Description:FIELD OF THE INVENTION
The present invention relates to 25 Hydroxy Vitamin D3 1.25% spray dried Feed composition for animals and to the process for preparation thereof.

BACKGROUND OF THE INVENTION
Vitamin D (D3 and D2), which is mostly produced in the skin when exposed to the sun, is a biologically inactive substance that is transformed into an active form in the body after passing through several stages. First, in the liver, vitamin D is turned into 25-hidroxyvitamin D (25(OH)D) – calcifediol. In the next stage, in the kidneys, calcifediol is transformed into the active form of the vitamin – calcitriol.
The activated vitamin acts through the vitamin D receptors that are located on the cell nuclei membranes of various organs. The decrease in calcitriol is the key indicator showing that vitamin D is not being assimilated properly (activated in the liver and kidneys) regardless of its adequate intake.

Vitamin 1.25-(OH)2-D3 is called calcitriol, biologically the most active form of vitamin D, regulates the assimilation of calcium and phosphorus in the small intestine, bone mineralization, and the synthesis and secretion of the parathyroid hormone (PTH). It is important for the formation and renewal of bones.

US7632518 discloses calcifediol 1.25% spray dried feed grade composition comprising encapsulated calcifediol in oil with an encapsulating agent and an antioxidant.

ROVIMIX® available in the market contains 25-Hydroxy Vitamin D3 1.25% along with an antioxidant, vegetable oil, modified food starch, sodium ascorbate, maltodextrin and silicon dioxide. The composition is described in the PCT publication no. WO2023139249. The said WO’249 discloses a feed premix composition for intake by aquatic animals.
25-hydroxy Vitamin D3 is unstable under conventional storage conditions that can adversely affect intake of 25-hydroxy Vitamin D3 when incorporated in to animal feed.

The present inventors therefore felt a need to provide 25-Hydroxy Vitamin D3 1.25% composition as animal feed which would not adversely affect intake of 25-hydroxy Vitamin D3 when incorporated in to animal feed and would be effective in reducing the bone disorders and help in weight gain. This remains the objective of the invention.

SUMMARY OF THE INVENTION
In accordance to the above, the present invention provides a stable 25 Hydroxy Vitamin D3 1.25% spray dried feed composition comprising;
i. Maltodextrin as encapsulating agent in an amount of 10-20%w/w;
ii. Modified corn starch as encapsulating agent in an amount of 65-75%w/w;
iii. Sodium Ascorbate/Ascorbic acid as preservative in an amount of 1-3%w/w;
iv. Calcifediol as active in an amount of 1-2%;
v. Antioxidant in an amount of 1-6%;
vi. Vegetable oil selected from Sunflower oil or Corn Oil in an amount of 2-8%w/w;
vii. and
viii. Silicon dioxide in an amount of 0.5-5%w/w.

In an aspect, the feed composition of the present invention provides an emulsion comprising an aqueous composition and an oil composition, wherein the oil composition contains 25 Hydroxy Vitamin D3 dissolved in Sunflower oil or Corn Oil in an amount of 2-8% by weight based on the total weight of the oil composition and antioxidant.

In another aspect, the aqueous composition of the present invention comprises the encapsulating agents like modified corn starch in an amount of 65-75%, maltodextrin in an amount of 10-20% along with preservative sodium ascorbate or ascorbic acid dissolved in water.

In yet another aspect, the present invention provides a premix feed composition comprising an encapsulated oil composition of the present invention admixed with the nutritional additives selected from vitamins, carbohydrates, proteins, lipids (for example fish meal, fish oil, feather meal or other types of meal produced from other slaughterhouse waste); amino acids, antimicrobial peptides; polyunsaturated fatty acids; and/or at least one enzyme, probiotics, prebiotics, mineral premix, medicinal additives, starches alone or mixture thereof.

In yet another aspect, the present invention provides a method of reducing bone disorders and increasing the body weight in animals comprising feeding the animals with feed composition or the premix feed of the present invention.

DESCRIPTION OF THE FIGURES
Fig 1: Depicts the Manufacturing Process flow chart of Calcifediol 1.25% spray dried.

