COATED COMPOSITIONS OF BIOLOGICALLY ACTIVE INGREDIENTS FOR ORAL ADMINISTRATION TO RUMINANTS

Abstract

A coated composition for feeding a ruminant may include: (A) a core of a biologically active ingredient selected from (A-i) amino acids, N-acylamino acids, N-guanylamino acids, and/or salts thereof and/or 2-hydroxy-4-(methyl)butyric acid calcium salt; (A-ii) proteins; (A-iii) peptides; (A-iv) carbohydrates; (A-v) vitamins, vitamin A (acetate or palmitate), B, D2, D3, E, thiamine (HCl), riboflavin, nicotinic acid (amide), calcium pantothenate, choline pantothenate, pyridoxine hydrochloride, choline chloride, cyano-cobalamin, biotin, folic acid, and p-aminobenzoic acid; (A-vi) probiotic microorganisms; (A-vii) prebiotic foods; (A-viii) choline and salts thereof; (A-ix) polyunsaturated fatty acids, and salts thereof; and x) any combination of (A-i) to (A-ix); and (B) a coating, encapsulating the core, constituting 1 to 15% of total coated composition weight and including 85 to 95% wt./wt. of a film-forming agent, which is triethyl citrate, and 5 to 15% wt./wt. of an additive selected from an acetic ester of monoglycerides and triethyl citrate.

Claims

1. A coated composition suitable for feeding a ruminant, the composition comprising: (A) a core comprising a biologically active ingredient comprising (A-i) an amino acid, amino acid salt, N-acylamino acid, N-acylamino acid salt, N-guanylamino acid, N-guanylamino acid salt, and/or 2-hydroxy-4-(methyl)butyric acid calcium salt, (A-ii) protein, (A-iii) peptide, (A-iv) carbohydrate, (A-v) vitamin, (A-vi) probiotic microorganism, (A-vii) prebiotic food, (A-viii) choline and/or choline salt, and/or (A-ix) polyunsaturated fatty acid and/or polyunsaturated fatty acid salt; and (B) a coating, encapsulating the core, constituting 1 to 15 wt. % of total coated composition weight, wherein the coating comprises (B-i) 85 to 95% wt./wt. of ethyl cellulose and (A-ii) 5 to 15% wt./wt. of an additive, and wherein additive comprises a monoglyceride acetate and/or triethyl citrate acetate.

2. The coated composition of claim 1, wherein the coating is in a range of from 2 to 8 wt. % of the total coated composition weight.

3. The coated composition of claim 1, wherein the coating comprises in a range of from 86 to 93% wt./wt. of the ethyl cellulose and in a range of from 7 to 14% wt./wt. of the additive.

4. The coated composition of claim 1, wherein the additive is triethyl citrate.

5. The coated composition of claim 1, wherein the coating is a one-layer coating.

6. The coated composition of claim 1, comprising a first layer and a second layer of the coating, wherein the first layer and the second layer have a different composition.

7. The coated composition of claim 1, having a particle size in a range of from 1 to 6 mm.

8. A method for preparing the coated composition of claim 1, the method comprising: heating a core particles a biologically active ingredient in a fluidized bed coater to a temperature in a range of from 40° C. to 90° C., to obtain heated particles; applying a mixture, comprising a solvent, the film-forming agent, and the additive, onto the heated particles via a spray nozzles of a fluidized bed coater to give a coated product; drying the coated product, to obtain a composition: and cooling the composition or allowing the composition to cool down.

9. The method of claim 8, wherein the heating and applying are repeated with the composition obtained in the drying, to obtain the coated composition comprising two or more layers.

10. The method of claim 8, wherein the temperature in the heating is in a range of from 45° C. to 80° C.

11. The method of claim 8, wherein aqueous or organic solvents are used.

12. The method of claim 8, wherein the solvent comprises a saturated hydrocarbon, an unsaturated hydrocarbon, aromatic hydrocarbon, short-chain alcohol, and/or ketone.

