MXPA00005830A - Granule with hydrated barrier material - Google Patents

Abstract

A granule having high stability and low dust is described. The granule includes a hydrated barrier material having moderate or high water activity. Also described are methods of producing the granules.

Description

GRANULO WITH HYDRATED PROTECTIVE MATERIAL Background of the Invention Recently, the use of enzymes, especially of microbial origin, has become more and more common. Enzymes are used in various industries that include, for example, the industry. starch, the dairy industry and the detergent industry. It is well known in the detergent industry that the use of enzymes, particularly proteolytic enzymes, has created industrial hygiene concerns for workers in detergent factories, particularly because of the health risks associated with the dustiness of available enzymes.

Since the introduction of enzymes in the detergent business, many developments in the granulation and coating of enzymes have been offered by the industry. See for example the following patents related to the granulation of enzymes: The patent of E. U. 4,106,991 describes an improved formation of enzymatic granules by including within the REF .: 120476 composition that undergoes in granulation finely divided cellulose fibers in an amount of 2-40% W / W based on the dry weight of the entire composition. Furthermore, this patent discloses that waxy substances can be used to coat the particles of a granulated material.

US Patent 4,689,297 describes particles containing enzymes comprising a core of particles and dispersible in water having 150-2,000 microns in its longest dimension, a uniform layer of enzyme around the core particle giving a total of 10 % -35% by weight of the weight of the core particle, a layer of a macro-molecular, film-forming, water-soluble or dispersible coating agent that uniformly surrounds the enzyme layer wherein the combination of the enzyme and the agent Cover is from 25-55% of the weight of the core particle. The core material described in this patent includes clay, a sugar crystal encased in layers of corn starch which is coated with a layer of dextrin, agglomerated potato starch, particulate salt, agglomerated trisodium citrate, NaCl flakes, bread crystallized granules or bentonite beads converted from a current flow, granules containing bentonite, kaolin and diatomaceous earth with sodium citrate crystals. The film-forming material may be an ester of fatty acids, an alkoxylated alcohol, a polyvinyl alcohol or an ethoxylated alkyl phenol.

U.S. Patent 4,740,469 describes a granular enzymatic composition consisting essentially of 1-35% by weight of an enzyme and from 0.5-30% by weight of a synthetic fibrous material having an average length of 100-500 microns and a fineness in the range from 0.05-0. 7denier, with the balance being a filler or diluent. The granular composition may further comprise a fused waxy material, such as polyethylene glycol, and optionally a dye such as titanium dioxide.

U.S. Patent 5,254,283 describes a particulate material that has been coated with a continuous layer of a water-insoluble polymer of deformation size. U.S. Patent 5,324,649 describes granules containing enzymes having a core, an enzymatic layer and an outer coating layer. The enzyme layer and, optionally, the core and the outer coating layer containing a vinyl polymer.

WO 91/09941 describes a preparation containing enzymes wherein at least 50% of the enzymatic activity is present in the preparation as enzymatic crystals. The preparation can be either a granulated material or a thick paste.

WO 97/12958 discloses a microgranular enzymatic composition. The granules are made by agglomeration of fluid bed resulting in granules with various carrier or seed particles coated with an enzyme and joined together by a binder.

Nevertheless, even in light of these developments offered by the industry (as described above) there is a continuing need for enzymatic granules that produce little amount of powder and have additional beneficial characteristics. The additional beneficial features needed in the granulation enzymatic industry are formulations of low-residue granules (where low residues are defined as a reduced tendency to leave apparent undissolved residues in clothing or other material), and improved stability formulations . Achieving all these desired characteristics simultaneously is a particularly challenging task since, for example, many agents that produce a small amount of powder and prolonged release such as fibrous cellulose or deformation size polymers leave behind insoluble residues.

Therefore, it is an object of the present invention to provide highly soluble enzymatic granules which produce little residue, and which produce little amount of powder having increased stability. It is another object of the present invention to provide processes that allow the formation of these improved granules.

Brief Description of the Invention One embodiment of the present invention is a granule that includes a protein core and a hydrated protective material with moderate or high water activity. The hydrated protective material may be one or more layers and / or may be included in the protein core.

Another embodiment of the present invention is a granule that includes an enzymatic core of a hydrated protective material with moderate or high water activity. The hydrated protective material can be in one or more layers and / or can be included in the enzymatic core.

Another modality of his method to produce the previous granule.

Detailed Description of the Invention The present invention provides a granule with improved stability and producing little amount of powder. The granule includes a protein core of a hydrated protective material with moderate or high water activity.

