Description:
FIELD OF THE INVENTION

The present invention relates to a milk-soluble Vitamin E composite formulation with Tocotrienol Rich Fraction (TRF), a natural Vitamin E system. In particular, the present invention relates to a milk-soluble Vitamin E composite formulation with Tocotrienol Rich Fraction (TRF) to be incorporated in Milk of either liquid and/or powdered form. The present invention further relates to a process of preparation of a milk-soluble Vitamin E composite formulation with Tocotrienol Rich Fraction which can be incorporated homogenously in Milk thereby enhancing the taste, texture and stability of the liquid as well as the powdered milk. The present invention further relates to a milk formulation comprising said milk-soluble Vitamin E composite formulation with Tocotrienol Rich Fraction (TRF) with enhanced taste, texture and stability.

BACKGROUND OF THE INVENTION

Vitamin E is an essential micronutrient for humans and animals. Vitamin E is a very good antioxidant. Various studies suggest that Vitamin E offers a variety of positive effects. This includes personal care and cosmetic application for hair, skin and scalp rejuvenation and protection against harmful oxidants. Studies in humans and animals show a positive effect on cholesterol metabolism and other lipid-based and lipid-independent risk factors for cardiovascular diseases. This relates to the reduction in the risk of developing cardiovascular diseases, lowering blood cholesterol, regulation of blood coagulation, inhibiting the growth of tumours and optimizing the immune status. They improve the blood-flow characteristics.

Vitamin E has eight isomers divided into Tocopherols and Tocotrienols, four tocopherols (the alpha, beta, gamma and delta) and four Tocotrienols (the alpha, beta, gamma and delta). Tocopherol is the most known and most used Isomer of Vitamin E. While Tocotrienols are the least used isomer, several studies showed that Tocotrienol has better antioxidant properties and greater health benefits than Tocopherol. Tocopherols have a longer tail, whereas Tocotrienols have a shorter, agile tail for heightened mobility. This small difference in molecular structure allows Tocotrienol to cover a larger surface area more quickly, hence making them up to 50 times more effective as antioxidants, protecting cells from oxidative stress. Researchers and scientists are continuously studying, developing and increasing the bioavailability of Tocotrienol for various applications in nutraceuticals and cosmeceuticals.

Furthermore, products that contain fats and oils turn rancid and deteriorate in other ways when they are exposed to air. The action, known as autoxidation, imparts offensive flavours and odours to fats and foods containing them. For a long time, investigators spent a vast amount of effort trying to find ways to prevent autoxidation. Specifically, the basic problem was the mechanism of autoxidation. It is important to prevent autoxidation because fat and fat containing foods valued at several billion dollars are produced and marketed annually. Again, measures that were moderately effective in civilian foods availed little for food that had to be transported long distances and stored in unfavourable climates. In different fats, under different conditions, autoxidation produces tallow, burned, fishy, grassy, and other off-flavours and odours and can be detected quite readily. Natural antioxidants, such as Vitamin E are added to prevent autoxidation.

Tocotrienol Rich Fraction (TRF) is an extract from Palm Oil. Depending on the source and extraction process, TRF normally consists of 75% tocotrienols (alpha-tocotrienol, beta-tocotrienol, gamma-tocotrienol, delta-tocotrienol) and 25% alpha-tocopherol. There are several studies as such, TRF is a more potent antioxidant than alpha-tocopherol alone.

Furthermore, natural Vitamin E is fatty in nature and is not readily soluble in water, as well as in milk.

Milk in human nutrition has a nutritional, protective and detoxification function. It represents inter alia an important dietary resource of minerals and vitamins. Vitamin E Tocotrienol Rich Fraction enhances the fat solubility of milk and sweetness.

Although Vitamin E has been known as an essential nutrient for almost 80 years, the complete understanding of all the aspects related to bioavailability and its effects on health and milk quality has not been achieved.

According to different researches, adding Vitamin E in animal’s diet helps to enhance animal health and productivity. When dietary selenium is adequate, it significantly reduces the occurrence of intermammary infections and clinical mastitis. It is well known that Vitamin E supplementation has a positive effect not only on animal health, but also on the quality of the milk they produce. Furthermore, as an antioxidant, Vitamin E helps to slow lipid peroxidation, maintain oxidative stability and enhance the flavour of milk. The positive effects by adding Vitamin E in Milk were seen as a result of its high antioxidant activity.

