CN116669565A - Animal-free formula comprising fat-soluble vitamins - Google Patents

Abstract

The present invention relates to a milk replacer comprising an animal-free preparation comprising a) vitamin A or its C _1‑20 alkyl esters and optionally vitamin D; b) in an amount ≤ 25% by weight Gum Arabic; c) at least one ingredient selected from modified food starches, starch hydrolysates, monosaccharides and disaccharides; d) at least one antioxidant; e) anti-caking agents; f) optional oils; g) optional residual moisture; wherein all amounts of a) to f) add up to 100% by weight and are based on the sum of the weights of a) to f), wherein said compound a) is encapsulated in said compound b ) and c) in the matrix formed. The invention further relates to said animal-free preparation, and its use in milk substitutes, especially for young non-human mammals, such as calves, foals, young goats, young sheep and camel calves. A further object of the invention is a method for the manufacture of such animal-free preparations and such formulas.

Description

Animal-free milk substitutes containing fat-soluble vitamins

Summary of the invention

The present invention relates to a milk substitute, the milk substitute comprising an animal-free preparation, the animal-free preparation comprising

a) vitamin A or its C _1-20 alkyl ester and optionally vitamin D;

b) gum arabic in an amount ≤ 25% by weight;

c) at least one ingredient selected from the group consisting of modified food starch, starch hydrolysates, monosaccharides and disaccharides;

d) at least one antioxidant;

e) anti-caking agents;

f) optional oil;

g) optionally residual moisture;

wherein all amounts of a) to f) add up to 100% by weight and are based on the sum of the weights of a) to f), wherein compound a) is encapsulated in a matrix formed by compounds b) and c).

"Encapsulation" means that compound a), i.e. vitamin A or its C _1-20 alkyl ester and optionally vitamin D, is embedded in a matrix of gum arabic b) and compound c), thereby being protected from oxidation and degradation. The precursor of the preparation, i.e. the dispersion obtained after step C) of the method for manufacturing the preparation according to the present invention has been performed, forms an oil-in-water dispersion, wherein compound a) is an oil located in the internal phase and gum arabic acts as an emulsifier. Compound c) is a filler and additional emulsifier in the matrix that contributes to its stability. After drying the dispersion in the presence of an anti-caking agent e), i.e. preferably "spray drying" or "powder capturing" the dispersion, the matrix containing the compound a) is coated with an anti-caking agent. "Coating" in the context of the present invention means that the anti-caking agent surrounds the matrix.

The invention further relates to said animal-free preparation, and to its use in milk replacers, in particular for young non-human mammals, such as calves, foals, young goats, young sheep and camel calves. In this context, "young" means from the birth of a mammal to the time when said mammal should be suckled by its mother.

Milk replacers are specifically powders that are dissolved in water and administered to young non-human mammals, such as preferably calves, foals, young goats, young camels and young sheep, instead of the animal's mother's milk. Milk replacers preferably contain essential nutrients, including proteins, amino acids, carbohydrates, fats, vitamins and minerals.

Preferably, the compound a) is first encapsulated, i.e. embedded in a matrix of gum arabic b) and compound c), in particular according to the method disclosed below, to obtain an animal-free formulation, which is then mixed with other ingredients (e.g. proteins, amino acids, carbohydrates, fats, water-soluble vitamins and minerals) to obtain the formula according to the invention.

Other fat-soluble vitamins may be encapsulated together with compound a) or separately and then mixed with other ingredients to obtain the formula according to the present invention.

Besides compound a), other fat-soluble vitamins can also be encapsulated in other hydrocolloids.

A further object of the invention is a process for producing such an animal-free formulation and such a milk replacer.

Background Art

Vitamin A is essential for the development of young and growing animals. Without it, animals will grow poorly and eventually die. Vitamin A deficiency can lead to four major different physiological changes:

- Loss of vision due to failure of rhodopsin formation in the retina

-Defects in bone growth and structure

- Reproductive defects

-Defects in growth and differentiation of epithelial tissues.

Therefore, milk replacers, especially for non-human mammals such as calves, foals, young goats, camels and young sheep, contain formulations of vitamin A or its derivatives to ensure that the required optimum levels thereof are absorbed by the young mammals.

Such preparations are preferably animal-free, ie they do not contain gelatin or any other ingredients of animal origin.

Since formulas have a color similar to milk, i.e. white or beige or yellow, when dissolved in water, such formulations preferably also have such a color. In order to ensure a uniform distribution of the formulation in the formula, it is advantageous if the formulation has the same or similar bulk density as the formula without such formulation. The bulk density of formulas is typically in the range of 0.4 g/cm ³ to 0.9 g/cm ³ .

Therefore, there is a need to provide an animal-free formulation of vitamin A, vitamin A derivatives and any mixtures thereof having a bulk density that is the same or similar to that of a formula not containing such formulations, i.e., a bulk density in the range of 0.4 g/cm ³ to 0.9 g/cm ³ , thereby ensuring uniform distribution within the formula.

DETAILED DESCRIPTION

Therefore, the present invention satisfies this need and is directed to a formula substitute comprising an animal-free formulation comprising

a) vitamin A or its C _1-20 alkyl ester and optionally vitamin D;

b) gum arabic in an amount ≤ 25% by weight;

c) at least one ingredient selected from the group consisting of modified food starch, starch hydrolysates, monosaccharides and disaccharides;

d) at least one antioxidant;

e) anti-caking agents;

f) optional oil;

g) optionally residual moisture;

wherein all amounts of a) to f) add up to 100% by weight and are based on the sum of the weights of a) to f), wherein compound a) is encapsulated in a matrix formed by compounds b) and c).

Preferably, the amount of gum arabic b) is at most 50% by weight relative to the sum of the amounts of components b) and c). More preferably, the weight ratio of gum arabic to component c) is <1.

In a preferred embodiment of the present invention, the animal-free formulation consists of ingredients a) to g). The term "consisting of" means that the formulation does not contain any other ingredients than a) to g).

Based on the total weight of the formulation, i.e. the sum of the weights of a), b), c), d), e), f) and g), the amount of residual moisture in the final formulation is preferably in the range of 0.1-8.0 wt %, more preferably in the range of 0.5-7.0 wt %, and most preferably in the range of 1.0-6.0 wt %. By disclosing these ranges, any combination of any lower value with any higher value of a range is also disclosed, i.e. 1.0-8.0 wt %, 0.5-6.0 wt %, etc.