A "protein core" or an "enzyme core" may be homogeneous as described in the patent application of E. U. 08 / 995,457 or layered as described in U.S. Patent No. 5,324,649.

Proteins that are within the scope of the present invention include pharmaceutically important proteins such as hormones or other therapeutic proteins and industrially important proteins such as enzymes.

Any enzyme or combination of enzymes can be used in the present invention. Preferred enzymes include those enzymes capable of hydrolyzing substrates, for example, dyes. These enzymes are known as hydrolases that include, but are not limited to, proteases (bacterial, fungal, acidic, neutral or alkaline), amylases (alpha or beta), lipases, cellulases and samples thereof. Particularly preferred enzymes are subtilisins or cellulases. More preferred are subtilisins such as those described in US Patent 4,760,025, and EP 130 756 Bl and EP Patent Application WO 91/06637, which are incorporated herein by reference, and cellulases such as Multifect L250 ™ and Puradax ™, commercially available from Genencor International. Other enzymes that can be used in the present invention include oxidases, transferases, dehydratases, reductases, hemicellulases and isomerases.

As noted, the protective material can be coated on the protein core in one or more layers or made part of the protein core in order to isolate or prevent the transport of water by protein inactivating substances. When the protective material is part of the protein core, it can disperse along the nucleus or as a layer in the nucleus.

Suitable hydrated protective materials with a moderate or high affinity to water may include salts of an organic or inorganic acid, sugars, polysaccharides, lipids, proteins or synthetic polymers; preferably salts.

The term 'water activity', which is symbolized as aw, refers to the relative humidity in a fraction of an atmosphere in equilibrium with a solid or liquid phase material, that is, the proportion of the partial pressure of water to which it is present above pure water at the same temperature.Of all the phases between which the distribution of water has reached an equilibrium, it is by definition the same.The term 'relative humidity' is generally used to describe water in an atmosphere or Gas phase in equilibrium with the solid, and is expressed as a percentage, with 100% as the relative humidity of the pure water in an enclosed system. Therefore, for any water activity value, there is a corresponding relative humidity given by% RELATIVE HUMIDITY = 100 * a ".

Water activity can be quantified by methods known in the art, typically by placing a sample of temperature controlled material from a water activity meter, such as the Model D2100 Water Activity System available from Rotronic Instrument Corp. (Huntington, NY), and allow the quantification to reach its equilibrium as indicated on the display.

A "hydrated" protective material contains water in free or bound form, or a combination of both.The water of hydration can be added either during or after the coating process.The degree of hydration is a function of the material itself and the temperature, humidity and drying conditions under which they are applied.

The "moderate or high" water activity includes a water activity of at least 0.25, preferably greater than 0.30, more preferably greater than 0.35, the water activity referred to here is that of a granule itself once it has the protective material - but without additional coatings - and coated in it.The subsequent coatings can mask the exact quantifications of the water activity of the protective material as an indistinct layer.

Without wishing to be bound by theory, materials with a water activity greater than 0.25 are expected to have a reduced driving force to collect water under storage conditions in which the relative humidity is greater than 25%. Most climates have a relative humidity greater than 25%. Many detergents have water activities in the range of about 0.3-0.4. if the activity of the granular water is in fact greater than that of the surrounding detergent or of the storage climate, the driving force for collecting the water by the granule must be eliminated, and in fact the water can be given by the granule to its surroundings . The use without activity of the water of the granule is less than that of the detergent or the corresponding relative humidity, the water present in the protective layer can act as a shield that limits the amount of water and therefore inactivate the substances that are collected by the granule and affect the protein core.

In the case of hydrates of salts, the hydrated material and its crystalline hydrate of salt with water (s) joined of crystallization. The hydrate should be chosen and applied in such a way that the resulting coated granule has an activity of water in excess of 0.25, or as high as possible while providing a granule that is dry to the touch. By applying a salt hydrate, or any other suitable hydrated protective material, such as noted above, one expects this to eliminate any driving force to further collect the water by the granule. As an important consequence, the driving force for the transport of substances may be detrimental to the enzymatic activity such as perborate or peroxide anion, is removed. Without water as a vehicle, these substances are less likely to penetrate the enzymatic core. The empirical data show that the enzymatic activity in the granule substantially intensifies the covering of the enzymatic nucleus with stable salt hydrates.

