COMPOSITION FOR CONTROLLED RELEASE OF PHYSIOLOGICALLY ACTIVE SUBSTANCES AND PROCESS FOR ITS PREPARATION

Abstract

The present invention relates to a rumen-resistant composition in the form of microgranules, a process for its production and a feedstuff containing such composition.

Claims

1. A rumen-resistant composition in the form of microgranules, each microgranule comprising: i) a core comprising: a) one or more physiologically active substances selected from the group consisting of amino acids, vitamins, enzymes, proteins, carbohydrates, probiotic microorganisms, prebiotic foods, mineral salts, choline derivatives of choline and organic acids; and b) a matrix comprising substances selected from the group consisting of binding substances, inert substances and extrusion adjuvants; and ii) at least one core coating layer; wherein each core further comprises at least one disintegrant agent in an amount by weight of between 1.5% and 6.5%.

2. The composition according to claim 1, wherein said disintegrant agent is a substance that modifies its own state or configuration in presence of a post-ruminal aqueous environment and, following this modification, is apt to determine a disintegrating action of the core, or wherein said disintegrant agent is apt to adsorb or recall water in a post-ruminal aqueous environment so as to favour a disintegration of the core.

3. The composition according to claim 2, wherein said disintegrant agent is selected from the group consisting of emulsifiers, thickeners, effervescent mixtures, polysaccharides and methacrylate polymers.

4. The composition according to claim 3, wherein said disintegrant agent is selected from the group consisting of amides in dry form, vegetal lecithins, ethoxylated oils, mono- and diglycerides of fatty acids, agar-agar or Arabic rubber in dry form, effervescent mixtures comprising carbon dioxide of an alkali metal or carbon dioxide of ammonium and a polycarboxylic acid, cellulose in dry form, and methacrylate polymers, or a combination thereof.

5. The composition according to claim 4, wherein the amount by weight of said disintegrant agents is between 1.75% and 5.75%, and more preferably between 2% and 5% with respect to said core weight.

6. The composition according to claim 5, wherein said disintegrant agent comprises a substance selected from the group including soy lecithin, a combination of citric acid and sodium bicarbonate; a combination of soy lecithin, and citric acid and sodium bicarbonate; sunflower lecithin; carboxymethyl cellulose (CMC) in dry form; corn starch in dry form; ethoxylated castor oil; palmitic acid, oleic acid, linoleic acid, linolenic acid and/or stearic acid; agar agar or gum Arabic in dry form; and polymethyl methacrylate (PMMA) or a combination thereof.

7. The composition according to claim 1, wherein said disintegrant agent includes soy lecithin.

8. The composition according to claim 1, wherein said disintegrant agent includes a combination of citric acid and sodium bicarbonate.

9. The composition according to claim 1, wherein said disintegrant agent includes a combination of soy lecithin, and citric acid and sodium bicarbonate.

10. The composition according to claim 1, wherein said disintegrant agent includes sunflower lecithin.

11. The composition according to claim 1, wherein said disintegrant agent includes carboxymethyl cellulose (CMC) in dry form.

12. The composition according to claim 1, wherein said disintegrant agent includes corn starch in dry form.

13. The composition according to claim 1, wherein said disintegrant agent includes ethoxylated castor oil.

14. The composition according to claim 1, wherein said disintegrant agent includes palmitic acid, oleic acid, linoleic acid, linolenic acid and/or stearic acid or a combination thereof.

15. The composition according to claim 1, wherein said disintegrant agent includes agar agar or gum Arabic in dry form.

16. The composition according to claim 1, wherein said disintegrant agent includes polymethyl methacrylate (PMMA).

17. The composition according to claim 1, wherein said core has a cylindrical shape, the height of which is comprised between 0.5 mm and 2 mm or a spheroidal shape, the diameter of which is comprised between 0.5 mm and 2 mm.

18. The composition according to claim 1, wherein the total weight of the coating layers is between 10% and 70% of the microgranule weight.

19. The composition according to claim 1, wherein said coating layer comprises at least one polluting substance selected from the group of emulsifying substances, fatty acids, and methacrylate polymers.

20. The composition according to claim 19, wherein said emulsifying substance is selected from the group consisting of soy or sunflower lecithin, ethoxylated castor oil, alkali metal and magnesium alginates or a combination thereof.