No animal products

The formulation is described in more detail below. Further disclosed are essential ingredients and their amounts, as well as ingredients not present in the formulation.

Therefore, the present invention encompasses any combination of any preferred option with another preferred option, even though a specific combination thereof may not be explicitly disclosed.

Ingredients not present

The formulation is animal-free, ie, the formulation does not contain proteins or other compounds, such as gelatin, from animal sources.

In contrast to the formulation disclosed in EP 494 417A2, the matrix of the formulation of the present invention does not contain any of the following salts: water-soluble carboxylates, sodium carbonate, potassium carbonate, calcium sulfate and calcium phosphate. Examples of such water-soluble carboxylates that are not present in the matrix of the formulation of the present invention are: basic aluminum acetate, sodium tartrate, sodium glutarate, sodium acetate, calcium acetate, sodium propionate, calcium propionate and sodium benzoate. However, some of these salts can be used as anti-caking agents.

An advantage of the process of the present invention is that no organic solvent other than water is used, so that the preparation according to the present invention is substantially free of organic solvents. "Substantially free" means that the amount thereof is ≤5 wt. %, preferably ≤3 wt. %, more preferably ≤1 wt. %, even more preferably ≤0.5 wt. %, most preferably ≤0.1 wt. %, based on the total weight of the preparation.

Another component that is not present in the formulation according to the invention is a polyol. Examples of polyols that are not present are glycerol, monoesters of glycerol with C _1-5 monocarboxylic acids, glycerol monoethers, diglycerol, triglycerol, polyglycerol, propylene glycol, dipropylene glycol, 1,3-butylene glycol, ethylene glycol, polyethylene glycol, sorbitol, xylitol, maltitol, erythritol, mannitol, etc.

Furthermore, no cross-linking agents, such as acetaldehyde, glutaraldehyde or glyoxal, are present in the formulations of the invention. As the formulations of the invention must be water soluble, no cross-linking is required.

Phenols having bulky alkyl groups, such as butylated hydroxyanisole ("BHA") and butylated hydroxytoluene ("BHT"), are also absent from the formulations of the present invention.

BHA is preferably a mixture of 2-tert-butyl-4-hydroxy-anisole and 3-tert-butyl-4-hydroxy-anisole. BHT is preferably 2,6-di-tert-butyl-p-cresol (IUPAC name = 2,6-di-tert-butyl-4-methylphenol).

Ethoxyquin, also known as "EMQ" (IUPAC name: 6-ethoxy-2,2,4-trimethyl-1,2-dihydroquinoline), is also absent from the formulations of the present invention.

Basic Ingredients

Fat-soluble vitamins

For the purposes of the present invention, the term "fat-soluble vitamins" includes vitamins A and/or D and the corresponding derivatives, such as esters, especially _C1 - _C20 alkyl esters, and any mixtures thereof.

"Vitamin D" refers to vitamin _D3 (cholecalciferol) or vitamin _D2 (ergocalciferol) or both.

"Vitamin D derivative" refers to any derivative of vitamin D, such as 25-hydroxyvitamin D ₃ (so-called "HyD"), 1,25-dihydroxyvitamin D ₃ , or 24,25-dihydroxyvitamin D ₃ .

Vitamin K may also be present. "Vitamin K" means vitamin _K1 or vitamin _K2 or vitamin _K3 or any mixture thereof. Vitamin _K1 is also known as phylloquinone, and the systematic name is all-E-2-methyl-3-(3,7,11,15-tetramethylhexadec-2-enyl)naphthalene-1,4-dione. Vitamin _K2 is a mixture of homologue molecules based on the naphthoquinone structure and isoprenoid chains of varying lengths. These compounds are known as menaquinones. Menaquinones have side chains consisting of all-E polyprenyl residues; usually they are represented as MK-n, where n specifies the number of isopentenyl repeating units. The minimum value of n is 2 (the value of n is usually 2-11). The most preferred menaquinone is MK-7 (with 7 isoprenyl groups). Vitamin _K3 is also known as menadione. Vitamin _K3 is the most preferred vitamin K in the formulations and formulas of the present invention.

Particularly preferred examples of fat-soluble vitamins are vitamin A, vitamin A acetate, vitamin A propionate, vitamin A butyrate, vitamin A palmitate, vitamin D ₃ and 25-hydroxy-vitamin D, and any mixture thereof. More preferred are vitamin A acetate, vitamin D ₃ and any mixture thereof. Even more preferably, the fat-soluble vitamin is vitamin A acetate or a mixture of vitamin A acetate and vitamin D3, preferably the weight ratio of vitamin A acetate to vitamin D3 is in the range of 1:1 to 100:1, more preferably in the range of 10:1 to 85:1. The most preferred fat-soluble vitamin is vitamin A acetate.

The amount of vitamin A or its derivative is selected in such a way that the final amount of vitamin A or its derivative in the formulation is preferably in the range of 10% to 29% by weight, more preferably in the range of 12% to 24% by weight, even more preferably in the range of 15% to 23% by weight, and most preferably in the range of 17% to 22% by weight, based on the sum of the weights of a) to f). By disclosing these ranges, any combination of any lower value of a range with any other value is also disclosed, i.e., 10-22% by weight, 12-29% by weight, 15-24% by weight, etc.

If a mixture of vitamin A acetate and vitamin D3 is used, the amount of vitamin A acetate is as given above for the vitamin A derivative, and the amount of vitamin D3 is selected in such a way that its final amount in the formulation is preferably in the range of 0.01% to 10% by weight, more preferably in the range of 0.05% to 5% by weight, even more preferably in the range of 0.1% to 3.5% by weight, and most preferably in the range of 0.3% to 2.5% by weight, based on the total weight of a) to f). By disclosing these ranges, any combination of any lower value with any higher value of a range is also disclosed, i.e. 0.01-5% by weight, 0.05-10% by weight, 0.1-5% by weight, etc.

When vitamin D, preferably vitamin D _3, is present in the formulation of the invention, oil is advantageously also present.