Preferred salts include magnesium sulfate heptahydrate, zinc sulfate heptahydrate, copper sulfate pentahydrate, sodium dibasic phosphate heptahydrate, magnesium nitrate hexahydrate, sodium borate decahydrate, sodium citrate dihydrate and magnesium acetate tetrahydrate.

The granules of the present invention may also comprise one or more coating layers. For example, such coating layers can be one or more intermediate coating layers, or such coating layers can be one or more coating layers or a combination thereof. The coating layers can serve a variety of functions in a granular composition, depending on the final use of the granule. For example, the coatings can produce a resistance of the protein to oxidation by chlorine, remove the desired rates of dissolution upon introduction of a granule into the aqueous medium, or provide a barrier against environmental humidity in order to intensify the storage stability of the enzyme and reduce the possibility of microbial growth within the granule.

Suitable coatings include polyvinyl alcohol (PVA), polyvinyl pyrrolidone (PVP), cellulose derivatives such as methyl cellulose, hydroxypropylmethyl cellulose, hydroxycellulose, ethylcellulose, carboxymethyl cellulose, hydroxypropyl cellulose, polyethylene glycol, polyethylene oxide, chitosan, gum arabic, xanthenes, carrageenan, latex polymers and enteric coatings. In addition, the coating agents can be used in conjunction with other active agents of the same or different categories.

Suitable PVAs for incorporation into the granule coating layer (s) include partially hydrolyzed PVA, fully hydrolyzed or intermediately hydrolyzed having low or high viscosity grades. Preferably, the outer coating layer comprises partially hydrolyzed PVA having low viscosity. Other vinyl polymers that may be useful include polyvinyl acetate and polyvinyl pyrrolidone. Useful copolymers include, for example, PVA-methylmethacrylate copolymer and PVP-PVA copolymer.

The coating layers of the present invention may further comprise one in addition to the following: plasticizers, diluents, lubricants, pigments, and optionally additional enzymes. Suitable plasticizers useful in the coating layers of the present invention are plasticizers including, for example, polyols such as sugars, sugar alcohols, or polyethylene glycols (PEG), urea, glycol, propylene glycol or other known plasticizers such such as triethyl citrate, dibutyl or dimethyl phthalate or water. Suitable pigments useful in the coating layers of the present invention include, but are not limited to, finely divided whiteners such as titanium dioxide or calcium carbonate or pigments with color or dyes or a combination thereof. Preferably these pigments are pigments that produce little residue upon dissolution. Suitable diluents include sugars such as sucrose or starch hydrolyzates such as maltodextrin and corn syrup solids, clays such as kaolin and bentonite, and talc. Suitable lubricants include nonionic surfactants such as Neodol, bait alcohols, fatty acids, salts of fatty acids such as magnesium stearate and fatty acid esters.

The outer coating layer of the present invention preferably comprises between about 1-25% by weight of the coated granule.

The adjunct ingredients can be added to the granules of the present invention. The attached ingredients may include: metal salts; solubilizers, activators, antioxidants; dyes; inhibitors; agglutinators; fragrances; agents / scavengers enzyme protecting such as ammonium sulfate, ammonium citrate, urea, guanidine hydrochloride, guanidine carbonate, sulfonate, guanidine, thiourea dioxide, monoethanolamine, diethanolamine, triethanolamine, amino acids such as glycine, sodium glutamate and the similar, proteins such as bovine serum albumin, casein and the like, etc .; surfactants including anionic surfactants, ampholytic surfactants, non-ionic surfactants, cationic surfactants and salts of long-chain fatty acids; builders; alkalis or inorganic electrolytes; bleaching agents; indigo agents, fluorescent dyes and bleaches; and blockage inhibitors.

The grains described herein can be made by methods known to those skilled in the art of enzymatic granulation, including pan coating, bed fluid coating, fluid agglomeration in bed, transformation to spherical pellets from the free fluid, disc granulation, drying. by atomization, extrusion, centrifugal extrusion, spherolization, drum granulation, high shear agglomeration, or combinations of these techniques.

The following examples are representative and are not intended to be limiting. One skilled in the art can choose other proteins, protein cores, enzymes, enzyme cores, seed particles, methods and coating agents based on the teachings herein.