21. The composition according to claim 20, wherein said polluting fatty acid is selected from the group consisting of palmitic acid, oleic acid, linoleic acid, linolenic acid, and stearic acid or a combination thereof.

22. The composition according to claim 1, wherein the composition comprises at least two core coating layers, wherein each coating layer comprises at least one polluting substance selected from the group of emulsifying substances, fatty acids, and methacrylate polymers.

23. The composition according to claim 20, wherein the two coating layers have a different composition of said polluting substance.

24. The composition according to claim 1 comprising: at least one coating layer comprising one or more hydrophobic substance selected from the group consisting of fats, fatty acids, hydrogenated oils, mono- and di-glycerides of fatty acids, esters of fatty acids and fatty alcohols; and at least one coating layer comprising one or more hydrophobic substances selected from the group consisting of microcrystalline waxes, paraffin waxes, vegetal waxes and synthetic edible waxes.

25. A process for the preparation of the composition according to claim 1 comprising the steps of: extruding a mixture comprising one or more physiologically active substances selected from the group consisting of amino acids, vitamins, enzymes, proteins, carbohydrates, probiotic microorganisms, prebiotic foods, mineral salts, choline derivatives of choline and organic acids; binding substances, inert substances and extrusion adjuvants, and disintegrant agents; optionally subjecting the microgranule to spheronization; and forming one or more coating layers.

26. A premixture for animal feedstuff comprising the composition according claim 1.

27. A feedstuff comprising the premixture according to claim 26.

Description

EXAMPLES

[0108] For the following examples, the following analysis and inspection tools were used: [0109] Sieves, arranged at the end of each single production step, for the microgranule dimension line check; [0110] Microscope for visual checks; [0111] Melting point gauge for thermal resistance check; [0112] Penetrometer for hardness and mechanical resistance check; [0113] Automatic titrator or HPLC for the quantitative determination of the active ingredients (concentration); [0114] Pharmaceutical dissolver for determining the degree of ruminal bypass. The experimental conditions of use were 39° C., 15 rpm, 8 hours presence in the solution with “ruminal” pH of about 6.8. [0115] Daisy.sup.II ANKOM: commercially available laboratory artificial rumen, for determining the degree of ruminal bypass. The experimental conditions of use were 39° C., 8 hours presence in ruminal conditions, buffer solution at “ruminal” pH with insertion of “ruminal inoculum” and re-creation of ruminal anaerobiosis according to a university procedure known to those skilled in the art. The pH normally tested in the quality control phase is 6.8. Pharmaceutical dissolver for determining the degree of post-ruminal digestibility. The experimental conditions of use followed the procedure known to experts in the field as “Boisen Test” and, in particular, they were: [0116] 39° C., 30 rpm, 2 h, buffer solution with “gastric” pH (2.0)+pepsin inoculum. [0117] 39° C., 30 rpm, 4 h, buffer solution with “intestinal” pH (6.8)+pancreatine inoculum. [0118] 39° C., 30 rpm, 18 h, buffer solution with “intestinal” pH (6.8)+lipase and bile extract inoculum.

Example 1—Preparation of Microgranules Containing Choline Chloride

Example 1.1

[0119] 425 kg of choline chlorine with a purity of 99% were mixed with 10 kg of spray rice wax, 15 kg of zinc stearate, 10 kg of soy lecithin and 40 kg of silica. The mixture was extruded using an extruder with sectors at different temperature gradients according to the following program:

TABLE-US-00001 Sector 1 Sector 2 Sector 3 Sector 4 Sector 5 Sector 6 85° C. 85° C. 50° C. 50° C. 45° C. 65° C.

[0120] The cores thus obtained had a concentration of 85% in choline chloride. The cores were subsequently subjected to coating in a pan.

[0121] A first coating layer was formed, coating 400 kg of microgranules with 120 kg of a coating mixture comprising: [0122] 65% by weight of hydrogenated palm oil; [0123] 32% by weight of palmitic acid; [0124] 3% by weight of soy lecithin.

[0125] A second coating layer was then formed, coating the microgranules coated by the first layer with 40 kg of a coating mixture comprising: [0126] 50% by weight of hydrogenated palm oil; [0127] 50% by weight of rice wax.