Furthermore, it is known that Vitamin E, being fatty in nature, is not readily soluble in water-based and dairy-based solutions. While it is easy to develop a Vitamin E mixture for water-based beverages, it is a challenge to develop a unique Vitamin E composite that can be easily mixed in the Milk. The focus of the present invention is to incorporate Vitamin E in Milk.

Reference is being made to US10493055B2. This patent document discloses a formulation in the form of self-emulsifying drug delivery formulation for improved delivery of tocotrienols comprising a fat-soluble compound, at least one emulsifier, and an oil carrier. The formulation shows an improved bioavailability of tocotrienols. However, this document does not teach explicitly that said formulation is able to dissolve in the milk homogeneously.

Therefore, there remain a need to formulate a milk-soluble Vitamin E composite formulation with Tocotrienol Rich Fraction (TRF) to incorporate Vitamin E, more specifically, the Tocotrienol Rich Fraction in Milk homogenously and conveniently.

SUMMARY OF THE INVENTION

The following disclosure presents a simplified summary of the invention in order to provide a basic understanding of some aspects of the invention. This summary is not an extensive overview of the present invention. It is not intended to identify the key/critical elements of the invention or to delineate the scope of the invention. Its sole purpose is to present some concept of the invention in a simplified form as a prelude to a more detailed description of the invention presented later.

The object of the present invention is to formulate a milk-soluble Vitamin E composite formulation with Tocotrienol Rich Fraction (TRF) to incorporate Vitamin E, more specifically, the Tocotrienol Rich Fraction in Milk homogenously and conveniently.

Another object of the present invention relates to a formulation of a Vitamin E composite of Tocotrienol Rich Fraction applicable for Milk, wherein the Milk can be in liquid or powdered form and classification as raw milk and pasteurized milk.
First aspect of present invention relates to a milk-soluble vitamin E composite formulation comprising;
a. Tocotrienol Rich Fraction (TRF); wherein the amount of the Tocotrienol Rich Fraction (TRF) is about 4% to about 14%, by weight of the formulation;
b. a fatty acid carrier, wherein the amount of fatty acid carrier is about 60% to about 85%, by weight of the formulation; and
c. a solubilizer, wherein the amount of solubilizer is about 4% to about 28%, by weight of the formulation; wherein the ratio of solubilizer and Tocotrienol Rich Fraction (TRF) is from 2:1.

In preferred embodiments, the solubilizer was selected from the group of polysaccharides like xanthan gum or starch or maltodextrin, polyol like glycerine or glycerol or sorbitol, and glycerophospholipids such as lecithin and/or its hydrolysed form.

The second aspect of present invention relates to a process of preparation of a milk-soluble vitamin E composite formulation, comprising:
Phase 1:
1. heating a blending tank to 40 degree centigrade;
2. initiating Nitrogen flow in the blending Tank;
3. adding a solubilizer in the tank after 60 seconds, once nitrogen is thoroughly flushed in the tank;
4. adding 20% of total required Tocotrienol Rich Fraction (TRF) volume in the tank;
5. stirring the solution of step (4) at 80 rpm constantly and continuously;
6. adding another 60% of said TRF in the tank after 300 seconds with constant stirring;
7. blending for another 300sec;
8. adding the remaining 20% TRF and blending for 300sec;
9. removing the blended concoction of step (8)
10. switching off the nitrogen and the heater.

Phase 2:
1. charging a fatty acid carrier in a separate vessel,
2. stirring said fatty acid carrier at a speed of 50 rpm;
3. adding the resulting solution of step (2) in Phase 1 slowly and gradually;
4. allowing homogenous blending of phase 1 and phase 2 for about 300 sec;
5. remove the blended concoction of step (4).

The third aspect of present invention relates to a milk formulation comprising said milk-soluble vitamin E composite formulation, with enhanced sweetness, taste, texture, mouth-feel and shelf-life of milk.

The fourth aspect of present invention relates to a process of preparation of said milk formulation comprising said milk-soluble vitamin E composite formulation, comprising
mixing 0.02-0.20% of said vitamin E composite formulation with milk at 40-80°C using a homogenizer/ stirrer at 20 rpm for 10-30 minutes.