13. The method of claim 11, wherein the alcohol is ethanol, iso-propanol, or a mixture thereof.

14. A feed, feed material, or premix of feed additive, suitable for feeding a ruminant, the feed, feed material, or premix comprising: the coated composition of claim 1.

15. A method of supplementing the diet of a ruminant with a biologically active ingredient, the method comprising: providing the ruminant with the coated composition of claim 1 and/or with a feed, feed material, or premix of feed additive comprising the coated composition.

16. The coated composition of claim 1, wherein the vitamin (A-v) comprises vitamin A, vitamin A acetate, vitamin A palmitate, vitamin B, thiamine, thiamine hydrochloride, riboflavin, nicotinic acid, nicotinic acid amide, calcium pantothenate, choline pantothenate, pyridoxine hydrochloride, choline chloride, cyano-cobalamin, biotin, folic acid, p-aminobenzoic acid, vitamin D2, vitamin D3, and/or vitamin E,

17. The method of claim 8, wherein the temperature in the heating is in a range of from 45 to 70° C.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0038] In context of the present invention, the term “biologically active ingredient” is used to denote any compound which has a functional or nutritive activity to a biological system, such as the organism of an animal, in particular of a ruminant, e.g. cattle, sheep or goat. Typically, a biologically active ingredient exhibits low stability and thus a reduced or even lost bio-effectiveness when exposed to unfavorable conditions, for example moisture, elevated temperature, oxygen and acidic and/or basic pH. When the biologically active ingredient is exposed to such conditions, it can for example decompose, dissociate, deactivate, and/or lose viability.

[0039] The term “biologically active ingredient” therefore refers to, for example, i) amino acids, such as lysine, methionine, tryptophan, arginine, histidine, isoleucine, leucine, phenylalanine, valine, and threonine, N-acylamino acids, e.g. N-acylmethionine, and N-guanylamino acids, e.g. N-guanylglycin also known as guinidinoacetic acid, and/or salts of thereof, such as lysine sulfate, lysine hydrochloride, and the salts of lysine with polyunsaturated fatty acids, and/or 2-hydroxy-4-(methylthio)butyric acid calcium salt, also known as the calcium salt of methionine hydroxy analogue (calcium MHA), ii) proteins, such as casein, corn proteins, and potato proteins, iii) peptides, such as oligo-polypeptides, e.g. methionyl-methionine (Met-Met) and/or polypeptides, iv) carbohydrates, such as starch, cane sugar, and glucose, v) vitamins, vitamin A, vitamin A acetate, vitamin A palmitate, vitamin B, thiamine, thiamine hydrochloride, riboflavin, nicotinic acid, nicotinic acid amide, calcium pantothenate, choline pantothenate, pyridoxine hydrochloride, choline chloride, cyano-cobalamin, biotin, folic acid, p-aminobenzoic acid, vitamin D2, vitamin D3, and vitamin E, vi) probiotic microorganisms, vii) prebiotic foods, viii) choline and salts thereof, and ix) polyunsaturated fatty acids (PUFAs), such as omega-3 fatty acids, e.g. alpha-linolenic acid (ALA), eicosapentanoic acid (EPA), and docosahexanoic acid (DHA) and salts thereof.

[0040] Preferably, the biologically active ingredient of the composition according to the present invention is an amino acid, such as lysine, methionine, tryptophan, arginine, histidine, isoleucine, leucine, phenylalanine, valine, and threonine, an N-acylamino acid, e.g. N-acylmethionine, and an N-guanylamino acid, e.g. N-guanylglycin also known as guinidinoacetic acid, and/or a salt thereof, such as lysine sulfate, lysine hydrochloride, and the salts of lysine with polyunsaturated fatty acids, and/or 2-hydroxy-4-(methylthio)butyric acid calcium salt, also known as the calcium salt of methionine hydroxy analogue (calcium MHA) or a peptide, such as an oligopolypeptide, e.g. methionyl-methionine (Met-Met) and/or polypeptides. In particular, the biologically active ingredient of the composition according to the present invention is lysine, methionine, guinidinoacetic acid, methionyl-methionine (Met-Met), and/or a salt thereof.

