HK1196038A - Bioactive compounds protection method and compositions containing the same - Google Patents

Description

Method for protecting biologically active compounds and compositions containing same

The present application is a divisional application of chinese patent application 200480023845.7 entitled "method of protecting bioactive compounds and compositions containing the same" filed 6/17/2004.

Technical Field

The present invention relates generally to methods for delivering health-enhancing compounds to mammals, and more particularly to methods for encapsulating and embedding bioactive ingredients in mammalian food or feed formulations.

Background

Biologically active proteins include EGF (epidermal growth factor), insulin and insulin-like growth factor, insulin-like growth factor binding proteins, immunoglobulins (e.g., helicobacter pylori (h. pyrori) antibodies), proline-rich polypeptides, lactoferrin, proteases, whey proteins, interleukins, lysozyme, TGFA (transforming growth factor a), and PDGF (platelet-derived growth factor).

These proteins may have been shown to have a positive effect on one or more of the following health-promoting advantages: protein efficiency ratio, weight gain, immune system function, normal growth of organs and cells, systemic and local DNA regulation, maintenance of good health, amelioration of the disease state, and recovery and cure from the disease state.

Most industrial processes involve production conditions that are highly damaging to the potential durability of biologically active compounds such as functional proteins. These conditions include high temperature and pressure, low and high pH, light, drying and other similar adverse production conditions. Furthermore, the demand for shelf-life increases due to supply chain constraints, where long term storage under adverse conditions causes a loss of the biological activity of these proteins. Thus, it is difficult to include such compounds in commercially produced feeds and foods (e.g., complete animal rations or infant formulas).

This is critical because mammalian newborns, such as human infants and many livestock newborns, often lose the opportunity to consume natural breast milk immediately after birth or shortly after birth, rely primarily on milk substitutes (milk substitets) for their nutrition, and semi-solid and solid mixtures and pellet diets after weaning.

This generally means that the desired positive health benefits provided by the original natural breast milk are largely absent from natural milk substitute products and subsequent post-weaning mixes and pelleted foods used to feed important animal infants in agriculture.

For example, the immune components of colostrum and subsequent mature milk include IgG, IgM, and IgA. These components confer passive immunity to the neonate, protecting them from infection early after parturition. The presence of insulin (and possibly IGF-1, IGF-2 and EGF) in the colostrum and subsequent mature milk has been shown to allow the development of the gastrointestinal system to be faster and better in mammalian newborns fed with this natural breast milk compared to mammalian newborns fed with artificial replacement food or feed. In addition, the presence of insulin in colostrum and subsequently in mature milk has been shown to have a positive effect on reducing the probability of later development of type I diabetes (insulin dependent diabetes mellitus) in mammalian infants. However, insulin and insulin-like proteins are very sensitive to production conditions and environmental storage conditions such as light, radiation, low/high temperature, low/high pressure, gastrointestinal digestive compounds and the presence of chemical compounds.

Disclosure of Invention

In one embodiment of the present invention, there is provided a method of encapsulating and embedding a bioactive ingredient in a mammalian neonatal formula, comprising the steps of: (i) mixing the bioactive ingredient with a food-grade or feed-grade encapsulating material to form a liquid mixture, (ii) drying the liquid mixture to form a dry blend, and (iii) adding the dry blend to a mammalian neonatal formulation, thereby being a method of encapsulating and embedding a bioactive ingredient in a mammalian neonatal formulation.

In another embodiment of the present invention, there is provided a method of encapsulating and embedding a bioactive ingredient in a mammalian neonatal formula, comprising the steps of: (i) mixing the bioactive ingredient with a food-grade or feed-grade encapsulating material, thereby forming a liquid mixture, (ii) drying the liquid mixture, thereby forming a dry blend, (iii) coating the dry blend with at least one layer of another food-grade or feed-grade encapsulating material, and (iv) adding the dry blend to the mammalian neonatal formula, thereby becoming a method of encapsulating and embedding a bioactive ingredient in a mammalian neonatal formula.

In another embodiment of the invention, a neonatal formulation is provided comprising a bioactive ingredient encapsulated or embedded in a food-grade or feed-grade encapsulating material.

In one embodiment of the invention, a method is provided for improving the health, growth and development of a mammal comprising the step of administering to said mammal a newborn formulation comprising a bioactive ingredient encapsulated or embedded in a food-grade or feed-grade encapsulating material, said compound being capable of improving the health, growth and development of a mammal.

In another embodiment of the invention, a method of enriching an infant formula or formula is provided comprising the step of admixing to the infant formula or formula a bioactive ingredient encapsulated or embedded in a food-grade or feed-grade encapsulating material.

In one embodiment of the present invention, there is provided a method of encapsulating or embedding a bioactive ingredient in a solid or semi-solid mammalian feed formulation comprising the steps of: (i) mixing the bioactive ingredient with a food-grade or feed-grade encapsulating material to form a liquid mixture, (ii) drying the liquid mixture to form a dry blend, (iii) coating the dry blend with at least one layer of another food-grade or feed-grade encapsulating material, and (iv) adding the dry blend to the solid or semi-solid mammalian feed formulation.

