JP2001078718A - Production of active ingredient preparation, active ingredient preparation and food or feed - Google Patents

Abstract

PROBLEM TO BE SOLVED: To avoid the use of an aldehyde as a cross-linking agent and obtain an active ingredient preparation excellent in safety and useful for a food, a feed, etc. by carrying out an atomization of a cross-linkable protein, a transglutaminase and the active ingredient under specific conditions. SOLUTION: (A) A cross-linkable protein selected from the group of gelatin, casein, soybean proteins, corn proteins and collagen is completely mixed with a transglutaminase derived from a microorganism and an aqueous solution of an active ingredient such as a vitamin, a human food additive or an animal feed additive and the ratio of the active ingredient to the protein is (1:10) to (4:1) expressed in terms of weight. Furthermore, an atomization of the resultant mixture is then carried out in an inert gas atmosphere loaded with a hydrophobic silica, corn starch or a hydrophobic corn starch to produce an active ingredient preparation. The resultant preparation is preferably further dried until the residual moisture content is below 10 wt.% subsequently to the atomization.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

FIELD OF THE INVENTION The present invention relates to a method of embedding one or more active ingredients in a protein matrix,
The protein relates to an active ingredient preparation which is cross-linked with transglutaminase. The active ingredient preparation is, for example, in the form of a stable, dry powder. Further, the present invention relates to human food or animal feed having the above-mentioned preparation,
It relates to a method for the production of dry powders and the special use of crosslinked proteins. The invention particularly relates to dry powders having vitamins, human food additives and animal feed additives, such as carotenoids. Further, various additives can be embedded.

[0002]

BACKGROUND OF THE INVENTION Vitamin and carotenoid products in powder form are generally known and are used extensively in the pharmaceutical industry and in the human food and animal feed industries. Thus, various production methods for the production of suitable products are described in the literature.

[0003] A variety of manufacturing methods, especially spray methods, describe the preparation of preparations in powder form which can protect oxidizable substances, such as oil-soluble vitamins or carotenoids, against oxidative effects.

[0004] DE 1035319 describes the atomizing of oily vitamin dispersions in a large excess of powdered starch having a low moisture content (up to 8%). Water is removed from the atomized particles by dry starch powder. This powder solidifies, in which case a large amount of starch on the surface of the particles remains adhesive. Further excess starch content must be removed and subsequently returned to the process.

[0005] Swiss Patent No. 488,455 describes:
It is described that a mixture of a water-repellent inorganic substance and a water-absorbing substance is used as a dusting agent. The danger of explosion caused by finely dispersed dried starch is thereby avoided.

[0006] The specification of Swiss Patent No. 389,505 is
It describes atomizing a dispersion of the active ingredient in a cooled gaseous medium and solidifying the atomized particles in the medium by cooling. 15 for this method
A drop height of up to m is required, and the temperature must be kept distinctly below room temperature.

Another possible method for solidifying the atomized particles is trapping in powders consisting of metal salts of higher fatty acids. This method is described in Swiss Patent 43
No. 1252.

Another method for preparing stable active ingredient-containing preparations is described in EP 0 618 001. In this case, the spherical particles, in which the active ingredient is embedded in a mixture of various carrier substances, consist of first adding the active ingredient, a substance in the form of an oil, protein and water to a solvent which is immiscible with water. It is formed by controlled distribution of spheroids from an oil-in-water emulsion and is then produced by separating the resulting spheroids. A special mixing system is required for the production of this spheroid. The particles obtained in this case are then treated with an aldehyde, for example acetaldehyde, glutaraldehyde or glyoxal, with a chemical cross-linking, which is also evident from the fact that the obtained material is insoluble in water. ), And additionally stabilization of the active ingredient is achieved.

US Pat. No. 4,670,247 discloses that
Other methods of making crosslinked particles are described. This is done by first converting an emulsion consisting mainly of oil-soluble vitamins, protective colloids such as gelatin and reducing sugars into particles in powder form by a spraying and drying process. The particles are then heated in a thermal process to a temperature of 105-180C. The Maillard reaction between the amino group of the protein and the oxo group of the reducing sugar achieves the insolubility of the dry powder resulting from the cross-linking of the matrix components.

[0010] EP 782883 describes edible microcapsules having a capsule wall,
It is produced by salting out the protein with edible salt and crosslinking the capsule wall with transglutaminase.