DETAILED DESCRIPTION OF THE INVENTION
The invention will now be described in detail in its various preferred and optional embodiments so that the various aspects will be clearly understood however without limiting the scope of the invention.

Definitions:
The term “Animal” refers to the pet animals, poultry, cattle, small and large farm animals and swine.

The term “feed” refers to any compound, preparation, or mixture suitable for, or intended for intake by animals.

25 Hydroxy Vitamin D3 1.25% or calcitriol is the active form of vitamin D, normally made in the kidney.

The term “Premix Feed” relates to the composition comprising of 25 Hydroxy Vitamin D3 1.25% spray dried animal feed mixed with the premix consisting of the nutritive additives as described herein in the entire specification.

In an embodiment, the present invention relates to a stable 25 Hydroxy Vitamin D3 1.25% spray dried animal feed comprising;
i. Maltodextrin in an amount of 10-20%w/w;
ii. Modified corn starch in an amount of 65-75%w/w;
iii. Sodium Ascorbate/Ascorbic acid in an amount of 1-3%w/w;
iv. Calcifediol in an amount of 1-2%;
v. Antioxidant in an amount of 1-6%;
vi. Sunflower oil or Corn Oil in an amount of 2-8%w/w; and
vii. Silicon dioxide in an amount of 0.5-5%w/w.

The active ingredient “25 Hydroxy Vitamin D3 1.25%” of the present composition is embedded in an oil carrier selected from vegetable oil such as sunflower oil or corn oil.

In an embodiment, the active 25 Hydroxy Vitamin D3 1.25% is dissolved in the vegetable oil in an amount of 2-8% by weight based on the total weight of the composition and the amount of the vegetable oil is less than 70% by weight based on the total weight of the oil composition.

The microencapsulated oil composition may optionally contain any other suitable material including, but not limited to preservatives, dissolution agents, surfactants, pH adjusting agents or buffers, humectants, and mixtures thereof in suitable amounts.

Once formed, the oil composition of the invention may be incorporated in various other useful compositions, for example, emulsions may be formed, which may be converted into various encapsulated oils having superior stability properties.

A variety of emulsions may be prepared by combining the oil compositions described above with an aqueous composition. The emulsion may be of any type. Suitable emulsions include oil-in-water emulsions, water-in-oil emulsions, anhydrous emulsions, solid emulsions, and micro emulsions. The emulsions may be prepared by any convenient method. Preferably, the emulsion is oil in water.

The microencapsulated 25 Hydroxy Vitamin D3 1.25% in the oil of the present composition contains the antioxidant selected from the group consisting of butylated hydroxy toluene (BHT), butylated hydroxy anisole, natural antioxidants and the like; preferably the antioxidant is BHT.

In an embodiment, the encapsulating agent is selected from the group consisting of gelatin, a modified food starch such as modified corn starch, plant protein, a maltodextrin, a cellulose derivative, a saccharide, a gum, and mixtures thereof; preferably modified corn starch and maltodextrin.

The encapsulated oil composition further comprises preservatives selected from sodium ascorbate/ascorbic acid and silicon dioxide to improve the flowability.

In an embodiment, the feed composition of the present invention may be added as a feed to the “premix” that may contain nutritive ingredients selected from vitamins, carbohydrates, proteins, lipids (for example fish meal, fish oil, feather meal or other types of meal produced from other slaughterhouse waste); amino acids, antimicrobial peptides; polyunsaturated fatty acids; and/or at least one enzyme, probiotics, prebiotics, mineral premix, medicinal additives, starches alone or mixture thereof.

In an embodiment, the feed composition of the present invention comprises calcifediol in the range of 1250ppm (1.25%), preferably in the range of 1250ppm to 1500ppm (i.e.1.25% to 1.5%). The final concentration of 25-hydroxyvitamin D, preferably 25-hydroxy vitamin D3 (25-OH D3), in the feed is determined by HPLC according to standard methods.

