. The test reliably provides confidence of the quality of the protein supplement such as whey protein.

The method of the invention can be performed as follows:
Requirements:
1. Analytical Weighing Balance that can weigh up to ± 0.1g. The protein supplement amount can be the suggested serving as per product recommended usage.
2. Horizontal shaking machine (SK300 Dasin or as any other shaker machine as available).
3. Shaker bottle - Approximately 825 ml capacity, measurement marking to 600 ml. External diameter 70-80 mm and height 170-180 mm. Water or water-based liquid as applicable is added with a calibrated measuring cylinder.
4. Thermometer - 0 to 100 °C ± 0.5 °C.
5. General Lab equipment - Laboratory spatula, weighing boats.
6. Glass measuring cylinder.
Method:
An automatic shaker is used to mix the protein supplement powder in water-based liquid or water taken in a flask/shaker and mixing them at constant, specific speed and time. An automatic shaking machine contains an oscillating board that is used to hold the flask/shaker at the time of shaking.
1. Add water or water-based liquid (8 to 15 °C) in the shaker (suggested dilution as per product recommended usage) using a measuring cylinder in ml.
2. Add powder to the shaker as per applicable suggested serving recommended usage in g ±0.1g.
3. Mix the contents of the shaker by the horizontal shaking machine. Set the shaking machine at speed of 6 for 15 seconds.

4. Once shaking is done transfer the contents immediately into a 500 ml measuring cylinder through the side walls of the cylinder. Keep the cylinder still without disturbing it on a plane surface, read the height of the foam layer on top of the liquid in mL.

5. Take the readings after 1 and 2 minutes intervals respectively without disturbing the measuring cylinder.

As a precaution, for mixing of product on the shaking machine, use one shaking arm at a time. When mixing is done remove the shaker gently and pour it immediately and gently in the measuring cylinder through the side walls of it.

Calculation
Foam % = [A/(A+B)]/100; wherein
A = Foam Volume (ml)
B = Liquid Volume (ml) without foam layer

Precautions to be taken during performing the method:
1. Add water first before adding powder.
2. Calibrated stopwatch must be used.
3. The content of the shaker should be completely transferred through the side walls gently avoiding any vigorous pouring, to mitigate the risk of formation of extra foam due to air entrapping.
4. Measuring cylinder should not be disturbed and kept still before and while taking the readings on the plane surface (at eye level is recommended).
Note - Shaking is required approximately 180-200 counts per 60 seconds. This can be ensured by using a Tachometer or applicable device.

Table 1: Foam Percentage of Some Commercial Protein Supplements Determined by the Method of the Invention.
S. No. Product Details Foaming (%)
1 minute 2 minutes
1 Whey Gold Rich Milk Chocolate (B. No. : AGWCF421) 19.61 15.15
2 Whey Gold Rich Milk Chocolate
(B. No. : AGWCF446) 5.95 6.02
3 Whey Gold Rich Milk Chocolate (B. No. : AGWCF449) 28.57 19.56
4 Whey Protein Cookies and Cream (B. No. : AWPCCF043) 5.21 3.95
5 Whey Hydro Chocolate Flavour (B. No. : AWHCF031) 7.32 5.45
6 Whey Protein Cafe Mocha (B.No. : AWPCM062) 2.44 2.01
7 Iso Zero Low Zero Carb (B. No. : AIZLCF019) 9.7 7.75
8 ISO ZERO Low Carb Chocolate Flavour (B. No. : AIZLCF022) 9.09 8.23
9 Iso Zero Strawberry (B. No. : AIZSF040) 14.61 5.95
10 WPC 80 ( B. No. : ML8219A4) 2.35 1.19
,CLAIMS:1. A method for the determining the foaming percentage in a protein supplement wherein the method comprises shaking a predetermined quantity of the protein supplement suspended in a water-based liquid with a mechanical shaker at a fixed vibration for a predetermined time and transferring the suspension in a measuring apparatus and measuring the percentage of foam at the top of the suspension.
2. The method as claimed in claim 1, wherein water-based liquid is selected from the group consisting of water and milk.
3. The method as claimed in claim 1, wherein the suspension of protein supplement in water-based liquid is shaken in a shaker bottle of about 825mL capacity.
4. The method as claimed in claim 1, the suspension is in a quantity as per the recommended dosage/serving.
5. The method as claimed in claim 1, wherein the predetermined quantity of the protein supplement is a unit dose amount for the protein supplement.
6. The method as claimed in claim 1, wherein the suspension is transferred in a 500 ml measuring cylinder.
7. The method as claimed in claim 1, wherein the protein supplement is whey protein supplement.
8. The method as claimed in claim 1, wherein the mechanical shaker is a horizontal mechanical shaker.