When oxidizable compounds, especially fat-soluble vitamins and carotenoids, come into contact with air, they react with oxygen and are converted into undesirable compounds.
This leads to loss of active ingredients. To avoid this oxidation,
For example, certain additives may be added to the preparation that make it slightly permeable to oxygen by reacting with reactive groups in the protein, thus providing stabilizing protection to the active ingredient. This can be done by reacting the reducing sugar with the protein in a Maillard-type reaction, for example, to prevent proteins soluble in water. This can also achieve cross-linking by reacting proteins with aldehydes which also increase the stability with respect to the loading matrix.

[0012]

However, this method has certain drawbacks, for which it is apparent that it would be desirable to seek an improved stabilization method. in this way,
The use of the Maillard reaction between the protein and the reducing sugar in any case implies thermal stress and therefore at least some degradation of the active ingredient. In addition, the product tends to have a brownish hue.

The use of aldehydes as crosslinkers is in this case associated with the various disadvantages of using highly reactive additives which are extremely harmful to health. The products produced by this method are not unconditionally acceptable to consumers.

[0014]

In contrast, the use of enzymatic cross-linking as a principle of stabilization avoids thermal processes and adds in all respects a harmless addition of substances which are natural components of all food chains. Means both. The purified product has no more brownish hue.

The process according to the invention for preparing the active ingredient preparation comprises the steps of: a) thoroughly mixing the aqueous solution of the crosslinkable protein, transglutaminase and the active ingredient, and b) atomizing the active ingredient. The ratio of to protein is 1 to 10 to 4 to 1 by weight.

The preferred ratio of active ingredient to protein is from 1: 4 to 2: 1, particularly preferably from 1: 3 to 1: 1, by weight.

Preferred crosslinkable proteins are gelatin, casein, soy protein, corn protein and collagen.

An advantageous method according to the invention is a method of atomizing an emulsion or dispersion containing the active ingredient into an inert gas atmosphere loaded with hydrophobic silica.

A further preferred method is a method which, following atomization, is dried to a residual moisture content of less than 10% by weight, preferably less than 6% by weight.

The temperature of the process according to the invention is mainly kept below 80 ° C., preferably below 60 ° C.

The invention furthermore relates to an active ingredient preparation obtained by the process according to the invention, and in a further aspect,
The present invention relates to an active ingredient preparation containing a release agent or a mixture of release agents at 0.025 to 4 times the weight of the active ingredient, and to human food or animal feed with an active ingredient preparation of said type.

Preferred release agents are hydrophobic silica, corn starch, corn starch which has been rendered hydrophobic by chemical treatment, metal salts of higher fatty acids and other vegetable starch.

In the case of hydrophobic silica, the content relative to the active ingredient is in the range from 0.025 to 0.4, preferably from 0.05 to 0.2. In the case of corn starch, the corresponding proportion is preferably from 0.25 to 2, particularly preferably from 0.5 to 1.5.

The active ingredient preparation according to the invention produces a dispersion mainly consisting of active ingredients, proteins and other carriers and fillers, for example from the group of carbohydrates and / or natural or chemically modified starches. It is obtained by doing. It may also contain other additives, such as stabilizers or emulsifiers. It also contains enzymes that can link protein molecules together in a variety of ways. The cross-linking achieved thereby imparts reduced water solubility and increased stability to the protein in which the active ingredient is embedded and thus to the matrix.

The active ingredient preparation obtained is substantially homogeneous with respect to the distribution of the crosslinked matrix.

Preferred crosslinkable proteins are gelatin, such as bone gelatin, bovine gelatin, fish gelatin, milk proteins, such as casein, soy protein, corn protein and collagen, particularly preferred proteins are milk protein and soy protein.
The cross-linking enzyme according to the present invention is a transglutaminase,
Particularly, transglutaminase obtained from a microorganism, particularly transglutaminase derived from a microorganism.

Preferred active ingredients are vitamins, human food additives and animal feed additives. Hydrophobic active ingredients are particularly advantageous, especially those which are easily oxidized. Vitamins of the groups A, D, E and K are such substances. Preferred human food and animal feed additives are carotenes and carotenoids, such as β-carotene and, for example, astaxanthin, astaxin, ethyl apo-8'-carotenoate, citranaxanthin, canthaxanthin, zeaxanthin, apo-8'-.
Carotenal, lutein, capsanthin, lycopene and mixtures thereof.