In another embodiment, a solid or semi-solid mammalian feed formulation is provided, wherein the solid or semi-solid formulation comprises a bioactive ingredient encapsulated or embedded in a food-grade or feed-grade encapsulating material.

In one embodiment of the invention, a method for improving the health, growth and development of a mammal is provided comprising incorporating into the solid or semi-solid mammal feed formulation a bioactive ingredient encapsulated or embedded in a food-grade or feed-grade encapsulating material.

In another embodiment of the present invention, there is provided a method of enriching a solid or semi-solid mammalian feed formulation comprising the step of incorporating into the solid or semi-solid mammalian feed formulation a bioactive ingredient encapsulated or embedded in a food-grade or feed-grade encapsulating material, the method of preparation comprising the steps of: (i) mixing the bioactive ingredient with a food-grade or feed-grade encapsulating material to form a liquid mixture, (ii) drying the liquid mixture to form a dry blend, (iii) coating the dry blend with at least one layer of another food-grade or feed-grade encapsulating material, and (iv) adding the dry blend to the solid or semi-solid mammalian feed formulation.

Detailed Description

In one embodiment of the invention, there is provided a method of encapsulating a bioactive material in a food-grade or feed-grade glassy matrix, the method comprising the steps of: (i) uniformly and intimately mixing at least one bioactive material with at least one food-grade or feed-grade wall-forming encapsulating material to form a mixture, (ii) mixing the mixture with a suitable plasticizer, (iii) rapidly removing the plasticizer while inhibiting crystallization of the wall-forming material, thereby completing encapsulation of the bioactive material in a food-grade or feed-grade glassy matrix.

In another embodiment of the present invention, a method of encapsulating a bioactive material is provided, comprising the steps of: (i) encapsulating at least one bioactive material in a food-grade or feed-grade glassy matrix is accomplished by mixing the at least one bioactive material with at least one molten food-grade or feed-grade wall-forming encapsulating material, and (ii) rapidly cooling the at least one molten wall-forming material.

For example, in the food and pharmaceutical industries, microencapsulation is used to stabilize the core material, to control the timing and rate of release of the core material, and to isolate reactive or incompatible ingredients in multi-component formulations and to control chemical reactions therebetween. Thus, microencapsulation enables protection of sensitive food ingredients to ensure that the nutritional value is not lost and that taste and odor are masked or preserved. Microencapsulation also increases the stability of vitamin additives that are generally sensitive to, for example, UV radiation, light, oxygen, metals, humidity, and temperature. Microencapsulation is also used in the pharmaceutical industry to protect the interior of the mouth and esophagus from the harsh oral drug which is released in the stomach upon exposure of the microcapsule coating to gastric acid.

Microencapsulation is a process in which one or more active ingredients are coated or embedded in another material or system. Encapsulation of heat sensitive compounds such as nutrients, enzymes or biologically active proteins in edible substrates is not usually easy for a number of reasons, one of which is the important reason that conventional encapsulation processes expose the substrate material to high temperatures encountered in extrusion, which leads to thermal destruction or loss of biological tolerance of the encapsulate. Thus, unless a large excess of encapsulates is required, which is not only expensive but potentially dangerous, the encapsulates will not support the encapsulation process at all. If the encapsulate can be encapsulated in a matrix at a sufficiently low temperature, the resulting product is a solid, characterized as a hard glassy solid, which can be further processed to give a free-flowing powder, which can be further processed. Furthermore, although the temperature at which the particles are consumed, or referred to as the consumption temperature, is typically below 50 degrees celsius, which is well below the glass transition temperature t.sub., care is taken to design the glassy matrix to enable the encapsulates to be released under the desired conditions of temperature, humidity and pH. They can also be used in the form of dense pellets in a variety of processing applications where controlled release of heat sensitive encapsulated mist is desired. The physical hardness of the product and its mechanical stability are advantageous for many processing applications.

In one embodiment, plasticizers, as used herein, refer to other compounds that are capable of increasing the free volume of a liquid encapsulate without affecting the total cumulative volume of the encapsulated matrix and plasticizing compound.

In one embodiment of the invention, there are provided protected bioactive ingredients, including but not limited to proteins used in dietary formulations.

In another embodiment of the invention, methods of preparing the protected bioactive components are provided to retain the biological activity of these proteins.

The present invention may be used to retain the biological activity of a biologically active ingredient against any of the following or similar damaging factors: adverse temperature, pressure, humidity, pH, osmolarity, ionic concentration, chemical degradation, presence of metals, surfactants and chelating agents, radiation (including but not limited to UV, IR, visible light), enzymatic and microbial degradation. In addition, the present invention may also be used to protect bioactive components against physical changes, including but not limited to primary and secondary phase changes.

In one embodiment of the present invention, there is provided a protected bioactive ingredient comprising at least one protective layer encapsulating a bioactive ingredient present in mammalian breast milk.