[0128] All the above indicated percentages are percentages by weight based on the total weight of the covering layer.

[0129] The microgranules thus obtained had a concentration of 60% by weight of choline chloride with respect to the total weight of the microgranules.

[0130] Then the microgranules were subjected to an evaluation of the degree of ruminal by-pass, post-ruminal digestibility and total bioavailability using the Boisen method. For a complete description of the Boisen method, reference is made to the publication S. Boisen, J. A. Fernàndez “Prediction of total tract digestibility of energy feedstuffs and pig diets by in vitro analyses” Animal Feed Science Technology 68, 277-286, 1997.

[0131] The in vitro results are highlighted in Table 1.

TABLE-US-00002 TABLE 1 Degree of Content ruminal Digestibility Bioavailability Sample (%) bypass (*) (**) (**) 1 59.4 80.3 89.3 71.7

Example 1.2

[0132] 425 kg of choline chlorine with a purity of 99% were mixed with 10 kg of spray rice wax, 15 kg of zinc stearate, 10 kg of soy lecithin, 5 kg of citric acid, 5 kg of sodium bicarbonate and 30 kg of silica. The mixture was extruded using an extruder with sectors at different temperature gradients according to the following program:

TABLE-US-00003 Sector 1 Sector 2 Sector 3 Sector 4 Sector 5 Sector 6 70° C. 75° C. 80° C. 80° C. 80° C. 85° C.

[0133] The cores thus obtained had a concentration of 85% in choline chloride. The cores were subsequently subjected to coating in a pan.

[0134] A first coating layer was formed, coating 400 kg of microgranules with 100 kg of a coating mixture comprising: [0135] 75% by weight of hydrogenated palm oil; [0136] 23% by weight of palmitic acid; [0137] 2% by weight of ethoxylated castor oil.

[0138] A second coating layer was then formed, coating the microgranules coated by the first layer with 60 kg of a coating mixture comprising: [0139] 50% by weight of hydrogenated rapeseed oil; [0140] 45% by weight of carnauba wax; [0141] 5% by weight of polymethyl methacrylate.

[0142] All the above indicated percentages are percentages by weight based on the total weight of the covering layer.

[0143] The microgranules thus obtained had a concentration of 60% by weight of choline chloride with respect to the total weight of the microgranules.

[0144] Then the microgranules were subjected to an evaluation of the degree of ruminal by-pass, post-ruminal digestibility and total bioavailability using the Boisen method. For a complete description of the Boisen method, reference is made to the publication S. Boisen, J. A. Fernàndez “Prediction of total tract digestibility of energy feedstuffs and pig diets by in vitro analyses” Animal Feed Science Technology 68, 277-286, 1997.

[0145] The in vitro results are highlighted in Table 2.

TABLE-US-00004 TABLE 2 Degree of Content ruminal Digestibility Bioavailability Sample (%) bypass (*) (**) (**) 2 59.7 78.2 92 71.9

Example 1.3

[0146] The extruded cores obtained with the procedure of Example 1.2 were spheronized with the use of an aqueous 75% choline chloride solution as an adjuvant for spheronization and subsequently they were coated in a pan.

[0147] A first coating layer was formed, coating 400 kg of spheroidal microgranules with 125 kg of a coating mixture comprising: [0148] 72% by weight of hydrogenated palm oil; [0149] 25% by weight of palmitic acid; [0150] 3% by weight of ethoxylated castor oil.

[0151] A second coating layer was then formed, coating the microgranules coated by the first layer with 20 kg of a coating mixture comprising: [0152] 50% by weight of hydrogenated palm oil; [0153] 50% by weight of rice wax.

[0154] Finally, a third coating layer of 20 kg containing 100% of polymethyl methacrylate was formed.

[0155] All the above indicated percentages are percentages by weight based on the total weight of the covering layer.

[0156] The microgranules thus obtained had a concentration of 60% by weight of choline chloride with respect to the total weight of the microgranules.

[0157] The in vitro results are highlighted in Table 3.