The emulsion may be a hydrophobic release agent such as hydrophobic silica, a natural starch such as corn starch,
Hydrophobic starch derivatives, such as hydrophobic corn starch,
Atomization is performed using a salt of a long-chain fatty acid or a mixture of these substances. However, the release agent is first introduced in a spray chamber, for example with an inert gas (eg
May be present as hydrophobic silica particles therein. Subsequent drying of the atomized particles, optionally after removal of the release agent, optionally up to 80 ° C., preferably
It is carried out by heating in a stream of air or a protective gas, particularly preferably at room temperature, with mild heating to 0 ° C.

[0029]

EXAMPLES Example 1 81.8 g of gelatin (type A 100 Bloom) and 50.7 g of Isosweet (manufactured by Amylim) were mixed with 36 parts of water.
Dissolution was induced by dispersing in 0 g and stirring at 60 ° C for 30 minutes. Next, 62.6 g of constarch was added and stirring was continued until all components were uniformly dispersed. Subsequently, Ethoxyquin (100mg /
10 ⁶ IU) and BHT (4.5 mg / 10 ⁶ IU)
62.9 g of vitamin A acetate stabilized by the addition of Vitamin A was emulsified into the aqueous phase by using a highly efficient stirrer. Finally, the resulting emulsion is concentrated to 10%
PH by adding aqueous sodium hydroxide solution
It was adjusted to 8.0 and mixed with 20 ml of an aqueous solution of 200 units of bacterial transglutaminase.

The emulsion obtained was atomized at 55 ° C. through a one-component nozzle into a nitrogen atmosphere loaded with hydrophobic silica under a pressure of 4.2 bar. The resulting product was then dried in a fluid bed dryer in a stream of nitrogen within 15 hours to a residual moisture content of 4.2% at room temperature.

Example 2 A dry powder was prepared as in the above example, but without the addition of transglutaminase. The product had a residual moisture content of 3.9% after drying.

Example 3 A portion of the dry powder from Example 2 was subjected to a heat treatment in a rotary flask at 120 ° C. for 20 minutes in an oil bath, whereby a brown discoloration resulted from the Maillard reaction. The residual moisture content of the product was reduced in this way to 1.9%.

Example 4 An astaxanthin preparation was applied to EP-A-0065.
It was prepared by the method described in detail in the specification of Japanese Patent No. 193.

30 g of astaxanthin were suspended together with 1.1 g of ascorbyl palmitate and 6.4 g of ethoxyquin in 240 g of isopropanol and continuously mixed with 370 g of isopropanol in the mixing chamber with the pressure regulating valve set at 30 bar. . A mixing temperature of 173 ° C. was set in the mixing chamber with a feed rate of 6 l / h on the suspension side and a feed rate of 9 l / h on the solvent side. Residence time 0.
After 3 seconds, the molecular dispersion is placed in another mixing chamber with 400 ml of water.
P of 38.6 g of gelatin and 105 g of glucose in 0 g
The solution adjusted to H9.5 was mixed at a processing speed of 80 l / h. A colloidally dispersed active ingredient suspension was obtained. Particle size analysis showed an average of 0.15 μm with a distribution width of ± 31%.

The colloidal dispersion suspension has a solids content of about 25%.
Concentrated in a thin-layer evaporator. The concentrated product was then melted at 60 ° C., 50 ml of an aqueous solution of 250 units of bacterial transglutaminase were added and stirred in a four-necked flask using a paddle stirrer.

The finished dispersion was put into an autoclave, and atomized into a container containing Sipernat D17 as a spray agent dispersed in air through a nozzle. The dry powder thus produced was dried in a fluid bed dryer at room temperature within 20 hours to a moisture content of <6%.

Example 5 A fraction having a particle size of 250-355 μm was screened from the obtained dry powder and subjected to a stability test in a standard premix. For this, approximately 100 mg of test sample are weighed into test tubes (4 weighed for each sample and test time), 60% bran, 3050% choline chloride on silica and 37.43% CuSO _4. × 5 H ₂
O; 46.78% FeSO ₄ × 7H ₂ O; 11.79
% ZnO; 3.61% MnO and 0.39% Co
4 g of a premix consisting of 10% of a trace element mixture consisting of CO ₃ was added and subsequently mixed by hand with care.

The test sample was placed in an open container in a cabinet at a constant temperature and humidity (40 ° C. and 70% rel. Humidity).
For 6 weeks. At the start of storage and after 4 and 6 weeks, four test samples prepared at each test point were taken out and tested for the residual content of vitamin A active ingredient.