Oil

Oil can be natural, modified or synthetic.If oil is natural, they are preferably vegetable oils.Therefore, term "oil" encompasses any vegetable oil, such as corn oil, sunflower oil, soybean oil, safflower oil, rapeseed oil, peanut oil, palm oil, palm kernel oil, cottonseed oil, olive oil, coconut oil, canola oil, sesame oil, hazelnut oil, almond oil, cashew oil, macadamia oil, mongongo tree (mongongo) nut oil, bacas fruit oil, pecan oil, pine nut oil, pistachio oil, Sacha inchi (Inca Inchi (Plukenetia volubilis)) oil, walnut oil and medium chain triglyceride (" MCT ") and any mixture thereof.Preferably, corn oil, peanut oil, safflower oil or sunflower oil are used.

Preferably, the oil or fat is not from an animal source.

The weight ratio of vitamin D to oil ranges preferably from 1:1 to 1:10, more preferably from 1:2 to 1:5.

Gum Arabic

Gum Arabic, also known as "acacia gum," is a sticky exudate from the stems and branches of the Acacia tree when the stems and branches are stressed. The gum is collected from the Acacia senegal tree and, to a lesser extent, the Acacia seyal tree in several countries in sub-Saharan Africa. The gum Arabic is preferably used in the form of a spray-dried powder.

Preferably gum arabic is used, the viscosity of a 25 wt% aqueous solution of which, when measured with a Brookfield viscometer according to method PM GA 02, is in the range of 60 to 150 mPa·s.

The term "gum arabic" encompasses "natural gum arabic" as well as "modified gum arabic". Natural gum arabic means that it has not been chemically modified. Gum arabic from Acacia with a tannin content of >700 ppm (w/w) may also be used, as disclosed in more detail in WO 2017/017248.

"Modified gum arabic" refers to gum arabic which has been hydrolyzed to degrade the polysaccharides and, where appropriate, the protein fraction, as described, for example, in WO 2006/053761. Preferably, the modified gum arabic is a gum arabic whose protein fraction has been hydrolyzed to an extent of up to 30%, preferably to an extent of 1 to 30%, more preferably to an extent of 1 to 10%, most preferably to an extent of 1 to 5%. In certain embodiments of the invention, a modified gum arabic is used, the protein fraction of which has been hydrolyzed by between 0.05% and 1%.

Preferably, the term "gum arabic" does not include "heat-treated gum arabic" and in particular means gum arabic which has been treated according to the method as disclosed in WO 2008/110225, i.e. at a temperature in the range of 100-115°C, preferably in the range of 105-115°C, more preferably in the range of 108-113°C, most preferably 110°C, for 13-38 hours, preferably more than 15 hours and less than 24 hours, more preferably 16-18 hours, or at a temperature in the range of 55-85°C, preferably in the range of 65-85°C, more preferably in the range of 70-85°C, most preferably in the range of 70-80°C, for 17-65 hours, preferably 38-65 hours, more preferably 42-65 hours, most preferably 42-50 hours.

The amount of gum arabic b) in the formulation of the present invention is selected in such a way that the final amount of gum arabic b) in the formulation preferably ranges from 10 to 25% by weight, more preferably from 15 to 25% by weight, most preferably from 19 to 24.5% by weight, based on the sum of the weights of ingredients a) to f).

Ingredient c)

Ingredient c) is selected from modified food starches, starch hydrolysates, monosaccharides and disaccharides, and any mixtures thereof. At least one ingredient c) is used.

The amount of ingredient c) in the formulation of the present invention is selected in such a way that the final total amount of ingredient c) in the formulation preferably ranges from 35% to 58% by weight, more preferably from 38% to 55% by weight, most preferably from 40% to 52% by weight, based on the sum of the weights of ingredients a) to f).

In a preferred embodiment of the present invention, the weight ratio of gum arabic to component c) is in the range of 1:1.3 to 1:3, preferably in the range of 1:1.5 to 1:2.6.

Preferably, ingredient c) is a starch hydrolysate, or a mixture of modified food starch and a disaccharide, such as sucrose, with the preferences defined below.

If a mixture of modified food starch and a disaccharide (e.g. sucrose) is used as ingredient c), the weight ratio of modified food starch to disaccharide is preferably in the range of 1:2 to 2:1, more preferably in the range of 1:1.5 to 1.5:1, most preferably in the range of 1:1.3 to 1.3: 1. One or more modified food starches and/or one or more disaccharides may be used herein.

Starch hydrolysate

Starch hydrolysates are classified according to their DE values.

The term "dextrose equivalent" (DE) indicates the degree of hydrolysis and is a measure of the amount of reducing sugars calculated as D-glucose on a dry weight basis; the scale is based on native starch with a DE close to 0 and glucose (= dextrose) with a DE of 100.

Dextrin has DE=1-13; maltodextrin has DE=3-20, and glucose syrup has DE>20.

Preferably starch hydrolysates with DE≤40, more preferably DE≤30, even more preferably DE≤25 are used.

In a preferred embodiment of the present invention, dry glucose syrup with a DE of 20-23, which is commercially available under the name "Maltodextrin 2023", has been used.

Modified food starch

Modified food starch is a food starch that has been chemically modified by known methods to have a chemical structure that provides it with a hydrophilic part and a lipophilic part. Preferably, the modified food starch has long hydrocarbon chains as part of its structure (preferably _C5 - _C18 ).

At least one modified food starch is preferably used as ingredient c), but mixtures of two or more different modified food starches can be used for the formulation and thus also for the formula of the invention.

Starch is hydrophilic and therefore has no emulsifying ability. However, modified food starch is made from starch that has been substituted with hydrophobic moieties by known chemical methods. For example, the starch may be treated with a cyclic dicarboxylic anhydride (such as succinic anhydride) to replace it with a hydrocarbon chain. A particularly preferred modified food starch has the following formula (I):

Wherein St is starch, R is an alkylene group, and R' is a hydrophobic group. Preferably, R is a lower alkylene group, such as dimethylene or trimethylene. R' can be an alkyl or alkenyl group preferably having 5 to 18 carbon atoms. A preferred compound of formula (I) is "OSA starch" (sodium octenyl succinate starch). The degree of substitution, i.e. the ratio of the number of esterified hydroxyl groups to the number of free non-esterified hydroxyl groups, generally varies in the range of 0.1% to 10%, preferably in the range of 0.5% to 4%, more preferably in the range of 2% to 3%.

The term "OSA starch" means any starch from any natural source treated with octenyl succinic anhydride (OSA). The degree of substitution, i.e. the ratio of the number of hydroxyl groups esterified by OSA to the number of free non-esterified hydroxyl groups, generally varies in the range of 0.1% to 10%, preferably in the range of 0.5% to 4%, more preferably in the range of 2% to 3%. "Modified food starch" is a synonym often used for OSA starch.