TABLE-US-00005 TABLE 3 Degree of Content ruminal by- Digestibility Bioavailability Sample (%) pass (*) (**) (**) 3 61.4 86.4 85.0 73.4

Example 2—Preparation of Microgranules Containing Lysine HCl

Example 2.1

[0158] 400 kg of micronized lysine hydrochloride were mixed with 60 kg of granulated rice wax, 48 kg of powdered powder, and 12 kg of soy lecithin. The mixture was extruded using an extruder with sectors at different temperature gradients according to the following program:

TABLE-US-00006 Sector 1 Sector 2 Sector 3 Sector 4 Sector 5 Sector 6 85° C. 65° C. 50° C. 50° C. 50° C. 50° C.

[0159] The cores thus obtained had a concentration of 75% in lysine hydrochloride. The cores were subsequently subjected to coating in a pan.

[0160] A first coating layer was formed, coating 350 kg of microgranules with 100 kg of a coating mixture comprising: [0161] 95% by weight of hydrogenated palm oil; [0162] 5% by weight of sunflower lecithin.

[0163] A second coating layer was then formed, coating the microgranules coated by the first layer with 75 kg of a coating mixture comprising: [0164] 50% by weight of hydrogenated rapeseed oil; [0165] 48% by weight of carnauba wax; [0166] 2% by weight of sunflower lecithin.

[0167] All the above indicated percentages are percentages by weight based on the total weight of the covering layer.

[0168] The microgranules thus obtained had a concentration of 50% by weight of lysine hydrochloride with respect to the total weight of the microgranules.

[0169] Then the microgranules were subjected to an evaluation of the degree of ruminal by-pass, post-ruminal digestibility and total bioavailability using the Boisen method. For a complete description of the Boisen method, reference is made to the publication S. Boisen, J. A. Fernàndez “Prediction of total tract digestibility of energy feedstuffs and pig diets by in vitro analyses” Animal Feed Science Technology 68, 277-286, 1997.

[0170] The in vitro results are highlighted in Table 4.

TABLE-US-00007 TABLE 4 Degree of Content ruminal Digestibility Bioavailability Sample (%) bypass (*) (**) (**) 4 50.2 73.4 86 63.1

Example 2.2

[0171] 324 kg of micronized lysine hydrochloride were mixed with 64 kg of rice wax spray and 12 kg of soy lecithin. The mixture was extruded using an extruder with sectors at different temperature gradients according to the following program:

TABLE-US-00008 Sector 1 Sector 2 Sector 3 Sector 4 Sector 5 Sector 6 85° C. 85° C. 80° C. 80° C. 80° C. 80° C.

[0172] The cores thus obtained had a concentration of 80% lysine hydrochloride. The cores were spheronized, using a 50% lysine hydrochloride solution in water as a spheronization adjuvant, and subsequently subjected to coating in a pan.

[0173] A first coating layer was formed, coating 350 kg of microgranules with 90 kg of a coating mixture comprising: [0174] 75% by weight of hydrogenated palm oil; [0175] 23% by weight of palmitic acid; [0176] 2% by weight of ethoxylated castor oil.

[0177] A second coating layer was then formed, coating the microgranules coated by the first layer with 50 kg of a coating mixture comprising: [0178] 48% by weight of hydrogenated rapeseed oil; [0179] 52% by weight of rice wax; [0180] 2% by weight of sunflower lecithin.

[0181] A third coating layer was then formed, coating the microgranules coated by the second layer with 40 kg of a coating mixture identical to that of the first layer.

[0182] A fourth coating layer was then formed, coating the microgranules coated by the third layer with 30 kg of a coating mixture identical to that of the second layer.

[0183] All the above indicated percentages are percentages by weight based on the total weight of the covering layer.

[0184] The microgranules thus obtained had a concentration of 50% by weight of lysine hydrochloride with respect to the total weight of the microgranules.

[0185] Then the microgranules were subjected to an evaluation of the degree of ruminal by-pass, post-ruminal digestibility and total bioavailability using the Boisen method. For a complete description of the Boisen method, reference is made to the publication S. Boisen, J. A. Fernàndez “Prediction of total tract digestibility of energy feedstuffs and pig diets by in vitro analyses” Animal Feed Science Technology 68, 277-286, 1997.

[0186] The in vitro results are highlighted in Table 5.