Other aspects, advantages, and salient features of the invention will become apparent to those skilled in the art from the following detailed description, which, taken in conjunction with the annexed drawings, discloses exemplary embodiments of the invention.

BRIEF DESCRIPTION OF THE ACCOMPANYING DRAWINGS

The above and other aspects, features and advantages of the embodiments of the present disclosure will be more apparent in the following description taken in conjunction with the accompanying drawings, in which:
Fig. 1 (a) illustrates comparison chart between raw milk without present formulation and raw milk with present formulation obtained from Sensorial Evaluation Study as disclosed in one of the embodiment of present invention.

Fig. 1 (b) illustrates comparison chart between pasteurized milk without present formulation and pasteurized milk with present formulation obtained from Sensorial Evaluation Study as disclosed in one of the embodiments of present invention.

Fig. 2 illustrates comparison chart of raw milk and pasteurized milk with and without present formulation in view of (a) and (c) pH stability and (b) and (d) viscosity stability evaluation as disclosed in one of the embodiments of present invention.

Fig. 3 illustrates the comparison data of antioxidant activity of present formulation, Rosehip oil and Tocopherol acetate as disclosed in one of the embodiments of present invention.

Persons skilled in the art will appreciate that elements in the figures are illustrated for simplicity and clarity and may not have been drawn to scale.

DETAILED DESCRIPTION OF THE INVENTION

The following description with reference to the accompanying drawings is provided to assist in a comprehensive understanding of various embodiments of the present disclosure as defined by the claims and their equivalents. It includes various specific details to assist in that understanding, but these are to be regarded as merely exemplary. Accordingly, those of ordinary skill in the art will recognize that various changes and modifications of the various embodiments described herein can be made without departing from the scope and spirit of the present disclosure. In addition, descriptions of well-known functions and constructions may be omitted for clarity and conciseness.

The terms and words used in the following description and claims are not limited to the bibliographical meanings, but, are merely used by the inventor to enable a clear and consistent understanding of the present disclosure. Accordingly, it should be apparent to those skilled in the art that the following description of various embodiments of the present disclosure is provided for illustration purpose only and not for the purpose of limiting the present disclosure as defined by the appended claims and their equivalents.

All terms (including technical and scientific terms) used herein have the same meaning as commonly understood by one of ordinary skill in the art to which various embodiments belong. Further, the meaning of terms or words used in the specification and the claims should not be limited to the literal or commonly employed sense but should be construed in accordance with the spirit of the disclosure to most properly describe the present disclosure.

The terminology used herein is for the purpose of describing particular various embodiments only and is not intended to be limiting of various embodiments. As used herein, the singular forms "a," "an" and "the" are intended to include the plural forms as well, unless the context clearly indicates otherwise. It will be further understood that the terms "comprises" and/or "comprising" used herein specify the presence of stated features, integers, steps, operations, members, components, and/or groups thereof, but do not preclude the presence or addition of one or more other features, integers, steps, operations, members, components, and/or groups thereof.

As used herein, the term "antioxidant" is recognized in the art and refer to synthetic or natural substances that prevent or delay the oxidative of free radical or photo induced deterioration of a compound. Exemplary stabilizers include tocopherols, tocotrienols flavonoids, catechins, superoxide dismutase, lecithin, gamma oryzanol; vitamins, such as vitamins A, C (ascorbic acid) and E (tocotrienol and tocopherol and tocopherol homologues and isomers, especially alpha and gamma-tocopherol) and beta-carotene; natural components such as camosol, carnosic acid and rosmanol found in rosemary and hawthorn extract, proanthocyanidins such as those found in grapeseed or pine bark extract, and green tea extract

The present invention relates to the incorporation of Vitamin E in Milk by developing a unique formulation of Vitamin E composite of Tocotrienol Rich Fraction that can be easily incorporated in Milk, wherein the composition comprises of Tocotrienol Rich Fraction, solubilizers, and plant-origin fatty acid carriers. The formulation of the present invention solubilizes the fat content of the milk, and enhances the sweetness, taste, texture, mouthfeel and shelf-life of the milk.