In another embodiment of the present invention, mammalian breast milk as used herein refers to colostrum.

In another embodiment of the invention, the protected bioactive ingredient is present in natural mammalian milk, but at a very low, absent, unavailable or absent concentration in commercially processed milk or formula.

In another embodiment of the invention, concentrations as used herein refer to molarity and its fraction or percentage relative to molarity present in colostrum.

The term "very low" as used herein refers in one embodiment to an amount of bioactive ingredient in commercially processed milk that is from about 0.1% to about 50% of the amount present in colostrum.

In one embodiment of the invention, the amount of bioactive ingredient in the commercially processed milk is at most 50% of the amount present in the colostrum.

In another embodiment of the invention, the amount of bioactive ingredient in the commercially processed milk is up to 25% of the amount present in the colostrum.

In another embodiment of the invention, the amount of bioactive ingredient in the commercially processed milk is at most 10% of the amount present in the colostrum.

In another embodiment of the invention, the amount of bioactive ingredient in the commercially processed milk is at most 1% of the amount present in the colostrum.

In another embodiment of the invention, the amount of bioactive ingredient in the commercially processed milk is at most 0.1% of the amount present in the colostrum.

In another embodiment, the amount of milk processed is undetectable using methods known to those skilled in the art.

In one embodiment of the invention, the at least one protective layer is capable of maintaining the bioactive properties of the bioactive ingredient in a "resting state", which in one embodiment of the invention is the protected protein in a dry or substantially dry state, such as in powdered infant formulas, dairy products and semi-solid/solid mixtures and pellets.

In another embodiment, the resting state of a protein as used herein refers to the retention of the native tertiary and quaternary structure of the protein in the anhydrous state.

In one embodiment of the invention, the at least one protective layer provides protection for the encapsulated bioactive ingredient, so that the protein will substantially maintain its bioactive properties under adverse conditions such as high temperature, high pressure, humidity, adverse osmotic pressure, high or low pH, strong enzymatic degradation, high solvent concentrations, etc., which typically result in protein denaturation. Then, in another embodiment, upon an initiating event, the outer protective layer dissolves and the "dormant" bioactive component will be released and exhibit physiological activity.

In one embodiment, the protected bioactive ingredient is designed such that release of the bioactive ingredient occurs prior to entry into the gastrointestinal system of a mammalian neonate consuming the formulation.

In another embodiment of the invention, the release is upon contact with a different site of the gastrointestinal tract.

In another embodiment of the invention, the encapsulated bioactive ingredient is protected from conditions encountered in commercial extrusion processes including, but not limited to, cold extrusion or hot extrusion under or different from standard temperature and pressure conditions.

In another embodiment of the invention, the encapsulated bioactive ingredient is protected from conditions encountered in commercial comminution processes including, but not limited to, colloid mills, including frusto-conical and gear-type (cron and tooth type) stator rotors, ball mills, impact mills, jet impact mills, homogenizing mills, sonication, high speed mixers, and membrane emulsification devices.

In one embodiment of the invention, the encapsulated bioactive ingredient is protected from the conditions encountered in commercial baking processes.

In another embodiment of the invention, the protected bioactive ingredient is protected from the conditions encountered during commercial freezing.

In one embodiment of the invention, a newborn formulation is provided comprising a bioactive ingredient encapsulated or embedded in an edible ingredient.

In one embodiment of the invention, the newborn formulation may be an infant formula or milk replacer/replacer (milk replacer/subltite) for consumption by a mammalian newborn.

In another embodiment, a milk replacer/good as used herein refers to any milk replacer/good of a newborn of a mammal, wherein the mammal is a cow, a horse, a porcine animal, such as calf, lamb, pig, cow, sheep, goat, yaez, cat, dog and horse.

In one embodiment of the invention, the milk replacer is any milk replacer for newborns of mammals, wherein the mammals are felines and canines.

In another embodiment, the bioactive ingredient is encapsulated in a plastic matrix material that can be plasticized with a liquid plasticizer or liquid encapsulating component at low temperatures, which can be a plastic biopolymer.

In one embodiment, the plasticized material includes, but is not limited to, carbohydrate polysaccharides such as pentosans, physically or chemically modified starches or cyclodextrins, and mixtures thereof.

In another embodiment, the plasticized material is a polymer such as polyvinylpyrrolidone (PVP, povidone) or other non-hydrophobic polymers such as N-vinyl pyrrolidone (NVP) and (vinyl) acetate copolymers, (polyvinyl) alcohol chitosan or mixtures thereof. In one embodiment, the plasticized material is a cellulose ester, a cellulose ether, and a polyethylene glycol. In another embodiment, the plasticized material is a hydrocolloid such as xanthan gum, carrageenan, alginate, gum arabic, acacia gum, tragacanth gum, gum conjac and mixtures thereof.

In one embodiment, the plasticized material is glutenin and gliadin, such as vital wheat gluten or separated gluten, zein, vegetable or dairy protein from soy or milk, and mixtures thereof.