[0041] Preferably, the core of the composition according to the present invention consists of the biologically active ingredient itself. In this way, the loading of the composition according to the present invention with the biologically active ingredient may be further increased. However, there may be still cases where the core does not only consist of the biologically active ingredient itself. This is the case for example when further additives are necessary in order to provide granulates or spherical particles of the biologically active ingredient or when the biologically active ingredient in pure form is only poorly soluble in water and therefore has be converted into its acid addition salt such as lysine hydrochloride or lysine hydrosulfate.

[0042] Advantageously, the coating constitutes 2 to 8% of the total weight of the coated composition and/or the coating comprises or consists of 86 to 93% wt./wt. of the film-forming agent and 7 to 14% wt./wt. of the additive.

[0043] Deviations from any specific values, in particular values for weight percentages, explicitly mentioned herein, are also encompassed by the present invention, provided that they also lead to the effects achieved by the present invention.

[0044] In the context of the present invention, the terms “additive” and “plasticizer” are used interchangeably and have the same meanings. The terms “additive” and “plasticizer” refer to biocompatible compounds having the ability of increasing the plasticity of the film-forming agent. Plasticizers achieve through physical interaction with a polymer a reduction in the glass transition temperature and promote film formation, depending on the added amount. Suitable substances usually have a molecular weight of between 100 and 20 000 and comprise one or more hydrophilic groups in the molecule, e.g. hydroxyl, ester or amino groups.

[0045] The additive may comprise one or more of the additives indicated above.

[0046] However, in one specific embodiment according to the present invention, the coating comprises a single additive.

[0047] Preferably, the additive is triethyl citrate (TEC).

[0048] In one specific embodiment according to the present invention, the biologically active ingredient is selected from the group consisting of amino acids, N-acylamino acids, N-guanylamino acids, salts of amino acids and/or 2-hydroxy-4-(methyl)butyric acid calcium salt; the film-forming agent is ethyl cellulose (EC) and the additive is triethyl citrate (TEC).

[0049] As indicated above, the coating according to the present invention comprises or consist of a film-forming agent and an additive. The coating comprises or consist of 85 to 95% wt./wt., in particular 86 to 93% wt./wt. of the film-forming agent, and between 5 and 15 wt.-%, in particular between 7 and 14 wt.-% of the additive, each based on the total weight of the coating. The coating as a whole constitutes 1 to 15%, in particular 2 to 8% of the total weight of the coated composition.

[0050] In one specific embodiment according to the present invention, the film-forming agent is ethyl cellulose, and the additive is triethyl citrate. Preferably, ethyl cellulose is present in an amount of 85 to 95% wt./wt., in particular 86 to 93% wt./wt.; and triethyl citrate is present in an amount of between 5 and 15 wt.-%, in particular between 7 and 14 wt.-%, each based on the total weight of the coating. The coating as a whole constitutes 1 to 5%, in particular 2 to 8% of the total weight of the coated composition.

[0051] The coating in the composition according to the present invention is not subject to any limitation regarding the number of layers. It may be desirable to apply more than one, e.g. two, three, or multiple layers of a coating as taught herein to prevent or conceal defects, e.g. cracks, and pores, formed in the coating during the preparation of the products. In addition, a mechanical impact on the products according to the present invention during their further handling may lead to micro-fissures or cracks in the outer layer. However, an overlap of the two or more layers significantly reduces or even avoids the danger of a potential leakage of the biologically active ingredient during the residence time of the products according to the present invention in the rumen. Thus, the presence of two or more layers in the coating of the products according to the present invention may also contribute to high yield of the rumen protected fraction of the biologically active ingredient. In one embodiment, the two or more layers of the coating each have a different composition, provided that the coating as such, comprising the two or more layers, comprises the amounts of film-forming agent and additive according to the present invention.