In another embodiment, the present invention discloses a process for preparation of said feed composition comprising;
i. Preparing an emulsion of Calcifediol i.e. 25 hydroxy Vitamin D3, antioxidant selected from BHT (Butylated Hydroxy toluene) in the oil selected from Sunflower Oil or Corn oil;
ii. Preparing a water phase of Sucrose, Maltodextrin, Modified starch, Sodium Ascorbate or ascorbic acid, in purified water;
iii. Homogenizing the emulsion phase of step (i) together with water phase of step (ii) forming a uniform emulsion and optionally adding purified water to maintain the viscosity of the emulsion in the range of 30 centipose to 120 centipose;
iv. Spray drying the homogenized emulsion of step (iii) to obtain spray dried encapsulated product;
v. Blending the spray dried product of step (iv) with silicon dioxide, sifting and packing under inert atmosphere.

In an embodiment, the spray dried product may be in the form of powder, pellets, granules, flakes and may optionally be formulated as liquids using the acceptable feed excipients. Preferably, the feed composition of the present invention is in form of powder.

In an embodiment, the water content of the spray dried feed composition of the present invention is less than 5%.

In an embodiment, the active ingredient Calcifediol i.e. 25 hydroxy Vitamin D3, of the present feed is embedded in an oil carrier which is a coating oil i.e. the vegetable oil selected from sunflower or corn oil; preferably sunflower oil.

In another embodiment, the active ingredient Calcifediol i.e. 25 hydroxy Vitamin D3, of the present feed is used for increasing the levels of vitamin D3 such that the animal is vitamin D3 replete.

In an embodiment, the feed composition of the present invention improve the feed conversion ratio (FCR), Immunity, feed efficiency (FE) protein efficiency ratio (PER) and/or the growth performance (nutrient utilization, body protein content, protein efficiency ratio (PER) and/or the growth performance (specific growth rate SGR, Final body weight and/or carcass yields) in animals.

The present feed composition provides vitamin D3 to the animals in an amount of 12.5 to 15g/ kg of the feed, preferably in an amount not less than 12.5g/kg of the feed.

In an embodiment, the present invention discloses a method of reducing bone disorders and increasing the body weight in animals comprising feeding the animals with feed composition of the present invention.

In an embodiment, the feed composition of the present invention thus increased 25-hydroxy Vitamin D3 retention over a wide range of temperatures and times of storage, even in premixes containing components which promote degradation of 25-hydroxy Vitamin D3. The active 25 hydroxy Vitamin D3 is in powder form which is dissolved in Oil (Sunflower or corn oil) and added to water phase to form emulsion which is spray dried by evaporating water at high temperatures. Modified Starch encapsulates D3 and makes it stable.

The feed composition of the present invention is used to reduce the bone disorders and increasing the body weight of the animals thus providing complete nutrition to the animals. The feed composition of the present invention provides a supra-physiological amount of the components such that the animal is not vitamin or nutrient deficient.

In another embodiment, the present invention relates to the use of the feed composition as a feed in the premix. The premix comprises the nutritive additives selected from vitamins, carbohydrates, proteins, lipids (for example fish meal, fish oil, feather meal or other types of meal produced from other slaughterhouse waste); amino acids, antimicrobial peptides; polyunsaturated fatty acids; and/or at least one enzyme, probiotics, prebiotics, mineral premix, medicinal additives, starches alone or mixture thereof.

In yet another embodiment, the present invention discloses a feed composition comprising;
A. An encapsulated oil composition which include ;
i. Maltodextrin as encapsulating agent in an amount of 10-20%w/w;
ii. Modified corn starch as encapsulating agent in an amount of 65-75%w/w;
iii. Sodium Ascorbate/Ascorbic acid as preservative in an amount of 1-3%w/w;
iv. 25 Hydroxy Vitamin D3 (Calcifediol) as active in an amount of 1-2%;
v. Antioxidant in an amount of 1-6%;
vi. Vegetable oil selected from Sunflower oil or Corn Oil in an amount of 2-8%w/w; and
vii. Silicon dioxide in an amount of 0.5-5%w/w; and
B. One or more nutritive additive.

The nutritive additive selected from vitamins, carbohydrates, proteins, lipids (for example fish meal, fish oil, feather meal or other types of meal produced from other slaughterhouse waste); amino acids, antimicrobial peptides; polyunsaturated fatty acids; and/or at least one enzyme, probiotics, prebiotics, mineral premix, medicinal additives, starches alone or mixture thereof.