In another embodiment of the present invention, the starch useful in the present invention is a physically or chemically modified starch having an amylose/amylopectin ratio of from about 1 to about 0.001, and is derived from corn, wheat, rice, potato, tapioca, yuka, and kudzu (arrow root).

In one embodiment, starch sources that may be used also include powders from cereals such as corn, wheat, durum wheat, rice, barley, oats or rye and mixtures thereof.

In another embodiment, only wall materials approved by FDA or similar regulatory agencies in europe and elsewhere are used, since the microcapsules formed will be used in neonatal formulations or in solid or semi-solid feed formulations.

In one embodiment, the GRAS list provides a list of compounds that can be used to form the capsule wall.

In one embodiment of the invention, any other food-grade or feed-grade encapsulating material approved by regulatory agencies for approved human and/or animal consumption (the applicable party) will be used as encapsulating material in the process.

In one embodiment of the invention, the wall material used is poly (DL-lactide-co-glycolide).

In another embodiment of the invention, the food-grade or feed-grade encapsulating material used in the neonatal formulation comprises a polysaccharide, maltodextrin, milk powder, whey protein, lipid, gum arabic or microcrystalline cellulose or a combination thereof.

In one embodiment of the invention, the encapsulated or embedded bioactive ingredient retains or substantially retains its bioactive function and properties during the formulation of the neonatal formula.

In one embodiment of the invention, the encapsulated or embedded bioactive ingredient retains or substantially retains its bioactive function and properties during the normal shelf life of the base (undersying) newborn formulation or solid or semi-solid feed formulation in which it is incorporated.

In one embodiment of the invention, the biologically active ingredient may be a glycoprotein, an immunoglobulin, a peptide, a polypeptide, a hormone or an enzyme.

In another embodiment of the invention, the bioactive ingredient is insulin, IGF-I, IGF-2 or EGF.

In one embodiment of the invention, the biologically active ingredient includes, but is by no means limited to, alpha-1 protease inhibitors, alkaline phosphatase, angiogenin, antithrombin III, chitinase, extracellular superoxide dismutase, factor VIII, factor IX, factor X, fibrinogen, glucocerebrosidase, glutamate decarboxylase, human serum albumin, myelin basic protein, lactoferrin, lactoglobulin, lysozyme, lactalbumin, proinsulin, soluble CD4, components and complexes of soluble CD4, tissue plasminogen activator, and variants, combinations, and pharmaceutically acceptable salts thereof.

In another embodiment of the invention, insulin or any other protein may be a recombinant, synthetic, purified from natural sources, biologically active, and peptide or polypeptide having the amino acid sequence of a human or mammalian protein, such as human or bovine or porcine insulin.

In one embodiment, depending on their method of preparation, crude extracts may be used, e.g. synthetic proteins may be prepared using chemical peptide synthesis or any other similar suitable method.

In one embodiment of the invention, the neonatal formulation comprises an encapsulated bioactive ingredient of uniform size, wherein the particles have a radius of about 0.001 to about 5000 microns.

In one embodiment of the present invention, a solid feed formulation as used herein refers to a formulation capable of retaining its density and supporting its own weight at room temperature.

In another embodiment of the present invention, a semi-solid formulation, as used herein, refers to a formulation having a viscosity of from about 1 to about 600000 pascal-seconds that is capable of flowing under its own weight.

In one embodiment of the invention, the formulation is for post-weaning mammals.

In another embodiment, post-weaning mammals as used herein refer to the age at which full-growth (intense growth) mammals typically break breast milk. For example, lambs that grow at full speed are typically weaned 25-35 days after birth. Piglets growing at full speed are usually weaned 30-50 days after birth. Calves growing at full speed are usually weaned 40-70 days after birth.

In all these newborn animals, in one embodiment of the invention, the amount of milk replacer containing the bioactive ingredient provided is gradually reduced, while the amount of mixture, pellets or other semi-solid or solid feed is increased.

According to another embodiment of the invention, it is advantageous to integrate the bioactive ingredient in the mixture/pellet for up to 1-9 months after weaning.

In one embodiment of the invention, the solid or semi-solid feed formulation may be in the form of, but is not limited to, a pasty food, pellets, granules, briquettes, extrudates or a combination thereof.

In another embodiment of the invention, the encapsulated or embedded bioactive ingredient retains or substantially retains its bioactive function during the formulation of the solid or semi-solid mammalian feed formulation.

In another embodiment of the invention, the encapsulated or embedded bioactive ingredient retains or substantially retains its bioactivity during digestion of the feed.

In one embodiment of the invention, the encapsulated or embedded bioactive ingredient is released upon contact with a liquid.

In one embodiment of the invention, the solid or semi-solid feed formulation is a protein glycoprotein, immunoglobulin, peptide, polypeptide, hormone or enzyme alone or in combination.

In another embodiment of the present invention, the bioactive ingredient may be insulin, IGF-I, IGF-II or EGF alone or in combination.

In one embodiment of the invention, the biologically active ingredient is encapsulated or embedded in a polysaccharide, such as maltodextrin.