Starch can be derived from natural sources such as corn, waxy corn, wheat, tapioca, peas, and potatoes, or it can be synthetic.

The term "OSA starch" also encompasses starches that are commercially available, for example, under the trade names HiCap 100, Capsul (octenyl succinate starch dextrin), Capsul HS, Purity Gum 2000, Cleargum COA1, Cleargum CO03, UNI-PURE, HYLON VII; commercially available from Ingredion and Roquette, respectively; commercially available from Cargill under the trade name C*EmCap, or commercially available from Tate & Lyle.

The terms "modified starch" and "OSA starch" also encompass modified starch/OSA starch which is partially hydrolyzed enzymatically, for example by glycosylases (EC 3.2; see http://www.chem.qmul.ac.uk/iubmb/enzyme/EC3.2/), and modified starch/OSA starch which is partially hydrolyzed chemically by known methods (so-called acid degradation).

Enzymatic hydrolysis is conventionally carried out at a temperature of about 5°C to about <100°C, preferably at a temperature of about 5°C to about 70°C, more preferably at a temperature of about 20°C to about 55°C.

Glycosylases can be derived from fruits, animal sources, bacteria or fungi. Glycosylases can have endo-activity and/or exo-activity. Thus, any mixture of enzyme preparations of endo-glycosylases and exo-glycosylases or mixtures thereof can be used. Preferably, the glycosylase has pectinase activity and/or hemicellulase activity. Glycosylases also typically exhibit unknown side activities, but the side activities are not important for the manufacture of the desired product.

Examples of glycosylases are commercially available enzyme preparations from the suppliers Novozymes, Genencor, AB-Enzymes, DSM Food Specialities, Amano and others.

Glycosylase is added to provide a concentration of about 0.01 wt % to about 10 wt %, preferably about 0.1 wt % to about 1 wt %, based on the dry weight of the modified starch/OSA starch. In a preferred embodiment of the method of the present invention, the enzyme is added once. Enzymatic hydrolysis can also be performed stepwise. For example, a glycosylase or a glycosylase mixture is added to the incubation batch in an amount of, for example, 1%, and then, for example, after 5 to 10 minutes (at a temperature of 35° C.), another glycosylase or glycosylase mixture that may be the same or different from the glycosylase or glycosylase mixture added for the first time is added (for example, added in an amount of 2%), and then the incubation batch is hydrolyzed at 35° C. for 10 minutes. Using this procedure, a starting modified starch/OSA starch with a degree of hydrolysis of about zero can be used.

The duration of the hydrolysis can vary between about a few seconds and about 300 minutes. The exact duration of the enzymatic treatment can be determined empirically based on the desired properties of the modified starch/OSA starch, such as emulsion stability, emulsifying capacity, droplet size of the emulsion, which depends largely on parameters such as enzyme activity or substrate composition. Alternatively, it can be determined by measuring osmolality (W. Dzwokak and S. Ziajka, Journal of food science, 1999, 64 (3) 393-395).

Inactivation of the glycosylase is suitably achieved by heat denaturation, for example by heating the incubation batch to about 80°C to 85°C for 5 to 30 minutes, especially 5 to 10 minutes.

OSA starch modified according to the methods as disclosed in WO 2020/093962, WO 2020/093919, WO 2020/093960 and CN-A 109 517080 can also be used as ingredient c) of the formulations and milk substitutes according to the invention.

Monosaccharides and disaccharides

Examples of monosaccharides are fructose, glucose (= dextrose), mannose, galactose, sorbose, and any mixtures thereof.

The term "glucose" in the context of the present invention is intended not only to mean the pure substance, but also glucose syrup with a DE ≥ 90 (DE as defined above). This also applies to other monosaccharides.

Examples of disaccharides are sucrose (=cane sugar), isomaltose, lactose, maltose and nigerose, and any mixtures thereof.

Preferably sucrose is used.

Antioxidants d)

The antioxidant may be a water-soluble antioxidant or a fat-soluble antioxidant, or any mixture thereof. Therefore, mixtures of water-soluble antioxidants, mixtures of fat-soluble antioxidants, and mixtures of one or more water-soluble antioxidants with one or more fat-soluble antioxidants are also included in the term "antioxidant". Fat-soluble antioxidants and mixtures thereof, and mixtures of water-soluble antioxidants with fat-soluble antioxidants are preferred. It is particularly preferred to use a single fat-soluble antioxidant. Inorganic antioxidants may also be present.

Examples of inorganic antioxidants are NaBH ₄ , Na ₂ SO ₃ and/or Na ₂ S ₂ O ₃ .

Examples of fat-soluble antioxidants are ascorbyl palmitate, polyphenols, flavonoids substituted with one or more hydroxyl groups, isoflavones substituted with one or more hydroxyl groups, tocotrienols and analogs thereof, tocopherols and analogs thereof, or any mixtures thereof. Preferred examples of fat-soluble antioxidants are ascorbyl palmitate, tocotrienols and analogs thereof, tocopherols and analogs thereof, or any mixtures thereof.

Analogs of tocopherol and tocotrienol are compounds having shorter side chains, especially at the 2-position, compared to tocopherol and tocotrienol.

Examples of flavones substituted with one or more hydroxyl groups are: 6-hydroxyflavone, 5,7-dihydroxyflavone (=chrysin), 4',5,7-trihydroxyflavone (=apigenin), 3',4',5,7-tetrahydroxyflavone (luteolin) and 4',5,6,7,8-pentamethoxyflavone (citrus flavonoids).

Examples of isoflavones substituted by one or more hydroxyl groups and optionally methoxy groups are daidzein (=4',7-dihydroxyisoflavone), genistein (=4',5,7-trihydroxyisoflavone), safflower (=4',5-dihydroxy-7-methoxyisoflavone), biochanin A (=5,7-dihydroxy-4'-methoxy-isoflavone), olopol (=3',4',5,7-tetrahydroxyisoflavone), santol (=3',4',5-trihydroxy-7-methoxy-isoflavone) and safflower (=3',5,7-trihydroxy-4'-methoxy-isoflavone).

Examples of water-soluble antioxidants are ascorbic acid and its salts (eg, sodium ascorbate), citric acid and its salts (eg, sodium citrate), and any mixtures thereof.