TABLE-US-00009 TABLE 5 Degree of Content ruminal Digestibility Bioavailability Sample (%) bypass (*) (**) (**) 5 49.5 78 91 71

Example 3—Preparation of Microgranules Containing DL-Methionine

Example 3.1

[0187] 430 kg of DL-methionine were mixed with 30 kg of rice wax spray, 25 kg of hydrogenated palm oil, 5 kg of silica, 5 kg of citric acid and 5 kg of sodium bicarbonate. The mixture was extruded using an extruder with sectors at different temperature gradients according to the following program:

TABLE-US-00010 Sector 1 Sector 2 Sector 3 Sector 4 Sector 5 Sector 6 85° C. 45° C. 40° C. 40° C. 35° C. 60° C.

[0188] The cores thus obtained had a concentration of 85% DL-methionine. The cores were subsequently subjected to coating in a pan.

[0189] A first coating layer was formed, coating 400 kg of microgranules with 40 kg of a coating mixture comprising: [0190] 70% by weight of hydrogenated rapeseed oil; [0191] 30% by weight of linolenic acid.

[0192] A second coating layer was then formed, coating the microgranules coated by the first layer with 35 kg of a coating mixture comprising: [0193] 35% by weight of hydrogenated palm oil; [0194] 65% by weight of carnauba wax.

[0195] A third coating layer was then formed, coating the microgranules coated by the second layer with 10 kg of a coating mixture identical to that of the first layer.

[0196] All the above indicated percentages are percentages by weight based on the total weight of the covering layer.

[0197] The microgranules thus obtained had a concentration of 70% by weight of DL-Methionine with respect to the total weight of the microgranules.

[0198] Then the microgranules were subjected to an evaluation of the degree of ruminal by-pass, post-ruminal digestibility and total bioavailability using the Boisen method.

[0199] The in vitro results are highlighted in Table 6.

TABLE-US-00011 TABLE 6 Degree of Content ruminal Digestibility Bioavailability Sample (%) bypass (*) (**) (**) 6 69.3 82.3 75.0 61.7

Example 3.2

[0200] Further 400 kg of extruded granules obtained following the same procedure described in the first part of the previous example were coated in a pan using 3 coating layers.

[0201] A first coating layer was formed, coating 400 kg of microgranules with 35 kg of a coating mixture comprising: [0202] 82% by weight of hydrogenated palm oil; [0203] 18% by weight of linolenic acid.

[0204] A second coating layer was then formed, coating the microgranules coated by the first layer with 35 kg of a coating mixture comprising: [0205] 52% by weight of hydrogenated rapeseed oil; [0206] 45% by weight of carnauba wax; [0207] 3% by weight of sunflower lecithin.

[0208] A third coating layer was then formed, coating the microgranules coated by the second layer with 15 kg of a coating mixture comprising: [0209] 50% by weight of hydrogenated palm oil; [0210] 49% by weight of rice wax; [0211] 1% of ethoxylated castor oil.

[0212] All the above indicated percentages are percentages by weight based on the total weight of the covering layer.

[0213] The microgranules thus obtained had a concentration of 70% by weight of DL-Methionine with respect to the total weight of the microgranules.

[0214] Then the microgranules were subjected to an evaluation of the degree of ruminal by-pass, post-ruminal digestibility and total bioavailability using the Boisen method.

[0215] The in vitro results are highlighted in Table 7.

TABLE-US-00012 TABLE 7 Degree of Content ruminal Digestibility Bioavailability Sample (%) bypass (*) (**) (**) 7 69.2 78.8 90 70.9

Example 4—Preparation of Microgranules Containing L-Lysine, Nicotinic Acid and DL-Methionine

Example 4.1

[0216] 180 kg of 99% choline chloride were mixed with 240 kg of 99% L-lysine hydrochloride, 65 kg of nicotinic acid (vitamin PP) and 145 kg of D, L-methionine. 20 kg of citric acid, 20 kg of sodium bicarbonate, 5 kg of sunflower lecithin and 225 kg of rice wax spray were added. The mixture was extruded.

[0217] The cores thus obtained had a concentration of 20% of choline chloride, 26.7% of L-lysine hydrochloride, 7.2% of nicotinic acid, 16.1% of D, L-methionine.

[0218] The nuclei were subjected to coating in a pan.

[0219] A first coating layer was formed, coating 400 kg of microgranules with 200 kg of a coating mixture comprising: [0220] 66% by weight of hydrogenated rapeseed oil; [0221] 34% by weight of palmitic acid.