In another embodiment of the invention, the solid or semi-solid mammalian feed formulation comprises uniformly sized particles of the encapsulated bioactive ingredient, the particles having an average particle size of from about 10 to about 4000 microns.

The formulation used in one embodiment of the invention is effective in increasing the rate of weight gain in mammals, preventing diarrhea and other gastric disorders, and for extending the life expectancy of mammals after birth.

The products containing the protected bioactive ingredients of another embodiment of the present invention may be consumed by a variety of individuals, including but not limited to preterm infants, preterm infants after discharge, term infants, toddlers, adolescents, adults, elderly, non-human mammalian infants or adults, including but not limited to cows, pigs, sheep, cats, dogs, or horses, and non-mammalian infants or adults.

In another embodiment of the present invention, there is provided a method of encapsulating or embedding a bioactive ingredient in a mammalian neonatal formula, comprising the steps of: (i) mixing the bioactive ingredient with an edible food-grade or feed-grade encapsulating material to form a liquid mixture, (ii) drying the liquid mixture, (iii) coating the dry blend with at least one layer of another food-grade or feed-grade encapsulating material, and (iv) adding the dry blend to the mammalian neonatal formulation.

In one embodiment, the mammalian newborn formulation may be an infant formulation or a milk replacer/grade. Such a formulation, in another embodiment, is in the form of a powder, solution, suspension, emulsion, ointment, liquid, semi-solid, or solid cream.

In another embodiment of the invention, a post-weaning mammalian formulation is provided, which is solid or semi-solid, comprising an encapsulated and embedded bioactive ingredient, and which is prepared by the following method: (i) mixing the bioactive ingredient with a food-grade or feed-grade encapsulating material, thereby forming a liquid mixture, (ii) drying the liquid mixture, thereby forming a dry blend, (iii) coating the dry blend with at least one other food-grade or feed-grade encapsulating material, and (iv) adding the dry blend to the solid or semi-solid mammalian feed formulation. The solid or semi-solid formulation may be in the form of pellets or a paste/mixture.

Furthermore, according to one embodiment of the invention, the step of mixing the bioactive material with food-grade or feed-grade wall-forming material comprises the addition of a liquid such as, but not limited to, water, saline, alcohol, molasses, or similar food-grade or feed-grade encapsulating material solvent.

In another embodiment of the invention, the ratio of the food-grade or feed-grade material to the food-grade or feed-grade encapsulating material solvent may be from about 1:1 to about 1:1000 in one embodiment of the invention.

In one embodiment of the invention, the ratio of the food-grade or feed-grade material to the food-grade or feed-grade encapsulating material solvent is about 1:3 to 1: 100.

In another embodiment of the invention, the dry blend is further milled.

In one embodiment, the encapsulated bioactive ingredient may continue to be encapsulated with at least one other protective layer, which may be formed of the same food-grade or feed-grade encapsulating material in another embodiment, or may be formed of a different food-grade or feed-grade encapsulating material in another embodiment.

In one embodiment, the dry blend is further mixed with the food-grade or feed-grade encapsulating material to form at least one further layer of food-grade or feed-grade encapsulating material encapsulating the active ingredient.

In one embodiment of the invention, the biologically active ingredient may be a-1 protease inhibitor, alkaline phosphatase, angiogenin, antithrombin III, chitinase, extracellular superoxide dismutase, factor VIII, factor IX, factor X, fibrinogen, glucocerebrosidase, glutamate decarboxylase, human serum albumin, myelin basic protein, lactoferrin, lactoglobulin, lysozyme, lactalbumin, proinsulin, soluble CD4, components and complexes of soluble CD4, tissue plasminogen activator, or variants, pharmaceutically acceptable salts or combinations thereof.

In another embodiment of the invention, the food-grade or feed-grade encapsulating material is a polysaccharide, milk powder, whey protein, lipid, acacia, microcrystalline cellulose, analogs thereof, or combinations thereof.

In one embodiment of the invention, the food-grade or feed-grade encapsulating material is solid at a temperature of up to 70 ℃.

In another embodiment of the present invention, the method used in the step of drying the food-grade or feed-grade encapsulated material and the bioactive material includes, but is not limited to: freeze drying, vacuum drying, spray drying, osmotic dehydration, fluidized bed dehydration, solvent evaporation dehydration, ultrasound assisted dehydration, microwave assisted dehydration, RF assisted dehydration, alone or in commercially acceptable combinations.

In one embodiment of the invention, the liquid mixture is lyophilized after introducing the at least one bioactive ingredient and the at least one food-grade or feed-grade encapsulating material ingredient.

In one embodiment, lyophilization produces droplets in the glassy state containing at least one protected bioactive ingredient and at least one food-grade or feed-grade encapsulating material.

In one embodiment, a flash freezer is used to dry the liquid mixture by using a liquefied gas including, but not limited to, nitrogen, carbon dioxide, and the like.

In one embodiment, the droplet is from about 10 to about 5000 microns in size.