Examples of mixtures of water-soluble antioxidants and fat-soluble antioxidants are tocopherol and sodium ascorbate, tocopherol and ascorbic acid, wherein the tocopherol may be α-tocopherol, β-tocopherol, γ-tocopherol or δ-tocopherol, preferably wherein the tocopherol is α-tocopherol or δ-tocopherol, more preferably wherein the tocopherol is α-tocopherol, most preferably wherein the tocopherol is DL-α-tocopherol.

Tocopherol, tocotrienol and their analogs

Examples of suitable tocopherols and their analogs are compounds of formula (II)

wherein R ^1a and R ^2a are independently H or C _1-11 alkyl or (CH ₂ ) _n -OH, wherein n is an integer from 1 to 4, or R ^1a and R ^2a together represent a keto group, A is CHR ^3a or C(＝O), and wherein R ^3a , R ^4a and R ^6a are independently H or C _1-4 alkyl, and wherein R ^5a is H or OH or C _1-4 alkyl or C _1-4 alkoxy, as disclosed in WO 2019/185894.

Another suitable tocopherol is a compound of formula (II), wherein one of the two substituents R ^1a and R ^2a is C _12-21 alkyl and the other of the two substituents R ^1a and R ^2a is hydrogen or C _1-5 alkyl or (CH ₂ ) _n -OH, wherein n is an integer from 1 to 5, and wherein A is CH(R ^3a ), and wherein R ^3a , R ^4a and R ^6a are independently of each other H or C _1-4 alkyl, and wherein R ^5a is H or OH or C _1-4 alkyl or C _1-4 alkoxy, as disclosed in WO 2019/185938.

Compounds of formula (II) wherein A is CH ₂ , R ^1a is C _1-5 -alkyl, R ^2a is H or C 1-2 -alkyl, _R ^5a is H or C _1-4 -alkoxy or C _1-4 -alkyl, and R ^4a and R ^6a are independently of each other H or C _1-4 -alkyl, with the preferences as disclosed in WO 2019/185900, are also suitable antioxidants in the formulations of the present invention.

Preferred examples of antioxidants of formula (II) as disclosed in WO 2019/185894 are compounds of the following formulae (1) to (11), wherein "Me" is a methyl group:

Other examples of suitable antioxidants that can be used in the formulations of the present invention are compounds of formula (III) and formula (IV),

wherein R ^1b and R ^2b are independently H or C _1-11 alkyl or CH ₂ ) _n -OH, wherein n is an integer from 1 to 6, or R ^1b and R ^2b together represent a keto group, and wherein R ^3b , R ^4b , R ^5b , and R ^6b are independently H or C _1-6 alkyl or C _1-6 alkoxy, and R ^7b is H or C _1-6 alkyl, as disclosed in WO 2019/185898.

Thus, "alkyl" and "alkoxy" include both straight-chain and branched-chain alkyl groups, and both straight-chain and branched-chain alkoxy groups, respectively.

Preferred examples of the compounds of formula (III) and formula (IV) are the following compounds (12) to (19):

Another suitable antioxidant is a compound of formula (V), wherein R ¹ , R ² and R ³ are independently H or linear C _1-6 alkyl or branched C _3-8 alkyl, wherein preferably R ¹ is H or methyl or ethyl or n-propyl or isopropyl or tert-butyl, and R ² and R ³ are independently H or methyl or ethyl, wherein further preferences are as disclosed in WO 2019/185940.

In addition, compounds of formula (VI) as disclosed in WO 2019/185904, wherein n is 1 or 2, R ^1b and R ^3b are independently H or C _1-5 alkyl, and R ^2b is H or C _1-5 alkyl or C _1-5 alkoxy, preferably with the proviso that at least one of R ^1b , R ^2b and R ^3b is H.

Therefore, the compounds of the following formulae (VI-1) and (VI-2) are particularly preferred:

An asterisk * marks each chiral/stereomeric center, ie all possible isomers with any configuration at said center are encompassed by the terms "compound of formula (VI-1)" and "compound of formula (VI-2)", respectively.

Likewise, suitable antioxidants are compounds of the following formulae (VII) and (VIII) as disclosed in WO 2019/185942 and WO 2019/185888, respectively, wherein R ^1c , R ^2c and R ^3c are independently H or C _1-4 -alkyl. Preferred examples thereof are tocotrienols and tocopherols of formulae (20) to (27) as shown below.

An asterisk * marks each chiral/stereomeric center. The term "compound of formula (VII)/(VIII)" encompasses all possible isomers with any configuration at the center in question.

Examples of particularly preferred compounds of formula (VII) are compounds of the following formulae (20) (=α-tocotrienol), (21) (=β-tocotrienol), (22) (=γ-tocotrienol) and (23) (=δ-tocotrienol), including all possible diastereomers and enantiomers.

Examples of particularly preferred compounds of formula (VIII) are compounds of the following formulae (20) (=α-tocopherol), (21) (=β-tocopherol), (22) (=γ-tocopherol) and (23) (=δ-tocopherol), including all possible diastereomers and enantiomers.

An asterisk * marks each chiral/stereomeric center. The term "compound of formula (20)/(21)/(22)/(23)/(24)/(25)/(26)/(27)" encompasses all possible isomers having any configuration at the center.

Polyphenols

Examples of suitable polyphenols are 2,4,5-trihydroxybutyrophenone, epigallocatechin gallate ("EGCG"), epigallocatechin, epigallocatechin gallate, hydroxytyrosol, resveratrol, carnosol, 2-(3,4-dihydroxyphenyl)acetic acid and _C1-6 alkyl esters thereof, and any mixtures thereof.

Further suitable polyphenols are derivatives of hydroxybenzoic acid, such as gallic acid (=3,4,5-trihydroxybenzoic acid) and syringic acid (=4-hydroxy-3,5-dimethoxy-benzoic acid), preferably esters and alkali (earth) metal salts. Examples of preferred esters are C _1-20 alkyl esters of gallic acid, such as propyl gallate, octyl gallate or dodecyl gallate, and C _1-20 alkyl esters of syringic acid.