In another embodiment, the present invention discloses the use of premix feed composition to reduce bone disorders and increase the body weight in animals.

In yet another embodiment, the present invention discloses a method of reducing the bone disorders and increasing the body weight in animals comprising feeding the animal with the feed composition of the present invention.

Examples:
Example 1: Feed Composition
Ingredients Amount (%w/w)
Maltodextrin 10-20%
Modified corn starch 65-75%
Sodium Ascorbate/Ascorbic acid 1-3%
Calcifediol 1-2%
Butylated Hydroxy Toluene 1-6%
Sunflower oil/ Corn Oil 2-8%
Silicon dioxide 0.5-5%

Process:
Step 1: Emulsion
To a stainless steel or a glass container was added Calcifediol i.e. 25 hydroxy Vitamin D3, oil (Sunflower Oil or Corn oil) and Butylated Hydroxy toluene and the mixture was stirred for 30 to 150minutes at a temperature in the range of 60-90°C until a uniform emulsion is formed.

Step 2: Water Phase
To another stainless steel container or a glass container was added Sucrose, maltodextrin, (Modified starch), sodium ascorbate or ascorbic acid, in purified water and the mixture was stirred for 30 to 90°C for 30-120 minutes to obtain the water phase.

Step 3: Homogenization (Emulsion Formation)
The emulsion phase of step 1 and the water phase of step 2 were mixed and homogenized together with a high-speed homogenizer at a rate of 4000 to 10000RPM for a period of 15 to 90 minutes to obtain a uniform emulsion. The viscosity was intermittently checked which should be around 30 centipose to 120 centipose. Optionally purified water was added to maintain the viscosity of 30 centipose to 120 centipose.

Step 4: Spray Drying
The homogenized emulsion of step 3 was spray dried through a nozzle equivalent to 1.0 /1.2/1.5/2.0/2.5 mm size using a peristaltic pump with an inlet temperature of 130°C to 230°C and an outlet temperature of 70°C to 120°C. The spray dried encapsulated product was collected in product receivers of spray drier. For flowability silicon dioxide was blended with the product, sifted through a 30 mesh or 40 mesh or 80 mesh and packed under nitrogen/vacuum blanketing to avoid degradation.
Inlet Temperature: 130 to 230 degrees
Outlet Temperature: 70 to 120 degrees
Pump Speed: 20 – 80 L/hour
Product Moisture: <5% LOD
The microencapsulated granular powder has the following characteristics:
Loose Bulk Density: ~ 0.3 g/L to 0.5 g/L
Tapped Bulk Density: ~ 0.3 g/L to 0.7 g/L
Loss on Drying : <5% LOD
The microencapsulated composition showed a high degree of homogeneity as measured by HPLC.

Example 2A: Stability data
Calcifediol 1.25% SD
Name of Product Calcifediol 1.25% SD Batch No. Lab#003
Mfg. Date March,2024 Stability
Initiated on March,2024
Stability Condition 25± 2°C and 60 ± 5 % RH Sample Kept
For 12M
Type of Study Long Term Reference Lab trial
TEST LIMITS Time Intervals (Months)
Initial 1M 3M 6M 12M
Assay – % NLT 1.25% 1.44 1.47 1.36
Appearance White to off white
powder White powder White powder White powder

Example 2B:
Calcifediol 1.25% SD
Name of Product Calcifediol 1.25% SD Batch No. Lab#003
Mfg. Date March,2024 Stability
Initiated on March,2024
Stability Condition 40± 2°C and 75 ± 5 % RH Sample Kept
For 6M
Type of Study Accelerated Reference VDSD0124002
Test Limits Time Intervals (Months)
1M 3M 6M
Assay – % NLT 1.25% ND 1.41
Appearance White to off white
powder White to off white
powder White to off white
powder

It will be understood that the above description is intended to be illustrative and not restrictive. The embodiments will be apparent to those in the art upon reviewing the above description. The scope of the invention should therefore, be determined not with reference to the above description but should instead be determined by the appended claims along with full scope of equivalents to which such claims are entitled.