In another embodiment, the droplet size distribution is dependent on a variety of parameters including, but not limited to, the frozen nebulizer nozzle size, liquefied gas temperature, cell temperature, mixture component ratios and gas flow rates, food grade or feed grade encapsulating material concentration, plasticizer type and amount, and frozen cell wall geometry.

In one embodiment of the invention, the size distribution of the glassy droplets resulting from this process is from 50 microns to 1000 microns.

In one embodiment, such treatment results in glass-like frozen droplets, wherein each droplet contains at least one protected bioactive ingredient, at least one food-grade or feed-grade encapsulating material, and a food-grade or feed-grade solvent.

In another embodiment, once such frozen microdroplets are placed at a temperature above the melting point of the mixture, the liquid mixture from the previous stage of the process is rehydrated.

In one embodiment of the invention, the method further comprises freeze-drying the combination of at least one bioactive ingredient and at least one food-grade or feed-grade encapsulating material.

In another embodiment, the liquid mixture of at least one protected bioactive ingredient and at least one food-grade or feed-grade encapsulating material, or the frozen glassy droplets described herein above, may be freeze-dried.

In one embodiment, this freeze-drying process results in a dried glassy material comprising at least one food-grade or feed-grade encapsulating material and the at least one protected bioactive ingredient.

In another embodiment, the liquid mixture is freeze-dried, which results in a bulk dried material, which is naturally porous, comprising a glassy matrix of dried food-grade or feed-grade encapsulating material encapsulating the bioactive material.

In one embodiment, the product of the rapid freeze spray process is freeze dried to yield glass-like droplets comprising food-grade or feed-grade encapsulating material incorporating the bioactive ingredient.

In another embodiment, the combination of at least one bioactive ingredient and at least one food-grade or feed-grade encapsulating material is subjected to low temperature spray drying.

In one embodiment, the bioactive material is dispersed in the food-grade or feed-grade encapsulating material and atomized at a temperature of up to 45 ℃.

In another embodiment, the maximum temperature is 37 ℃ to prevent denaturation of the bioactive ingredient. In one embodiment, a liquid mixture of at least one protected bioactive ingredient, at least one food-grade or feed-grade encapsulating material, and at least one chaperone-like protective protein may be spray dried, resulting in a dried material comprising the food-grade or feed-grade encapsulating material and the at least one protected bioactive ingredient.

In one embodiment of the invention, when the bioactive component is a protein, the food-grade or feed-grade encapsulating material and the bioactive material are dehydrated at a temperature preferably below the denaturation temperature of the bioactive component.

In another embodiment, the dehydration of the food-grade or feed-grade encapsulating material and the bioactive material is performed at a temperature below a threshold temperature at which the bioactive material begins to degrade.

In one embodiment of the invention, the dehydration process of the food-grade or feed-grade encapsulated material and said biologically active material is carried out at a temperature of at most 50 ℃.

In another embodiment of the invention, the step of drying the liquid mixture results in glass-like freeze-dried droplets comprising the bioactive ingredient and at least one food-grade or feed-grade encapsulating material.

In one embodiment of the invention, the freeze-drying step is preceded by a step of spraying the liquid mixture from an atomizer in the presence of a liquefied gas.

In one embodiment, extrusion is used as the encapsulation process, wherein the core material is dispersed in a liquid of the bioactive ingredient and at least one food-grade or feed-grade encapsulation material, and the microcapsules are finally formed.

In another embodiment of the invention, encapsulating or embedding the bioactive ingredient in the above formulation comprises the step of premixing a small amount of the mixture with the mammalian neonatal formula or food grade or feed grade encapsulating material or semi-solid or solid formulation to ensure homogeneity before it is mixed with the entire formulation.

In one embodiment of the invention, the protection methods suitable for use herein include, but are not limited to, methods for producing protected bioactive components in the form of: powders, microencapsulated powders, nanocapsule powders, liquids, microemulsion liquids, nanoemulsions, solutions, microemulsion solutions, nanoemulsion solutions, paste-like foodstuffs, pastes, ointments, microdroplets, nanodroplets, tablets and solids such as pellets.

In another embodiment of the invention, the microencapsulation process comprises two phases (duplex), W/O/W, O/W/O, double or multiple emulsions.

In one embodiment of the invention, the comminution is achieved by suspending a mixture of at least one bioactive material and at least one food-grade or feed-grade encapsulating material and at least one surfactant having an HLB value substantially below 7 in the immiscible food-grade or feed-grade encapsulating material and further mixing in the above mentioned process.

In another embodiment, the milled emulsion is further mixed with food-grade or feed-grade materials miscible with said food-grade or feed-grade encapsulating material and a food-grade or feed-grade surfactant having an HLB value substantially higher than 7, and further comminuted in the above mentioned manner.

According to an embodiment of the present invention, after the bioactive ingredient is formulated, a micro-or nano-emulsification of the bioactive ingredient is performed.

In one embodiment, the formulated bioactive ingredient is mixed with an emulsion incorporating an aqueous phase, an oil phase, and a surfactant. The result of this mixing is a redistribution of the bioactive ingredient molecules into the dispersed phase of the emulsion.