The present invention specifically relates to a milk-soluble vitamin E composite formulation comprising;
a. Tocotrienol Rich Fraction (TRF); wherein the amount of the Tocotrienol Rich Fraction (TRF) is about 4% to about 14%, by weight of the formulation;
b. a fatty acid carrier selected from the fatty acids having carbon numbers C6- C12, wherein the amount of fatty acid carrier is about 60% to about 85%, by weight of the formulation; and
c. a solubilizer, wherein the amount of solubilizer is about 4% to about 28%, by weight of the formulation;
wherein the ratio of solubilizer and Tocotrienol Rich Fraction (TRF) is 2:1.

In another preferred embodiment, the solubilizers are selected from one or combination of the polysaccharides, polyol, glycerophospholipids.

In a most preferred embodiment, the solubilizer was selected from the group of polysaccharides including, but not limited to, xanthan gum or starch or maltodextrin, polyol including, but not limited to, glycerine or glycerol or sorbitol, glycerophospholipids including, but not limited to, lecithin and/or its hydrolysed form, sodium stearoyl lactylate, and the combinations thereof.

In a further embodiment, said fatty acid carriers include, but not limited to, plant-origin fatty acid.

Another embodiment of present invention relates to a process of preparation of a milk-soluble vitamin E composite formulation, comprising:
Phase 1:
1. heating the blending tank to 40 degree centigrade;
2. initiating Nitrogen flow in the blending Tank;
3. adding a solubilizer in the tank after 60 seconds, once nitrogen is thoroughly flushed in the tank;
4. adding 20% of total required Tocotrienol Rich Fraction (TRF) volume in the tank;
5. stirring the solution of step (4) at 80 rpm constantly and continuously;
6. adding another 60% of said TRF in the tank after 300 seconds with constant stirring;
7. blending for another 300sec;
8. adding the remaining 20% TRF and blending for 300sec;
9. removing the blended concoction of step (8)
10. switching off the nitrogen and the heater.

Phase 2:
1. charging a fatty acid carrier in a separate vessel,
2. stirring said fatty acid carrier at a speed of 50 rpm;
3. adding the resulting solution of step (12) in Phase 1 slowly and gradually;
4. allowing homogenous blending of phase 1 and phase 2 for about 300 sec;
5. removing the blended concoction of step (14).

According to yet another embodiment of present invention, there is provided a homogeneous milk formulation comprising 0.02-0.20% of said milk-soluble vitamin E composite formulation, with enhanced sweetness, taste, texture, mouth-feel and shelf-life.

In a preferred embodiment, said milk formulation may be in liquid form or in powder form.

In a most preferred embodiment, said liquid milk may include raw milk as well as pasteurized milk.

According to further embodiment of present invention, there is provided a process of preparation of said homogeneous milk formulation comprising said milk-soluble vitamin E composite formulation, comprising
mixing 0.02-0.20% of said vitamin E composite formulation with milk using a homogenizer/ stirrer at 20 rpm for 10-30 minutes.

STUDY AND OBSERVATIONS

Example 1 – Solubility of Milk-Soluble Vitamin E Composite Formulation in Liquid Raw Milk and Pasteurized Milk

The study was conducted to determine the ideal ratio of solubilizer to TRF for a homogenous incorporation of Vitamin E Composite in Milk.

Observations 1.A: Solubility and Stability of Raw Milk Incorporated with Vitamin E Composite Formulation

Table 1.A-A Solubilizer A and TRF Ratio (S: TRF)
1:1 2:1 3:1
Parameters RT 70°C 20°C RT 70°C 20°C RT 70°C 20°C
Colour White White White White White White White White White
Odour None None None None None None None None None
Granulation Yes Yes Yes Yes slight Yes slight slight slight
Film Formation Yes No Yes Yes No Yes Yes Yes Yes
Milk separation Yes Yes Yes Yes Yes Yes Yes Yes Yes

Legend: RT = Room Temperature, 25°C
Solubilizer A: Polyols like Glycerol

Table 1.A-B Solubilizer B and TRF Ratio (S: TRF)
1:1 2:1 3:1
Parameters RT 70°C 20°C RT 70°C 20°C RT 70°C 20°C
Colour White White White Yellowish Yellowish Yellowish Yellowish Yellowish Yellowish
Odour None None None None None None None None None
Granulation Yes slight Yes Yes slight Yes slight slight slight
Film Formation Yes No Yes Yes No Yes Yes No Yes
Milk separation Yes Yes Yes Yes No Yes Yes No Yes