The test results are shown in the following table: Vitamin A residue (%) Test product

[0040]

[Table 1]

Example 6 As in Example 1, the emulsion was prepared by adding 420 g of water and gelatin 10
0Bloom A 102.3 g, Isosuite 63.
4 g, vitamin A acetate (2160000 IU / g;
Ethoxyquin 100 mg / million U. And BH
T 14.5 mg / million U. 7)
6.5 g and 80.6 g of corn starch, then pH adjusted with 10% strength aqueous sodium hydroxide solution.
It was adjusted to 8.0. This emulsion was divided into three equal parts and treated as follows: A: no additives B: concentration of transglutaminase ("TG") in polysaccharide 1% Addition of 2.05 g of preparation C: transglutaminase in polysaccharide Addition of 5.11 g of a 1% strength ("TG") preparation. These emulsions were atomized as in Example 1 into a nitrogen atmosphere loaded with hydrophobic silica.

Batch A was dried in a stream of nitrogen at room temperature until the residual moisture content was 3.1%. A portion was additionally heated at 120 ° C. for 20 minutes.

Batch B was dried at room temperature and after storage for 15 hours, likewise in a fluidized bed at room temperature.

Batch C was treated as batch B.

The product obtained was subjected to a stability test as in Example 5. The measured data is summarized in the following table:

[0046]

[Table 2]

[0047]

[Table 3]

Gelatin is a mixture of protein molecules of various sizes. All molecules were labeled with a fluorescent dye. During the observation time, in the case of significant cross-linking, only the smallest molecules could escape through the network of cross-linked gelatin. In addition, relatively small molecules were less likely to be directly affected by crosslinking. Observation by FCS of the molecular size in the supernatant of gelatin that was significantly cross-linked by transglutaminase revealed a high diffusion rate (tau (μ
Sec) is smaller than the control)-typical of smaller molecular sizes. It could also be observed that the intensity (I, kcps) was sufficiently comparable between all samples.

Example 7 In another test, various amounts of transglutaminase were added to the astaxanthin dispersion as in Example 4 and processed into a dry powder.

[0050]

[Table 4]

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat ゛ (Reference) A23K 1/16 303 A23K 1/16 303F 304 304B 1/165 1/165 C A23L 3/3463 A23L 3/3463 (72) Inventor Laurent Betz Niederkirchen im Bruhl 5 (72) Inventor Erik Rüdeck Germany Mutterstadt Thomas-Mann-Strasse 27 (72) Inventor Thomas Friedrich Darmstadt Saarbau, Germany Strase 22-24 F-term (reference) 2B150 CC11 CC14 CE06 CE08 DC23 DD11 DE02 DE14 DE15 DF11 4B018 LE03 MD23 MD24 MD26 MF02 MF06 MF07 MF12 4B021 LA02 MC03 MC07 MP05

Claims (12)

[Claims] 1. A method for producing an active ingredient preparation, comprising: a) a step of completely mixing an aqueous solution of a crosslinkable protein, transglutaminase and an active ingredient; b) a step of atomizing, wherein the active ingredient and the protein are Is 1 to 10 to 4 to 1 by weight. 2. The method according to claim 1, wherein the active ingredient is a vitamin, a human food additive or an animal feed additive. 3. The method according to claim 1, wherein the active ingredient is hydrophobic. 4. The crosslinkable protein is selected from the group consisting of gelatin, casein, soy protein, corn protein and collagen.
The method according to any one of the preceding claims. 5. The method according to claim 1, wherein transglutaminase obtained from a microorganism is used. 6. The process as claimed in claim 1, wherein the atomization is carried out in an inert gas atmosphere loaded with hydrophobic silica, corn starch or hydrophobic corn starch. 7. The process as claimed in claim 1, wherein the drying is effected subsequent to the atomization until the residual water content is below 10% by weight. 8. The method according to claim 1, wherein the temperature of the active ingredient preparation is maintained mainly at 80 ° C. or lower. 9. An active ingredient preparation obtained by the method according to any one of claims 1 to 8. 10. active ingredients for the following groups: selected from carotenoids Vitamin A Vitamin E Vitamin D _3, the active ingredient preparations according to claim 9. 11. The release agent or release agent mixture content of 0.025 to 4 times the weight of the active ingredient.
Or the active ingredient preparation according to 10. 12. A human food or animal feed comprising the active ingredient preparation according to any one of claims 9 to 11.