In another embodiment, the microemulsion or nanoemulsion provides protection to the bioactive ingredient in relation to temperature contact protection and increases the solubility of the bioactive ingredient after release from its encapsulation in the food or feed in which it is located, but before consumption and/or during digestion.

In another embodiment, the bioactive component in the nanoemulsion or microemulsion is initially protected in the liquid microemulsion or liquid nanoemulsion.

One of ordinary skill in the art will readily recognize that the invention is not limited to the details of construction and arrangement of components set forth herein below. It will be understood that various modifications may be made without substantially changing the scope or spirit of the invention. It should be noted that practicing the invention is not limited to the applications mentioned herein below, and that many other applications and variations may be made without departing from the scope of the invention. Also, it is to be understood that the terminology used herein is for the purpose of description and should not be regarded as limiting.

In one embodiment of the present invention, there is provided a method of encapsulating a bioactive material in a food grade glassy matrix, the method comprising the steps of: (i) uniformly and intimately mixing at least one bioactive material with at least one food-grade or feed-grade wall-forming encapsulating material to form a mixture, (ii) mixing the mixture with a suitable plasticizer, (iii) rapidly removing the plasticizer while inhibiting crystallization of the wall-forming material, thereby completing encapsulation of the bioactive material in a food-grade or feed-grade glassy matrix.

In another embodiment of the present invention, a method of encapsulating a bioactive material is provided, comprising the steps of: (i) mixing at least one bioactive material with at least one molten food-grade or feed-grade wall-forming encapsulating material, and (ii) rapidly cooling the at least one molten wall-forming material, thereby completing encapsulation of the bioactive material in a food-grade or feed-grade glassy matrix.

In another embodiment of the present invention, there is provided a method of encapsulating and embedding a bioactive ingredient in a mammalian neonatal formula, the method comprising the steps of: (i) mixing the bioactive ingredient with a food-grade or feed-grade encapsulating material, thereby forming a liquid mixture, (ii) drying the liquid mixture, thereby forming a dry blend, (iii) coating the dry blend with at least one layer of another food-grade or feed-grade encapsulating material, and (iv) adding the dry blend to the mammalian neonatal formula, thereby becoming a method of encapsulating and embedding a bioactive ingredient in a mammalian neonatal formula.

In another embodiment of the invention, a mammalian neonatal formulation is provided which comprises a bioactive ingredient encapsulated or embedded in a food-grade or feed-grade encapsulating material.

In another embodiment of the present invention, there is provided a method of encapsulating and embedding a bioactive ingredient in a solid or semi-solid mammalian feed formulation comprising the steps of: (i) mixing the bioactive ingredient with a food-grade or feed-grade encapsulating material, thereby forming a liquid mixture, (ii) drying the liquid mixture, thereby forming a dry mix, (iii) coating the dry mix with at least one layer of other food-grade or feed-grade encapsulating material, and (iv) adding the dry mix to the solid or semi-solid mammalian feed formulation, thereby being a method of encapsulating and embedding a bioactive ingredient in a solid or semi-solid mammalian feed formulation.

In another embodiment of the invention, a solid or semi-solid mammalian feed formulation is provided comprising a bioactive ingredient encapsulated or embedded in a food-grade or feed-grade encapsulating material.

The following examples are provided to more fully illustrate some embodiments of the invention. They should in no way be construed as limiting the scope of the invention.

Examples

Example 1

Effect of dietary insulin on weight gain in newborn cattle

Materials and methods

100 Friesy and Charole' calves of 12-15 days after birth were used. Each calf received no more than 6 liters of milk replacer per day until day 37 after birth. From day 37, the amount of milk replacer was reduced by 0.5 liters every two days while the amount of pellets was increased until the calves were 57 days old, at which time they were considered to be completely weaned. The test group of calves received 600 micro units/cc of liquid milk replacer. Control group calves received no insulin. The grams of daily growth per kilogram of birth weight per calf was measured and calculated.

Results

The group receiving insulin showed a 26% increase in weight from day 17 to day 67 after birth, compared to the control group. At the end of the study period, the average daily weight gain in grams/kg of birth weight was 22.47, and the control group was 18.71 grams/kg of birth weight over the same period. These results demonstrate that insulin is an important factor in weight gain in calves from 12-15 to 68 days after birth.

Example 2

Effect of encapsulated bioactive ingredient-enriched feed formulations on neonatal sheep health

Materials and methods

More than 1000 Assaf specie lambs are used for 2-3 days after birth for 12 months. The growth phase is divided into two phases: feeding milk replacer for lambs 3-28 days after birth; postnatal 29-75 days during which lambs are fed the post-weaning mixture/slurry. Lambs in the study group received between 600 and 3000 micro units of insulin per ml of milk replacer and between 1600 and 5000 micro units of insulin per g of mixture per slurry. Control lambs received no insulin.