Legend: RT = Room Temperature, 25°C
Solubilizer B: Maltodextrin

Table 1.A-C Solubilizer C and TRF Ratio (S: TRF)
1:1 2:1 3:1
Parameters RT 70°C 20°C RT 70°C 20°C RT 70°C 20°C
Colour White White White White White White White White White
Odour None None None None None None None None None
Granulation Yes Slight Yes No No No slight slight slight
Film Formation Yes No Yes No No No Yes No Yes
Milk separation No No No No No No No No No

Legend: RT = Room Temperature, 25°C
Solubilizer C: Glycerophospholipids

Observations 1.B: Solubility and Stability of Pasteurized Milk Incorporated with Vitamin E Composite Formulation

Table 1.B-A Solubilizer A and TRF Ratio (S: TRF)
1:1 2:1 3:1
Parameters RT 70°C 20°C RT 70°C 20°C RT 70°C 20°C
Colour White White White White White White White White White
Odour None None None None None None None None None
Granulation Yes Yes Yes Yes slight Yes slight slight slight
Film Formation Yes No Yes Yes No Yes Yes Yes Yes
Milk separation Yes Yes Yes Yes Yes Yes Yes Yes Yes

RT = Room Temperature, 25°C
Solubilizer A: Polyols like Glycerol

Table 1.B-B Solubilizer B and TRF Ratio (S: TRF)
1:1 2:1 3:1
Parameters RT 70°C 20°C RT 70°C 20°C RT 70°C 20°C
Colour White White White Yellowish Yellowish Yellowish Yellowish Yellowish Yellowish
Odour None None None None None None None None None
Granulation slight slight Yes slight slight Yes slight slight slight
Film Formation Yes No Yes Yes No Yes Yes No Yes
Milk separation Yes Yes Yes Yes No Yes Yes No Yes

RT = Room Temperature, 25°C
Solubilizer B: Maltodextrin

RT = Room Temperature, 25°C
Solubilizer C: Glycerophospholipids

Table 1.B-C Solubilizer C and TRF Ratio (S: TRF)
1:1 2:1 3:1
Parameters RT 70°C 20°C RT 70°C 20°C RT 70°C 20°C
Colour White White White White White White White White White
Odour None None None None None None None None None
Granulation slight slight slight No No No No No No
Film Formation No No No No No No Yes No Yes
Milk separation No No No No No No No No No

Inference:
The tables in Observations 1.A-A, 1.A-B and 1.A-C showed that formulation with Solubilizer C is the most stable of all formulations. Similarly, the tables in Observations 1.B-A, 1.B-B and 1.B-C showed that formulation with Solubilizer C is the most stable of all formulations. There were no Film Formation and Milk Separation on both formulation A-C and B-C when added in Raw and Pasteurized Milk.

It was also observed that the most stable ratio of formulation A-C and B-C is 2:1.

Based on the observations and gathered data, it was concluded that Solubilizer C to TRF Ratio in the Vitamin E Composite Formulation should be in 2:1 ratio. It was observed that the formulation ratio of 3:1 and 1:1 is not stable in milk and hence is not applicable to the intended focus of the present invention of incorporating Vitamin E Composite of Tocotrienol Rich Fraction in Milk.

Example 2 – Solubility of Milk-Soluble Vitamin E Composite Formulation in Powdered Milk

The study was conducted to determine the ideal ratio of solubilizer to TRF for a homogenous incorporation of Vitamin E Composite in Powdered Milk. The powdered milk was diluted with water and was observed for any instability.

Observations 2.A: Stability of Powdered Milk Incorporated with Vitamin E Composite Formulation
Table 2.A Solubilizer and TRF Ratio (S:TRF)
1:1 2:1 3:1
Parameters RT 70°C 25°C RT 70°C 25°C RT 70°C 25°C
Colour White White White White White White White White White
Odour None None None None None None None None None
Granulation None None None None None None Slight None None
Film Formation No No No No No No Yes No No
Milk separation No No No No No No No No No

Legend: RT = Room Temperature

Inference:
The table in Observations 2.A showed instability of the 3:1 Vitamin E Composite Formulation in Raw Milk. There were film formations and granulation observed in the 3:1 formulation.