[0052] In one specific embodiment according to the present invention, the coating is a one-layer coating.

[0053] The core in the composition according to the present invention is not subject to any limitation regarding its shape. For example, the core may have a cylindrical or a spherical shape. Preferably, the core particles have a uniform shape and an even surface. Furthermore, a narrow particle size distribution of the core particles is advantageous.

[0054] Depending on the number of coating layers applied onto the core or particle comprising or consisting of the biologically active ingredient, the particle size of the compositions may be varied to obtain a given or desired particle size of the final product. It is preferred that the size of the compositions according to the present invention are such that they are not regurgitated or vomited by a ruminant upon ingestion.

[0055] Therefore, the average particle size of the compositions according to the present invention may be in the range of ca. 1 mm to ca. 6 mm, ca. 1.2 mm to ca. 5 mm, ca. 1.2 mm to ca. 4 mm, ca. 1.4 mm to ca. 3 mm, ca. 1.2 mm to ca. 2.8 mm, ca. 1.4 mm to ca. 2.6 mm, ca. 1.6 mm to ca. 2.4 mm, ca. 1.6 mm to ca. 2.2 mm or in the range of ca. 2 mm. Preferably, the compositions according to the present invention have an average particle size of at least 2 mm for reducing the chance of regurgitation or vomiting by a ruminant upon ingestion.

[0056] When preparing the product according to the present invention, it may be advantageous to add one or more additional ingredients to the coating as taught therein. In one specific embodiment according to the present invention, the coating consists only of the film-forming agent and an additive but does not comprise any additional ingredient. Preferably the film-forming agent according to this embodiment is ethyl cellulose and the additive according to this embodiment is triethyl citrate. Preferably, triethyl citrate is present in an amount of between 5 and 15 wt.-%, in particular between 7 and 14 wt.-% based on the total weight of the coating. The coating as a whole constitutes 1 to 5%, in particular 2 to 8% of the total weight of the coated composition.

[0057] It may be advantageous to add further ingredient(s) to the core comprising the active ingredient, such as one or more ingredients selected from binding substances (e.g. cellulose derivatives such as hydroxy-propyl cellulose, methyl cellulose, sodium carboxymethylcellulose, vinyl derivatives such as polyvinyl alcohol or polyvinylpyrrolidone, gum arabicum, guaiac gum, sodium polyacrylate, and the like), filling substances (e.g. starch, proteins, crystalline cellulose and the like), inert ingredients (e.g. silica and silicate compounds), flow-control substances that help the formation of pellets, preservative agents (propionic acid or its salt, sorbic acid or its salt, benzoic acid or its salt, dehydro-acetic acid or its salt, para-hydroxybenzoic acid esters, imazalil, thiabendazole, ortho-phenyl phenol, sodium ortho-phenyl phenol, diphenyl, and others compounds and mixtures thereof), antibacterial agent, and other compounds. The skilled person is familiar with techniques and compounds which are useful to achieve these purposes, and which are compatible with the production of the product according to the present invention.

[0058] It may also be advantageous to further enhance the nutritional value and/or the therapeutic value of the product according to the present invention by adding further feed ingredients (e.g. nutritional ingredients, veterinary or medicinal agents etc.) or other ingredients to the compositions as taught herein.