In addition, derivatives, preferably esters and alkali (earth) metal salts of cinnamic acid and hydroxycinnamic acids, such as ferulic acid (=3-(4-hydroxy-3-methoxyphenol)prop-2-enoic acid), caffeic acid (=3,4-dihydroxycinnamic acid), dihydrocaffeic acid (=3-(3,4-dihydroxyphenyl)propionic acid), chlorogenic acid (=ester of caffeic acid and (-)-quinic acid), o-coumaric acid, m-coumaric acid, p-coumaric acid (=2-hydroxycinnamic acid/3-hydroxycinnamic acid/4-hydroxycinnamic acid), rosmarinic acid (=caffeic acid ester of 3-(3,4-dihydroxyphenyl)lactic acid) or sinapinic acid (=3,5-dimethoxy-4-hydroxycinnamic acid) can be used as antioxidants in the present invention.

Examples of the derivatives of cinnamic acid are Z-ethoxyethyl p-methoxycinnamate, ethylhexyl p-methoxycinnamate, 2-ethylhexyl 4-methoxycinnamate, methyl diisopropylcinnamate, isoamyl 4-methoxycinnamate, and diethanolamine 4-methoxycinnamate.

The most preferred antioxidants for use in the formulations of the present invention are DL-α-tocopherol, and mixtures of DL-α-tocopherol with sodium ascorbate, mixtures of DL-α-tocopherol with ascorbic acid, and mixtures of DL-α-tocopherol with ascorbyl palmitate.

The weight ratio of DL-α-tocopherol to sodium ascorbate and DL-α-tocopherol to ascorbic acid ranges preferably from 5:1 to 1:5, more preferably from 2:1 to 1:4.5, most preferably from 1.5:1 to 1:4.1.

Preferably, the total amount of antioxidant is selected in such a way that the final amount of antioxidant in the formulation is preferably ≤7.5 wt.-%, more preferably the final amount of antioxidant is in the range of 0.5 wt.-% to 7.5 wt.-%, based on the total weight of a) to f) in the formulation.

If DL-α-tocopherol is used as the sole antioxidant, it is preferably used in an amount of at least 1 wt %, more preferably in an amount ranging from 1.0 wt % to 5.0 wt %, based on the sum of the weights of ingredients a) to f).

It is assumed that the amounts of other tocopherols or their analogs and tocotrienols and their analogs are in a similar range as DL-α-tocopherol.

Anti-caking agent e)

Suitable organic anti-caking agents are talc, cellulose, microcrystalline cellulose, cellulose derivatives or fibers, ammonium ferric citrate, sodium salts of fatty acids (e.g. sodium stearate), potassium salts of fatty acids (e.g. potassium stearate), calcium salts of fatty acids (e.g. calcium stearate), magnesium salts of fatty acids (e.g. magnesium stearate), aluminum salts of fatty acids (e.g. aluminum stearate), ammonium salts of fatty acids (e.g. ammonium stearate), and any mixture of any of them.

Other suitable anti-caking agents are inorganic anti-caking agents, for example silicate H _2n+2 Si _n O _3n+1 and its alkali/alkaline earth metal salts, precipitated silicic acid, silica (=silicon dioxide), modified silica, hydrophobically modified silica, precipitated silica, magnesium oxide, dicalcium diphosphate, tricalcium phosphate, magnesium phosphate, sodium carbonate, potassium carbonate, calcium carbonate, magnesium carbonate, calcium oxide, magnesium oxide, potassium silicate, calcium silicate, magnesium silicate, magnesium trisilicate, aluminum silicate, sodium aluminum silicate, potassium aluminum silicate, calcium aluminum silicate, zeolites (aluminosilicates), disodium sulfate, or mixtures thereof.

Likewise, mixtures of organic and inorganic anti-caking agents may be used.

Preferred anti-caking agents for use in the formulations of the present invention are those having a particle size D (v, 50%) of 100 nm to 10 μm, preferably 100 nm to 9 μm, more preferably 150 nm to 5 μm, as measured as a dry dispersion with a Malvern MasterSizer 3000 (laser diffraction).

The particle size of the anti-caking agent can be measured with a laser diffraction system (e.g. Malvern MasterSizer 3000) as a dry dispersion or as a wet dispersion in oil or Volasil (a mixture of volatile and cyclic silicones, such as Octamethylcyclotetrasiloxane and Decamethylcyclopentasiloxane). The particle size can also be measured with an electron microscope.

More preferred anti-caking agents are silicic acid H _2n+2 Si _n O _3n+1 , silicon dioxide, microcrystalline cellulose, and any mixture thereof.

The most preferred anti-caking agents are hydrophilic precipitated silicic acid H _2n+2 Si _n O _3n+1 , hydrophilic precipitated silica, and any mixtures thereof.

The term "anti-caking agent" therefore preferably does not include titanium dioxide.The animal-free formulation according to the invention as well as the milk replacer according to the invention therefore do not contain titanium dioxide.

The amount of anti-caking agent e) preferably ranges from 0.01 wt% to 10 wt%, based on the sum of the weights of ingredients a) to f). The more preferred amount of anti-caking agent depends on the drying technique and is disclosed below.

Characteristics of the formulation of the present invention

The formulations of the present invention preferably exhibit a bulk density ranging from 0.4 g/cm ³ to 0.9 g/cm ³ , more preferably ranging from 0.6 g/cm ³ to 0.7 g/cm ³ .

Furthermore, the formulations of the present invention preferably exhibit a tap density ranging from 0.7 g/cm ³ to 0.85 g/cm ³ .

Bulk density and tap density were measured as follows:

Fill a 250 ml glass cylinder with the sample. Measure the volume and weight. Bulk density is the weight divided by the volume.

To measure the tap density, the samples were tapped with a Stampfvolumeter JEL STAV II (J. Engelsmann AG) at 2000 taps. The tapped volume was measured.

Tap density is weight divided by tap volume.

Particle size and measurement

To determine the particle size of the animal-free formulations of the present invention, a sieving test can be performed using filters having 850 μm pores (mesh 20), 425 μm pores (mesh 40), and 150 μm pores (mesh 100).

Additionally, the particle size of animal-free formulations can also be determined by laser diffraction analysis, wherein a dry dispersion of the sample is measured using a Malvern Mastersizer 2000 or 3000 and Fraunhofer calculations.

When measured by laser diffraction using a Malvern Mastersizer 3000 and Fraunhofer calculations, the particle size of the formulation was as follows:

D(v,10%)＝1-60μm;

D(v,50%)＝80-140μm;

D(v,90%)＝180-240μm.

Synonyms for “D(v, 10%)” are “d(v, 10%)”, “d(v, 10)”, “d(v, 0.1)”, “D(v, 10)”, and “D(v, 0.1)”. This applies in a similar manner to “D(v, 50%)” and “D(v, 90%)”.