[0222] A second coating layer was then formed, coating the microgranules coated by the first layer with 100 kg of a coating mixture comprising: [0223] 48% by weight of hydrogenated palm oil; [0224] 50% by weight of rice wax; [0225] 2% by weight of soy lecithin.

Example 5

[0226] Using the above mentioned analytical methods (pharmaceutical dissolver), in vitro tests were carried out to determine the degree of by-pass and the bioavailability of choline chloride: [0227] a product A consisting of choline chloride, 99% pure; [0228] a product B consisting of microencapsulated chlorine choline granules obtained by means of spray-cooling technology, containing 25% by weight of choline chloride with respect to the total weight of the granule; [0229] a product C consisting of microgranules obtained according to Example 4 of the patent EP1791532, containing 50% by weight of choline chloride with respect to the total weight of the granule; [0230] a product D consisting of microgranules obtained according to Example 1.3 described above.

[0231] Test results are shown in the following Tables 8 and 9.

[0232] Table 8 shows the results obtained considering a theoretical administration of 200 g of product.

TABLE-US-00013 TABLE 8 Choline Choline Chloride Bio- Chloride lost in Bypassed available Choline admin- the Choline Choline Chloride istered rumen in Chloride Chloride in faeces Product (g) 8 h (g) (g) (g) (g) A 198 198 0 0 0 B 50 37.5 12.5 12.5 0 C 100 20 80 40 40 D 120 17 103 88 15
Table 8 9 shows the results obtained with a theoretical administration of 100 g of choline chloride.

TABLE-US-00014 TABLE 9 Product Choline Bio- to be Chloride Bypassed available Choline admin- lost in Choline Choline Chloride istered the rumen Chloride Chloride in faeces Product (g) in 8h (g) (g) (g) (g) A 101 101 0 0 0 B 400 75 25 25 0 C 200 20 80 40 40 D 167 13.5 86.5 73 13.5

Example 6

[0233] Using the above mentioned analytical methods (pharmaceutical dissolver), in vitro tests were carried out to determine the degree of by-pass and the bioavailability of choline chloride: [0234] a product A consisting of choline chloride, 99% pure; [0235] a product B consisting of microencapsulated chlorine choline granules obtained by means of spray-cooling technology, containing 25% by weight of choline chloride with respect to the total weight of the granule [0236] a product C consisting of microgranules obtained according to Example 4 of the patent EP1791532, containing 50% by weight of choline chloride with respect to the total weight of the granule; [0237] a product D consisting of microgranules obtained according to Example 1.3 described above; [0238] a product E consisting of microgranules obtained according to the extruded form of Example 1.3, but coated with coating layers devoid of pollutants; [0239] a product F consisting of microgranules obtained according to the extruded form of Example 1.3, in which the portions of disintegrants were replaced by rice wax spray (binder), but the coating layers were “polluted” with twice the amount of pollutants compared to Example 1.3.

[0240] The results of the tests are shown in the following tables 10 and 11. The tables show that the mere presence of the disintegrant is effective in making the choline bioavailable in the post-ruminal phase.

[0241] Table 10 shows the results obtained considering a theoretical administration of 200 g of product.

TABLE-US-00015 TABLE 10 Choline Choline Bio- Chloride Chloride Bypassed available Choline admin- lost in Choline Choline Chloride istered the rumen Chloride Chloride in faeces Product (g) in 8 h (g) (g) (g) (g) A 198 198 0 0 0 B 50 37.5 12.5 12.5 0 C 100 20 80 40 40 D 120 17 103 88 15 E 120 12 108 54 54 F 120 45 75 55 20

[0242] Table 11 shows the results obtained considering a theoretical administration of 100 g of choline chloride.

TABLE-US-00016 TABLE 11 Product Choline Bio- to Chloride Bypassed available Choline be admin- lost in Choline Choline Chloride istered the rumen Chloride Chloride in faeces Product (g) in 8 h (g) (g) (g) (g) A 101 101 0 0 0 B 400 75 25 25 0 C 200 20 80 40 40 D 167 13.5 86.5 73 13.5 E 167 9.6 90.4 47 43.4 F 167 36.9 63.1 45.3 17.8