[0059] For instance, one or more ingredients selected from grain products, plant products, animal products, proteins (e.g. protein ingredients as obtained from sources such as dried blood or meat meal, meat and bone meal, cottonseed meal, soybean meal, rapeseed meal, sunflower seed meal, canola meal, safflower meal, dehydrated alfalfa, corn gluten meal, soybean protein concentrate, potato protein, dried and sterilized animal and poultry manure, fish meal, fish and poultry protein isolates, crab protein concentrate, hydrolyzed protein feather meal, poultry byproduct meal, liquid or powdered egg, milk whey, egg albumen, casein, fish solubles, cell cream, brewer's residues, and the like), mineral salts, vitamins (e.g. thiamine HCl, riboflavin, pyridoxine HCl, niacin, inositol, choline chloride, calcium pantothenate, biotin, folic acid, ascorbic acid, vitamin B12, p-aminobenzoic acid, vitamin A acetate, vitamin K, vitamin D, vitamin E, and the like), sugars and complex carbohydrates (e.g. water soluble and water-insoluble monosaccharides, disaccharides, and polysaccharides), veterinary compounds (e.g. promazine hydrochloride, chloromedoniate acetate, chloro-tetracycline, sulfamethazine, monensin, sodium monensin, poloxaline, oxytetracycline, BOVATEC, and the like), antioxidants (e.g. butylated hydroxyanisole, butylated hydroxytoluene, tertiary-butylhydroquinone, tocopherols, propyl gallate and ethoxyquin), trace element ingredients (e.g. compounds of cobalt, copper, manganese, iron, zinc, tin, nickel, chromium, molybdenum, iodine, chlorine, silicon, vanadium, selenium, calcium, magnesium, sodium and potassium and the like), and other compounds or ingredients, may be added to the product according to the present invention.

[0060] The skilled person is familiar with methods and ingredients that are suitable to enhance the nutritional and/or therapeutic or medicinal value of ruminant feeds, feed materials, premixes, feed additives, and feed supplements, and knows how to enhance the nutritional and/or therapeutic or medicinal value of the product according to the present invention.

[0061] In addition, or as an alternative, it is also possible to combine the product according to the present invention with a feed, feed material, or premix for feeding a ruminant. In context of the present invention the term premix or nutrient premix is used as known to the person skilled in the art and denotes a mixture comprising one or more ingredients such as vitamins, trace minerals, medicaments, feed supplements and diluents. The use of premixes has the advantage that a farmer who uses his own grain can formulate his own rations and be assured his animals are getting the recommended levels of minerals and vitamins.

[0062] Another object of the present invention is therefore a feed, feed material, or premix of feed additive for feeding a ruminant comprising the composition according to the present invention. Preferably, the premix further comprises a vitamin, trace mineral, feed supplements diluents, and/or medicaments, such as antibiotics, probiotics and/or prebiotics.

[0063] In principle, the application of the coating according to the present invention around a core comprising or consisting of a biologically active ingredient may be performed according to any suitable method known in the art, such as drum coating or fluidized bed coating. However, it was found that the best method of providing a biologically active ingredient with the coating according to the present inventions is fluidized bed coating.

[0064] A further object of the present invention in the provision of a method for preparing a coated composition as described above, the method comprising the following steps: [0065] a) providing core particles containing or consisting of a biologically active ingredient in a fluidized bed coater, [0066] b) providing a coating mixture comprising a solvent, the film-forming agent and the additive, [0067] c) heating the particles of step a) to a temperature in the range of 40° C. to 90° C., in particular to 45° C. to-80° C., preferably to 45° C. to 70° C.; [0068] d) applying the mixture of step b) onto the heated particles of step c) via the spray nozzles of the fluidized bed coater to give a coated product, [0069] e) drying the coated product obtained in step d), and [0070] f) cooling the composition obtained in step e) or allowing the composition obtained in step e) to cool down.

[0071] In case the coated composition to be prepared is to have two or more layers, steps c) to d) may be repeated with the composition obtained in step e) accordingly.

[0072] The term “core particles” is to be understood as defined above.

[0073] In order to obtain the coating solution of step b), the film-forming agent, the additive and optionally further auxiliary ingredients are admixed with in a suitable solvent. Both, aqueous and organic solvents may be used.

[0074] In one embodiment, the solvent is an organic solvent. Said organic solvent may be selected from saturated, non-saturated and aromatic hydrocarbon compounds, short-chain alcohols, ketones or mixtures of the aforementioned solvents. Suitable alcohols are e.g. ethanol and iso-propanol, or mixtures thereof. An example for a suitable ketone compound is acetone. Preferably, the solvent to be used is a short-chain alcohol, in particular ethanol.