Preferred embodiments of the preparation of the present invention

Table I and Table II show preferred formulations according to the invention and their composition. Instead of vitamin A acetate, another vitamin C _1-20 alkyl ester, preferably another vitamin AC _2-16 alkyl ester, can also be used. The formulation preferably contains only the ingredients cited in the corresponding table and does not contain further ingredients. Therefore, the formulation preferably consists only of the ingredients cited in the corresponding table. All amounts in Table I and Table II, except residual moisture, are given in weight %, totaling up to 100 weight % and based on the total weight of all ingredients except residual moisture. The amount of moisture in Table I and Table II is given in weight % and based on the total weight of the formulation.

The formulation according to Table I comprises vitamin A acetate and optionally vitamin D3, gum arabic; starch hydrolysate with DE≤40, preferably DE≤30, more preferably DE≤25; one or more antioxidants, anti-caking agents and optionally residual moisture. When vitamin D3 is present, preferably oil is also present in the amounts given above. Preferably, the weight ratio of gum arabic to starch hydrolysate with DE≤40 ranges from 1:1.3 to 1:3, more preferably it ranges from 1:1.5 to 1:2.6.

The formulation according to Table II comprises vitamin A acetate and optionally vitamin D3, gum arabic, modified food starch, disaccharides, one or more antioxidants, an anti-caking agent and optionally residual moisture. When vitamin D3 is present, preferably oil is also present in the amounts given above. Preferably, the weight ratio of modified food starch to disaccharides ranges from 1:2 to 2:1, more preferably from 1:1.5 to 1.5:1, most preferably from 1:1.3 to 1.3:1.

Table I:

Table II

Method for manufacturing the formulation

The formulation of the present invention is preferably manufactured according to a method comprising the following steps:

A) dissolving gum arabic b), ingredient c) and water-soluble antioxidant d) (if present) in water to obtain a matrix;

B) heating the fat-soluble vitamins a), the fat-soluble antioxidants d) and the oil f) (if present) to obtain an active phase;

C) emulsifying the active phase obtained in step B) into the matrix obtained in step A) to obtain a dispersion;

D) Drying the dispersion obtained in step C), optionally in the presence of an anti-caking agent, to obtain the formulation.

The individual steps are disclosed in more detail below. Further details are given in the examples, which may also be generalized.

Step A)

The amounts of gum arabic b), ingredient c) and water-soluble antioxidant c) (if present) are chosen so that after steps A) to D) have been carried out, the final amounts of these compounds in the animal-free formulation are as described above.

Step A) is preferably performed at a temperature ranging from 45 to 80°C, more preferably ranging from 50 to 78°C, most preferably ranging from 55 to 75°C.

Step B)

The amounts of fat-soluble vitamins a), fat-soluble antioxidants d) and oil f) (if present) are chosen so that the final amounts of these compounds in the animal-free formulation after steps A) to D) have been carried out are as described above.

Step B) is preferably carried out at a temperature at which components a) and d) become liquid.

When vitamin D, preferably vitamin D _3, is present in the formulation of the present invention, vitamin D is preferably added to other fat-soluble vitamins and fat-soluble antioxidants as an oil suspension, whereby the weight ratio of vitamin D to oil ranges preferably from 1:1 to 1:10, more preferably from 1:2 to 1:5.

Step C)

Preferably, this step is performed at a mixing temperature ranging from 40 to 78°C, more preferably ranging from 50 to 75°C, even more preferably ranging from 55 to 70°C to obtain a dispersion.

Homogenization can be achieved by using a rotor-stator device or a high pressure homogenizer or both. Other devices known to those skilled in the art can also be used.

Step D)

Separation and drying of the mixture of the oil-in-water preparation to produce the animal-free formulation according to the invention can be carried out in any conventional manner, for example spray cooling, modified spray cooling, spray drying, a combination of spray drying and fluidized bed granulation, modified spray drying or sheet drying and crushing, see for example WO 91/06292 A1.

Furthermore, the conversion to an animal-free formulation can be achieved by powder capture technology, whereby the sprayed dispersion droplets are captured by an anti-caking agent (the so-called "capture medium") and dried.

If spray drying is carried out as the drying step, an anti-caking agent is preferably added in an amount such that the amount in the final formulation is up to 1.5 wt %, based on the total weight of ingredients a) to f) together, preferably the amount ranges from 0.01 wt % to 1.0 wt %.

If the powder capture process is carried out as a drying step, an anti-caking agent is preferably added in an amount such that the amount in the final formulation is at least 5.0 wt %, based on the total weight of ingredients a) to f) together, preferably the amount is in the range of 5.0 wt % to 10.0 wt %, more preferably the amount is in the range of 5.0 wt % to 7.0 wt %.

Other embodiments of the present invention

use

The invention also relates to the use of a preparation according to the invention with the preferences given above as an additive to milk replacers, in particular for young non-human mammals, such as calves, foals, young goats, young camels and young sheep.

Formula according to the invention

The invention also relates to a milk replacer comprising the formulation according to the invention with the preferences given above. "Milk replacer" means any substance or product intended for use in replacing milk of non-human mammals, such as calves, foals, young sheep, camels and young goats, including processed, partially processed or unprocessed additives.

Such formulas preferably further comprise essential nutrients including proteins, amino acids, carbohydrates, fats, vitamins and minerals.

The amount of animal-free products and fat-soluble vitamins in these milk replacers follows the regulatory guidelines in the region, depending on the specific animal species and its age.

In supplementation guidelines, amounts of vitamin A and _D3 are given in international units ("IU").

To ensure that the active ingredients in the feed are delivered in a systemic manner, "I.U." is used as a universal unit for fat-soluble vitamins, since there are different forms of vitamins with varying amounts of fat-soluble vitamins.

The preparation according to the invention is usually added to the milk replacer in the form of a premix (i.e. a mixture with other micronutrients such as other vitamins or their preparations and minerals). For many species, the premix in the feed contains <1% by weight. However, it can also be added as is.

The amount of the formulation according to the invention that needs to be included in the formula is calculated based on the active content of the formula and the target dose of the fat-soluble vitamins taking into account the inclusion level.