Based on the observations and gathered data, it was concluded that Solubilizer to TRF Ratio in the Vitamin E Composite Formulation should be kept below the 3:1 ratio. It was observed that the formulation ratio of 3:1 is not stable in milk and hence is not applicable to the intended focus of the present invention of incorporating Vitamin E Composite of Tocotrienol Rich Fraction in Milk.

Hence, the chosen Vitamin E Composite Formulation is with Solubilizer to TRF ratio of 2:1.

Example 3 – Effects of Adding Milk-Soluble Vitamin E Composite Formulation in Liquid Raw Milk and Pasteurized Milk

The Milk-Soluble Vitamin E Composite Formulation was further tested of its impact in Raw Milk and Pasteurized Milk. The formulation was incorporated in Milk at room temperature, and the resulting solution was then heated and cooled. This process was done for four consecutive days.

Observations 3.A: Milk-Soluble Vitamin E Composite Formulation in Raw Milk

Table 3.A 1st Day 3rd Day 4th Day
Parameters at RT After Heating After cooling
1 Colour Off White Off White White Off White Off White
2 Odour Milky Creamy Milky Creamy Milky Creamy Milky Creamy Milky Creamy
3 Granulation NO NO NO NO NO
4 Film Formation NO NO O NO O
5 Milk separation NO NO NO NO NO
6 Flavour Sweet
Mouthful Sweet
Mouthful Sweet
Mouthful Sweet
Mouthful Sweet
Mouthful

Legend: RT= Room Temperature; NO = Not Observed; O = Observed

Observations 3.B: Milk-Soluble Vitamin E Composite Formulation in Pasteurized Milk

Table 3.B 1st Day 3rd Day 4th Day
Parameters at RT After Heating After cooling
1 Colour White White Bright White White White
2 Odour Milky Creamy Milky Creamy Milky Creamy Milky Creamy Milky Creamy
3 Granulation NO NO NO NO NO
4 Film Formation O NO O NO O
5 Milk separation NO NO NO NO NO
6 Flavour Sweet
Mouthful Sweet
Mouthful Sweet
Mouthful Sweet
Mouthful Sweet
Mouthful

Legend: RT= Room Temperature; NO = Not Observed; O = Observed

Inference:
Based on the observations and gathered data, it was concluded that the Vitamin E Composite Formulation was stable in both Raw Milk and Pasteurized Milk. There was no milk separation in the solutions.

Example 4 – Sensorial Evaluation Study of Milk-Soluble Vitamin E Composite Formulation in Milk

The sensorial effects of adding Milk-Soluble Vitamin E Composite Formulation in Milk were analysed using the magnitude-estimation method. A group of 15 professional taste tester participated in the study. The result was shown on a scale of 0 to 5 where 5 is the best and 0 is the worst.

The Magnitude Estimation Method is psychophysical method of evaluating stimuli above threshold. The subject assigns numbers according to the apparent magnitudes of the stimuli. The results relating the magnitude of sensation S and the stimulus intensity, I, usually follows the Stevens' Power Law, that is S = kIn, where k is a constant and n is the exponent which depends on the sensory modality.

Observations 4.A: Comparison of Raw Milk without Milk-Soluble Vitamin E Composite and Raw Milk with Milk-Soluble Vitamin E Composite

Table 4.A-1 : Raw Milk without Milk-Soluble Vitamin E Composite
Subjects Parameters
Taste Texture Colour Mouthfeel Sweetness
Group 1 2 3 Off White 3 3
Group 2 3 4 Off White 3 3
Group 3 4 3 Off White 3 4
Group 4 3 3 Off White 2 3
Group 5 2 3 Off White 3 3
Mean Ave. 2.8 3.2 --- 2.8 3.2

Table 4.A-2 : Raw Milk with Milk-Soluble Vitamin E Composite
Subjects Parameters
Taste Texture Colour Mouthfeel Sweetness
Group 1 4 5 Off White 5 4
Group 2 3 4 Off White 3 3
Group 3 3 4 Off White 4 4
Group 4 3 4 White 4 4
Group 5 3 3 Off White 4 4
Mean Ave. 3.2 4 --- 4 3.8