[0075] In one specific embodiment according to the present invention, the film-forming agent and the additive were dissolved in ethanol. Preferably the film-forming agent according to this embodiment is ethyl cellulose and the additive according to this embodiment is triethyl citrate.

[0076] It was further found that administering to a ruminant the composition according to the present invention, a composition obtained or obtainable by a process according to the present invention and/or a feed, feed material, premix or feed additive according to the present invention is suitable to compensate amongst others for a lack of nutrients and essential amino acids in a ruminant diet.

[0077] Yet a further object of the present invention is therefore a method of supplementing the diet of a ruminant with a biologically active ingredient, comprising the step of providing the ruminant with a composition according to the present invention and/or with a feed, feed material, premix or feed additive according to the present invention.

[0078] In the following, the present invention will be more particularly elucidated below on the basis of exemplary and non-limiting embodiments.

EXAMPLES

I. Material and Methods

[0079] The below coating experiments were carried out in conventional fluidized bed coating units for laboratory scale (test series 1, 2 and 4) and for production scale (test series 3), respectively.

[0080] Chemicals and Reagents:

[0081] Active Ingredients

[0082] Biolys®; Methionine (Evonik Industries)

[0083] Film-Forming Agent

[0084] Ethyl cellulose (PHT International Inc.)

[0085] Additives

[0086] Acetic ester of monoglycerides (commercial product Acetem 95 Co, Danisco)

[0087] Triethyl citrate (Sigma Aldrich)

[0088] Analytical Methods

[0089] For assessing the coating quality, the rumen protection rate (RPR) was determined for the test samples.

[0090] In vitro test

[0091] The RPR corresponds to the content of still protected biologically active ingredient after an incubation time of 6 h to 24 h in a test medium (water pH 5.5 or buffer solution). The 6 h value is a qualitative measure for the protection rate. The 24 h value does not reflect the real processes in the animal but is an indication whether the tested sample may be over-protected.

[0092] The coated composition is shaken in desalinated water at 37° C./pH 5.5 in a shaking water bath.

[0093] Desired value for RPR:

TABLE-US-00001 for 6 h (rumen) high (≥70%) for 24 h (rumen and further digestive tract) low (approx. ≤50%)

[0094] After the particular period of time in the vibrating water bath, the solutions were filtered, the biologically active ingredient released from the coated composition was determined quantitatively in the filtrate and the protection rate was calculated according to the following formula:

RPR[%]=1−(amount of released active ingredient/starting amount of active ingredient)*100

[0095] In Situ Method for Testing Digestibility of Coated Active Ingredients in the Rumen

[0096] Nylon bags: ANKOM, type R510 (5×10 cm) for concentrates.

[0097] Preparation: After labeling, the bags were washed before used for the first time by using a washing machine choosing the cold wash program within a laundry net (the same program that will be used later for the incubated bags). After washing, the bags were dried at 60° C. for 12 hours in a forced air oven, then cooled down in a desiccator and subsequently weighted to get their empty weight.

[0098] Sealing the bags: Heat-sealing

[0099] Preparation of Feeds and Initial Weight

[0100] Dry Matter (DM) content was measured first (103° C. for 4 hours).

[0101] The initial weight was 1.5 g for active ingredient per bag. The ratio of initial weight to bag surface should be 12.5 mg DM/cm.sup.2 on average.

[0102] Incubation of the Bags in the Rumen

[0103] The incubation period was 8 h (4 bags per animal). The determination of wash-out losses in the washing machine is done with 3 bags per feed.

[0104] All bags were incubated simultaneously at the same time and removed at the same time (subsequent incubation sequences). The bags were equally distributed at the cylinder or item that is put into the rumen.

[0105] Incubation started directly before morning feeding. Before incubation, bags were put for 1 min in warm water (40° C.) for conditioning.