4 kg of maltodextrin, 40 l of 0.9% saline and 100000IU (International Unit) of insulin are mixed to obtain a liquid mixture (e.g. 25IU of insulin per 1 g of maltodextrin). The liquid mixture was then freeze dried. The product of this process is maltodextrin encapsulated insulin. The bioactive properties of insulin were retained by 97% after the freeze-drying process. A10 Kg premix was prepared from standard milk powder and the maltodextrin encapsulated insulin. This premix was then mixed with 740Kg of a standard milk substitute to give 750Kg of a commercial insulin-rich milk substitute.

Results

Weight gain: on average, lambs in the study group receiving insulin gained 5% -7% more weight than the control group, whether the insulin was in the milk replacer or in the mixture/slurry. Lambs from the study group that received insulin only in the milk replacer (not in the mixture/slurry) gained 3% -5% more weight than the control group.

Incidence of disease: the incidence of diarrhea and/or pneumonia in the study group was 10% -25% lower in lambs compared to the control group. In addition, the recovery of lambs with this disease was compared to that of the control groupThe lamb phase is 5% -20% faster.

Mortality rate: the mortality rate in the study group was 20% -80% lower than in the control group, depending on the particular study. For example, in one study, the insulin-fed group started with 70 newborn lambs, whereas 68 lambs survived at the point of sale (e.g., 150 days after birth) (mortality rate 2.8%). The control group received no insulin, starting with 69 newborn lambs, whereas only 61 lambs survived at the time of sale (e.g., mortality rate 11.6%). This means that the mortality rate in the group fed insulin was 74.8% lower than that in the control group.

These results demonstrate that insulin is an important factor in lamb weight gain and health from days 2-3 to 75 after birth. Furthermore, the results show that the encapsulated insulin retains biological activity during the preparation of the solid feed and during transport in the digestive system of the lambs.

Claims (19)

1. A method of encapsulating and embedding a heat sensitive bioactive ingredient in a mammalian neonatal formula, comprising the steps of:

(i) mixing a heat sensitive bioactive ingredient with an encapsulating material to form a liquid mixture, wherein the encapsulating material is a food grade material and/or a feed grade material,

(ii) drying the liquid mixture to form a dry blend,

(iii) coating said dry mix with at least one other food-grade or feed-grade coating layer, and

(iv) adding the dry blend to the mammalian newborn formulation,

whereby the heat sensitive bioactive ingredient is encapsulated and embedded in the mammalian neonatal formulation, wherein the steps of mixing all ingredients and drying are all performed at a temperature below 50 ℃ such that the activity of the heat sensitive bioactive ingredient is retained.

2. The method of claim 1, wherein the mammalian newborn food is a human infant formula.

3. The method of claim 1, wherein the mammalian newborn food is a milk replacer, milk replacer or a combination thereof.

4. The method of claim 1, wherein the mammalian neonatal formulation is in a form selected from the group consisting of a powder, a solution, a liquid, a paste, a semi-solid, or a solid.

5. The method of claim 1, wherein the heat sensitive bioactive component is selected from the group consisting of a glycoprotein, an immunoglobulin, a peptide, a polypeptide, a hormone, or an enzyme.

6. The method of claim 1, wherein the heat sensitive bioactive ingredient is selected from the group consisting of insulin, IGF-I, IGF-II, or EGF.

7. The method of claim 1, wherein the drying is performed by freeze drying, low temperature vacuum heat drying, or low temperature spray drying.

8. The method of claim 7, wherein said freeze-drying is preceded by a freeze-spraying step.

9. The method of claim 8, wherein said freeze-drying is preceded by an extrusion step.

10. The method of claim 1, wherein drying the liquid mixture produces glassy freeze-dried droplets containing the heat-sensitive bioactive ingredient and at least one encapsulating material, wherein the encapsulating material is a food-grade material and/or a feed-grade material.

11. The method of claim 1, wherein the food-grade encapsulating material is maltodextrin.

12. An edible newborn formulation comprising a heat sensitive bioactive ingredient encapsulated by the method of claim 1.

13. The newborn formulation of claim 12, wherein said formulation is selected from the group consisting of human infant formulations and formulations for consumption by primates, bovines, porcines, ovines, canines, felines, and caprines.

14. The newborn formulation of claim 12, wherein said encapsulated heat sensitive bioactive ingredient is released upon contact with a liquid.

15. The newborn formulation of claim 12, wherein said heat sensitive bioactive ingredient is selected from the group consisting of glycoproteins, immunoglobulins, peptides, polypeptides, hormones or enzymes.

16. The newborn formulation of claim 12, wherein said heat sensitive bioactive ingredient is selected from the group consisting of insulin, IGF-I, IGF-II, or EGF.

17. The newborn formulation of claim 12, comprising uniformly sized particles of said encapsulated heat sensitive bioactive ingredient, wherein said particles have a radius of 1-1000 microns.

18. A method for improving the health of a mammal comprising the step of administering to said mammal the newborn formulation of claim 12, thereby improving the health of the mammal.

19. The method of claim 18, wherein the health condition is growth, development, or a combination thereof of the mammal.