Examples Example 1. Stability of Protease Granules Coated with Magnesium Sulfate A. In a Deseret 60 fluid bed coater apparatus, 54.1 kg of non-parell sucrose / starch seeds are loaded and fluidized. In these cores, 75.8 kg of protease UF concentrate containing 62.9 g / kg of protease subtilisin were spread under the following conditions. (The ranges indicate the initial and final values over the course of time at a specified slope): Slope time: 80 minutes Feed speed 0.6-1.0 liter / min fluid Atomization pressure 75 psi Inlet air temperature 85-92 ° C Outlet air temperature 50 ° C Air fluidization speed 18m3 / min Se prepare a solution of magnesium sulfate by adding 22.2 kg of magnesium sulfate heptahydrate in 22.2 kg of water, and this is atomized in the nuclei coated with the enzyme under the following conditions in order to provide that 20% of the final granule is sulphate of magnesium heptahydrate, taking care to keep the temperature of the bed close to, but slightly below, 50 ° C: Slope time: 40 minutes Fluid feed speed 0.6-1.7 liters / min Atomizing pressure 45 psi Inlet air temperature 70-84 ° C Outlet air temperature 48-50 ° C Air fluidization speed 18m3 / min Finally, a solution for coating polymers is prepared by dissolving 6.35 kg of polyvinyl alcohol Elvanol 51-05, 7.94 kg of titanium dioxide and 1.59 kg of nonionic surfactant Neodol 23-6.5T in 50.12 kg of water and atomizing it on the enzymatic nuclei. coated with salt under the following conditions: Slope time: 10 minutes, then constant for 100 minutes Feed speed 0.6 liter / min fluid Atomization pressure 75 psi Inlet air temperature 50 ° C Outlet air temperature 75-80 ° C Air fluidization speed 18m3 / min The granules that are harvested have an enzymatic concentration of approximately 40 g / kg.

Accelerated Stability Analysis B. The stability of various enzyme granules formulated in detergents containing bleach is generally excellent, generally showing no more than about 10 to 20% loss in activity during 6 weeks of storage at 30 to 37 ° C and 70% to 80% R. H: However, to aid in the development and detection of granular formulations, it is desirable to have an accelerated medium to determine the relative stability of the granule. The conditions of accelerated stability analysis (AST) are much more severe than enzymatic granules or detergents will ever find in realistic storage or transport. The AST is a "stress analysis" designed to discriminate differences between formulations that would not otherwise be evident for weeks or months.In this analysis, a detergent base is made from the analysis made of the following ingredients: 72% detergent (WFK, Forsshunginstitut fuer base WFK-1 Reinigungstechnologie E.V., Krefeld, Germany) % perborate (Degussa Corp., Allendale Sodium Monohydrate Park, New Jersey) 3% activator (War ick International, from the bleach TAED Mostyn, United Kingdom) (= tetraacetylethylethyl amine; For each enzyme sample to be analyzed, 3 identical tubes are prepared by adding one gram of the analysis base and 30 mg of enzymatic granules to a 15 ml conical tube and mixing by inverting the capped tube 5-8 times with the hand. Drill a hole in the tube cap with a 1/16 inch drill bit. One of the three tubes is immediately evaluated and the other two are stored in a humid bed set at 50 ° C and 70% RH. One of the two tubes that are stored is valued after one day of storage; the second, after three days of storage. Storage stability is reported for Day 1 and Day 3 by dividing the remaining activity by the original activity on Day zero, which is expressed as a percentage.

Enzyme activity is determined by adding to each tube 30 ml of 0.25M MES buffer pH 5.5 containing 20 μl of HP L5000 Catalase (Genencor International, Rochester, NY) and incubated for 40 minutes to inactivate perborate. After this, the enzyme is titrated by adding 10 μl from the analysis tube mixture and 10 μl of protease substrate sAAPF to 980 μl of 0.1 M Tris pH 8.6, and then incubated at 25 ° C for 3 minutes, and the optical absorbance at 410 nm is quantified. Then the slope of the absorbance against time is multiplied by the dilution factor and the extinction coefficient that is known for the specific protease to obtain an enzymatic activity co or in concentration in mg / ml.

The process described in A above is repeated three more times, the only difference is that the temperature of the exhaust air is controlled at a certain point of 40, 60 and 70 ° C in each of the three separate passes. The samples are removed from all four batches after the magnesium sulfate protective coating is applied, and the water activities of the granules are quantified in a Rotronic Water Activity System, as reported in Table 1. Two of the granules, after the application of the final coating with polymer, are placed in the WFK-1 detergent formula and stored in tubes with perforated lids for 3 days at 50 ° C and 70% relative humidity, according to the method of accelerated stability analysis described above. The tubes are removed from the humid chamber and evaluated after one day and three days. The retained percentage activities are reported in Table 1. The results indicate that the granules of which are coated with magnesium sulfate at 50 ° C and exit temperature are significantly more stable than those that are coated at 70 ° C, and the more stable granules have a water activity greater than 0.35, while less stable granules have significantly less water activity.