[0106] Removing and Treatment of the Bags After the Incubation

[0107] After removing the bags from the rumen, they were put into iced water, afterwards roughly rinsed with cold tap water and then washed in the washing machine (in a laundry net). The rinsed bags can also be frozen or stored for a maximum of 24 hours before they are washed. After the washing machine, the bags were dried or stored frozen until drying.

[0108] In the final step, the incubated bags were dried at 40° C. for 24 hours in a drying oven with circulating air (and cooled down in a desiccator for weighting).

II. Coating Experiments

[0109] Test Series 1

[0110] An ethanolic solution of the film-former ethyl cellulose (EC) was prepared with a solids content of 7.1 wt. %. To this solution, either triethyl citrate (TEC, Example 1) or the acetic ester of monoglycerides (commercial product Acetem 95 Co, Danisco; Example 2) was added under stirring in order to obtain the coating composition as a sprayable solution. The amount of additive was 10% of the dry ethyl cellulose mass. Thereby, a coating comprising 9 wt. % TEC additive and 91 wt. % of EC (film forming agent) was obtained.

[0111] A coating solution without additive was prepared for comparison purposes.

[0112] Subsequently, 3 kg Biolys ® granules (sieve fraction 1.6-2 mm) were introduced into the fluidized bed coater and heated to a starting temperature of T=50° C. After having reached the starting temperature, the coating process was started, and the respective coating solutions were sprayed on the granules. In total, a coating amount of 10 wt.-% (based on the total weight of the coated composition) was applied. Accordingly, a coating comprising 9 wt. % TEC additive and 91 wt. % of EC (film forming agent) was obtained. After having reached the desired coating amount, the spraying was finished, the coated product was dried and then removed from the coater.

[0113] The RPR was determined for the thus-obtained samples via the above-described in vitro test. The results are summarized in Table 1 and show the lysine retention after 6h and 24 h, respectively.

TABLE-US-00002 TABLE 1 RPR after RPR after Sample 6 h [%] 24 h [%] Comparative Example 1 67 35 (without additive) Example 1 76 39 (additive: TEC) Example 2 71 36 (additive: Acetem 95 Co)

[0114] As derivable from the results presented in Table 1, the addition of an additive in the above-specified amounts enables a recognizable improvement of the rumen protection rate, especially for the 6 h value where PRPs>70% could be achieved. A slightly higher protection rate is achieved with triethyl citrate as additive. The 24 h value shows that, although higher protection rates were achieved, no over-protection of the granules occurred.

[0115] Test Series 2

[0116] In test series 2, the examples were carried out in an analogous manner as in test series 1, except that the amount of the ethyl cellulose-based coating was varied in the range of 5-10 wt.-% (based on the total weight of the coated composition). The additive portion in the coating was adjusted again to 10% (in relation to dry ethyl cellulose). Thereby, a coating comprising 9 wt. % TEC additive and 91 wt. % of EC (film forming agent) was obtained. Analyses were carried out in the same way as in test series 1. The results were compiled in table 2.

TABLE-US-00003 TABLE 2 Amount of ethyl cellulose- Sample based coating [%] 5 7, 5 10 Comparative RPR after 6 h [%] 43 55 65 Example 1 RPR after 24 h [%] 18 24 30 (without additive) Example 1 RPR after 6 h [%] 49 64 76 (additive: TEC) RPR after 24 h [%] 24 32 39 Example 2 RPR after 6 h [%] 47 61 71 (additive: RPR after 24 h [%] 22 31 36 Acetem 95 Co)

[0117] Based on the results shown in Table 2, it is evident that with the addition of an additive in the above-specified amounts, the protection of the active ingredient can be improved, even at lower coating amounts. A slightly higher protection rate is achieved with triethyl citrate as additive.

[0118] In case the ethyl cellulose-based coating constitutes more than 15% of the total weight of the coated composition, too little of the protected active ingredient will be released and available to the animal (“overprotection”).