Observations 4.B: Comparison of Pasteurized Milk without Milk-Soluble Vitamin E Composite and Pasteurized Milk with Milk-Soluble Vitamin E Composite

Table 4.B-1 : Raw Milk without Milk-Soluble Vitamin E Composite
Subjects Parameters
Taste Texture Colour Mouthfeel Sweetness
Group 1 3 3 White 3 3
Group 2 3 4 Off White 3 3
Group 3 3 3 White 4 3
Group 4 3 4 White 3 4
Group 5 3 3 White 3 3
Mean Ave. 3 3.4 --- 3.2 3.2

Table 4.B-2 : Raw Milk with Milk-Soluble Vitamin E Composite
Subjects Parameters
Taste Texture Colour Mouthfeel Sweetness
Group 1 4 4 White 4 5
Group 2 4 5 White 4 4
Group 3 3 4 White 4 5
Group 4 4 4 White 5 4
Group 5 4 5 White 5 4
Mean Ave. 3.8 4.4 --- 4.4 4.4

Inference:
Based on the comparison evaluation, observations and gathered data, it was concluded that the Vitamin E Composite Formulation enhanced the properties, the taste, texture, mouthfeel, and sweetness of both Raw Milk and Pasteurized Milk.

Example 5 – Shelf-life Analysis of Milk Incorporated with Milk-Soluble Vitamin E Composite Formulation

The milk with Vitamin E Composite was subjected to shelf-life analysis using a thaw-freeze method. The milk sample incorporated with Vitamin E Composite was kept on a room temperature, then was kept inside the refrigerator, and back to the room temperature in a six-hour interval for 96-hours. The observations were recorded accordingly.

Observations 5.A: Shelf-Life Analysis of Raw Milk without Milk-Soluble Vitamin E Composite and Raw Milk with Milk-Soluble Vitamin E Composite

Table 5.A-1: Raw Milk without Milk-Soluble Vitamin E Composite
Parameters PERIOD, HOURS
6 hrs 12 hrs 24 hrs 36 hrs 74 hrs 96 hrs
Colour OW OW OW OW OW OW
Texture S S S S --- ---
pH 6.96 6.88 6.48 4.57 --- ---
Viscosity 500 550 1100 <2000 --- ---
Flow F F F F F NF
Phase separation NO NO NO NO O O

Table 5.A-2: Raw Milk with Milk-Soluble Vitamin E Composite
Parameters PERIOD, HOURS
6 hrs 12 hrs 24 hrs 36 hrs 74 hrs 96 hrs
Colour OW OW OW OW OW OW
Texture S S S S S S
pH 6.4 6.42 6.48 6.48 6.5 6.5
Viscosity 500 550 500 580 500 500
Flow F F F F F F
Phase separation NO NO NO NO NO NO

Legends: OW: Off-white; S: Smooth; F: Flowable; NF: Not Flowable; NO: Not Observed, O: Observed.

Observations 5.B: Shelf-Life Analysis of Pasteurized Milk without Milk-Soluble Vitamin E Composite and Raw Milk with Milk-Soluble Vitamin E Composite

Table 5.B-1: Pasteurized Milk without Milk-Soluble Vitamin E Composite
Parameters PERIOD, HOURS
6 hrs 12 hrs 24 hrs 36 hrs 74 hrs 96 hrs
Colour W W W W W W
Texture S S S S --- ---
pH 6.96 6.88 6.48 6.2 4.76 ---
Viscosity 500 550 1100 1800 <2000 ---
Flow F F F F F NF
Phase separation NO NO NO NO O O

Table 5.B-2: Pasteurized Milk with Milk-Soluble Vitamin E Composite
Parameters PERIOD, HOURS
6 hrs 12 hrs 24 hrs 36 hrs 74 hrs 96 hrs
Colour W W W W W W
Texture S S S S S S
pH 6.60 6.65 6.60 6.62 6.63 6.60
Viscosity 500 500 550 500 550 550
Flow F F F F F F
Phase separation NO NO NO NO NO NO

Legends: OW: Off-white; S: Smooth; F: Flowable; NF: Not Flowable; NO: Not Observed, O: Observed.

Inference:

A sudden pH dropped in milk, or the sudden viscosity increase of milk are signs of its expiration. In this study, the pH and viscosity were observed as they were the critical indicator of the milk's life.