The conversion factors for fat-soluble vitamins are as follows:

1I.U. vitamin A is equivalent to 0.344μg vitamin A acetate;

1I.U. Vitamin D ₃ is equivalent to 0.025μg Vitamin D ₃ ;

1 g of vitamin K ₃ (menadione) is equivalent to 2.0 g of menadione sodium bisulfite ("MSB") or 2.3 g of menadione nicotinamide sulfite ("MNB");

Table III below shows the amounts of fat soluble vitamins added. The exact amount depends on several factors such as the stage/age of the animal, the animal species and local legal restrictions.

Table III

1) The amount of milk replacer powder added per kg of calf.

2) Daily supplement amount per animal.

The invention will now be further illustrated in the following non-limiting examples.

Example

The following Examples 1 and 2 illustrate the preparation of the formulations of the present invention.

Example 1 and Example 2 (Table 1)

The base components, namely gum arabic, ingredient c) and a water-soluble antioxidant, are dissolved in water at about 70°C to obtain a "base".

Vitamin A acetate, fat-soluble antioxidant and oil (if present) are heated at about 65°C with stirring until the vitamin A acetate ("active phase") is completely melted. They are then emulsified into the base.

Therefore, the amounts of the ingredients were chosen in such a way that their concentrations in the final formulation were as disclosed in Table 1, respectively (n.d. = not determined).

After thorough mixing, the resulting dispersion is sprayed into a spray tower, optionally in the presence of an anti-caking agent, to form droplets of a desired size. The solidified droplets are then dried by dry air at different temperatures (5-75°C).

Example 1: If spray drying: The dried powder was sieved through a 1 mm sieve to discard agglomerates.

Example 2: If powder capture is performed: The dried powder is separated from the bulk of the anti-caking agent and sieved through 150 μm and 600 μm filters.

The particle size of the dried powder was determined by laser diffraction analysis using a Malvern Mastersizer 3000. The sample was thus dry dispersed. The particle size distribution of the sample was calculated by applying the Fraunhofer theory.

In addition, bulk density and tap density were measured according to the procedure described above.

Table 1

Claims (15)

1. A formula comprising an animal-free formulation comprising

a) Vitamin A or C _1-20 Alkyl esters and optionally vitamin D;

b) Gum arabic in an amount of 25 wt.% or less;

c) At least one ingredient selected from the group consisting of modified food starch, starch hydrolysates, mono-and disaccharides;

d) At least one antioxidant;

e) An anti-caking agent;

f) Optionally an oil;

g) Optionally residual moisture;

wherein all amounts of a) to f) add up to 100% by weight and are based on the sum of the weights of a) to f), wherein the compound a) is encapsulated in a matrix formed by the compounds b) and c).

2. Formula according to claim 1, wherein the formula is for a young non-human mammal, preferably for a young non-human mammal selected from calves, foal, young goats, young sheep and camels.

3. An animal-free formulation comprising

a) Vitamin A or C _1-20 Alkyl esters and optionally vitamin D;

b) Gum arabic in an amount of 25 wt.% or less;

c) At least one ingredient selected from the group consisting of modified food starch, starch hydrolysates, mono-and disaccharides;

d) At least one antioxidant;

e) An anti-caking agent;

f) Optionally an oil;

g) Optionally residual moisture;

wherein all amounts of a) to f) add up to 100% by weight and based on the sum of the weights of the components a) to f), wherein the compound a) is encapsulated in a matrix formed by the compounds b) and c).

4. The animal-free formulation of claim 3, wherein the formulation has a bulk density in the range of 0.4g/cm ³ To 0.9g/cm ³ 。

5. The animal-free formulation according to claim 3 and/or 4, wherein the amount of component c) ranges from 35 to 58 wt%, preferably the amount ranges from 38 to 55 wt%, more preferably the amount ranges from 40 to 52 wt%, based on the sum of the weights of components a) to f).

6. The animal-free formulation according to any one or more of claims 3 to 5, wherein the ingredient c) is a starch hydrolysate having DE ∈40 or a mixture of modified food starch and disaccharide, wherein the weight ratio of the modified food starch to the disaccharide preferably ranges from 1:2 to 2:1.

7. The animal free formulation of any one or more of claims 3-6, wherein the anti-caking agent e) has a particle size D (v, 50%) ranging from 100nm to 10 μιη, measured with Malvern MasterSizer 3000 (laser diffraction) as a dry dispersion.

8. The animal-free formulation of any one or more of claims 3-7, wherein vitamin a or C thereof, based on the sum of the weights of ingredients a) to f) _1-20 The amount of alkyl ester ranges from 10 wt% to 29 wt%, preferably the amount ranges from 12 wt% to 24 wt%, more preferably the amount ranges from 15 wt% to 23 wt%, most preferably the amount ranges from 17 wt% to 22 wt%.

9. The animal-free formulation according to any one or more of claims 3 to 8, wherein the amount of gum arabic b) ranges from 10 to 25 wt%, preferably the amount ranges from 15 to 25 wt%, most preferably the amount ranges from 19 to 24.5 wt%, based on the sum of the weights of ingredients a) to f).

10. The animal-free formulation according to any one or more of claims 3 to 9, wherein the amount of antioxidant d) is less than or equal to 7.5 wt%, preferably the amount ranges from 0.5 wt% to 7.5 wt%, based on the sum of the weights of the ingredients a) to f).

11. The animal-free formulation of any one or more of claims 3-10, wherein the amount of the anti-caking agent e) ranges from 0.01 to 10 wt% based on the sum of the weights of the ingredients a) to f).

12. Use of a formulation according to any one or more of claims 3 to 11 in a formula.

13. Use according to claim 12, wherein the formula is for calves, foal, young sheep, young goats and young camels.

14. A method for manufacturing a formulation according to any one or more of claims 3 to 11, the method comprising the steps of:

A) Dissolving gum arabic b), component c) and, if present, a water-soluble antioxidant d) in water to obtain a matrix;

b) Heating the fat-soluble vitamin a), the fat-soluble antioxidant d) and, if present, the oil f) to obtain an active phase;

c) Emulsifying the active phase obtained in step B) into the matrix obtained in step a) to obtain a dispersion;

d) Drying the dispersion obtained in step C), optionally in the presence of an anti-caking agent, to obtain the formulation.

15. A method for manufacturing a formula according to claim 1 and/or claim 2 by mixing an animal free formulation according to any one or more of claims 3 to 11 with essential nutrients including proteins, amino acids, carbohydrates, fats, vitamins and minerals.