Table 1. Stability of coated enzyme granules are Magnesium Sulphate Example 2. Stability of Protease Granules Resisted with Sodium Citrate A. In a Vector 60 Coating Apparatus, 25 kg of seeds without sucrose / starch pareil and 30.9 kg of subtilisin protease concentrate with a concentration of 65.9 g / L are fluidized and 18.3% of total solids are atomized in the fluidized cores under the following conditions: Slope time: 55 minutes Feed speed 0.5-0.9 liter / min fluid Atomization pressure 75 psi Inlet air temperature 60-95 ° C Outlet air temperature 50 ° C Fluidization speed 24m3 / min air A solution of trisodium citrate is prepared by adding 13.2 kg of trisodium citrate dihydrate in 19.7 kg of water, and this is atomized in the nuclei coated with the enzyme under the following conditions in order to provide that 25% of the final granules are dihydrated trisodium citrate, taking care to maintain the temperature of the bed near 50 ° C: Slope time: 23 minutes Feed speed 0.6-1.9 liter fluid / s Pressure of atomization 75 psi Inlet air temperature 60-95 ° C Exit air temperature 50 ° C Air fluidization speed 24m3 / min Finally, a polymeric coating solution is prepared by dissolving 2.94 kg of Methocel HPMC, 0.98 kg of polyethylene glycol, with a molecular weight of 600, 2.06 kg of titanium dioxide and 0.59 kg of the nonionic surfactant Neodol 23-6.5T at 55.88. kg of water and is atomized on the enzyme cores coated with salt under the following conditions: Slope time: 10 minutes, then constant for 80 minutes Feed speed 0.5-0.7 liter / min fluid Atomization pressure 75 psi Inlet air temperature 75-80 ° C Outlet air temperature 60 ° C Fluidization speed of air 18m3 / min The granules that are harvested have a weight of 49.5 kg and an enzymatic concentration of approximately 40 g / kg.

B. The above process is repeated under the same conditions, but the air outlet temperature is controlled at a certain point of 70 ° C. The samples are removed from both batches after the protective coating of sodium citrate has been applied, and the water activities of the granules are quantified in a Rotronic Water Activity System, as reported in Table 2. The two Granules, after the application of the final coating with the polymer, are placed in an automatic dish detergent base and stored in sealed tubes for 84 days at 37 ° C. The tubes are removed from the humid chamber and titrated after 14, 42 and 84 days. The percentage activities obtained are reported in Table 2. The results indicate that the granules in which the sodium citrate is coated at 50 ° C exit temperature are significantly more stable than those that are coated at 70 ° C, the granules more stable have a water activity greater than 0.25, while less stable granules have significantly less water activity.

Table 2. Stability of Enzymatic Granules Coated with Sodium Citrate It is noted that in relation to this date, the best known method for the applicant to carry out the aforementioned invention, is the conventional one for the manufacture of the objects or products to which it refers.

Having described the invention as above, property is claimed as contained in the following:

Claims (11)

Claims
1. A granule comprising an enzymatic core of a protective material, characterized in that the protective material comprises a hydrated protective material with a moderate or high water activity.
2. The granule according to claim 1, characterized in that the protective material is a salt.
3. The granule according to claim 1, characterized in that the salt is selected from the group consisting of magnesium sulfate heptahydrate, zinc sulfate heptahydrate, copper sulfate pentahydrate, sodium phosphate dibasic heptahydrate, magnesium nitrate hexahydrate, borate sodium decahydrate, sodium citrate dihydrate and magnesium acetate tetrahydrate.
4. The granule according to claim 1, characterized in that the protective material is part of a protein core.
5. The granule according to claim 1, characterized in that the protective material is coated on the protein core.
6. The granule according to claim 1, characterized in that it also comprises a layer of material between the protein core and the protective material.
7. The granule according to claim 1, characterized in that it also comprises a layer of material on the protective layer and the protein core.
8. The granule according to claim 1, characterized in that the protein is an enzyme.
9. The granule according to claim 1, characterized in that the activity of the water is greater than 0.25.
10. . A method for producing a granule and characterized in that it comprises: a) providing a protein core; b) coating a hydrated protective material with high or moderate water activity in the protein core.
11. 11. The method according to claim 10, characterized in that it also comprises a coating on the protective material.