, Claims:
1. A stable 25 Hydroxy Vitamin D3 1.25% spray dried animal feed comprising;
i. Maltodextrin as encapsulating agent in an amount of 10-20%w/w;
ii. Modified corn starch as encapsulating agent in an amount of 65-75%w/w;
iii. Sodium Ascorbate/Ascorbic acid as preservative in an amount of 1-3%w/w;
iv. 25 Hydroxy Vitamin D3 (Calcifediol) as active in an amount of 1-2%;
v. Antioxidant in an amount of 1-6%;
vi. Vegetable oil selected from sunflower oil or corn Oil in an amount of 2-8%w/w; and
vii. Silicon dioxide in an amount of 0.5-5%w/w.
2. The stable feed composition as claimed in claim 1, wherein the active is dissolved in the vegetable oil in an amount of 2-8% by weight based on the total weight of the composition and the amount of the vegetable oil is less than 70% by weight based on the total weight of the oil composition.
3. The stable feed composition as claimed in claim 1, wherein the feed composition is in powder form.
4. The stable feed composition as claimed in any one of the claims 1 to 3, prepared by the process comprising;
i. Preparing an emulsion of Calcifediol i.e. 25 hydroxy Vitamin D3, antioxidant selected from BHT (Butylated Hydroxy toluene) in the oil selected from sunflower Oil or corn oil;
ii. Preparing a water phase of Sucrose, Maltodextrin, (Modified starch), Sodium Ascorbate or ascorbic acid in purified water;
iii. Homogenizing the emulsion phase of step (i) together with water phase of step (ii) forming a uniform emulsion and optionally adding purified water to maintain the viscosity of the emulsion in the range of 30 centipose to 120 centipose;
iv. Spray drying the homogenized emulsion of step (iii) to obtain spray dried encapsulated product;
v. Blending the spray dried product of step (iv) with silicon dioxide, sifting and packing under inert atmosphere.
5. The feed composition as claimed in any one of the claims 1 to 4, wherein 25 hydroxy vitamin D3 is in an amount of 12.5to 15g/ kg of the feed, preferably in an amount of 12.5g/kg of the feed.
6. Use of feed composition as claimed in any one of the claims 1 to 5 to reduce bone disorders and increase the body weight in animals.
7. Use of feed composition as claimed in any one of the claims 1 to 5 as a feed in the premix comprising the nutritive additives selected from vitamins, carbohydrates, proteins, lipids (for example fish meal, fish oil, feather meal or other types of meal produced from other slaughterhouse waste); amino acids, antimicrobial peptides; polyunsaturated fatty acids; and/or at least one enzyme, probiotics, prebiotics, mineral premix, medicinal additives, starches alone or mixture thereof.
8. A method of reducing the bone disorders and increasing the body weight in animals comprising feeding the animal the stable feed composition as claimed in any one of the claims 1 to 5.
9. A premix feed composition comprising
A. An encapsulated oil composition which comprise;
i. Maltodextrin as encapsulating agent in an amount of 10-20%w/w;
ii. Modified corn starch as encapsulating agent in an amount of 65-75%w/w;
iii. Sodium Ascorbate/Ascorbic acid as preservative in an amount of 1-3%w/w;
iv. 25 Hydroxy Vitamin D3 (Calcifediol) as active in an amount of 1-20%;
v. Antioxidant in an amount of 1-6%;
vi. Vegetable oil selected from Sunflower oil or Corn Oil in an amount of 2-8%w/w; and
vii. Silicon dioxide in an amount of 0.5-5%w/w; and
B. One or more nutritive additive.
10. The premix feed composition as claimed in claim 9, wherein the nutritive additive selected from vitamins, carbohydrates, proteins, lipids (for example fish meal, fish oil, feather meal or other types of meal produced from other slaughterhouse waste); amino acids, antimicrobial peptides; polyunsaturated fatty acids; and/or at least one enzyme, probiotics, prebiotics, mineral premix, medicinal additives, starches alone or mixture thereof.
11. Use of premix feed composition as claimed in any one of the claims 9 and 10 to reduce bone disorders and increase the body weight in animals.
12. A method of reducing the bone disorders and increasing the body weight in animals comprising feeding the animal the premix feed composition as claimed in any one of the claims 9 and 10.