[0119] Comparing the RPR results for 6 h for the sample without additive addition versus those with admixture of triethyl citrate, a saving of coating agent can be estimated. With addition of triethyl citrate approx. 2% less coating agent are required, to obtain comparable protection rates. This results to a reduction of a specific amount in the range of 20-30 g ethyl cellulose/kg pellet. Accordingly, the amount of “non-natural” ingredients contained in the animal feed is significantly reduced.

[0120] Test Series 3

[0121] An ethanolic solution of the film-former ethyl cellulose (EC) was prepared with a solids content of 7.1 wt. %. To this solution, triethyl citrate (TEC) was added under stirring in order to obtain the coating composition as a sprayable solution. The amount of additive was 10% of the dry ethyl cellulose mass. Thereby, a coating comprising 9 wt. % TEC additive and 91 wt. % of EC (film forming agent) was obtained.

[0122] A coating solution without additive was prepared for comparison purposes.

[0123] Subsequently, 350 kg of uncoated pelleted methionine (cylindrical pellets having a diameter of 1.6 to 1.8 mm and a length of 2 to 3 mm) was introduced into the fluidized bed coater and heated to a starting temperature of T=55° C. Methods for pelleting of methionine may be found e.g. in EP 0 495 349. After having reached the starting temperature, the coating process was started and the respective coating solutions were sprayed on the granules. In total, a coating amount of 4.2% the above-described ethyl cellulose-based coating (based on the total weight of the coated composition) was applied. After having reached the desired coating amount, the spraying was finished, the coated product was dried and then removed from the coater.

[0124] The RPR was determined for the thus-obtained samples via the above-described in vitro and in situ tests. The results are compiled in table 3.

TABLE-US-00004 TABLE 3 in vitro in situ RPR after RPR after RPR after Sample 6 h/[%] 24 h [%] 8 h [%] Comparative 80 40 75 Example (without additive) Example 1 83 48 79 (additive: TEC)

[0125] In accordance with test series 1 and 2, the results for coated methionine indicate that by adding triethyl citrate (TEC), the rumen protection rate can be improved vis-à-vis the reference product (RPR after 6 h and 8 h, respectively). Both the laboratory results and the results of the in situ tests show a higher methionine active substance protection. At the same time, good release rates after 24 h are ensured (meaning that “overprotection” is avoided).

[0126] The skilled person will understand that due to its high hygroscopy/sensitivity towards water, lysine requires slightly higher amounts of the ethyl cellulose—based coating in order to achieve desirable RPR values.

[0127] Test Series 4

[0128] For comparison reasons the test series 1 was extended with investigation of the impact of an increased additive addition, esp. for triethyl citrate. Starting from a concentration of 10 wt. % TEC (based on the amount of dry ethyl cellulose, corresponding to a coating composition comprising 9 wt. % TEC and 91 wt. % EC), in the test series 4 the TEC amount was increased to 15 wt. % (corresponding to a coating composition comprising 13 wt. % TEC and 87 wt. % EC) and 20 wt. % (corresponding to a coating composition comprising 16.7 wt. % TEC and-83,3 wt. % EC) respectively.

[0129] During processing it was observed that with higher TEC concentration, the particles turned more sticky, meaning that more agglomerates were generated.

[0130] The resulting RPR values from the in vitro analyses are summarized in table 4.

TABLE-US-00005 TABLE 4 in vitro RPR after RPR after Sample 6 h/[%] 24 h [%] Example 1 76 39 9 wt. % TEC; 91 wt. % EC Example 2 71 33 13.5 wt. % TEC; 86.5 wt. % EC Example 3 66 27 18 wt. % TEC; 82 wt. % EC

[0131] As derivable from the results presented in Table 4, an almost linear decrease of the protection rate with increase of TEC concentration occurs. It is thus evident that an amount of additive 14 wt. % is particularly desirable.

[0132] The 24 h release rates are also efficient in each of Examples 1 to 3.