Based on the comparison evaluation, observations and gathered data, it was concluded that the Milk incorporated with Vitamin E Composite Formulation has better pH and viscosity stability. The data also shows that the Milk incorporated with Vitamin E Composite has extended shelf-life up to 96 hours as compared to the Milk without Vitamin E Composite.

Example 6 – Antioxidant Analysis of Milk-Soluble Vitamin E Composite Formulation
The study was conducted to analyse the antioxidant activity of the Vitamin E Composite Formulation. The analysis was done using DPPH Assay Method.

Table 6: Antioxidant Activity Comparison
Concentrations in Percentage Vitamin E Composite Formulation Rosehip Oil Tocopherol Acetate
0.01 32.15 7 1.5
0.025 80.37 17.49 3.7
0.05 >100 34.98 7.5
0.08 55.97 12
0.1 69.97 15
0.15 >100 22.5
0.2 29.6
0.25 37.5
0.3 45
0.37 51.8
0.5 74.9
0.65 97.4

Inference:
Based on the gathered data, it can be concluded that the Vitamin E Composite Formulation has a very strong antioxidant activity. At 0.05% concentration, the Vitamin E Composite Formulation was able to achieve 100% antioxidant activity. While, at the same concentration, Rosehip Oil has 34.98% antioxidant activity and Tocopherol Acetate has 7.5%.
, Claims:
1. A process of incorporating Tocotrienol Rich Fraction into milk. Wherein, the Tocotrienol Rich Fraction and Milk mixture is homogenous.

2. A milk-soluble vitamin E composite formulation comprising;
a. Tocotrienol Rich Fraction (TRF); wherein the amount of the Tocotrienol Rich Fraction (TRF) is about 4% to about 14%, by weight of the formulation;
b. a fatty acid carrier selected from the fatty acids having carbon numbers C6- C12, wherein the amount of fatty acid carrier is about 60% to about 85%, by weight of the formulation; and
c. a solubilizer, wherein the amount of solubilizer is about 4% to about 28%, by weight of the formulation;
wherein the ratio of solubilizer and Tocotrienol Rich Fraction (TRF) is 2:1.

3. The formulation as claimed in claim 2, wherein the solubilizer is a glycerophospholipids.

4. The formulation as claimed in claim 2, wherein said fatty acid carriers include, but not limited to, plant-origin fatty acid.

5. A process of preparation of a milk-soluble vitamin E composite formulation, comprising:
a. preparing Phase 1 comprising;
1. heating a blending tank to 40 degree centigrade;
2. initiating Nitrogen flow in the blending Tank;
3. adding a solubilizer in the tank after 60 seconds, once nitrogen is thoroughly flushed in the tank;
4. adding 20% of total required Tocotrienol Rich Fraction (TRF) volume in the tank;
5. stirring the solution of step (4) at 80 rpm constantly and continuously;
6. adding another 60% of said TRF in the tank after 300 seconds with constant stirring;
7. blending for another 300sec;
8. adding the remaining 20% TRF and blending for 300sec;
9. removing the blended concoction of step (8); and
10. switching off the nitrogen and the heater;
b. preparing Phase 2 comprising;
11. charging a fatty acid carrier in a separate vessel,
12. stirring said fatty acid carrier at a speed of 50 rpm;
13. adding the resulting solution of step (12) in Phase 1 slowly and gradually;
14. allowing homogenous blending of phase 1 and phase 2 for about 300 sec;
15. removing the blended concoction of step (14).

6. A homogeneous milk formulation comprising 0.02-0.20% of said milk-soluble vitamin E composite formulation as claimed in claims 1-4, with enhanced sweetness, taste, texture, mouth-feel and shelf-life.

7. The milk formulation as claimed in claim 6, wherein said milk formulation is in liquid form or in powder form.

8. The milk formulation as claimed in claim 9, wherein said milk is selected from raw milk or pasteurized milk or powder milk.

9. A process of preparation of said homogeneous milk formulation comprising a milk-soluble vitamin E composite formulation as claimed in claims 1-4, comprising mixing 0.02-0.20% of said vitamin E composite formulation with milk using a homogenizer/ stirrer at 20 rpm for 10-30 minutes.